Accueil / Information / Aller plus loin... / Latex et Venins / L’allergie au latex

L’allergie au latex

mercredi 26 novembre 2008, par Allerdata

L’allergie au latex d’Hevea brasiliensis a pris un essor considérable dans les années 1980 avec l’apparition puis l’expansion du SIDA. Si la prévalence d’allergie au latex est estimée à environ 1% dans la population générale, elle toucherait de 6 à 17% des personnels soignants et 30 à 60% des enfants ayant du subir des actes chirurgicaux répétés .

Les différentes formes cliniques d’allergie au latex ne sont pas revues ici : elles ont fait l’objet de nombreuses revues . On distingue :

  • les réactions respiratoires et/ou dermatologiques rencontrées chez les personnels soignants (PS) par suite de leur exposition professionnelle
  • le cas des patients poly-opérés, et notamment les enfants atteints d’un spina bifida (SB)
  • les anaphylaxies per-opératoires, a priori sans antécédents connus concernant le latex : le latex représente à lui seul environ 20% des causes d’anaphylaxie per-opératoire (et même plus de 50% chez les sujets de moins de 18 ans)
  • les allergies croisées favorisées par une allergie au latex, et notamment les allergies alimentaires (cf. Latex et aliments)

Sans revenir non plus sur les modes de sensibilisation dans le détail, on notera cependant :

  • que l’atopie joue un rôle important dans la sensibilisation professionnelle
  • tandis que l’âge et la pathologie traitée semblent influer sur la survenue d’une sensibilisation per-opératoire .

Les objets susceptibles de favoriser une sensibilisation au latex sont clairement identifiés dans le cadre de l’activité de soins, à commencer par les gants qu’ils soient d’examen ou chirurgicaux.

L’exposition à des objets contenant du latex d’Hevea en dehors d’un cadre d’activité de soins est plus mal cernée et pourrait constituer une forme de sensibilisation "cachée".

Schématiquement, on distingue 2 grandes catégories de produits en latex  :

  • ceux qui, comme les gants, sont obtenus à partir d’une fraction concentrée du latex récolté sur les arbres (la "rubber phase", cf. schéma)
  • et ceux qui résultent du latex naturel coagulé puis lavé et séché à forte température (le "dry rubber") : on utilise ce type de latex séché pour fabriquer des objets aussi divers que des pneus, des bandes élastiques, des tubes, des chaussures, etc..

Est-ce que le contact avec ces objets dans la "vie courante" peut initier une sensibilisation vis à vis des allergènes du latex chez des sujets prédisposés ?

Divers objets en latex semblent capables de relarguer des allergènes : matelas, gants de ménage, ballons, semelles, bandes élastiques, etc.. . Pour certains auteurs, il existe donc une exposition et celle-ci pourrait comporter un risque chez un sujet prédisposé. Les preuves d’une réelle sensibilisation par ces objets sont manquantes mais une telle exposition « cachée » pourrait être mise au crédit de primo-réactions per-opératoires chez des sujets a priori sans facteur de risque.

Il en est de même pour les tétines et autres sucettes destinées aux nourrissons  : des observations avalisent un lien entre l’usage de tels objets en latex et des symptômes allergiques . Des extraits de ces tétines/sucettes ont montré une IgE-réactivité, plutôt faible en général .

On connaît mal ce qu’il en est des élastiques et gouttières utilisées en orthodontie.

Tous ces objets sont, bien sûr, déconseillés en cas d’allergie connue au latex ou de réaction apparemment liée à leur contact . Mais on ne sait pas si ce contact est susceptible de générer une sensibilisation à bas bruit au latex.

La question du latex “ caché ” concerne aussi le transfert possible d’allergènes du latex à des aliments à la suite du contact de ces aliments par des objets en latex (ex. gants) . Bien que probablement très rare, cette cause peut constituer un piège diagnostique chez un patient exploré pour une réaction alimentaire. Sa réalité a été confirmée par TPODA .

Enfin, il a été rapporté des cas d’anaphylaxie vis à vis du latex contenu dans des produits destinés à fixer un apport capillaire

Les allergènes du latex

Le latex d’Hevea brasiliensis, comme le latex d’autres plantes, est une sécrétion à visée défensive : colmater une blessure, éviter que cette dernière soit la porte d’entrée pour des micro-organismes pathogènes.

Il n’est donc pas surprenant que de nombreux allergènes du latex soient des protéines de défense végétale . De plus, la sélection de clones d’Hevea plus productifs, la scarification répétée et l’utilisation de phyto-hormones ne font qu’accroître les concentrations des protéines de défense (ex. l’hévéine ).

Les protéines du latex sont très variées : on en différencie au moins 250 dans le latex ammoniaqué .

Toutes ne sont pas IgE-réactives. Cependant une résolution en blot bidimensionnel montre de l’ordre de 60 spots . Si un certain nombre d’entre eux sont des isoformes d’un même allergène, on est loin malgré tout d’avoir identifié la nature de tous ces spots .

L’IUIS reconnaît 13 allergènes différents, nommés de Hev b 1 à Hev b 13.

Allergène Fonction Glycosylé
Hev b 1 Facteur d’élongation (REF)
Hev b 2 Beta 1,3 glucanase Oui
Hev b 3 REF-like
Hev b 4 Lécithinase Oui
Hev b 5  ?
Hev b 6.01 (pro-hévéine) Barwin-like
Hev b 6.02 (hévéine) Lectine
Hev b 7 Patatine-like
Hev b 8 Profiline
Hev b 9 Enolase
Hev b 10 Superoxyde dismutase (Mn)
Hev b 11 Chitinase de classe 1
Hev b 12 Lipid transfer protein (LTP)
Hev b 13 Early Nodule Specific Protein Oui

D’autres protéines, non estampillées IUIS, sont cependant clairement IgE-réactives :

  • l’hévamine,
  • une citrate binding protein,
  • une isoflavone réductase ;
  • une rotamase,
  • une thiorédoxine,
  • une UDP-glucose pyrpphosphorylase .
  • une papaïne-like ?

Quelles particularités ont les allergènes du latex ?

Hev b 1

Cette protéine, aussi dénommée REF (Rubber Elongation Factor), est indispensable à la polymérisation des particules de caoutchouc contenues dans le latex natif. Elle représente 10 à 60% des protéines totales du latex et se trouve à la surface des particules de caoutchouc, dans ce qui est nommé la “ rubber phase ” après centrifugation du latex (cf. schéma ci-dessous)


Les fractions du latex après centrifugation :

Décrit par Czuppon dès 1993 , cet allergène apparaît sous forme de monomère et de tétramère (58 kDa).

Hev b 1 est particulièrement IgE-réactif chez les enfants multi-opérés : la part d’IgE-réactivité due à Hev b 1 dans le résultat pour l’extrait latex global était ainsi de plus de 50% pour 27 parmi 35 enfants avec spina bifida . Le nombre d’opérations faisait augmenter la réactivité à Hev b 1.

Hev b 1 est absent du C-sérum, de sorte que la méthode de préparation des extraits pour le diagnostic ne peut traduire exactement la proportion que cet allergène a dans le latex à l’état naturel.

Hev b 2

Cette béta 1,3-glucanase de 34 kDa ou de 36 kDa se présente sous au moins 3 isoformes, dont l’une d’entre elles n’est pas glycosylée.

Sa purification est délicate et il est possible que nHev b 2 soit parfois contaminé par Hev b 6 .

Hev b 2 est une béta 1,3-glucanase de classe 1, plus ou moins homologue de béta 1,3-glucanases présentes dans la tomate et le tabac.

Chez des sujets allergiques à l’olivier, le domaine N-terminal d’Ole e 9, la béta 1,3-glucanase du pollen d’olivier, est capable d’inhiber la réactivité à Hev b 2 . Mais dans ce même travail Hev b 2 ne montrait pas de réactivité croisée avec des béta 1,3-glucanases d’aliments.

Hev b 2 est capable de susciter des IgE par sa chaîne glucidique . Cet allergène est donc susceptible de se lier à des IgE anti-CCD et, sous l’action du periodate, environ la moitié des sujets dans un collectif d’allergiques au latex perdaient leur IgE-réactivité vis-à-vis d’Hev b 2 .

Ceci étant, toute l’IgE-réactivité d’Hev b 2 n’est pas due à son épitope glucidique car :

  • on peut obtenir des tests cutanés positifs avec nHev b 2
  • une des isoformes d’Hev b 2 n’est pas glycosylée
  • et en tests cellulaires, le periodate n’annule pas la réponse à nHev b 2 de manière systématique . Il est possible qu’une partie de la réactivité cellulaire soit le résultat d’une liaison mixte, peptidique + glucidique, selon les auteurs de ce travail.

Hev b 3

Hev b 3 est, comme Hev b 1, une protéine associée aux particules de caoutchouc dans le latex natif.

Sa ressemblance avec Hev b 1 lui confère une classification de “ protéine REF-like ”. Et une réactivité croisée entre Hev b 3 et Hev b 1 a été trouvée .

Alenius et, après lui d’autres auteurs, ont montré que Hev b 3 était très souvent IgE-réactif chez les enfants atteints de spina bifida .

Hev b 4

Cet allergène identifié par Sunderasan est en fait constitué d’un groupe de protéines réparties entre 50 et 57 kDa .

Hev b 4 provient des ces organelles appelées “ lutoîdes ” et en suspension dans le latex natif. Après centrifugation du latex ces lutoïdes sont rassemblés dans la phase inférieure, la “ bottom fraction ” ‘le B-sérum) (cf. schéma). Les lutoïdes renferment de nombreuses enzymes et Hev b 4 présente de fait une activité enzymatique de lécithinase et de glucosidase cyanogène .

Hev b 4 a 8 sites potentiels de glycosylation. Ces sites sont occupés de façon hétérogène, comme c’est souvent le cas : des chaînes différentes au même emplacement, etc.. . La glycosylation semble jouer un rôle important dans la réactivité d’Hev b 4 car la protéine une fois déglycosylée (ou le recombinant rHev b 4 préparé dans E. coli) voit son IgE-réactivité annulée .

Hev b 5

On ne sait pas quelle fonction biochimique possède Hev b 5. Cette protéine a été identifiée à partir d’un extrait de gant et du screening d’une bibliothèque d’ADNc du latex .

Hev b 5 présente une homologie modérée (47% d’identité) avec pKIWI501, une protéine du kiwi. La réactivité croisée entre Hev b 5 et pKIWI501 ne semble pas avoir été étudiée.

Hev b 6

On assimile souvent Hev b 6 à une chitinase du fait de sa responsabilité dans les réactions croisées du latex avec des chitinases alimentaires. Cependant, Hev b 6 n’est pas une chitinase : c’est une protéine de défense végétale du type “ barwin ”. Et c’est seulement une partie de cette protéine, le domaine N-terminal ou “ domaine hévéine ”, qui est impliqué dans les réactions croisées avec les chitinases alimentaires (cf. Syndrome latex-aliments).

En fait, on a l’habitude de nommer Hev b 6.01 la protéine entière (ou “ pro-hévéine ”), Hev b 6.02 le domaine hévéine et Hev b 6.03 le reste de la molécule (le domaine C-terminal).

Dans le latex naturel la pro-hévéine se scinde aisément en hévéine et domaine C-terminal et le rapport hévéine/pro-hévéine est très en faveur de l’hévéine. Cette dernière jouera donc un rôle important dans l’IgE- réactivité observé vis-à-vis du latex au cours des tests diagnostiques.

L’allergénicité de l’hévéine est plus marquée que celle du domaine C-terminal : par exemple, au sein d’un même groupe de patients allergiques au latex, 56% étaient positifs pour Hev b 6.02 contre seulement pour Hev b 6.03 .

En règle générale il existe un fort parallélisme entre la réactivité sérique vis-à-vis d’Hev b 6.02 et celle vis-à-vis de la pro-hévéine Hev b 6.01 . Ce qui est logique. La fréquence des patients mono-positifs est faible : par exemple, 3 cas pour Hev b 6.01 parmi 61 patients . Les rares discordances rencontrées sont dans des zones d’IgE-réactivité basse (zone de kU/l proche de la limite de détection).

On comprend d’autant mieux cette corrélation Hev b 6.01/Hev b 6.02 que la majorité des épitopes d’Hev b 6.01 semblent être concentrés sur le domaine hévéine . On retient habituellement la présence de 2 épitopes sur Hev b 6.02 , celui le plus N-proximal étant le plus important (cf. vue en 3D).

Ces épitopes sont conformationnels , ce qui justifie la fragilité de l’IgE-réactivité en cas de chauffage de ce type de molécule.

Hev b 7

Une bande de 46 kDa a été repérée comme IgE-réactive par Beezhold et identifiée plus tard comme une protéine patatine-like .

Hev b 7, qui est constitué de plusieurs isoformes, provient du cytosol dans le latex natif et présente une activité enzymatique de phospholipase A2 et d’acyl transférase.

Hev b 7 est modérément homologue de Sola t 1, la patatine de pomme de terre (env. 40% d’identité), et contrairement à Sola t 1 n’est pas glycosylé.

De fait, l’allergène naturel a une IgE-réactivité équivalente à celle du recombinant rHev b 7 préparé dans Pichia pastoris .

Hev b 8

L’IgE-réactivité de la profiline du latex a été repérée par Vallier dès 1995 . Hev b 8 a finalement été cloné en 2001 par Ganglberger .

Hev b 8 est présent dans le cytosol du latex et cette solubilité semble justifier les faibles concentrations de profiline retrouvées dans les extraits de gants, consécutivement aux lavages extensifs que subissent ces produits au cours de leur fabrication .

Hev b 9

Cet allergène, une énolase, a été identifié en 1997 par Posch . Bien que présentant environ 60% d’identité avec des énolases fungiques comme Alt a 11 ou Cla h 6, Hev b 9 ne semble pas croiser avec celles-ci .

Hev b 10

C’est aussi dans le même travail que Posch a montré qu’une superoxyde dismutase à manganèse (Mn SOD), Hev b 10, était aussi un allergène dans le latex.

Si les 49% d’identité d’Hev b 10 avec des enzymes fungiques homologues (ex. Asp f 6) ne justifient pas une réactivité croisée latex-moisissures importante, cela mérite d’être approfondi. En effet, il a été constaté une fréquence inattendue de positivités simultanées pour le latex et pour Alternaria . Cela concerne également la noix qui possède également une Mn SOD IgE-réactive (cf. Latex et fruits à coque).

Hev b 11

Cet allergène est une chitinase de classe 1, c’est-à-dire possédant un domaine hévéine (cf. schéma).

Cette chitinase a été suspectée par Lavaud dès 1992 et confirmée plus tard comme participant aux réactivités croisées latex-aliments végétaux .

L’importance d’Hev b 11 dans le syndrome latex-aliments n’est pas bien cernée. Pour Wagner , Hev b 11 ne croise pas avec les chitinases de fruits.

Il faut remarquer cependant que l’homologie entre le domaine hévéine d’Hev b 11 et celui de chitinases d’avocat, de banane ou de châtaigne (72 à 81% d’identité) est plus forte que l’homologie entre l’hévéine Hev b 6.02 et ces domaines dans les fruits (ex. 58% s’agissant de l’avocat) .

L’écart entre Hev b 6.02 et Hev b 11 est donc peut-être à attribuer à des facteurs d’accessibilité des épitopes (cf. vue en 3D).

Hev b 12

Caractérisée récemment , cette LTP n’a pas encore une place bien définie parmi les protéines IgE-réactives du latex. Ni d’ailleurs au sein d’un syndrome LTP.

Sa réactivité croisée avec d’autres LTP mériterait d’être étudiée  ; et l’observation de quelques sujets positifs pour rHev b 12 (6 cas sur 48 ) chez des patients allergiques à des fruits fait plutôt penser à une réactivité croisée induite par une sensibilisation initiale aux LTP de ces fruits.

Hev b 13

Cette protéine de 43 kDa a longtemps été confondue avec Hev b 7 car elle migre dans la même zone en SDS-PAGE. Mais l’inhibition de la bande “ Hev b 7 ” par la patatine de pomme de terre était grandement due à une réactivité croisée de type CCD, la patatine (glycosylée) inhibant Hev b 13 (glycosylé) plutôt qu’Hev b 7 (non glycosylé).

Hev b 13 est une “ early nodule specific protein ”. Sa relevance clinique est mal cernée, tant du fait d’une réactivité en partie CCD ‘excès de positivités in vitro) que par suite de défauts de purification de cet allergène (contamination par du Hev b 6) .

De fait, la prévalence de positivité du recombinant rHev b 13 (dans E. coli = non glycosylé) est plus basse que celle de l’allergène naturel .

Autres allergènes

  • l’hévamine (ou hévamine A) n’a pas reçu de nom IUIS. Cette protéine a activité mixte chitinase/lysozyme a cependant été montrée IgE-réactive depuis de nombreuses années . Cela tient notamment à sa faible prévalence de positivité
  • une papaïne-like IgE-réactive est très probable dans le latex d’Hevea : Baur observe 8 sujets positifs pour la papaïne parmi 24 allergiques au latex et 6 positifs au latex parmi 12 cas d’allergie professionnelle à la papaïne .
    (Voir aussi Figue, ficus et latex)
  • une chitinase de classe 2, c’est-à-dire dénuée de domaine hévéine, a été suggérée dans des travaux anciens . Mais sa faculté d’IgE réactivité doit être confirmée.
  • une protéine de 55 kDa, retrouvée dans le B-sérum et hydrophobe, positive in vitro chez une certaine proportion de personnels soignants (14%) et d’ouvriers de fabrication des gants (33%), mais dont la relevance clinique semble très limitée

Devenir des allergènes : de l’arbre aux objets et aux outils diagnostiques

En théorie, les outils diagnostiques sont sensés explorer les mêmes allergènes que ceux auxquels le patient a été exposé. Cette équivalence vaut :

  • qualitativement : toutes les sortes d’allergènes sont présentes dans le test diagnostique et ont les mêmes épitopes que ceux présentés au patient lors du contact avec le produit
  • quantitativement : la répartition des différents allergènes dans le test est superposable à celle dans le produit entrant en contact avec le patient.

On conçoit aisément que pour un aliment ces prérequis soient difficiles à maintenir du fait, notamment, des transformations dues au processus digestif et à une éventuelle cuisson.

Et on pourrait espérer que dans un produit comme le latex les allergènes soient moins atteints par ce problème de représentativité dans les tests diagnostiques.

Hélas, il n’en est rien. En effet, de multiples écueils compliquent cet objectif :

  • la voie de sensibilisation est au moins double, respiratoire pour certains sujets et cutanéo-muqueuse pour d’autres sujets. Un même extrait peut-il répondre à ces cadres très différents d’immunogénicité ?
  • les procédés de fabrication des produits manufacturés à base de latex sont d’au moins 2 catégories, avec des étapes de chauffage et de lavage, ainsi que des modes de contact, très différents : certains produits proviennent d’un latex d’abord coagulé, tandis que pour d’autres on procède en premier lieu à une concentration des particules de caoutchouc (ex. gants)
  • les extraits pour tests diagnostiques sont tantôt réalisés sur un latex natif, tantôt sur un latex additionné d’ammoniaque (NH4)
  • les particules aérosolisées, comme la poudre des gants, et les faces internes et externes des gants en latex contiennent des répartitions différentes d’allergènes
  • les extraits pour TC et ceux pour tests in vitro ont, au moins en France, une origine différente, ce qui favorise d’autant mieux des discordances entre leurs résultats
  • les protéines initialement présentes dans le latex naturel peuvent être modifiées du fait du processus de vulcanisation (ex. Hev b 5 ou Hev b 1 et Hev b 6.02 qui forment des fragments et/ou des réagrégats )
  • plus ou moins relarguées et/ou stables selon l’addition ou non d’NH4 (ex. Hev b 3 ou Hev b 5 )
  • plus ou moins éluées du produit fini selon la qualité et l’intensité des processus de lavage, et donc dépendre en partie des sources commerciales
  • enfin, les données sylvicoles influent également sur l’intensité de production des allergènes dans le latex des arbres : sélection de cultivars, stimulation par des phyto-hormones

Il n’est donc pas surprenant :

  • que des travaux n’aient pu trouver de corrélation entre les taux d’allergènes en atmosphère de travail et ceux mesurés dans des extraits aqueux de gants , même si l’usage de gants non poudrés améliore clairement l’exposition globale et les données cliniques des professionnels exposés
  • que les conditions-mêmes d’obtention des extraits de gants (ex. avec ou sans agent tensio-actif) aient pu donner des résultats très disparates
  • que des écarts de contenus soient donc constatés entre un extrait à visée diagnostique et un extrait de gants . Fuchs observe 16 patients positifs/56 pour un extrait de gant mais négatifs pour un extrait de latex ammoniaqué ; et vice versa pour 6 autres patients
  • que les allergènes repérés comme immuno-dominants à partir d’un extrait de gants varient selon les travaux : Hev b 5 et Hev b 13 , Hev b 6.02 , ..

Il faut ajouter à cela :

  • le rôle joué par les particules d’amidon des gants poudrés : celles-ci semblent posséder en soi un pouvoir immuno-adjuvant , capter Hev b 5 ou Hev b 1 beaucoup mieux que Hev b 6.02
  • les écarts de répartition des allergènes entre face externe et face interne des gants , Hev b 1 et Hev b 3 étant plutôt majoritaire en face externe et Hev b 5 et Hev b 6.02 en face interne .
    Cette différence, résultant peut-être d’un lavage plus poussé de la face externe au cours du process de fabrication des gants, est avancée pour expliquer la sensibilisation particulière à Hev b 1 et Hev b 3 des patients multi-opérés du fait d’un contact muqueux avec les gants .

La question de la représentativité des extraits pour tests diagnostiques est donc très complexe à résoudre dans le cas du latex. Il s’y rajoute, bien sûr, l’influence des éventuelles IgE anti-CCD en ce qui concerne les tests sérologiques (cf. Latex et CCD).

Tous les allergènes n’ayant pas forcément le même pouvoir allergisant, des travaux ont également cherché à repérer quels allergènes étaient importants à retrouver présents dans les extraits pour diagnostic.

Ainsi, Yeang a mesuré le seuil de réactivité en prick-test pour différents allergènes du latex et constaté qu’Hev b 13 était l’allergène donnant un TC positif avec la plus faible dose. Après Hev b 13, Hev b 4, 7, 2, 5, 3 et 6 demandaient une concentration-seuil 10 à 20 fois plus élevée ; quant à Hev b 1 il fallait environ 200 fois plus élevée.

Ces résultats sont cependant à pondérer pour au moins 2 raisons :

  • dans le latex naturel, et dans les extraits aussi, Hev b 6 est quantitativement surreprésenté par rapport à la plupart des autres allergènes . La réponse en prick-test vis-à-vis d’un extrait global de latex est, du coup, largement influencée par la réactivité du patient pour Hev b 6
  • les travaux réalisés avec des allergènes purifiés peuvent également souffrir d’une contamination par Hev b 6

Prévalences de positivité pour les allergènes du latex


Les tableaux ci-dessous reprennent les résultats de fréquence de positivité pour les différents allergènes du latex selon le cadre clinique.

Prévalences

Ces résultats ont été obtenus à l’aide de techniques in vitro dans leur quasi-totalité.

Des écarts importants apparaissent parfois pour un même allergène entre les études. Cela est du à plusieurs facteurs :

  • la sélection des patients
  • le type de méthode utilisée : qualitative (ex. immunoblot) ou quantitative (RAST, EILSA, CAP) avec des seuils de sensibilité variables d’une technique à une autre
  • certains résultats ont été obtenus avec des allergènes naturels purifiés, tandis que d’autres étaient basés sur des allergènes recombinants (cela est noté par un chiffre en italique dans les tableaux ci-dessous)
  • la pureté des allergènes purifiés :
    • Palosuo obtient des fréquences de positivité plus basses avec nHev b 2 et nHev b 13 quand ces allergènes sont hautement purifiés . Une contamination par des allergènes souvent positifs, comme Hev b 5 ou Hev b 6.01 est possible.
    • Cela pourrait expliquer les prévalences très élevées observées par Raulf-Heimsoth avec nHev b 2 et nHev b 13, puisque ces dernières ne pouvaient, dans cette étude, être attribuées à une réactivité de type CCD (seulement 9% des sujets étaient positifs pour la peroxydase de raifort).

Quels sont les allergènes cliniquement importants ?

Comme pour d’autres produits allergisants, la relevance clinique de certains des « allergènes » du latex n’est pas établie : si pour la plupart d’entre eux on a pu montrer qu’ils étaient, au moins, capables de réponses positives en tests cutanés, pour les autres une dénomination de « protéines IgE-réactives in vitro » serait plus conforme à ce que l’on sait au mieux. C’est le cas notamment pour Hev b 9, Hev b 10, Hev b 11 et Hev b 12.

On distingue deux grands cadres pour la sensibilisation au latex : l’exposition professionnelle et le contact avec des objets en latex, notamment au cours des actes chirurgicaux.

Parce que l’allergie au latex des professionnels de santé (PS) comporte, entre autres, une rhinite et/ou un asthme, il a été suggéré que la sensibilisation était respiratoire pour ces personnes.

Inversement, les sujets polyopérés, tels les spina bifida (SB) se sensibiliseraient par contact muqueux avec les objets en latex, comme les gants, les sondes, etc..

Ces 2 formes de sensibilisation pourraient s’expliquer par :

  • une composition en allergènes différente entre la surface externe et la surface interne des gants.
  • Le poudrage interne des gants, susceptible d’aérosoliser les allergènes dans l’atmosphère des blocs opératoires
  • Un profil d’IgE-réactivité différent pour les SB et pour les PS

De fait, certains travaux ont noté des taux d’allergènes extractibles depuis la surface des gants très différents selon la face testée : Hev b 1 et Hev b 3 sont principalement relargués par la face externe des gants et Hev b 5 par la face interne. Les choses étant égales pour Hev b 6.02 .

Et le profil d’IgE-réactivité des SB corrobore cette observation, ces patients ayant une prévalence d’IgE-réactivité très élevée pour Hev b 1 et Hev b 3, ce que l’on n’observe pas pour les PS (cf. le tableau des prévalences).

Mais ceci n’est peut-être qu’une simplification séduisante des choses.

Hev b 1 semble, en effet, avoir une grande affinité pour les grains d’amidon des gants poudrés et les taux d’allergènes mesurés dans l’air ne suivent pas ceux trouvés dans les gants .

Il faut, de plus, ajouter une autre voie de sensibilisation au latex : celle des contacts non spécifiques, tels ceux de la vie courante ou dans le cadre de métiers sans rapport avec la santé.

Chez les sujets qui se sont ainsi sensibilisés à leur insu, on pourra rencontrer des anaphylaxies per-opératoires.

Les moyens diagnostiques

En dehors des tests réalistes, on peut faire appel à des tests de réactivité cellulaire :

  • il est parfois utile de combiner plus d’un extrait en prick-test . Et cela ne suffit pas toujours
  • des tests en cytométrie de flux ont été proposés, le plus souvent avec une activation des basophiles repérée à l’aide d’un marquage de CD63 : c’est le « basophil activation test » (BAT) . Des résultats variés ont été trouvés, la corrélation entre ce test et le prick étant plutôt limitée (r = 0,41) , du même ordre que pour le CAP (ex. )
    Le choix du cut-off désignant une activation positive peut avoir influencé ces discordances entre travaux car chaque équipe définit son seuil. Cette technique mériterait d’autres travaux car elle explore une réactivité cellulaire, plus proche de la réalité clinique que les tests sérologiques.

S’agissant de ces derniers, la panoplie des outils diagnostiques a été améliorée :

  • par l’ajout de rHev b 5 dans l’extrait servant à préparer les CAP latex : un gain de sensibilité d’environ 7% a été obtenu avec ce CAP surchargé , avec une légère baisse de spécificité  : des CAP latex négatifs auparavant, malgré un TC positif, ont ainsi été trouvés positifs avec le nouveau CAP latex
  • la disponibilité de plusieurs allergènes recombinants : rHev b 1, 3, 5, 6.01, 6.02, 7, 8, 9, 10 et 11.

Malgré tout, un extrait contient une variété de protéines (et donc d’allergènes) qui est difficilement reproductible avec une combinaison de recombinants : ainsi, entre 8 et 13% des patients restent négatifs avec tous les recombinants malgré un TC et/ou un CAP latex global positif .

Par ailleurs les recombinants distribués par Phadia sont des protéines de fusion comprenant l’allergène et une autre protéine qui lui est « attachée ». Dans le cas présent il s’agit de MBP (maltose binding protein) et l’on teste donc des MBP-rHev b 1, etc...

Bien sûr, la validité de ces MBP-recombinants implique que les résultats ne soient pas dus à une réactivité des IgE du patient pour la MBP elle-même. Peu de travaux ont inclus une vérification de cette absence de MBP-réactivité chez les patients étudiés . Dans l’ensemble, une absence de réactivité était constatée , mais cette interférence ne peut être totalement éludée.
Par exemple, Rozynek a trouvé 8 sujets positifs pour la MBP parmi 33 patients . Ceci étant, les tests MBP positifs restaient faibles (<1,3 kU/l) et n’étaient pas vus chez les patients négatifs pour les recombinants (pas de faux positifs).

Les allergènes recombinants peuvent-ils remplacer les tests globaux pour le latex ?

Partiellement oui.

D’un côté il est peu recommandable d’effectuer une dizaine de tests (autant que d’allergènes individuels différents) au lieu d’un seul. Or aucun des allergènes du latex n’est suffisant en soi pour assurer une bonne sensibilité : au mieux, les prévalences de positivité sont de l’ordre de 80 % (Hev b 1, Hev b 6.02).

Ce n’est pas comme Bet v 1 (95 %) qui est un candidat recevable pour remplacer le test bouleau.

La positivité chez un même patient se répartit sur plusieurs allergènes. Cependant, l’utilisation d’une batterie de recombinants ne garantit pas une efficacité proche de 100 % :

  • 8 patients/61 (13 %) sont négatifs pour les 9 recombinants testés mais positifs pourtant avec le CAP latex
  • en tests cutanés, 8 recombinants différents ne donnent aucun positif chez 4 sujets/49 (8 %) .

D’un autre côté les recombinants actuellement testables in vitro ont l’avantage de ne pas être perturbés par la présence d’IgE anti-CCD. Ce qui est loin d’être le cas pour les tests globaux sur extrait de latex . Trois allergènes, au moins, concourent à une réactivité CCD dans le latex : Hev b 2, Hev b 4 et Hev b 13 (cf. Latex et CCD).

Place des recombinants

1 - Pour le diagnostic de l’allergie au latex

Hev b 1 et Hev b 3 sont beaucoup plus fréquemment positifs chez les SB et les poly-opérés que chez les PS.

Pour les autres allergènes principaux (Hev b 5, Hev b 6) il n’y a pas de différence suffisante pour être utile (cf. le tableau des prévalences).

Qu’apporterait la mesure de la réactivité pour Hev b 1 et/ou Hev b 3 ?

Comme on n’a pas besoin de tester Hev b 1 et/ou Hev b 3 pour savoir que l’enfant est SB ou poly-opéré, l’utilité de ces allergènes en pratique courante est encore à démontrer.

Par ailleurs, le parallélisme entre Hev b 1 et Hev b 3 chez le même patient inciterait à ne tester qu’un seul de ces 2 allergènes.


Les cas de mono-réactivité à 1 seul allergène du latex sont assez rares :

Hev b
Ref Recrutement Nb 1 5 6.01 8 11 13
CAP latex+ 61 1 3 3
CAP latex+ 21 1 1
profess. Santé 49 1
Allergie latex 21 2 1
poly-polliniques 28 14 1
spina bifida 31 3 1
profess. Santé 104 7 3 3 1


Pour Radauer, Hev b 8 pourrait avoir une relevance clinique chez les sujets mono-réactifs à cet allergène . Mais des résultats plus récents sont plutôt en faveur d’une trace immunologique due à la pollinose et dont l’impact clinique est loin d’être clair :

  • Beaudouin montre 14 mono-réactifs à rHev b 8 parmi 28 patients poly-polliniques asymptomatiques pour le latex . Ces sujets Hev b 8 positifs étaient aussi Bet v 2 positifs
  • parmi des patients positifs en CAP pour le latex : 11/21 sont positifs pour rBet v 2 mais n’ont pas d’allergie au latex, contre 0/17 en cas d’allergie au latex
  • une absence de pertinence clinique pour Hev b 8 a également été avancée à la suite d’études avec tests réalistes (gant en latex) . Ces résultats sont cependant plus ou moins critiquables car la présence de taux notables de profiline dans les gants est douteuse ...

De l’avis de la majorité des experts, les allergènes les plus importants à tester sont Hev b 5 et Hev b 6.01 (ou Hev b 6.02), ainsi que Hev b 1 (ou Hev b 3) chez les sujets avec un spina bifida.

Différentes combinaisons ont été évaluées :

  • rHev b 5 et rHev b 6.01 : sensibilité de 100% pour une positivité pour l’un ou les 2 allergènes dans un groupe de 21 allergiques au latex . Cependant cette combinaison est positive aussi chez 10/28 poly-polliniques non allergiques au latex (CAP+ latex)
  • on peut associer en plus rHev b 8 (ou rBet v 2) : avec les patients étudiés par Beaudouin aucun rHev b 8 n’est positif parmi les 19 rHev b 5 et/ou rHev b 6.01 positifs chez les sujets allergiques au latex (n=21), tandis que rHev b 8 est positif 15 fois sans réactivité pour rHev b 5 ni rHev b 6.01 chez les sujets poly-polliniques non allergiques au latex (n=28)
  • Raulf-Heimsoth recommande rHev b 5 + rHev b 6.01, en association avec Hev b 2 ou Hev b 13. Cependant ces deux derniers allergènes ne sont pas disponibles en routine actuellement. Par ailleurs, la distinction entre allergie et réactivité sans symptômes n’est pas assez claire : si 89% des spina bifida avec allergie sont positifs pour rHev b 5 et/ou 6.01 et/ou 2, 58% des spina asymptomatiques avec le latex sont eux aussi positifs

Le cas des patients ayant présenté un choc per-opératoire a été insuffisamment étudié sur le plan moléculaire. Bienvenu rapporte une série de 28 patients pour lesquels 8 recombinants différents ont été testés (cf. tableau des prévalences).

  • Il n’est pas possible, cependant, de tirer une enseignement en préopératoire avec la mesure de la réactivité pour tel ou tel allergène.
  • Rien de très spécifique n’est noté concernant ces patients, même si la positivité (a posteriori) pour rHev b 5 est un peu plus fréquemment observée en cas de choc.
  • Il faudrait peut-être réaliser des études comparant des sujets ayant subi un accident per-opératoire avec des sujets, positifs aussi pour le latex avant opération, mais n’ayant pas montré de réaction per-opératoire, à geste équivalent.

Au total, l’utilisation des différents allergènes disponibles in vitro est d’un intérêt limité pour avoir une vue plus précise sur une allergie au latex déjà caractérisée par l’enquête clinique et les TC.

Une exception pourrait être la discordance entre un TC négatif et une histoire clinique en faveur d’une allergie au latex.

  • Il semble, en effet, qu’un des allergènes importants, Hev b 5, soit plus difficile à récupérer dans les extraits que d’autres allergènes . Dans les rares cas où le patient se serait mono-sensibilisé à Hev b 5, on pourrait constater un TC négatif. C’est la raison pour laquelle le CAP latex a été surchargé récemment en rHev b 5. Cette addition permet d’augmenter (modestement) la sensibilité du CAP latex au détriment de la spécificité .

Au total, l’utilisation des allergènes recombinants ne permet pas, à l’heure actuelle, de différencier allergie et sensibilisation de façon nette.

Les allergènes recombinants du latex apportent cependant une aide pour confirmer un diagnostic, notamment en cas pollinose associée et/ou de présence d’IgE anti-CCD.

Contrairement à des tests comme rFel d 1 ou rBet v 1 dont l’utilisation peut se concevoir en première intention, les recombinants du latex ont plus leur place après une première approche comprenant déjà un test in vitro latex positif.

2 - Dans le cadre de réactions croisées avec des aliments, etc..

Certains allergènes du latex n’ont pas d’homologues IgE-réactifs connus (pour l’instant) dans d’autres produits : Hev b 1, Hev b 3, Hev b 4, Hev b 5.

C’est le cas aussi de protéines IgE-réactives nouvellement caractérisées comme l’UDPG-pyrophosphorylase, la rotamase ou la citrate-binding protein .

Pour d’autres protéines, la présence d’homologues alimentaires existe mais la réactivité croisée n’est pas connue : l’isoflavone réductase et la thioredoxine , la triose-P isomérase , l’hévamine.

La réactivité croisée d’Hev b 2 et celle d’Hev b 13 est a priori de nature CCD.

La profiline (Hev b 8), la LTP (Hev b 12), la patatine-like (Hev b 7), l’énolase (Hev b 9) et la superoxyde dismutase (Hev b 10) ont un intérêt mal défini tant pour l’allergie au latex que pour la réactivité croisée avec d’autres produits que le latex :

  • Hev b 8 : tester la profiline du latex n’apporte pas de renseignement particulier :
    • pour Ganglberger, la positivité pour Hev b 8 est la conséquence d’une sensibilisation pollinique (ou par un aliment végétal) .
    • Il semble donc plus pertinent de tester une profiline pollinique (rBet v 2 ou rPhl p 12) pour savoir si le patient est sensibilisé aux profilines.
  • Hev b 12 :
    • chez des sujets allergiques au latex, la positivité pour la LTP Hev b 12 ne s’accompagne pas fréquemment d’une allergie à des aliments “ de type LTP ” (ex. Rosacées) .
    • De même, pour Rihs, rHev b 12 est parfois positif chez des patients allergiques aux fruits, notamment en Espagne, mais cela représente la trace d’une sensibilisation alimentaire aux LTP
  • Hev b 7 : des prévalences non négligeables de positivité ont été relevées pour la patatine-like, Hev b 7, chez des patients allergiques au latex (cf. Tableaux de prévalences).
    • Cependant, la réactivité croisée entre Hev b 7 et la patatine de pomme de terre a une relevance clinique restant à démontrer : parmi des sujets sélectionnés sur la base d’une réactivité/allergie à la pomme de terre on ne trouve pas une fréquence inhabituelle d’allergiques au latex. (cf. Pomme de terre et latex)
  • Hev b 9 : cet allergène a des homologues dans les moisissures.
    • In vitro, la positivité pour rHev b 9 varie de 0 à 15% chez les sujets allergiques au latex et a été trouvée à 21-23% chez des sujets allergiques aux moisissures .
    • La réactivité pour rHev b 9 semble pas primitive, cet allergène inhibant mal ses homologues fongiques .
    • L’intérêt d’Hev b 9 dans l’exploration d’un sujet allergique au latex semble limité. On pourrait éventuellement tester rHev b 9 devant une réactivité in vitro pour le latex non confirmée en test cutané chez un patient allergique aux moisissures .
  • Hev b 10 : cet allergène a également des homologues dans les moisissures.
    • In vitro, la fréquence de positivité pour rHev b 10 a été trouvée le plus souvent proche de zéro chez des allergiques au latex et de 12% chez des allergiques aux moisissures .
    • rHev b 10 inhibe mal ses homologues fongiques .
    • Le cas d’Hev b 10 reste cependant ouvert car il a été constaté des fréquences inattendues de TC positifs pour Alternaria chez des sujets TC positifs latex (et vice versa), de même que pour la noix . Or, la noix, comme Alternaria, possède une Mn superoxyde dismutase IgE-réactive … (cf. Latex et fruits à coque).

C’est avant tout l’hévéine (Hev b 6.02), la pro-hévéine (Hev b 6.01) et la chitinase du latex (Hev b 11) qui sont concernées par les réactivités croisées latex-aliments végétaux (cf. Latex et aliments).

  • La réactivité croisée entre Hev b 6.02 et Hev b 6.01 est évidente, ces 2 protéines ayant un domaine identique (cf. schéma des chitinases).
  • Il est dès lors inutile de tester in vitro à la fois rHev b 6.01 et rHev b 6.02. Les rares cas de discordance entre ces 2 allergènes se rencontrent avec des réactivités toujours très faibles .
  • Hev b 6.02 est présent dans le latex naturel. Il provient du clivage d’Hev b 6.01.
  • Pour savoir si le patient est susceptible de réagir à des aliments par réactivité croisée avec le latex, le meilleur choix pourrait consister en rHev b 6.02. Mais certains auteurs préfèrent rHev b 6.01.

Pourquoi ne pas tester d’emblée rHev b11 ?

  • Si des chitinases alimentaires sont le support d’une association latex-aliment, la chitinase Hev b 11 devrait être la mieux adaptée. D’autant que le domaine hévéine d’Hev b 11 est plus homologue de ceux de banane, avocat ou châtaigne (72 à 81 % d’identité) que de l’hévéine elle-même (58 %).
  • Mais la relevance clinique de l’hévéine Hev b 6.02 s’avère très nettement supérieure à celle d’Hev b 11.
  • Et les sujets positifs pour rHev b 11 sont en règle générale positifs pour rHev b 6.02 et/ou rHev b 6.01 .
  • Enfin, quantitativement, la réactivité de rHev b 11 est le plus souvent inférieure à celle de rHev b 6.01 ou rHev b 6.02 .

Malgré tout, il existe des cas avec rHev b 11 positif et rHev b 6.01/6.02 négatif  ; et, parfois, l’allergie à des fruits exotiques débute avant celle pour le latex (cf. Syndrome latex-aliments). Des études complémentaires seraient nécessaires, particulièrement chez ces patients d’abord réactifs aux fruits, pour être en mesure d’attribuer un rôle primitif à la chitinase Hev b 11.

3 - Dans le suivi des patients

Les mesures d’éviction du latex ont fait l’objet de nombreuses études de suivi.

Le plus souvent ces études ont associé aux critères cliniques des résultats d’exploration par TC et/ou CAP latex, c’est-à-dire sans discrimination parmi les différents allergènes du latex . En règle générale, le prick-test reflétait mieux l’amélioration clinique que ne le faisait le CAP.

Une étude récente a recherché si l’évolution d’un allergène précis pourrait apporter plus de renseignements que l’évolution du test global latex  :

  • les tests cutanés effectués avec un extrait latex et 8 allergènes purifiés montrent, après 5 ans de mesures d’éviction du latex, une évolution qui est beaucoup moins nette que celle des paramètres cliniques ;
  • et sur les 8 allergènes purifiés testés aucun ne se distingue et pourrait constituer un témoin fidèle d’amélioration clinique.

4 – Pour l’estimation du risque de récidive per-opératoire

Rien de précis. Parfois il a été relevé une fréquence plus grande de résultats positifs pour rHev b 1 et/ou rHev b 3 en cas de choc per-opératoire (9 sujets/28) que pour d’autres situations d’allergie au latex (1/34) . A noter qu’il s’agissait de patients adultes.

5 - Les recombinants et l’immunothérapie

Le nombre des études portant sur l’immunothérapie (IT) d’une allergie au latex est restreint.

Une relation inverse (r = -0,72) a été notée entre le résultat pour rHev b 6.01 avant IT et la dose maximale tolérée pendant l’IT . Mais, dans une autre étude, rHev b 5 ou 6.01 ou 6.02 ne permettaient pas de prédire le risque de réaction clinique au cours de l’IT .

Quelques néo-réactivités en blot ont été observées après 6 mois d’IT : 1 pour Hev b 5 (sur 16 patients), 2 pour Hev b 6.01 et 2 pour Hev b 11. Toutes ces néo-réactivités étaient faibles.

Un autre travail a comparé différents recombinants avant le début de l’ITSL et 1 an après chez 9 patients adultes : globalement, les positivités se sont peu modifiées, notamment pour rHev b 5 et rHev b 6 . C’est aussi le cas pour la réactivité cellulaire (test BAT) vis à vis de rHev b 6.01/6.02 : après une baisse à +6 mois, cette réactivité revient dans la zone des valeurs initiales à +1 an .

Dans l’état actuel des choses, il semble difficile de tirer parti des recombinants dans ce cadre thérapeutique.

Latex et CCD

(voir aussi : Les CCD)

Le latex d’Hevea contient des glycoprotéines, dont des allergènes comme Hev b 2, Hev b 4 et Hev b 13. Les chaînes glucidiques de ces allergènes ont montré leur capacité d’IgE-réactivité .

La pertinence des tests in vitro pour le latex est fortement perturbée par la présence d’IgE anti-CCD dans le sérum des patients .

De très nombreuses observations le prouvent :

  • la prévalence des RAST latex positifs est de 7,3 % l’été contre 3,9 % l’hiver
  • l’excédent de tests latex in vitro positifs par rapport aux TC est très net dans les études de populations générales mais pas dans les cohortes de populations à risque
  • une corrélation est notée entre les résultats pour le latex et ceux pour le pollen de dactyle (r = 0,7)
  • parmi des sujets ayant un CAP latex positif, 10 % sont effectivement allergiques au latex s’ils sont par ailleurs polysensibilisés, alors que tous les mono-positifs latex sont allergiques au latex
  • parmi 1236 enfants Hourihane relève 4 % de CAP latex positifs mais seulement 0,47 % ont une véritable allergie au latex
  • des groupes contrôles s’avèrent CAP latex positifs de façon inattendue : par exemple 12/30 polliniques à l’armoise
  • les sujets allergiques aux hyménoptères présentent une proportion tout à fait anormale de résultats in vitro positifs pour le latex . Jappe relève 80% de CAP latex positifs dans une cohorte de patients avec double positivité abeille-guêpe due à des CCD (cf. Hyménoptères et latex).

Clairement, la pollinose (surtout la poly-réactivité pollinique ) et l’allergie aux venins d’hyménoptères génèrent des IgE capables de positiver faussement les tests in vitro pour le latex.

Dans une moindre mesure, l’alcoolisme peut également générer une IgE-réactivité de type CCD et entraîner des résultats latex faussement positifs .

Des glyco-reporters, comme la broméline ou la peroxydase de raifort (HRP), ont été testés et aboutissent à la même conclusion :

  • sur 23 sujets CAP positifs mais TC négatifs, le CAP broméline était positif 18 fois. Aucun des CAP et TC positifs n’était positif pour la broméline .
  • Parmi 23 enfants avec rhinite (souvent en lien avec des graminées) et un test in vitro positif pour le latex mais asymptomatiques au contact du latex, 18 étaient positifs pour un glyco-reporter .
  • Parmi 21 patients random avec CAP positif latex mais sans histoire clinique positive pour le latex, 5 sont trouvés CAP broméline positifs . Mieux 1 de ces 5 patients n’est positif pour aucun des 9 recombinants testés, le CAP latex étant donc apparemment positivé uniquement par une réactivité CCD.
  • dans une autre étude comportant aussi 9 recombinants, sur 6 patients négatifs pour les 9 recombinants 4 étaient positifs en broméline
  • une corrélation étroite entre latex et un glyco-reporter (HRP) existait chez des sujets doublement positifs abeille+guêpe

Avec une technique d’immuno-capture par la broméline et HRP, il est possible de montrer l’impact de cette réactivité CCD : elle représente parfois la totalité du résultat initial .

Enfin, l’interférence des IgE anti-CCD dans les tests sériques pour le latex est confortée par l’observation d’un nombre restreint de tests cellulaires (ex. le BAT) trouvés positifs en comparaison du nombre des bromélines positives chez les mêmes patients : 4 au lieu de 20 parmi 24 patients avec CAP positif mais TC négatif .

Devant cette difficulté majeure des tests in vitro basés sur les mélanges complexes que sont les extraits, on peut :

  • tester un glycoreporter (ex broméline ou HRP) : si le résultat est négatif, il faudra chercher pourquoi le test in vitro latex était positif contrairement aux autres données du dossier.
  • Si le résultat est positif, la valeur du test in vitro pour le latex est fragilisée. Mais on ne peut pas retrancher les kU/l trouvés pour la broméline des kU/l trouvés pour le latex. Aucun parallélisme n’existe entre les glycoreporters et les autres tests .
  • tester 1 ou 2 recombinants, si possible adaptés au cas du patient. Cela n’aura pas beaucoup d’intérêt pour les SB. Pour les patients non exposés professionnellement on pourrait suggérer de tester rHev b 5 et rHev b 6.02.
  • l’idéal serait d’épuiser le sérum de toutes ses IgE anti-CCD et d’effectuer la mesure in vitro avec un test latex classique. Mais cette technique de pré-traitement n’est pas disponible en routine pour le moment.

Latex et succédanés du latex

Si de nombreuses plantes sont capables de sécréter un latex (ex. le Ficus, des salades, etc..), les propriétés technologiques et les rendements à l’hectare du latex d’Hevea ne sont pas concurrencés par un autre latex jusqu’à présent.

L’écorce d’une plante tropicale, le Parthenium argentatum, fournit un latex qui possède des propriétés assez bonnes et qui ne semble pas montrer de réactivité croisée avec le latex d’Hevea . Des objets fabriqués avec ce latex, la guayule, pourraient donc convenir pour des patients allergiques au latex d’Hevea.

On manque cependant de travaux confirmant ces résultats préliminaires. Et absence de réactivité croisée n’implique pas absence d’immunogénicité.

Réactions croisées avec le latex (hors cadre alimentaire)

Gutta-percha

La gutta-percha, ou gomme gutte, est tirée du latex contenu dans les feuilles d’un arbre, le Palaquium gutta (Sapotacées). Cette gomme est utilisée notamment en soins dentaires.

Quelques travaux n’ont pu retrouver de réactivité croisée entre la gutta-percha et le latex d’Hevea  ; du moins avec le produit tel qu’utilisé en art dentaire et non la gomme native brute qui, elle, semble croiser avec le latex d’Hevea .

Un cas d’allergie à la gutta-percha chez un patient allergique au latex a été publié , mais dont l’exploration était insuffisante.

Poussières de bois

Certains asthmes professionnels ont pour cause les poussières de bois, notamment des bois exotiques.

Des travaux anciens avaient montré une réactivité croisée entre le bois d’abachi (Triplochiton scleroxylon, Sterculiacées) et celui de ramin (Gonystylus bancanus, Thyméléacées).

Le bois d’abachi a été plus étudié et une chitinase de classe 1 a été identifiée IgE-réactive dans ce bois . Une réactivité croisée existe entre le latex d’Hevea et le bois d’abachi . Mais la sensibilisation au bois d’abachi semble peu efficace pour induire une allergie au latex car les essais de réactivité croisée avec l’hévéine (Hev b 6.02) ont donné des résultats le plus souvent négatifs .

A noter un cas d’allergie à la tamarille (fruit du Cyphomandra betacea) chez un menuisier ayant un asthme au bois d’abachi et négatif en TC pour le latex .

Un autre bois, le jetulong (Dyera costulata) est également susceptible de réactivité croisée avec le latex d’Hevea .

Autres produits

Une réactivité croisée entre Ficus et latex d’Hevea est connue. D’autres plantes d’agrément peuvent avoir un lien avec le latex : ainsi parmi 82 patients consultant pour une allergie respiratoire et présentant un prick positif pour une plante d’intérieur, 7% étaient également TC positifs pour le latex (et négatifs pour le Ficus) .

Une réactivité croisée entre latex et Poinsettia a d’ailleurs été avancée .

Cela est le cas également pour la feuille de tabac , la réaction croisée pouvant concerner un homologue de la pro-hévéine, le CPB20, dans le tabac .

Avec l’écorce de condurango (Marsdenia condurango, Apocynacées) il est préparé des décoctions vendues pour des propriétés médicinales. Un cas d’anaphylaxie à ce type de produit a été décrit chez une patiente allergique au latex .

[1] - Braun JJ, de Blay F. La rhinite allergique au latex : du diagnostic à la prise en charge. Rev Fr Allergol Immunol Clin 2008;48:237-241
Malgré de nombreuses publications concernant la pathologie respiratoire professionnelle, l‚allergie au latex et les relations nezˆbronches, la rhinite allergique au latex (RAL) dans la littérature n‚est que très incomplètement individualisée dans ses aspects cliniques, épidémiologiques, évolutifs et thérapeutiques. À travers leur expérience et la revue de la littérature, les auteurs cherchent à situer la RAL dans son contexte environnemental et à préciser les principaux critères diagnostiques et modalités thérapeutiques. La précocité du diagnostic, avant la survenue de l‚asthme, et les mesures d‚éviction du latex permettent de limiter l‚impact socioéconomique de la RAL.
[5] - Mavale-Manuel S, Paty E, Scheinmann P, de Blic J. Allergie au latex chez l'enfant. Arch Pediatr 2003;10:700-706
The aim of our study was to determine the prevalence of latex allergy and the clinical features of children with latex allergy. PATIENTS AND METHODS: We prospectively investigated 243 children consulting in our allergy out-patients unit during 1 year. Parents answered a questionnaire, and children underwent skin prick tests with common allergens and latex. Latex-specific serum immunoglobulin E was determined by CAP test in children with latex sensitization. The results were compared in children with and without latex allergy . RESULTS: The prevalence of latex allergy was 1.3%. A family history of atopy (75%) and a personal history of previous surgery was associated with latex allergy (P < 0.0001). In children with latex allergy, the frequency of sensitization to inhaled and food allergens, atopic dermatitis, rhinitis and conjunctivitis was higher than in children without latex allergy (P < 0.05). Avocado allergy was the food allergy most commonly associated with clinical symptoms. Balloon was the most common latex product causing symptoms (60%) . CONCLUSIONS: Due to its potential severe consequences, latex allergy should be investigated in children who had undergone multiple surgical procedures and in the children with pollen-food allergy syndrome. Avoidance of latex is an important preventive measure.
[7] - Mertes PM, Malinovsky JM, Alla F, Studnicska D, Tréchot P, Laxenaire MC. Epidémiologie des réactions d'hypersensibilité immédiates peranesthésiques chez l'adulte et l'enfant: bilan de 8 années du Gerap. Rev Fr Allergol Immunol Clin 2008;48:217-221
Les réactions d‚hypersensibilité immédiate, qu‚elles soient médiée par les immunoglobulines E (HSI IgE) ou non (HSI non IgE) survenant au cours de l‚anesthésie demeurent un sujet de préoccupation majeure. Les auteurs rapportent les données de huit années de surveillance épidémiologique effectuée par le Gerap. Deux mille cinq cent seize patients ayant présenté une réaction clinique évocatrice d‚une hypersensibilité immédiate entre le 1er janvier 1997 et le 31 décembre 2004 ont été inclus dans l‚étude. Une réaction non IgE-médiée a été diagnostiquée dans 700 cas (27,82 %), un diagnostic de réaction à médiation IgE ayant été retenu dans 1816 cas (72,18 %). Les agents plus fréquemment incriminés étaient les curares (n =1067, 58,08 %), suivi du latex (n =361, 19,65 %) et des antibiotiques (n =236, 12,85 %). Chez l‚enfant, les résultats étaient significativement différents. Le latex étant impliqué dans plus de 50 % des cas, suivi des curares (39 %) et des antibiotiques (10,2 %). Chez l‚adulte, les résultats confirment l‚existence d‚une prédominance féminine, quelque soit le mécanisme de la réaction, cette prédominance n‚est pas retrouvée chez l‚enfant. Ces résultats confirment la nécessité d‚un bilan systématique en cas de survenue d‚une réaction d‚hypersensibilité immédiate survenant au cours de l‚anesthésie et l‚intérêt du développement de centres de consultation spécialisé en allergo-anesthésie capables d‚apporter une aide aux anesthésistes et aux allergologues.
[8] - Ebo DG, Stevens WJ. IgE-mediated natural rubber latex allergy: practical considerations for health care workers. Ann Allergy Asthma Immunol 2002;88:568-575
OBJECTIVE: To focus on some specific topics of major significance for health care workers confronted with natural rubber latex (NRL) allergy: 1) sensitization and routes of exposure; 2) threshold allergen exposure levels; 3) evaluation of occupational NRL allergy; 4) prevention of sensitization and allergic reactions to NRL-containing devices; 5) selecting the right alternative; and 6) regulatory responses to NRL allergy issues. DATA SOURCES: English and French language papers identified through a MEDLINE search and bibliographies of the identified papers and the National Institute for Occupational Safety and Health and Food and Drug Administration web sites . RESULTS: 1) The threshold levels of NRL exposure needed for sensitization and the precise way in which it comes about remain to be elucidated, but cutaneous, percutaneous, mucosal, and parenteral exposure can all give rise to symptoms. 2) Strengths and weaknesses of the questionnaire and currently available diagnostic techniques for NRL allergy are emphasized. 3) Prevention strategies should focus on dipped materials and stress upon the possibility of passive and active transmission of NRL aeroallergen. 4) Generally, vinyl gloves are an appropriate alternative for NRL gloves; however, when barrier integrity is a concern, nitrile gloves provide protection that is comparable with NRL. 5) Several government bodies, standards organizations, and regulatory agencies have issued regulations concerning the control and labeling of protein and allergen levels in NRL products . CONCLUSION: Accurate diagnosis and management of NRL allergy is essential because of the potential for severe hypersensitivity reactions. Major developments have been made in characterizing and cloning NRL allergens, and future development in this area may lead to better diagnostic tools and possible therapeutic agents for immunotherapy. However, at present, avoidance remains the only effective approach. We hope that additional well performed prospective incidence studies will bring more accurate data concerning sensitization doses and provide the valuable information to improve prevention strategies.
[9] - Nettis E, Micale G, Fanelli M, Serio G, Colanardi MC, Ferrannini A, et al. Atopy and Risk Factors for Latex Sensitization in a Selected Population. Allergy Asthma Proc 2003;24:185-191
Abstract: The value of recommending latex allergy screening in allergy departments of the Army's Hospital was studied. The purpose of the study was to evaluate whether atopy was a risk factor for latex sensitization in a specific population such as the young male soldiers of the Italian Army. The study was also aimed to assess the role of other risk factors. One thousand five hundred male subjects (1000 subjects who were atopic and 500 subjects who were nonatopic), visiting the Department of Allergology and Respiratory Physiopathology of the Army's Hospital in Bari, Italy, were enrolled into the study. The protocol included a questionnaire (symptoms of atopy, use of latex gloves and condoms and possible reactions previous surgical procedures), a clinical examination, a skin-prick test to latex and common allergens to evaluate atopy, and in part a latex challenge. Among the 1000 subjects who were atopic, 2.8% had evidence for sensitization to latex compared with 1.2% in the 500 subjects in the nonatopic group. The risk of latex sensitization was 19 times higher for subjects with a history of reactions to latex exposure and had a twofold increase for each surgical procedure and for each skin test positivity for inhalant allergens. Another risk factor was positivity to skin-prick tests for Artemisia vulgaris , cypress, and molds. Atopy significantly relates to an increased risk of latex sensitization. Screening is recommended in the Army's Hospital to identify latex-sensitized subjects and inform them about the risks connected with this condition.
[10] - Garabrant DH, Schweitzer S. Epidemiology of latex sensitization and allergies in health care workers. J Allergy Clin Immunol 2002;110(2 suppl.):S82-S95
Although it is often claimed that health care workers are at increased risk of latex sensitization and type I allergies, there has been no systematic analysis of the epidemiologic studies that are relevant to this conclusion. A systematic analysis of the epidemiologic literature found that, in the past 14 years, there have been 48 epidemiologic studies of type I latex allergy among health care workers. Of these, 2 cohort studies estimated the incidence of latex sensitization by skin prick testing at between 1% and 2.5% per year. Neither compared the risk to that in the general population. The prevalence of sensitization in health care workers varied between 0% and 30%, yet this large variation was unexplained. Increased risk of sensitization was not clearly associated with the duration of work in health care, the time spent wearing latex gloves, the frequency of exposure, the specific job categories, the use of powdered versus nonpowdered latex gloves, the use of latex versus nonlatex gloves, or any measurements of ambient exposure to latex proteins. The epidemiologic studies do not support a conclusion that health care workers are at clearly increased risk of latex sensitization or type I allergies compared to other occupations in the United States. The role of latex gloves in causing latex sensitization and type I allergic symptoms remains poorly defined because of the inconsistent results across studies. Future epidemiologic studies are needed that include measured exposures to latex antigens, that compare health care workers to appropriate referent groups, and that address confounding by atopy, age, sex, and race.
[11] - Mertes PM, Mouton C, Frémont S, Brugerolle B, Moneret-Vautrin DA, Lavaud F, et al. Latex hypersensitivity in spinal cord injured adult patients. Anaesth Intensive Care 2001;29:393-399
Latex hypersensitivity is a major cause of anaphylaxis during anaesthesia. Patients with spina bifida, health care or rubber industry workers have been considered at risk for latex sensitization. By analogy, the existence of other at-risk subsets of patients with latex exposure due to frequent surgical procedures has been suggested. The aim of this study was to evaluate the prevalence of latex sensitization in a cohort of adult patients with spinal cord injury and repeated latex exposure. Forty-two adult patients with spinal cord injury were studied and retrospectively compared to a group of 30 children with spina bifida evaluated using a similar protocol. Patients were administered a questionnaire concerning history of latex hypersensitivity, atopy, and surgical procedures. Latex sensitivity was investigated by skin prick-tests and latex-specific IgE assay. The search for atopy was based on in vivo and in vitro tests against a panel of environmental allergens. No chronic spinal cord injured patient had a history of latex allergy. When compared with spina bifida, the number of surgical procedures was not statistically different. Although not significantly different, the prevalence of atopy was higher in spina bifida patients. The high level of latex sensitization in spina bifida patients contrasted sharply with the absence of sensitization observed on both skin and in vitro tests in patients with spinal cord injury (P<0.0001). This study confirms that adult patients with chronic neurologic defects resulting from spinal cord injury exhibit a low risk of latex sensitization. These results suggest that considering adult patients with repeated surgical procedures as a group at risk for latex sensitization because of a high degree of latex exposure should be re-examined. Latex hypersensitivity is a major cause of anaphylaxis during anaesthesia. Patients with spina bifida, health care or rubber industry workers have been considered at risk for latex sensitization. By analogy, the existence of other at-risk subsets of patients with latex exposure due to frequent surgical procedures has been suggested. The aim of this study was to evaluate the prevalence of latex sensitization in a cohort of adult patients with spinal cord injury and repeated latex exposure. Forty-two adult patients with spinal cord injury were studied and retrospectively compared to a group of 30 children with spina bifida evaluated using a similar protocol. Patients were administered a questionnaire concerning history of latex hypersensitivity, atopy, and surgical procedures. Latex sensitivity was investigated by skin prick-tests and latex-specific IgE assay. The search for atopy was based on in vivo and in vitro tests against a panel of environmental allergens. No chronic spinal cord injured patient had a history of latex allergy. When compared with spina bifida, the number of surgical procedures was not statistically different. Although not significantly different, the prevalence of atopy was higher in spina bifida patients. The high level of latex sensitization in spina bifida patients contrasted sharply with the absence of sensitization observed on both skin and in vitro tests in patients with spinal cord injury (P<0.0001). This study confirms that adult patients with chronic neurologic defects resulting from spinal cord injury exhibit a low risk of latex sensitization. These results suggest that considering adult patients with repeated surgical procedures as a group at risk for latex sensitization because of a high degree of latex exposure should be re-examined.
[12] - Hilgert E, Jund F, Klemens C, Pfrogner E, Pauli C, Rasp G, et al. Latex allergy, a special risk for patients of otorhinolaryngology and head and neck surgery ? Am J Otolaryngol 2007;28:103-109
A total of 206 patients of the otorhinolaryngology (ORL) department and 204 of the visceral surgery department of the Ludwig Maximilians University of Munich were preoperatively evaluated for latex-specific sensitization. A prick test, a questionnaire, and an enzyme-linked immunosorbent assay immunoprecipitation for IgE antibodies were performed. Latex is a widely spread allergen, and it does not only concern healthcare populations. Within the ORL surgery group, 43 (20.9%) patients were sensitized against latex allergen, and 2 of them were reported to manifest symptoms in consorting with latex. In the cohort of visceral surgery patients, we detected only 23 patients (11.3%) with sensitization against latex. Moreover, most patients were positively detected with the skin prick test (86.4%), whereas the enzyme-linked immunosorbent assay method was less sensitive (18.3%). Patients of the ORL department were considerably more frequently exposed to latex protein particles than patients of the visceral surgery department. This difference attributes to their significant difference in mean age: 44 years in the ORL patients group versus 58 years in the visceral patients cohort. Furthermore, we did not find any correlation to the number of past operations--although undergoing any surgical procedures is a well-known risk factor in other studies about latex sensitization in surgical patients.
[13] - Eiwegger T, Dehlink E, Schwindt J, Pomberger G, Reider N, Frigo E, et al. Early exposure to latex products mediates latex sensitization in spina bifida but not in other diseases with comparable latex exposure rates. Clin Exp allergy 2006;36:1242-1246
BACKGROUND: The high prevalence of latex sensitization in patients with spina bifida (SB) has been attributed to repeated and early exposure to latex products. Other diseases such as gastroschisis/omphalocoele and post-haemorrhagic/congenital hydrocephalus are also associated with repeated and early latex exposure . OBJECTIVE: The aim of the study was to evaluate whether the high prevalence of latex sensitization in patients with SB is rather related to the underlying disease itself than to disease-associated known risk factors . METHODS: We compared children with SB (n=35), children with gastroschisis/omphalocoele (G/O, n=20) and children with post-haemorrhagic/congenital hydrocephalus (PH, n=45). All children with SB and PH had a ventriculo-peritoneal shunt since a very young age. Patients who underwent three or less surgical procedures matched in terms of age, number of operations, atopy and gender distribution, and were analysed for IgE sensitization rates to latex . RESULTS: In the SB group, 16 of 35 patients (46%) showed elevated latex-specific IgE antibodies in contrast to one of 20 patients (5%) in the G/O group and four of 45 patients (8.9%) in the PH group (P<0.0005 and P<0.005, Fisher's exact test). Comparing matched control groups (
[14] - Pires G, Morais-Almeida M, Gaspar A, Godinho N, Calado E, Abreu-Nogueira J, et al. Risk factors for latex sensitization in children with spina bifida. Allergol Immunopathol (Madr) 2002;30:5-13
BACKGROUND: Children with spina bifida represent the major risk group for latex sensitization.Purpose: To determine the prevalence of latex sensitization in these children and to identify risk factors. MATERIAL AND METHODS: We studied 57 patients with spina bifida. The mean age was 5.6 years and the male/female ratio was 0.8/1. In all patients a questionnaire, skin prick test (SPT) with latex (UCB-Stallergènes, Lofarma and ALK-Abelló), common aeroallergens and fruits (UCB-Stallergènes) and serum determination of total IgE (AlaSTAT) were performed . RESULTS: The prevalence of latex sensitization was 30 %; only two sensitized children (12 %) had symptoms after exposure. Risk factors for latex sensitization were age >/= 5 years (p = 0.008; OR = 6.0; 95 % CI = 1.7-22.1), having at least four previous surgical interventions (p < 0.0001; OR = 18.5; 95 % CI = 3.6-94.8), having undergone surgery in the first 3 months of life (p = 0.008; OR = 5.4; 95 % CI = 0.7-29.2) and total serum IgE >/= 44 IU/ml (p = 0.03; OR = 3.8; 95 %CI = 1.1-13.1). Multiple logistic regression analysis showed that only a history of four or more surgical interventions (p < 0.0001; OR = 26.3; 95 %CI = 2.9-234.2) and total serum IgE >/= 44 IU/ml (p = 0.02; OR = 8.6; 95 % CI = 1.4-53.4) were independently associated with latex sensitization. Sex, family and personal allergic history, hydrocephalus with ventriculoperitoneal shunt, cystourethrograms, intermittent bladder catheterization and atopy were not related to latex sensitization . CONCLUSIONS: In children with spina bifida, significant and independent risk factors identified for latex sensitization were multiple interventions and higher levels of total serum IgE. A prospective study will clarify the clinical evolution of assymptomatic children sensitized to latex.
[15] - Mazon A, Solera E, Alentado N, Oliver F, Pamies R, Caballero L, et al. Frequent IgE sensitization to latex, cow’s milk, and egg in children with short bowel syndrome. Pediatr Allergy Immunol 2008;19:180-183
Children with short bowel syndrome (SBS) undergo frequent operations, so they are at risk for sensitizing to latex. There have been isolated reports of sensitization to food in these children. In a cross-sectional study, we assessed sensitization to latex, cow's milk, and egg with skin prick tests (SPT) and serum-specific immunoglobulin E (IgE) in 14 children with SBS. Data were collected about the number of operations with latex devices, serum total IgE, and history of feeding with milk formula. Ten children were sensitized to latex (specific IgE median: 6.7 kU/l, range: 0.5-33). Compared with those non-sensitized, sensitized children had significantly (p < 0.05) higher levels of serum total IgE in z-units (mean rank 3.25 vs. 9.2, respectively), and more operations with latex devices (mean rank 3.75 vs. 9). Eight children were sensitized to cow's milk, one with only positive SPT, the other seven with serum-specific IgE (median: 3.5, range: 0.5-21.1 kU/l), and five to egg (specific IgE median: 0.68, range: 0.58-2.17 kU/l). Except for some isolated days with cow's milk formula, the children had been initially fed with a diet without intact cow's milk proteins. Sensitization to latex, cow's milk, and egg is very frequent in children with SBS. They should be treated in a latex-free environment since the very early stages of the disease, and should be routinely studied regarding food sensitization, as this might contribute as an added factor in the chronic diarrhea of these patients.
[16] - Yip E, Cacioli P. The manufacture of gloves from natural rubber latex. J Allergy Clin Immunol 2002;110(2 part 2):S3-S14
Gloves that will provide a barrier of protection from infectious organisms are an essential feature of medical practice for the protection of both patients and medical personnel. Natural rubber latex has consistently been the most satisfactory raw material for the manufacture of gloves. Certain latex proteins, carried over into the finished product by inadequate manufacturing processes, may pose a risk of provoking allergic reactions in some patients and medical workers. As with any allergy, the risk depends on the route of exposure and dose. Hence, the method of manufacture, including the means used to coat gloves to make donning easy, can influence the eventual exposure of sensitive people to latex allergens. In this article, we describe the several processes in use and their effects on latex protein content.
[17] - Crippa M, Belleri L, Mistrello G, Tedoldi C, Alessio L. Prevention of latex allergy among health care workers and in the general population: latex protein content in devices commonly used in hospitals and general practice. Int Arch Occup Environ Health 2006;79:1-8
Background: In this study the latex protein content in devices commonly used in hospitals and general practice were investigated. The main aim was to acquire information for preventing latex allergy in health care workers and in the general population. Methods: About 22 different types of medical devices and 23 devices commonly used in general practice were examined evaluating the total allergenic potency by a modified RAST-inhibition assay and quantitative determination of single allergens (Hev b1, Hev b5 and Hev b6.02) by using commercial ELISA kit. Results: A high level of inhibition was found in medical devices, such as elastic bandage (81.57%), tourniquet (74.09%), Foley urinary catheter (68.35%), Penrose drainage (67.25%) and taping (39.6%), and in common devices, such as rubber inner-sole (84.20%), toy balloon (78.62%), latex mattress (74.27%), household rubber gloves (49.10%), working gloves (38.25%), inflatable floating mattress (32.10%). Concentrations of latex extractable proteins and Hev b1, Hev b 5 and Hev b6.02 antigens were high in some medical and general devices. Conclusions: Latex exposure sources were found in hospitals and the home. These findings, though only preliminary and far from conclusive, could enable sensitized persons to avoid risky exposures and prevent allergic reactions. From the point of view of prevention, the time may come when every natural rubber object could be systematically labelled as "containing latex" together with the warning that "this item may cause allergic reactions in sensitized subjects.
[18] - Chardin H, Chen Z, Raulf-Heimsoth M, Mayer C, Senechal H, Desvaux FX, et al. Identification of Hev b 1 in natural latex mattresses. Int Arch Allergy Immunol 2000;121:211-214
BACKGROUND: Type 1 hypersensitivity to natural rubber latex proteins is a well-recognized health problem. Recent data have shown that allergens can be extracted from natural latex mattresses. As Hev b 1 (rubber elongation factor) and Hev b 6.02 (hevein) were described as major allergens, the present work was carried out to evaluate their presence in latex mattresses as well as in latex gloves. METHODS: Extracted proteins from latex mattresses and gloves were separated by SDS-PAGE or two-dimensional gel electrophoresis, transferred onto nitrocellulose and detected with monoclonal antibodies specific for Hev b 1 and Hev b 6.02. RESULTS: The results showed that various forms of Hev b 1, as well as degradation products of Hev b 1 were detected in latex mattresses and gloves, whereas Hev b 6.02 was not detected either in mattresses or in gloves. In a standardized latex extract, Hev b 1 and Hev b 6.01 (prohevein) were identified by the monoclonal antibodies. CONCLUSION: The fact that only Hev b 1 was detected by im munoblot in latex articles indicates that Hev b 1 may be the last protein to be washed out of latex products and that the Hev b 1 content may be used as a criterion for the estimation of the allergenicity of the latex products.
[19] - Untersmayr E, Lukschal A, Hemmer W, Harwanegg C, Breiteneder H, Jarisch R, et al. Exercise with latex sport bands represents a risk for latex allergic patients. Immunol Lett 2008;115:98-104
Based on two clinical observations of adverse reactions during exercise with latex sport bands, we aimed to assess the possible risk for allergic patients posed by this equipment by investigating allergen content and IgE binding potential. Protein extracts of three different latex sport bands were characterized with sera of latex allergic patients. The IgE recognition profile of the allergic patients was identified by component resolved diagnosis and the allergen composition of the extracts was characterized by inhibition assays with the recombinant latex allergens Hev b 1, 3, 5, 6.02, and 8. The sera showed pronounced IgE binding to all three blotted extracts, however with diverse patterns. Inhibition assays revealed the presence of Hev b 1, 3, 5, and 8 in latex sport band extracts. The clinical relevance of contained allergens was demonstrated by strong skin reactions when testing with latex sport bands. From our results we conclude that latex sport bands contain clinically relevant allergens and may cause latex allergic individuals to experience allergic symptoms, potentially amplified by exercise-induced mechanisms. Even though latex is labeled on products, it is important that patients as well as athletic trainers and physical therapists recognize the risk of adverse reactions with these bands.
[22] - Kimata H. Latex allergy in infants younger than 1 year. Clin Exp Allergy 2004;34:1910-1915
BACKGROUND: The prevalence of latex allergy in children is increasing worldwide. Previous multiple operations or atopic predisposition are known risk factors. In contrast, only sporadic cases of latex allergy have been reported in infants younger than 1 year, and the causative latex-containing products or symptoms in young infants have not been studied in detail . OBJECTIVE: The purpose of this study is to analyse the symptoms and risk factors of latex allergy in young infants . METHODS: Cases of latex allergy in infants younger than 1 year were studied in detail. Clinical course, causative latex-containing products were spotted and detailed analysis for latex allergy in patients and patients' parents was performed . CONCLUSION: We report nine cases of latex allergy in infants younger than 1 year. None of them have any abnormality or previous operations. Six patients had atopic eczema/dermatitis syndrome, one patient had bronchial asthma, whereas two patients had no overt allergic diseases. Symptoms of latex allergy were wheezing, swelling of face or lips, facial rash, or anaphylaxis, and causative latex-containing products were teat, pacifier, nose cleaner, teether, balloon, or enema tube. All of the nine patients had positive skin prick test to latex and extract from causative latex-containing products, whereas eight patients had positive serum latex-specific IgE. Study for family history revealed that latex allergy was noted in either father or mother in six patients, in both father and mother in one patient, whereas no latex allergy was noted in parents in two patients. It should be noted that all of these patients had latex-induced symptoms at home. Latex allergy in young infants may not be unusual. Physicians should be aware of latex allergy, and care should be taken to avoid contact with latex in young infants, especially when there is family history for latex allergy.
[23] - Mäkinen-Kiljunen S, Turjanmaa K, Palosuo T. Allergenicity of Baby Pacifiers and Bottle Teats Made from Natural Rubber Latex (NRL). J Allergy Clin Immunol 1997;99:156
Allergenicity of Baby Pacifiers and Bottle Teats Made from Natural Rubber Latex (NRL). S Mäkinen-Kiljunen, K Turjanmaa, T Palosuo, University Hospitals, Helsinki and Tampere, National Public Health Institute, Helsinki, Finland. NRL allergy has increasingly been recognized among children, but the sensitizer has not always been identified. Anaphylaxis has been reported from a baby pacifier, and variable NRL allergen levels have been measured in pacifiers and bottle teats. Previously, using immunoelectrophoretic methods we have demonstrated common allergens in a pacifier and NRL. The aim of the present study was to evaluate all pacifiers and bottle teats on the Finnish market for their NRL allergenicity. 31 pacifiers-including 12 teats, from 11 manufacturers were collected. For the analysis, samples were cut into pieces, extracted (1:5 w/v) and compared to NRL (100 000 AU/ml) in an ELISA inhibition test using sera from latex-allergic patients. 100-fold differences were seen between the pacifiers. In 12 samples-including 2 bottle teats, the allergenicity was below the detection limit of the assay (below 1 AU/ml). In 14 samples the allergenicity was low (1-10 AU/ml), and moderate (10-100 AU/ml) in 5-including one bottle teat. No sample exhibited a high NRL allergenicity (>100 AU/ml). In general, the allergenicity was low in baby pacifiers and teats made from NRL processed as dry rubber under high pressure and heat. In most items (n = 26) the activity was below 10 AU/ml, and all were far below the highest activity found in latex gloves (up to 1500 AU/ml). However, even moderate NRL allergen levels or lower might cause sensitization and elicit allergic reactions when in direct contact to mucous membrane in the mouth. In children with NRL allergy or at high risk to develop NRL allergy, a use of pacifiers and teats made from non-NRL should be considered.
[26] - Beezhold DH, Reschke JE, Allen JH, Kostyal DA, Sussman GL. Latex protein: a hidden ''food'' allergen ? Allergy Asthma Proc 2000;21:301-306
Avoidance of latex allergens is the primary method to prevent adverse reactions. Natural rubber latex is found in many different products in both the health care industry and in modern society, and consequently results in unexpected exposures of sensitized individuals. The use of latex gloves by food handlers provides one potential route for inadvertent exposure to latex allergens. In this study we have used two immunological methods to determine whether latex proteins are transferred to foods following contact with latex gloves. Direct transfer of latex protein to cheese was visualized using a modified immunoblot method. Sliced cheese was touched with a gloved finger. A nitrocellulose membrane was applied to lift the potential fingerprints and a rabbit anti-latex antiserum was used to visualize the transfer of any latex finger-prints. After handling lettuce with gloves, transferred protein was recovered by extracting the lettuce and quantified using an inhibition ELISA for latex proteins. Fingerprints of latex protein were readily detectable on cheese after contact with powdered latex gloves, but not with vinyl gloves. Furthermore, powdered latex glove use resulted in measurable amounts of latex protein on lettuce with an exposure-dependent increase in the latex protein levels. Lettuce alone or lettuce handled with vinyl gloves was negative for latex protein. The use of latex gloves by food handlers is the source of an indirect food additive in the form of latex proteins. It is recommended that food handlers avoid the use of latex gloves to eliminate inadvertent exposure of latex-sensitive individuals.
[28] - Cogen FC, Beezhold DH. Hair glue anaphylaxis: a hidden latex allergy. Ann Allergy Asthma Immunol 2002;88:61-63
Background: Increasingly popular cosmetic hair alterations use latex-containing bonding glue to attach hair to the scalp. Immunoglobulin E-mediated reactions to natural rubber latex may occur from unappreciated sources. Objective: Evaluation of a case of bonding glue anaphylaxis with immunochemical characterization of the glue. Methods and Results: A 37-year-old woman developed systemic anaphylaxis upon repeated exposure to hair bonding glue. Skin tests to the glue and latex RAST tests were positive. Her sera reacted to latex antigens including four recombinant Hev b allergens. Assays of the glue revealed antigen patterns resembling ammoniated latex. Antigen levels exceed those reported in other natural rubber latex-dipped products. Conclusions: Hair bonding glue contains high concentrations of soluble latex antigen and may cause anaphylaxis without mucosal contact. Repeated glue exposure may potentially sensitize consumers. Physicians, cosmetologists, and latex-allergic patients should be aware of bonding glue-induced immunoglobulin E-mediated reactions associated with hair alterations.
[29] - Wagner S, Breiteneder H. Hevea brasiliensis Latex Allergens: Current Panel and Clinical Relevance. Int Arch Allergy Immunol 2005;136:90-97
Latex allergy has been studied in detail in Europe and the US over the past two decades, resulting in specific guidelines that succeeded in reducing its incidence in high-risk populations within the medical field. How these developments have affected high-risk populations outside the health care scenario is an important unanswered question. In addition, a second wave of latex allergy may occur in nations that are striving to attain higher economic and technologic standards, including population-dense countries such as China. Therefore, the application of Hevea allergens in novel diagnostic assays and the development of specific latex immunotherapy will provide new opportunities for latex allergy research. In this review, we summarize current knowledge on the immunological properties of the 13 officially accepted Hevea brasiliensis latex allergens.
[30] - Yagami T, Sato M, Nakamura A, Komiyama T, Kitagawa K, Akasawa A, et al. Plant defense-related enzymes as latex antigens. J Allergy Clin Immunol 1998;101:379-385
BACKGROUND: Latex allergy is an increasing hazard to people who frequently come into contact with latex products. Of interest concerning this immediate-type allergy is the cross-reactivity to various vegetable foods and pollen. Despite its high prevalence, no adequate explanation has been provided for the cross-reactive antigens . OBJECTIVE: We have hypothesized that a series of plant defense-related proteins act as latex allergens, as well as vegetable food allergens. To evaluate this hypothesis, hydrolytic enzymes that are very likely to take on defensive roles in rubber trees were examined for their antigenicity . METHODS: By applying chromatographic procedures, defense-related enzymes were separated from nonammoniated latex (NAL). Their antigenicity was examined by immunoblotting and ELISA with sera containing IgE antibodies to crude latex proteins . RESULTS: Three kinds of hydrolytic enzymes (basic beta-1,3-glucanases [35, 36.5, and 38 kd], a basic chitinase/lysozyme [29.5 kd], and an acidic esterase [44 kd]) were separated from NAL. They were recognized by IgE antibodies from a significant number of patients allergic to latex. The basic beta-1,3-glucanases and the acidic esterase were also strongly recognized by IgE antibodies from several atopic subjects who were allergic to various vegetable foods rather than latex products . CONCLUSION: It was ascertained that the three defense-related enzymes separated from NAL constituted part of the latex antigens. Taking together the well-known serologic or immunologic relationships and amino acid sequence similarities of defense-related proteins coming from phylogenetically distant plant species, we can suspect their universal antigenicity and cross-reactivity.
[31] - Malandain H, Lavaud F. Allergénicité des protéines de défense végétale. Rev Fr Allergol Immunol Clin 2004;44:469-475
La synthèse de protéines de défense fait partie des réponses biochimiques que les plantes ont développées pour lutter contre leurs ennemis naturels et contre les stress environnementaux. De nombreuses protéines de défense végétale sont allergisantes : chitinases, protéines de transfert lipidique LTP, protéines Bet v 1-like. Cet article rappelle les principales familles de protéines de défense végétale, leur impact allergique et l'effet de certains procédés agricoles ou technoalimentaires sur l'allergénicité des aliments végétaux.
[32] - Broekaert W, Lee HI, Kush A, Chua NH, Raikhel N. Wound-induced accumulation of mRNA containing a hevein sequence in laticifers of rubber tree (Hevea brasiliensis). Proc Natl Acad Sci U S A 1990;87:7633-7637
Hevein is a chitin-binding protein that is present in laticifers of the rubber tree (Hevea brasiliensis). A cDNA clone (HEV1) encoding hevein was isolated by using the polymerase chain reaction with mixed oligonucleotides corresponding to two regions of hevein as primers and a Hevea latex cDNA library as a template. HEV1 is 1018 base pairs long and includes an open reading frame of 204 amino acids. The deduced amino acid sequence contains a putative signal sequence of 17 amino acid residues followed by a 187-amino acid polypeptide. This polypeptide has two striking features. The amino-terminal region (43 amino acids) is identical to hevein and shows homology to several chitin-binding proteins and to the amino termini of wound-inducible proteins in potato and poplar. The carboxyl-terminal portion of the polypeptide (144 amino acids) is 74-79% homologous to the carboxyl-terminal region of wound-inducible genes of potato. Wounding, as well as application of the plant hormones abscisic acid and ethylene, resulted in accumulation of hevein transcripts in leaves, stems, and latex but not in roots.
[33] - Sussman GL, Beezhold DH, Kurup VP. Allergens and natural rubber proteins. J Allergy Clin Immunol 2002;110(2 part 2):S33-S39
BACKGROUND: Allergy to natural rubber latex (NRL) results from exposure to proteins derived from Hevea brasiliensis. Type I latex hypersensitivity is observed in certain occupational and other high-risk groups with frequent exposure to NRL products. This includes health care workers (HCWs), workers in the latex industry, children with spina bifida, and atopic individuals . OBJECTIVES: Early reliable diagnosis and avoidance are required for better patient care. Standardized reagents are not presently available for in vitro and in vivo testing and treatment of patients with latex allergy. However, a number of allergens have been isolated and characterized from Hevea latex and NRL products. Currently, a total of 11 major and minor allergens are designated by the International Allergen Nomenclature Committee. This article reviews the structural and functional characteristics of these latex allergenic proteins . RESULTS: NRL-allergenic proteins include those involved in the biosynthesis of polyisoprene and coagulation of latex rubber elongation factor, small rubber particle protein, prohevein, and patatin. Pathogenesis-related proteins include beta-1,3-glucanases, chitinases, and hevamine; and the structural proteins include microhelix protein complex, proline-rich protein, profilins, enolases, and manganese superoxide dismutase. Recombinant allergens demonstrated skin test reactivity in patients with latex allergy. The minimal level of skin test reactivity was about 70 pg/mL for NRL and 1 ng/mL for recombinant allergens. The use of selected recombinant latex allergens (Hev b 5, Hev b 6, and Hev b 7) in skin prick tests identified 93% of allergic individuals, mainly health care workers . CONCLUSIONS: Recombinant latex allergens are clinically reactive and can be produced in a standardized manner, which could potentially provide safe and sensitive reagents for the diagnosis and treatment of type I latex allergy.
[35] - Chardin H, Raulf-Heimsoth M, Chen Z, Rihs HP, Mayer C, Desvaux FX, et al. Interest of Two-Dimensional Electrophoretic Analysis for the Characterization of the Individual Sensitization to Latex Allergens. Int Arch Allergy Immunol 2002;128:195-203
Background/Objective: Latex allergy is a type 1 hypersensitivity reaction that mainly affects high-risk populations such as health care workers, spina bifida-affected or multiply-operated children. Ten molecules have so far been identified and registered as latex allergens (Hev b 1 to Hev b 10). The aim of the present investigation was to identify the major latex allergens by an individual analysis of the IgE response of latex-allergic patients to latex proteins separated by two-dimensional (2-D) gel electrophoresis. Materials and Methods: Latex proteins from a sap or a glove extract were separated by 2-D electrophoresis and transferred to a nitrocellulose membrane. Each membrane was incubated with the serum of one latex-allergic patient. The most frequently recognized latex allergens were characterized in sap and glove extracts using monoclonal antibodies or amino acid microsequencing. Results: The one-dimensional screening of 54 patient sera revealed 4 major bands recognized by IgE. The 2-D analysis of the sensitization to latex allergens allows the identification of allergen isoforms and the characterization of an individual response diversity. Hev b 6.01 was recognized by 88.9% of the patients. Protein spots around 14 kD were recognized by 48.1% of the patients and corresponded to Hev b 6.03 as well as other proteins. A not yet characterized doublet of acidic proteins with molecular masses of 43 and 94 kD was recognized by 20.4% of the sera. Only 5.5% of the sera did not recognize any of these 4 major allergens. Hev b 1 is the main protein from the glove extract but was not constantly found in sap extracts. Conclusions: One-dimensional electrophoretic analysis of the allergen is usually not sufficient to characterize the individual specificity of the IgE response to latex allergens. Latex-glove proteins which are allergens can be absent from the sap extracts and the sensitization to these allergens could be underestimated. Individual 2-D analysis of the sensitization to latex allergens is useful to define the best allergen mixture required for diagnosis and needed for individual therapy monitoring.
[36] - Yagami T, Haishima Y, Tsuchiya T, Tomitaka-Yagami A, Kano H, Matsunaga K. Proteomic Analysis of Putative Latex Allergens. Int Arch Allergy Immunol 2004;135:3-11
BACKGROUND: Extensive analysis of allergenic proteins is generally time-consuming and labor-intensive. Accordingly, a rapid and easy procedure for allergen identification is required. As sequence information on proteins and genes is accumulated in databases, it is becoming easier to identify a candidate protein using proteomic strategies, i.e. two-dimensional gel electrophoresis, site-specific fragmentation, mass spectrometry and then database search. In this study, we evaluated the usefulness of a proteomic strategy for identifying putative allergens through its application to latex proteins . METHODS: Latex proteins were separated with two-dimensional gel electrophoresis, and putative allergens were visualized by IgE immunoblotting using pooled serum from latex-sensitive patients. The IgE-interactive proteins were cut out from the negatively stained two-dimensional gel and subjected to in-gel digestion by trypsin. Then the resulting peptides were analyzed with mass spectrometry. Based on the mass spectrometric data we obtained, the allergen candidates were assigned by a database search . RESULTS: Five previously reported allergens and five new allergen candidates were identified with the proteomic approach without isolating the individual proteins. Less than 1 mg of crude latex protein was sufficient for the entire protocol. Because plural proteins can be processed in parallel, analysis of about 50 IgE-interactive proteins was accomplished within 1 week . CONCLUSIONS: Analysis of putative allergens with proteomic strategies (allergenomics) is a promising avenue for rapid and exhaustive research. The high resolving power of two-dimensional gel electrophoresis is superior to conventional gel electrophoresis. Moreover, the notable sensitivity and speed of mass spectrometry have pronounced advantages over the N-terminal sequencing that has generally been used for protein identification.
[37] - Lee MF, Chen YH, Lin HC, Wang HL, Hwang GY, Wu CH. Identification of Hevamine and Hev b 1 as Major Latex Allergens in Taiwan. Int Arch Allergy Immunol 2006;139:38-44
BACKGROUND: Proteins from latex gloves have been documented to trigger occupational latex allergy among health care workers. Allergen characterization of latex glove extract has never been studied in Taiwan. This study aimed to identify allergenic proteins from latex gloves . METHODS: Crude extracts of latex gloves were prepared with phosphate-buffered saline and 20 medical workers with a history of latex allergy were enrolled in this study. The specific IgE antibody was determined by the Pharmacia CAP system and in-house enzyme-linked immunoassay and immunoblotting. The target proteins were excised from two-dimensional PAGE and analyzed by electrospray ionization tandem mass spectrometry . RESULTS: Immunoblotting of glove extracts revealed three IgE-binding proteins at a molecular mass of 45, 30 and 14 kDa. Peptide mass fingerprinting revealed that the protein at 45 kDa, which was recognized by 10% (2/20) of atopic sera tested, was an allergenic lipolytic esterase from Hevea brasiliensis (Hev b 13). The 30- and 14-kDa proteins, which were recognized by 55% (11/20) and 85% (17/20) of patients' sera, were found to be hevamine and rubber elongation factor (Hev b 1), respectively . CONCLUSIONS: Our results indicated that hevamine and Hev b 1 are the major allergens from latex gloves in Taiwan, which differs from the reports in Western countries.
[38] - Conti A, Giuffrida MG, Hoffmann-Sommergruber K, Wagner S, Amato S, Mistrello G, et al. Identification of latex UDP glucose pyrophosphorylase (Hev b UDPGP) as a novel cause of latex fruit allergy syndrome. Eur Ann Allergy Clin Immunol 2007;39:116-118
Based on a recently published unusual case of food allergy in a latex-allergic patients, the present study identifies Hev b UDPGP as a novel allergen in natural rubber latex able to cause latex-fruit allergy syndrome and as a novel, potential pan-allergen in vegetable foods.
[39] - Dennis MS, Light DR. Rubber elongation factor from Hevea brasiliensis. Identification, characterization, and role in rubber biosynthesis. J Biol Chem 1989;264:18608-18617
The presence of a protein, rubber elongation factor (REF), which is tightly bound to serum-free rubber particles purified from Hevea brasiliensis latex, is necessary for prenyltransferases from a number of sources to add multiple cis-isoprene units to rubber molecules. These prenyltransferases show normal farnesyl pyrophosphate synthase activity (two trans additions of isopentenyl pyrophosphate to dimethylallyl pyrophosphate) in the absence of REF bound to rubber particles. REF bound to rubber molecules can be highly purified from all other proteins in whole latex by treatment of rubber particles with low concentrations of detergent. Treatment of rubber particles with trypsin which hydrolyzes bound REF, removal of REF with high concentrations of various detergents, or treatment of whole latex with polyclonal antibodies specific for REF all prevent prenyltransferase from adding [14C]isopentenyl pyrophosphate to rubber molecules. However, we have not been successful using detergent-solubilized REF in the reconstitution of in vitro rubber biosynthesis with either REF-depleted rubber particles or allylic pyrophosphate primers. REF has a molecular mass of 14,600 Da and is associated specifically with rubber particles in whole latex. It makes up between 10-60% of the total protein in whole latex but is absent in C-serum, the supernatant fluid obtained when rubber particles are removed by centrifugation. The amount of REF in whole latex is proportional to the rubber content. Based on a number average molecular mass of 500,000 Da for rubber and the content of rubber and REF in whole latex or serum-free rubber particles, the stoichiometry of REF molecules to rubber molecules is 1:1 in both cases. There is sufficient REF to form a monomolecular protein layer coating large rubber particles (700-1,000 nm). In the electron microscope, serum-free rubber particle preparations contain particles with diameters from 800 to as small as 10 nm. In the presence of 1% sodium dodecyl sulfate no particles smaller than 100 nm are observed. We suggest that the smaller particles may be mainly composed of REF molecules.
[40] - Czuppon AB, Chen Z, Rennert S, Engelke T, Meyer HE, Heber M, et al. The rubber elongation factor of rubber trees (Hevea brasiliensis) is the major allergen in latex. J Allergy Clin Immunol 1993;92:690-697
BACKGROUND: Allergy to latex-containing articles is becoming more and more important because it can result in unexpected life-threatening anaphylactic reactions in sensitized individuals. METHODS: A protein of 58 kd with an isoelectric point of 8.45 was purified from raw latex and from latex gloves and identified as the major allergen, completely blocking specific IgE antibodies in the serum of latex-sensitized subjects. The allergen is a noncovalent homotetramer molecule, in which the 14.6 kd monomer was identified, by amino acid composition and sequence homologies of tryptic peptides, to be the rubber elongation factor found in natural latex of the Malaysian rubber tree. RESULTS: Competitive immunoinhibition tests showed that the starch powder covering the finished gloves is the airborne carrier of the allergen, resulting in bronchial asthma on inhalation. The purified allergen can induce allergic reactions in the nanogram range. CONCLUSION: The identification of the allergen (Hev b I) may help to eliminate it during the production of latex-based articles in the future
[41] - Chen Z, Cremer R, Posch A, Raulf-Heimsoth M, Rihs HP, Baur X. On the allergenicity of Hev b 1 among health care workers and patients with spina bifida allergic to natural rubber latex. J Allergy Clin Immunol 1997;100:684-693
BACKGROUND: Recent studies have caused much controversy about the prevalence of IgE antibodies to Hev b 1 among health care workers (HCWs) and patients with spina bifida (SB) who are allergic to latex. This investigation was carried out to verify the results reported. METHOD: Serum samples from 140 patients with SB as well as from 105 HCWs allergic to latex were tested by enzyme allergosorbest test (EAST) and EAST-inhibition assay to evaluate the rate and degree of sensitization to highly purified Hev b 1. RESULTS: Eighty-one percent of patients with SB who were allergic to latex had IgE antibodies against Hev b 1. The prevalence of anti-Hev b 1 antibodies among HCWs allergic to latex was 52.3%. In 15 of 33 serum samples from patients with SB that were randomly tested, the IgE binding to commercial latex allergens could be completely inhibited by Hev b 1; in only six cases was the maximum inhibition of IgE binding to latex by Hev b 1 less than 50%. Testing two monoclonal anti-Hev b 1 antibodies with extracts of five brands of latex gloves revealed a predominant presence of Hev b 1 protein as a monomer or its aggregates. Molecular analysis of human leukocyte antigen-D region genes DRB and DQB1 suggested no statistically significant correlation between the human leukocyte antigen alleles tested and IgE responsiveness to Hev b 1. CONCLUSIONS: Our results indicate that Hev b 1 not only makes significant contributions to the IgE binding to latex, but it is also the unique sensitizer in about 45% of patients with SB who are allergic to latex
[43] - Alenius H, Kalkkinen N, Lukka M, Reunala T, Turjanmaa K, Makinen-Kiljunen S, et al. Prohevein from the rubber tree (Hevea brasiliensis) is a major latex allergen. Clin Exp Allergy 1995;25:659-665
There is general agreement that proteins eluting from different natural rubber latex products can cause immediate type hypersensitivity reactions in latex-allergic patients. However, there is as yet no consensus as to what are the most important allergens in natural rubber latex. OBJECTIVE: We wanted to purify and characterize at the primary structure level three natural latex proteins, suggested to represent significant allergens. METHODS: Proteins were purified from ultracentrifuged bottom fraction of natural rubber latex using high performance liquid chromatography gel filtration and reversed phase chromatography. Purified proteins were subjected to tryptic cleavage, peptide separation and amino acid sequencing. Immunoblotting was used to demonstrate IgE antibodies to the purified proteins in sera from latex-allergic patients. RESULTS: A 20 kDa protein was identified by amino acid sequencing as prohevein, a major protein in the rubber tree Hevea brasiliensis, and a 30 kDa natural rubber latex protein as hevamine, another essential rubber tree protein. A third, previously undescribed natural rubber latex protein, showed high homology to several plant endo-1,3-beta-glucosidases. In immunoblotting, the purified prohevein bound IgE antibodies from 24/29 (83%) sera of latex-allergic patients including positive results in 4/6 latex-allergic children with spina bifida or other congenital anomalies. The purified prohevein elicited positive skin-prick test reactions in all six latex-allergic patients showing IgE to prohevein. The purified 36 kDa protein bound IgE from 6/29 (21%) latex-allergic sera, and the purified hevamine from only 1/29 patient sera. CONCLUSION: The observed high frequency of IgE antibodies to prohevein suggests that this protein is a major natural rubber latex allergen.
[45] - Palomares O, Villalba M, Quiralte J, Polo F, Rodriguez R. 1,3-beta-glucanases as candidates in latex-pollen-vegetable food cross-reactivity. Clin Exp Allergy 2005;35:345-351
Summary Background 1,3-beta-glucanases (group 2 of pathogenesis-related proteins) are enzymes widely distributed among higher plants and have been recently proven to be significant allergens. Objective The aim of this work was to study the potential implication of 1,3-beta-glucanases in cross-reactivities among latex, pollen and vegetable foods. Methods The cDNA encoding the N-terminal domain (NtD) of Ole e 9, a major allergenic 1,3-beta-glucanase from olive pollen, was amplified by polymerase chain reaction and produced as a recombinant protein in Pichia pastoris (recombinant N-terminal domain, rNtD). Circular dichroism, ELISA, immunoblotting and immunoblotting inhibition experiments were carried out. Sera from olive pollen allergic patients and a rNtD-specific polyclonal antiserum were used. Results The NtD of Ole e 9 has been produced at high yield in the yeast P. pastoris and possesses 1,3-beta-glucanase activity. The expressed polypeptide conserves IgE and IgG immunodominant epitopes of the whole Ole e 9. A rNtD-specific polyclonal antiserum and sera from olive pollen allergic patients allowed detection of IgG and IgE reactive peptidic epitopes common to 1,3-beta-glucanase Ole e 9 in extracts from ash and birch pollen, tomato, potato, bell-pepper, banana and latex. Conclusion rNtD and homologous glucanases are new molecules to be used in diagnostic protocols as they could help to identify allergic pollen patients who are at risk for developing allergic symptoms to fruits, vegetables and latex.
[46] - Yagami T, Osuna H, Kouno M, Haishima Y, Nakamura A, Ikezawa Z. Significance of Carbohydrate Epitopes in a Latex Allergen with beta-1,3-Glucanase Activity. Int Arch Allergy Immunol 2002;129:27-37
Background: One of the latex allergens, Hev b 2, has beta-1,3-glucanase activity. The entire sequence of this allergen is already known. There is one potential N-glycosylation site in this molecule (27Asn). Heterogeneous glycosylation of this Asn residue could be a source of the multiplicity of natural Hev b 2. Possible participation of the carbohydrate epitopes of latex beta-1,3-glucanase isoenzymes in their IgE-binding capacity and cross-reactivity was investigated in this study. Methods: beta-1,3-Glucanase isoenzymes were separated based on their affinities for concanavalin A. IgE-binding capacity and cross-reactivity were examined by immunoblotting and enzyme-linked immunosorbent assay (ELISA). Sequence heterogeneity among the isoenzymes was probed by peptide mass mapping after lysyl endopeptidase digestion. To clarify the relation to Hev b 2, N-terminal sequencing was performed on a fragmented peptide common to the separated isoenzymes. Results: Basic beta-1,3-glucanase was subdivided into two glycosylated isoenzymes (GI and GII) and one non-glycosylated isoenzyme (GIII). IgE antibodies in latex-positive sera chiefly recognized the glycosylated isoenzymes. Inhibition ELISA supported the significance of the carbohydrate epitopes for the IgE recognition and cross-reactivity. However, non-glycosylated GIII, as well as GI and GII, produced positive results in a skin prick test. The three beta-1,3-glucanase isoenzymes shared a partial sequence in common with Hev b 2. Conclusions: Our results suggest that the carbohydrate epitopes in Hev b 2 homologues are relevant to an in vitro diagnosis of latex allergy and the accompanying cross-reactivity. Carbohydrate epitopes do not necessarily provoke allergic symptoms. Therefore, the actual allergenicity of Hev b 2 and its homologues should be carefully evaluated not only by in vitro IgE tests but also by in vivo tests
[47] - Seifert U, Wagner S, Ebner C, Hafner C, Mari A, Arija M Arif S, et al. To what extent does glycosylation of nHev b 2, the b-1,3-glucanase from Hevea brasiliensis, contribute to IgE recognition of sera of latex sensitized patients ? EAACI 25th Congress, Vienna, 10-14 June, 2006, Poster n°1058
Background The b-1,3-glucanase of Hevea brasiliensis, Hev b 2, is regarded as a major latex allergen. Most isoforms of this protein are glycosylated with the MMXF structure, a common sugar motif of plant glycoproteins. The aim of this study was to elucidate the contribution of the carbohydrate moiety to IgE recognition. Methods Hev b 2 was purified from fresh Hevea latex. An aliquot was treated with periodate to destroy carbohydrate epitopes. An activity assay and ELISA experiments with antibodies to the carbohydrate moiety and to protein epitopes were performed for both samples. Sera from 105 latex sensitized patients were tested in IgE-ELISA experiments on nHev b 2 and periodate treated Hev b 2. Results A positive activity assay confirmed that the secondary structure of the periodate treated sample was not significantly altered. Of 105 sera from latex sensitized patients, 65 (62%) showed IgE binding to nHev b 2. 65% (42/65) of these patients showed allergic symptoms, the remaining patients (23/65) had no symptoms, but IgE to nHev b 2. 48% (20/42) of the allergic patients´sera lost IgE reactivity to the periodate treated Hev b 2.Of the sera from patients without symptoms, 70% (16/23) did not react with the periodate treated sample. Of all patients with IgE to Hev b 2 protein epitopes (29), 76% (22/29) show allergic symptoms to latex. Whereas only 56 % (20/36) of the patients with IgE to a glycan epitope suffer from symptoms. Conclusions Our data show that protein epitopes are more frequently recognized by allergic subjects´ IgE than by IgE from sensitized subjects. Furthermore, the data indicate that the carbohydrate moiety of this glycosylated allergen is of minor clinical importance. Experiments to elucidate the clinical relevance of the cross-reactive carbohydrate structure of Hev b 2 are in progress.
[48] - Yagami T, Osuna H, Kouno M, Haishima Y, Nakamura A, Ikezawa Z. Significance of Carbohydrate Epitopes in a Latex Allergen with beta-1,3-Glucanase Activity. Int Arch Allergy Immunol 2002;129:27-37
Background: One of the latex allergens, Hev b 2, has beta-1,3-glucanase activity. The entire sequence of this allergen is already known. There is one potential N-glycosylation site in this molecule (27Asn). Heterogeneous glycosylation of this Asn residue could be a source of the multiplicity of natural Hev b 2. Possible participation of the carbohydrate epitopes of latex beta-1,3-glucanase isoenzymes in their IgE-binding capacity and cross-reactivity was investigated in this study. Methods: beta-1,3-Glucanase isoenzymes were separated based on their affinities for concanavalin A. IgE-binding capacity and cross-reactivity were examined by immunoblotting and enzyme-linked immunosorbent assay (ELISA). Sequence heterogeneity among the isoenzymes was probed by peptide mass mapping after lysyl endopeptidase digestion. To clarify the relation to Hev b 2, N-terminal sequencing was performed on a fragmented peptide common to the separated isoenzymes. Results: Basic beta-1,3-glucanase was subdivided into two glycosylated isoenzymes (GI and GII) and one non-glycosylated isoenzyme (GIII). IgE antibodies in latex-positive sera chiefly recognized the glycosylated isoenzymes. Inhibition ELISA supported the significance of the carbohydrate epitopes for the IgE recognition and cross-reactivity. However, non-glycosylated GIII, as well as GI and GII, produced positive results in a skin prick test. The three beta-1,3-glucanase isoenzymes shared a partial sequence in common with Hev b 2. Conclusions: Our results suggest that the carbohydrate epitopes in Hev b 2 homologues are relevant to an in vitro diagnosis of latex allergy and the accompanying cross-reactivity. Carbohydrate epitopes do not necessarily provoke allergic symptoms. Therefore, the actual allergenicity of Hev b 2 and its homologues should be carefully evaluated not only by in vitro IgE tests but also by in vivo tests
[49] - Seifert U, Wagner S, Ebner C, Hafner C, Mari A, Arija M Arif S, et al. To what extent does glycosylation of nHev b 2, the b-1,3-glucanase from Hevea brasiliensis, contribute to IgE recognition of sera of latex sensitized patients ? EAACI 25th Congress, Vienna, 10-14 June, 2006, Poster n°1058
Background The b-1,3-glucanase of Hevea brasiliensis, Hev b 2, is regarded as a major latex allergen. Most isoforms of this protein are glycosylated with the MMXF structure, a common sugar motif of plant glycoproteins. The aim of this study was to elucidate the contribution of the carbohydrate moiety to IgE recognition. Methods Hev b 2 was purified from fresh Hevea latex. An aliquot was treated with periodate to destroy carbohydrate epitopes. An activity assay and ELISA experiments with antibodies to the carbohydrate moiety and to protein epitopes were performed for both samples. Sera from 105 latex sensitized patients were tested in IgE-ELISA experiments on nHev b 2 and periodate treated Hev b 2. Results A positive activity assay confirmed that the secondary structure of the periodate treated sample was not significantly altered. Of 105 sera from latex sensitized patients, 65 (62%) showed IgE binding to nHev b 2. 65% (42/65) of these patients showed allergic symptoms, the remaining patients (23/65) had no symptoms, but IgE to nHev b 2. 48% (20/42) of the allergic patients´sera lost IgE reactivity to the periodate treated Hev b 2.Of the sera from patients without symptoms, 70% (16/23) did not react with the periodate treated sample. Of all patients with IgE to Hev b 2 protein epitopes (29), 76% (22/29) show allergic symptoms to latex. Whereas only 56 % (20/36) of the patients with IgE to a glycan epitope suffer from symptoms. Conclusions Our data show that protein epitopes are more frequently recognized by allergic subjects´ IgE than by IgE from sensitized subjects. Furthermore, the data indicate that the carbohydrate moiety of this glycosylated allergen is of minor clinical importance. Experiments to elucidate the clinical relevance of the cross-reactive carbohydrate structure of Hev b 2 are in progress.
[50] - Banerjee B, Kanitpong K, Fink JN, Zussman M, Sussman GL, Kelly KJ, et al. Unique and shared IgE epitopes of Hev b 1 and Hev b 3 in latex allergy. Mol Immunol 2000;37:789-798
Of the several latex proteins cloned and expressed, the rubber elongation factor, Hev b 1, and the closely related Hev b 3, represent two major allergens associated with latex allergy. Although both allergens demonstrated IgE binding with sera from latex allergic patients, it was not known whether these two molecules shared any epitopes. Hence, in the present study using health care workers (HCW) and spina bifida (SB) patients with latex allergy, we investigated the IgE binding epitopes in Hev b 1 and Hev b 3. Recombinant Hev b 1 and Hev b 3 were expressed in a prokaryotic expression system, while overlapping decapeptides of Hev b 1 and Hev b 3 were synthesized on derivatized cellulose membrane. Eight IgE binding epitopes for Hev b 1 and eleven for Hev b 3 were identified using sera from latex allergic patients with SB. On further analysis of synthetic peptides encompassing these epitopes, similar IgE antibody reactivity was demonstrated with three Hev b 1 epitopes b1E3, b1E5, b1E6 and two Hev b 3 epitopes: b3E10 and b3E 11. For Hev b 1, a unique IgE binding epitope was identified in the region of amino acid residues 16-25. In competitive ELISA, peptides bIE2 and bIE4 together inhibited 58% of IgE binding of Hev b 1, while b3E5 showed 22% inhibition in the IgE binding of Hev b 3. The results of the present study suggest that the understanding of linear and conformational IgE epitopes in the major latex allergens may provide better insight into the structure-function relationship of the allergens, and may lead to the development of better patient care and management strategies in latex allergy.
[51] - Alenius H, Kalkkinen N, Lukka M, Turjanmaa K, Reunala T, Makinen-Kiljunen S, et al. Purification and partial amino acid sequencing of a 27-kD natural rubber allergen recognized by latex-allergic children with spina bifida. Int Arch Allergy Immunol 1995;106:258-262
We purified from natural rubber latex (NRL) by means of high- performance liquid chromatography a 27-kD protein, recognized characteristically by IgE in sera from latex-allergic children with spina bifida or other congenital anomalies and histories of multiple surgeries. N-terminal sequence analysis of the purified 27-kD protein was unsuccessful suggesting that its N-terminus is blocked. To obtain internal sequence information from the protein it was digested with trypsin and the purified tryptic peptides were subjected to sequence analysis. Thirteen of the 14 sequenced peptides revealed no significant homology to any of the published protein sequences indicating that the 27-kD protein is previously undescribed at the primary structure level. However, one of the 14 sequenced peptides showed significant homology to the rubber elongation factor, a 14.6-kD NRL protein. For the time being, the 27-kD NRL protein is the first molecularly characterized NRL allergen associated with defined clinical manifestations of latex allergy
[52] - Yeang HY, Ward MA, Zamri AS, Dennis MS, Light DR. Amino acid sequence similarity of Hev b 3 to two previously reported 27- and 23-kDa latex proteins allergenic to spina bifida patients. Allergy 1998;53:513-519
Separate studies have reported spina bifida patients to be especially allergic to proteins of 27 and 23 kDa found in the serum of centrifuged natural rubber latex. An insoluble latex protein located on the surface of small rubber particles, Hev b 3, has similarly been found to be allergenic to spina bifida patients. In this study, internal amino acid sequences of Hev b 3 showed similarity to the published sequences for the 27- and 23-kDa latex proteins. The latter allergens are hence identified as Hev b 3. Determination of the molecular weight of Hev b 3 revealed various species of 22-23 kDa. The consistent gaps of about 266 Da observed between various forms of the intact protein suggest that the protein undergoes post-translational modification. To determine whether Hev b 3 also occurs in a soluble form in the latex serum, its presence in molecular-filtered serum was checked by ELISA and Western blot. The results showed Hev b 3 to be largely absent in the C-serum from fresh latex. The protein is therefore in soluble in its native state. However, a small amount of the solubilized protein was detected in ammonia-stabilized latex (commonly used in the manufacture of latex products)
[53] - Lu LJ, Kurup VP, Hoffman DR, Kelly KJ, Murali PS, Fink JN. Characterization of a major latex allergen associated with hypersensitivity in spina bifida patients. J Immunol 1995;155:2721-2728
Hypersensitivity to latex proteins present in health care products, particularly in latex gloves, has emerged as an important public health problem in recent years. Most of these latex allergens have not been purified or characterized. Here we report the purification and characterization of a 23-kDa latex polypeptide from nonammoniated natural rubber latex. This purified 23-kDa polypeptide reacted with IgE from 13 of 17 (76%) latex-allergic spina bifida patients, but from only 1 of 5 health care workers with latex allergy. Furthermore, all the sera of patients with anaphylaxis to latex showed reactivity with the 23-kDa polypeptide. This allergen also induced significant lymphocyte proliferation by PBMC from spina bifida patients, but not from health care workers. The 90 amino acid residues sequenced from the 23-kDa polypeptide showed a 45.6% homology with a previously reported latex protein, rubber elongation factor. By using a mAb LAM-1, produced against the 23-kDa allergen, we could also detect this allerge n or its epitope in ammoniated latex and natural rubber latex products used in health care management, but not in non-latex extracts studied
[55] - Sunderasan E, Chow KS, Ward MA, Yeang HY. Hev b 4 Heavy Peptide Identified as Latex Cyanogenic Glucosidase. AAAAI 58th Annual Meeting, New York, 1-6 March, 2002, Poster n°1031
The latex protein Hev b 4 comprises a triplet of allergenic peptides of molecular weight 50 to 57 kDa. The three peptides share a common characteristic in remaining soluble only in the presence of high ionic concentration (e.g. 0.2M NaCl and higher). Dilution or dialysis induces precipitation of all three peptides that might have related biochemical functions in the latex, and that might be components of a protein complex. Hev b 4 has been shown to be allergenic by IgE-binding assays and in skin prick tests. The heaviest component of the Hev b 4 triplet, a 57 kDa peptide, was isolated from the latex B-serum by dialysis-induced protein precipitation and Sephacryl gel filtration. Further purification was carried out by preparative SDS-polyacrylamide gel electrophoresis. Following trypsin digestion, the 57 kDa peptide was scanned using a nano-electrospray tandem mass spectrometry (ESI-MS) and amino acid sequencing was carried out by MS/MS. Partial amino acid sequences of the protein matched published sequences of several plant glucosidases, including those of cyanogenic glucosidases. Enzymatic assays confirmed that the protein possessed glucosidase activity, the enzyme activity being further demonstrated by isozyme staining of the protein after electrophoretic separation on native polyacrylamide gels. Enzymic activity was also observed when linamarin was used as the specific enzymic substrate, indicating that the protein was a cyanogenic glucosidase (linamarase) as suggested by the partial amino acid sequences. A partial cDNA sequence with homology to plant glucosidases was obtained by reverse transcriptionˆPCR using sense and antisense strand degenerate primers. The 5´/3´RACE is being employed to generate the complete cDNA of H. brasiliensis latex glucosidase.
[56] - Sunderasan E, Bahari A, Arif SA, Zainal Z, Hamilton RG, Yeang HY. Molecular cloning and immunoglobulin E reactivity of a natural rubber latex lecithinase homologue, the major allergenic component of Hev b 4. Clin Exp Allergy 2005;35:1490-1495
BACKGROUND: Hev b 4 is an allergenic natural rubber latex (NRL) protein complex that is reactive in skin prick tests and in vitro immunoassays. On SDS-polyacrylamide gel electrophoresis (SDS-PAGE), Hev b 4 is discerned predominantly at 53-55 kDa together with a 57 kDa minor component previously identified as a cyanogenic glucosidase. Of the 13 NRL allergens recognized by the International Union of Immunological Societies, the 53-55 kDa Hev b 4 major protein is the only candidate that lacks complete cDNA and protein sequence information . OBJECTIVE: We sought to clone the transcript encoding the Hev b 4 major protein, and characterize the native protein and its recombinant form in relation to IgE binding . METHODS: The 5'/3' rapid amplification of cDNA ends method was employed to obtain the complete cDNA of the Hev b 4 major protein. A recombinant form of the protein was over-expressed in Escherichia coli. The native Hev b 4 major protein was deglycosylated by trifluoromethane sulphonic acid. Western immunoblots of the native, deglycosylated and recombinant proteins were performed using both polyclonal antibodies and sera from latex-allergic patients . RESULTS: The cDNA encoding the Hev b 4 major protein was cloned. Its open reading frame matched lecithinases in the conserved domain database and contained 10 predicted glycosylation sites. Detection of glycans on the Hev b 4 lecithinase homologue confirmed it to be a glycoprotein. The deglycosylated lecithinase homologue was discerned at 40 kDa on SDS-PAGE, this being comparable to the 38.53 kDa mass predicted by its cDNA. Deglycosylation of the lecithinase homologue resulted in the loss of IgE recognition, although reactivity to polyclonal rabbit anti-Hev b 4 was retained. IgE from latex-allergic patients also failed to recognize the non-glycosylated E. coli recombinant lecithinase homologue . CONCLUSION: The IgE epitopes of the Hev b 4 lecithinase homologue reside mainly in its carbohydrate moiety, which also account for the discrepancy between the observed molecular weight of the protein and the value calculated from its cDNA.
[57] - Kolarich D, Altmann F, Sunderasan E. Structural analysis of the glycoprotein allergen Hev b 4 from natural rubber latex by mass spectrometry. Biochim Biophys Acta 2006;1760:715-720
The lecithinase homolog (Hev b 4) from Hevea brasiliensis (Q6T4P0_HEVBR) is an important natural rubber latex allergen. Hev b 4 is a highly glycosylated protein and its carbohydrate moiety has been implicated in the binding of IgE from natural rubber latex allergic patients. The cDNA for Hev b 4 has recently been cloned and sequenced. Here, we have analyzed the post-translational modifications of natural Hev b 4 by liquid chromatography/electrospray ionization-mass spectrometry of tryptic peptides. Seven of the eight potential glycosylation sites were found to be occupied. One site, however, was only partially glycosylated. Asn224 was substituted by complex type N-glycans with fucose and xylose, whereas all other sites carried either oligomannose glycans or a mixture of oligomannose and complex N-glycans. Glycosylation site Asn308, the most C-terminal one of the eight sites, was only found in the non-glycosylated form. The complex type N-glycans apparently form the molecular basis for the immune reaction with patients' sera. A large fraction of Hev b 4 molecules contains two or more complex N-glycans and thus a physiological reaction against these polyvalent allergens on the basis of the carbohydrate is in theory possible. Aside from allowing glycosylation analysis, the mass spectrometric data defined the N-terminal cleavage site of Hev b 4. This study once more demonstrates the outstanding analytical potential of electrospray ionization-mass spectrometry coupled with liquid chromatographic separation.
[58] - Sunderasan E, Bahari A, Arif SA, Zainal Z, Hamilton RG, Yeang HY. Molecular cloning and immunoglobulin E reactivity of a natural rubber latex lecithinase homologue, the major allergenic component of Hev b 4. Clin Exp Allergy 2005;35:1490-1495
BACKGROUND: Hev b 4 is an allergenic natural rubber latex (NRL) protein complex that is reactive in skin prick tests and in vitro immunoassays. On SDS-polyacrylamide gel electrophoresis (SDS-PAGE), Hev b 4 is discerned predominantly at 53-55 kDa together with a 57 kDa minor component previously identified as a cyanogenic glucosidase. Of the 13 NRL allergens recognized by the International Union of Immunological Societies, the 53-55 kDa Hev b 4 major protein is the only candidate that lacks complete cDNA and protein sequence information . OBJECTIVE: We sought to clone the transcript encoding the Hev b 4 major protein, and characterize the native protein and its recombinant form in relation to IgE binding . METHODS: The 5'/3' rapid amplification of cDNA ends method was employed to obtain the complete cDNA of the Hev b 4 major protein. A recombinant form of the protein was over-expressed in Escherichia coli. The native Hev b 4 major protein was deglycosylated by trifluoromethane sulphonic acid. Western immunoblots of the native, deglycosylated and recombinant proteins were performed using both polyclonal antibodies and sera from latex-allergic patients . RESULTS: The cDNA encoding the Hev b 4 major protein was cloned. Its open reading frame matched lecithinases in the conserved domain database and contained 10 predicted glycosylation sites. Detection of glycans on the Hev b 4 lecithinase homologue confirmed it to be a glycoprotein. The deglycosylated lecithinase homologue was discerned at 40 kDa on SDS-PAGE, this being comparable to the 38.53 kDa mass predicted by its cDNA. Deglycosylation of the lecithinase homologue resulted in the loss of IgE recognition, although reactivity to polyclonal rabbit anti-Hev b 4 was retained. IgE from latex-allergic patients also failed to recognize the non-glycosylated E. coli recombinant lecithinase homologue . CONCLUSION: The IgE epitopes of the Hev b 4 lecithinase homologue reside mainly in its carbohydrate moiety, which also account for the discrepancy between the observed molecular weight of the protein and the value calculated from its cDNA.
[59] - Akasawa A, Hsieh LS, Lin Y. Comparison of latex-specific IgE binding among nonammoniated latex, ammoniated latex, and latex glove allergenic extracts by ELISA and immunoblot inhibition. J Allergy Clin Immunol 1996;97:1116-1120
BACKGROUND: Nonammoniated latex, ammoniated latex, and latex glove extracts have been used as source materials for the preparation of allergenic extracts for the diagnosis of latex allergy. These materials showed different patterns of protein bands and immunoreactive bands. However, their IgE-reactive repertoires were not compared. OBJECTIVE: The goals of this study were to compare the IgE reactivity and to define the common IgE-reactive epitopes among three latex allergenic extracts. METHODS: Two serum pools were obtained from adults and children with latex allergy to evaluate the IgE reactivity among three latex extracts. IgE reactivity and IgE-reactive proteins were compared by inhibition ELISA and inhibition immunoblot methods, respectively. RESULTS: In this study inhibition curves were similar for nonammoniated latex and ammoniated latex but were different when the latex glove extracts were used. Several protein bands of ammoniated latex and latex glove extracts could not be inhibited by the nonammoniated latex. The ammoniated latex and latex glove extracts were able to remove all the latex-specific IgE from the serum. CONCLUSION: The IgE-reactive proteins differ among different latex extracts. Ammoniated latex and latex glove extracts contain more complete immunoreactive repertoires for detecting IgE antibodies. Our study provides useful information for selecting the latex extract.
[60] - Slater JE, Vedvick T, Arthur-Smith A, Trybul DE, Kekwick RG. Identification, cloning, and sequence of a major allergen (Hev b5) from natural rubber latex (Hevea brasiliensis). J Biol Chem 1996;271:25394-25399
Proteins in commercial latex products, derived from the rubber tree Hevea brasiliensis, cause anaphylaxis in susceptible individuals, especially health care workers and children with spina bifida. To identify latex allergens, we utilized IgE from the serum of a latex-allergic health care worker to screen a cDNA library from Hevea latex. The identified cDNA clone, cDNA Hev b 5, encodes an open reading frame of 163 peptide residues. Hybridization analysis of cDNA Hev b 5 with RNA extracted from Hevea tissue indicates that the full-length transcript is about 1000 bases. The nucleotide and deduced protein sequences have significant homology to sequences from kiwi and potato, which are known to cause allergic reactions in some latex-allergic patients. Fifty-six percent of spina bifida patients and 92% of health care workers with latex allergy have IgE specific to the protein encoded by cDNA Hev b 5. A monoclonal antibody raised from a mouse immunized with Hev b 5 binds to a protein in Hevea latex with an Mr identical to that of the expressed and cleaved recombinant protein. Taken together, these results establish that the antigen Hev b 5 contains a major epitope for IgE-mediated reactions to H. brasiliensis latex products.
[61] - Alenius H, Kalkkinen N, Reunala T, Turjanmaa K, Palosuo T. The main IgE-binding epitope of a major latex allergen, prohevein, is present in its N-terminal 43-amino acid fragment, hevein. J Immunol 1996;156:1618-1625
Polypeptides of natural rubber latex (NRL) that elute from surgeon's gloves and other manufactured rubber products can sensitize exposed individuals and elicit severe hypersensitivity reactions. Previously, we showed that prohevein is a major allergen in NRL, the source material for rubber manufacturing. To analyze which region of the molecule carries the main IgE-binding epitope(s), we purified prohevein and its C-domain from NRL by gel filtration, reverse phase chromatography, and electroelution. In immunoblotting, prohevein bound IgE from 15 of 20 (75%), and the prohevein C-domain from 3 of 20 (15%) latex-allergic patient sera. In ELISA, 36 of 52 (69%) patient sera showed IgE binding to prohevein, whereas 11 of 52 (21%) sera had IgE to prohevein C-domain. We then purified from a brand of highly allergenic surgeon's gloves six hydrophilic peptides that revealed in amino- terminal sequencing 100% identity to the N-terminus of prohevein. In mass spectrometry, all purified peptides gave a molecular mass of 471 9 +/- 1.9 daltons, which corresponds to the molecular mass of hevein (4719.1 daltons), a 43-amino acid N-terminal fragment of prohevein. Purified hevein inhibited 72% of IgE binding from pooled sera of NRL- allergic patients to solid phase glove extract and 45% of IgE binding to solid phase NRL. Of the 43 NRL-allergic patient sera tested, 56% showed IgE Abs to purified hevein in ELISA. In skin prick testing, purified hevein elicited positive reactions in three-quarters of the latex-allergic patients tested. These results indicate that the majority of prohevein's IgE-binding ability resides in its N-terminal fragment, known as hevein. In one highly allergenic latex glove examined, the majority of IgE-binding ability was attributable to hevein molecules, suggesting that these peptides can be significant sensitizers in NRL allergy
[62] - Mari A, Scala E, D’Ambrosio C, Breiteneder H, Wagner S. Latex Allergy within a Cohort of Not-at-Risk Subjects with Respiratory Symptoms: Prevalence of Latex Sensitization and Assessment of Diagnostic Tools. Int Arch Allergy Immunol 2007;143:135-143
BACKGROUND: Immunoglobluin E (IgE)-mediated hypersensitivity to natural rubber latex (NRL) is a major problem in allergy practice. Currently, the use of skin prick tests (SPTs) with latex extracts and specific IgE detection for the diagnosis of NRL allergy in suspected patients is directed to identification of risk factors. Many cases of NRL allergy remain undiagnosed due to misreporting of symptoms by the patients or lack of proper questions asked by the physician. MATERIALS AND METHODS: A total of 6,126 subjects referred for respiratory symptoms underwent SPTs with NRL. Positive subjects were resurveyed for exposure to NRL, and specific IgE for NRL extracts and recombinant molecules was determined. Immunoblots of NRL extracts were performed to identify IgE patterns . RESULTS: Forty-six of 3,930 sensitized subjects had a positive SPT with NRL, displaying a prevalence of NRL sensitization of 0.75% for the general and 1.2% for the sensitized population. Eleven out of 46 (23.9%) subjects could be defined as NRL asymptomatic, whereas 35 (76.1%) developed symptoms upon exposure to NRL. Specific IgE to NRL was detected for 22 (75.86%) of 29 tested sera. Seventeen out of 22 (77%) sera displayed specific IgE to recombinant allergens with most reactions to Hev b 5, Hev b 6.01 and Hev b 6.02. Immunoblots of NRL extract fractions with patients' sera showed heterogeneous patterns . CONCLUSIONS: SPTs with NRL extract should be routinely performed in patients with respiratory symptoms. Hev b 5, Hev b 6.01 and Hev b 6.02 are the most important allergens, but further characterization of NRL extracts is needed to identify novel allergens and to clarify the role of crossreactive carbohydrate determinants.
[63] - Bienvenu F, Benoit Y, Robert O, Bouvier M, Pain V, Mezzarobba C, et al. Usefulness of recombinant allergens in the diagnosis of latex allergy. 16th European Congress of Immunology, Paris, September 6-9, 2006, poster n° 1975
Aim of the study: To investigate the contribution of recombinant allergens in the diagnosis and follow- up of latex allergy as previously done for birch and timothy pollens. Method: Selection of 61 sera with specific IgE against latex (k82), from adults who presented: peranaesthetic anaphylactic shocks (28), occupational allergies (28) and food allergies (5). Each serum was tested, using the ImmunoCAP 250 system, for specific IgE against each of the 9 latex recombinant allergens (rHev b 1, rHev b 2, rHev b 3, rHev b 5, rHev b 6.01, rHev b 6.02, rHev b 8, rHev b 9, rHev b 11) and bromelain. Results: rHev b 6.01, 6.02, rHev b2 et rHev b 5 were the most frequently positive allergens (52%, 49%, 48% and 43% respectively). rHev b 6.02 was strongly correlated with rHev b 6.01; the same is true for rHev b 2 with rHev b 6.01/02. rHev b 9 is slightly positive in only 2 patients/61. Positive results for rHev b 1 and/or rHev b 3 were observed only in the anaphylactic shock group (9/28). Two patients, with discrepant results between negative prick-test (PT) and recent evocative clinical symptoms of latex allergy had an isolated positive rHev b 5. In this series, 11 patients positive for k82 (range: 0,51 to 6.2 kU/L) had only a positive test for rHev b 8 which was always associated with a negative clinical history for latex together with a negative PT; these patients presented with pollinosis. In the 8/61 patients with negative results for all recombinants, the positivity of k82 was related to the presence of anti-CCD IgE (Carbohydrate Cross-reactive Determinants) detected by the bromelain test. Conclusion: These preliminary results provide interesting information which has to be confirmed in a larger population. The utilisation of three recombinants rHev b 5, rHev b 6.01 and rHev b 8 represents a new useful tool to confirm the clinical diagnosis of latex allergy and to understand cross-reactions. rHev b 1 and rHev b 3 are helpful in case of surgical procedures.
[64] - Wagner S, Breiteneder H. Hevea brasiliensis Latex Allergens: Current Panel and Clinical Relevance. Int Arch Allergy Immunol 2005;136:90-97
Latex allergy has been studied in detail in Europe and the US over the past two decades, resulting in specific guidelines that succeeded in reducing its incidence in high-risk populations within the medical field. How these developments have affected high-risk populations outside the health care scenario is an important unanswered question. In addition, a second wave of latex allergy may occur in nations that are striving to attain higher economic and technologic standards, including population-dense countries such as China. Therefore, the application of Hevea allergens in novel diagnostic assays and the development of specific latex immunotherapy will provide new opportunities for latex allergy research. In this review, we summarize current knowledge on the immunological properties of the 13 officially accepted Hevea brasiliensis latex allergens.
[65] - Beezhold DH, Kostyal DA, Sussman GL. IgE epitope analysis of the hevein preprotein; a major latex allergen. Clin Exp Immunol 1997;108:114-121
We have previously identified the hevein preprotein as a common allergen for latex allergic healthcare workers. The B cell epitopes in the hevein protein that are recognized by IgE of latex-allergic individuals have not been identified. In this study, we examined the hevein preprotein using epitope mapping. Overlapping synthetic peptides of 10 amino acids (two aa overlap) were synthesized on a derivatized cellulose membrane using Fmoc chemistry. The peptide spots were probed with pooled sera from 10 latex-allergic patients, and the IgE-reactive peptides identified with anti-IgE MoAbs. We identified six B cell epitopes within the full length hevein preprotein which bound IgE from latex-allergic patients. Two were located in the N-terminal 5-kD hevein domain and four were observed in the 14-kD C-domain. A broad epitope was located between the N-terminal amino acids 13-24. This epitope had nearly complete homology to wheat germ agglutinin (WGA). Immunological cross-reactivity to WGA was confirmed by Western blot analysis with purified WGA, and this reactivity could be inhibited by latex proteins or WGA. Of the five remaining epitopes, four had homologies to other proteins in the pathogenesis-related family of plant proteins (PR-4). The data demonstrate that hevein has multiple IgE epitopes. The significant homology of these epitopes to a broad family of plant defence proteins further explains the increased prevalence of food allergies in latex-allergic individuals.
[66] - de Silva HD, Gardner LM, Drew AC, Beezhold DH, Rolland JM, O'hehir RE. The hevein domain of the major latex-glove allergen Hev b 6.01 contains dominant T cell reactive sites. Clin Exp Allergy 2004;34:611-618
Summary Background Sensitization to natural rubber latex (Hevea brasiliensis) is a major cause of occupational asthma and rhinitis affecting frequent latex-glove users. Hev b 6.01, a known major latex allergen, is cleaved naturally into hevein (4.7 kDa) and a C-terminal fragment (14 kDa). Hevein is an abundant protein in latex-glove extracts. As the immune response to allergens is initiated by activation of allergen-specific CD4(+) T cells, identification of dominant T cell epitopes is crucial for the development of specific immunotherapy. Objective To identify dominant T cell epitopes of Hev b 6.01 in latex-allergic glove users. Methods Ten latex-allergic frequent glove users and six non-latex-allergic atopic control subjects were selected, based on clinical symptoms and positive latex-specific serum IgE. Serum IgE reactivity to glove extract and recombinant Hev b 6.01 (rHev b 6.01) were analysed by ELISA. Latex-specific short-term oligoclonal T cell lines were generated from peripheral blood of latex-allerg ic subjects. These lines were tested for proliferative responses to overlapping 20-mer peptides of the Hev b 6.01 molecule. CD4(+) T cell intracellular cytokines, IL-4 and IFN-gamma were assessed following stimulation with immobilized anti-CD3 in the presence of IL-2. Results All ten of the latex-allergic patients showed serum IgE binding to glove extract while eight of these also showed IgE binding to rHev b 6.01 by ELISA. Western blotting confirmed reactivity with rHev b 6.01 at around 20 kDa. T cell proliferation assays showed that latex-specific T cell lines from all subjects responded to one or more peptides, with greatest frequency of reactivity to peptides Hev b 6.01 p(10-29) and Hev b 6.01 p(19-38) in the hevein domain. An allergic-type cytokine profile with considerable IL-4 in addition to IFN-gamma was evident from intracellular cytokine staining. Conclusion Hevein is an important T cell as well as B cell immunogen and contains dominant T cell reactive sites
[67] - Reyes-Lopez CA, Pedraza-Escalona M, Mendoza G, Hernandez-Santoyo A, Rodriguez-Romero A. A single amino acid substitution on the surface of a natural hevein isoform (Hev b 6.0202), confers different IgE recognition. FEBS Lett 2006;580:2483-2487
Decreased immune reactivity of isoforms of major allergens has been reported. However, such claims have always been based on experiments with recombinant proteins. This work describes the molecular and physicochemical characterization of a hevein (Hev b 6.0201) natural isoform (Hev b 6.0202), which is present in rubber latex from Hevea brasiliensis. The isoallergen has a single substitution Asn14Asp, which gives rise to local differences in the surface potential, as observed from the crystal structure presented here. Besides, ELISA inhibition using serum pools of adult and pediatric patients showed reduced IgE-binding capacity (~27%) with the isoallergen. Overall, these results are relevant to delineate crucial residues involved in this dominant discontinuous epitope
[68] - Reyes-Lopez CA, Hernandez-Santoyo A, Pedraza-Escalona M, Mendoza G, Hernandez-Arana A, Rodriguez-Romero A. Insights into a conformational epitope of Hev b 6.02 (hevein). Biochem Biophys Res Commun 2004;314:123-130
Hevein (Hev b 6.02) is a major IgE-binding allergen in natural rubber latex and manufactured products. Both tryptophans (Trp(21) and Trp(23)) of the hevein molecule were chemically modified with BNPS-skatole (2-nitrophenylsulfenyl-3-methyl-3(')-bromoindolenine); derivatized allergen failed to significantly inhibit binding of serum IgE in ELISA assays. Similarly, skin prick tests showed that hevein-positive patients gave no response with the modified allergen. Dot blot experiments carried out with anti-hevein mono- and polyclonal antibodies confirmed the importance of Trp(21) and Trp(23) for antibody-recognition, and demonstrated the specific cross-reactivity of other molecules containing hevein-like domains. We also report the structure of Hev b 6.02 at an extended resolution (1.5A) and compare its surface properties around Trp residues with those of similar regions in other allergens. Overall our results indicate that the central part of the protein, which comprises three aromatic and other acidic and polar residues, constitutes a conformational epitope.
[69] - Pedraza-Escalona M, Becerril-Luján B, Agundis C, Domínguez-Ramírez L, Pereyra A, Riaño-Umbarila L, et al. Analysis of B-cell epitopes from the allergen Hev b 6.02 revealed by using blocking antibodies. Mol Immunol 2009;46:668-676
Hev b 6.02 (hevein), identified as a major allergen from natural rubber latex (NRL), is involved in the latex-fruit syndrome and also acts as a pathogenesis defense-related protein. Its 3D structure has been solved at high resolution, and its linear epitopes have already been reported. However, information about conformational epitopes is still controversial, even though it is relevant for an accurate diagnosis and treatment, as well as for the study of allergen-antibody molecular interactions. We sought to analyze the B-cell epitopes of Hev b 6.02 at a molecular and structural level, using specific recombinant antibodies. We obtained a murine monoclonal antibody (mAb 6E7) and three human single chain fragments (scFvs A6, H8, and G7) anti-Hev b 6.02 that were able to compete for hevein binding with serum IgEs from latex allergic patients. In vitro assays showed that the mAb 6E7 and scFv H8 recognized the area of Hev b 6.02 where the aromatic residues are exposed; while the scFv G7 defined the amino and carboxy-terminal regions that lie close to each other, as a different epitope. The structural modeling of the Hev b 6.02-scFv H8 and Hev b 6.02-scFv G7 complexes revealed the putative regions of two conformational epitopes. In one of these, the aromatic residues, as well as polar side chains are important for the interaction, suggesting that they are part of a dominant conformational epitope also presented on the Hev b 6.02-IgE interactions. Antibodies recognizing this important allergen have potential to be used to diagnose and ultimately treat latex allergy.
[70] - Beezhold DH, Sussman GL, Kostyal DA, Chang NS. Identification of a 46-kD latex protein allergen in health care workers. Clin Exp Immunol 1994;98:408-413
Latex allergy is an occupational hazard for health care workers. Extractable latex proteins are known to be allergenic, but most latex allergens have not been specifically identified. The purpose of this study was to characterize the IgE response of latex-allergic patients to latex proteins and to identify common protein allergens. Serum was obtained from 40 individuals who were skin test-positive to latex; 85% were health care workers. Western blots for IgE reactivity were performed using both ammoniated (AL) and non-ammoniated (NAL) latex proteins and IgE-reactive NAL proteins were analysed by microsequence analysis. The patients were grouped according to common patterns of reactivity. Pattern 1, the most common pattern of reactivity (9/40 patients) recognized two protein bands in both NAL and AL at 46 and 110 kD. A second, heterogeneous pattern of reactivity (pattern 2) recognized a diffuse pattern of polypeptides in the AL preparation. The n-terminal amino acid sequences for allergens at 14, 18, 29, 46 and 110 kD were determined. Sequence analysis identified the 14-kD and 18-kD allergens as the hevein proprotein. The 46-kD and 110-kD had identical sequences which were unique from known latex proteins. We conclude that multiple latex proteins are allergens with hevein preprotein and a previously unidentified 46/110-kD protein being commonly recognized in health care workers.
[71] - Kostyal DA, Hickey VL, Noti JD, Sussman GL, Beezhold DH. Cloning and characterization of a latex allergen (Hev b 7): homology to patatin, a plant PLA2. Clin Exp Immunol 1998;112:355-362
We previously identified a 46-kD protein allergen in latex as having amino acid sequence homology to the patatin gene family. The objective of this study was to characterize this protein by molecular techniques. RNA was isolated from the latex or leaf material from Hevea brasiliensis and from potato tubers. Specific polymerase chain reaction (PCR) primers were designed from the amino acid sequence and reverse transcriptase (RT)-PCR amplified a specific product from latex RNA that was subsequently cloned and sequenced. This product was 1493 bp in length with an 1167 bp open reading frame. The deduced amino acid sequence encodes for a 389 aa protein, pI 4.82 with 43% homology to tobacco patatin. Northern analysis of potato, Hevea leaf, and latex RNA demonstrated the message to be most abundant in latex, weakly present in Hevea leaf, but no hybridization occurred with potato RNA. Patatin has lipid acyl-transferase and PLA2-like activity, suggesting it plays a role as a defence-related protein. Other defence-related proteins in latex such as hevein, glucanase, and hevamine are also allergens. Increased production of defence-related proteins as a result of increased tapping of the rubber trees to meet the demand for latex may explain the increased allergenicity of latex.
[72] - Sowka S, Wagner S, Krebitz M, Arif SAM, Yusof F, Kinaciyan T, et al. cDNA cloning of the 43-kDa latex allergen Hev b 7 with sequence similarity to patatins and its expression in the yeast Pichia pastoris. Eur J Biochem 1998;255:213-219
IgE-mediated hypersensitivity to latex proteins present in health care products, particularly in latex gloves, has become an important public health problem in recent years. We purified natural Hev b 7, a 43-kDa patatin-like allergen from the latex of Hevea brasiliensis and determined several internal peptide sequences. A heterologous hybridization probe of a patatin gene of potato, to which these peptides could be aligned best, was used to screen a latex cDNA library. The cDNA encoded an acidic protein of 388 amino acids with a molecular mass of 42.9 kDa. The deduced amino acid sequence had 39-42% identity to patatins from Solanum tuberosum. The purified recombinant Hev b 7 expressed in the yeast Pichia pastoris displayed, similarly to patatins from S. tuberosum, esterase activity. Both natural and recombinant Hev b 7 were recognized by IgE from sera of latex-sensitized allergic individuals. In contrast to patatins from S. tuberosum and Nicotiana tabacum, natural Hev b 7 lacked an N-terminal leader peptide for targeting to the endoplasmatic reticulum and was not glycosylated. These results establish the 43-kDa patatin-like protein as a latex allergen and raise the possibility of different cellular localization and function compared to S. tuberosum patatins.
[73] - Jekel PA, Hofsteenge J, Beintema JJ. The patatin-like protein from the latex of Hevea brasiliensis (Hev b 7) is not a vacuolar protein. Phytochemistry 2003;63:517-522
Upon centrifugation, rubber latex is divided into a layer of rubber particles, the cytosol, and the lutoid-body fraction, which is of vacuolar origin. One of the proteins isolated from the lutoid-body fraction is a protein with a molecular mass of 43 kDa, which has esterase activity on p-nitrophenylpalmitate and which shows significant sequence similarity with patatin, a vacuolar protein with esterase activity from potato (Solanum tuberosum). This protein is a major allergen in rubber latex products (Hev b 7) and can also be isolated from the cytosol fraction of rubber latex. The mature protein isolated from lutoid-bodies has no structural features expected for a vacuolar protein: the N-terminal methionine in the cDNA-derived sequence is cleaved off, the second residue is N-acetylated, and the C-terminal sequence is identical to that in the cDNA-derived sequence. Thus the patatin-like protein in Hevea brasiliensis is not a vacuolar protein, but may be associated with not yet characterized particles in the cytoplasm, which either sediment with lutoid-bodies or remain in the cytosol fraction, depending on the centrifigation conditions.
[74] - Breiteneder H, Sowka S, Wagner S, Krebitz M, Hafner C, Kinaciyan T, et al. Cloning of the patatin-like latex allergen Hev b 7, its expression in the yeast Pichia pastoris and its immunological characterization. Int Arch Allergy Immunol 1999;118:309-310
The 43-kD latex allergen Hev b 7 was purified from the latex of Hevea brasiliensis and identified by N-terminal and internal peptide sequences as highly homologous to patatins. Patatins are storage proteins encoded by a multigene family found in plants such as potato and tomato. We have obtained a cDNA clone coding for a cytoplasmic form of Hev b 7. The recombinant protein was expressed in the methylotrophic yeast Pichia pastoris at 10 mg/l culture supernatant. Both natural Hev b 7 and rHev b 7 were recognized by IgE in 11% of the latex-allergic patients. rHev b 7 inhibited binding to its counterpart in natural rubber latex extracts. Purified rHev b 7 used at concentrations of 10 micrograms/ml in skin prick tests produced wheal-and-flare reactions of sizes equal to those produced by nHev b 7. Furthermore, we were able to show that rHev b 7 possessed esterase activity. A plant expression system for the production of larger quantities of recombinant latex allergens as an alternative to the preparation from H. brasiliensis sap is discussed.
[75] - Vallier P, Balland S, Harf R, Valenta R, Deviller P. Identification of profilin as an IgE-binding component in latex from Hevea brasiliensis: clinical implications. Clin Exp Allergy 1995;25:332-339
Considering the high occurrence of profilin as an allergen in many plant species, the assumption was made that profilin might be an allergen in Hevea brasiliensis, a member of the latex producing Euphorbiaceae family Using IgE-binding inhibition by purified profilins we demonstrated that profilin is an IgE-binding component in the cytosolic fraction of natural latex and, to a lower extent, in the rubber fraction. Thirty-five out of 36 sera containing IgE to ragweed-profilin reacted with profilin from latex, indicating structural homologies between profilins from latex and ragweed. A large percentage (59%) of these sera were found to be positive in CAP latex assay. The preincubation of these sera with purified ragweed profilin greatly inhibited the CAP latex. Because profilin is also present in banana extract, it is likely to be involved in cross-sensitivity to banana and latex. In a group of 19 individuals allergic to latex only two had anti-profilin IgE antibodies. Profilin was barely detectable on glove extract immunoblots, whereas some sera from patients allergic to latex reacted with a 15 kDa allergen which was not profilin. Consequently, IgE antibodies to latex-profilin is a questionable factor for sensitization of occupationally-exposed patients; however, sensitization to profilin should be taken into account when interpreting the results of latex IgE antibody assays.
[76] - Ganglberger E, Radauer C, Wagner S, O'Riordain G, Beezhold DH, Brehler R, et al. Hev b 8, the Hevea brasiliensis latex profilin, is a cross-reactive allergen of latex, plant foods and pollen. Int Arch Allergy Immunol 2001;125:216-227
BACKGROUND: Plant profilins are important pan-allergens. They are responsible for a significant percentage of pollen-related allergies. Limited information is available about their involvement in the latex-fruit syndrome and the cross-reactivities between latex and pollen. We aimed to clone and express the Hevea brasiliensis latex profilin to investigate its allergological significance and serological cross-reactivities to profilins from plant foods and pollens . METHODS: A DNA complementary to messenger RNA (cDNA) coding for the Hevea latex profilin, Hev b 8, was amplified by polymerase chain reaction from latex RNA. Recombinant (r)Hev b 8 was produced in Escherichia coli and used to screen sera from 50 latex- allergic health care workers (HCWs) with well-documented histories of food and pollen allergy and 34 latex-allergic spina bifida (SB) patients. The cross-reactivity of natural Hev b 8 and rHev b 8 with other plant profilins was determined by ELISA inhibition assays. A three-dimensional homology model of Hev b 8 was constructed based on known profilin structures . RESULTS: The cDNA of Hev b 8 encoded a protein of 131 amino acids with a predicted molecular mass of 14 kD. Twelve of the 50 HCWs and 2 of the 34 SB patients were sensitized to Hev b 8. All Hev b 8-sensitized patients showed allergic symptoms to pollen or plant foods. Cross-reactivities between profilins of latex, pollen and plant food were illustrated by their ability to inhibit IgE binding to rHev b 8. Homology modeling of Hev b 8 yielded a structure highly similar to Bet v 2, the birch pollen profilin, with the most distinct differences located at the N-terminus . CONCLUSIONS: We conclude that primary sensitization to latex profilin in the majority of cases takes place via pollen or food profilins. Additionally, pollinosis and food-allergic patients with profilin-specific IgE can be at risk of developing latex allergy
[77] - Vallier P, Balland S, Harf R, Valenta R, Deviller P. Identification of profilin as an IgE-binding component in latex from Hevea brasiliensis: clinical implications. Clin Exp Allergy 1995;25:332-339
Considering the high occurrence of profilin as an allergen in many plant species, the assumption was made that profilin might be an allergen in Hevea brasiliensis, a member of the latex producing Euphorbiaceae family Using IgE-binding inhibition by purified profilins we demonstrated that profilin is an IgE-binding component in the cytosolic fraction of natural latex and, to a lower extent, in the rubber fraction. Thirty-five out of 36 sera containing IgE to ragweed-profilin reacted with profilin from latex, indicating structural homologies between profilins from latex and ragweed. A large percentage (59%) of these sera were found to be positive in CAP latex assay. The preincubation of these sera with purified ragweed profilin greatly inhibited the CAP latex. Because profilin is also present in banana extract, it is likely to be involved in cross-sensitivity to banana and latex. In a group of 19 individuals allergic to latex only two had anti-profilin IgE antibodies. Profilin was barely detectable on glove extract immunoblots, whereas some sera from patients allergic to latex reacted with a 15 kDa allergen which was not profilin. Consequently, IgE antibodies to latex-profilin is a questionable factor for sensitization of occupationally-exposed patients; however, sensitization to profilin should be taken into account when interpreting the results of latex IgE antibody assays.
[79] - Posch A, Chen Z, Wheeler CH, Dunn MJ, Raulf-Heimsoth M, Baur X. Characterization and identification of latex allergens by two-dimensional electrophoresis and protein microsequencing. J Allergy Clin Immunol 1997;99:385-395
Proteins of natural rubber latex cause IgE-mediated sensitization in 3% to 18% of health care workers and in up to 50% of patients with spina bifida. OBJECTIVE: This study was aimed at the generation of a comprehensive latex protein database by two-dimensional electrophoresis (2-DE). METHODS: Proteins extracted from fresh Hevea brasiliensis latex were separated by 2-DE. IgE-reactive proteins were analyzed by immunoblotting with sera of health care workers with latex allergy. Protein microsequencing and monoclonal antibodies were used to identify the latex allergens. RESULTS: The latex C-serum 2-DE map was very complex and exhibited about 200 distinct polypeptides. The proteins eluted from the latex particles consisted primarily of two groups of acidic proteins located in the 8 to 14 kd and 22 to 24 kd areas of the 2-DE map. Major IgE-reactivity was detected with C-serum proteins in the 56, 45, 30, 20, 14, and <6.5 kd areas of the immunoblots. The 8 to 14 kd particle proteins exhibited distinct IgE reactivity, whereas the 22 to 24 kd proteins were not stained. Seven of the soluble IgE-reactive protein spots showed high homology with enolase, superoxide dismutase, triosephosphate isomerase, proteasome subunit, and chitinase and represent previously undescribed latex allergens; whereas nine protein spots corresponded to known latex allergens, namely prohevein, hevein, prohevein C-domain, and hevamine. As identified by monoclonal antibodies, the IgE-reactive latex particle proteins mainly represent the allergenic rubber elongation factor. CONCLUSIONS: Two-dimensional electrophoresis, followed by immunoblotting and protein microsequencing, can rapidly identify a large number of IgE-binding latex proteins. The 2-DE latex maps generated will provide valuable information for the development of strategies to isolate the relevant latex allergens. Because the novel latex allergens are common plant enzymes, they may also act as cross-reacting proteins in various foods.
[80] - Sussman GL, Beezhold DH, Kurup VP. Allergens and natural rubber proteins. J Allergy Clin Immunol 2002;110(2 part 2):S33-S39
BACKGROUND: Allergy to natural rubber latex (NRL) results from exposure to proteins derived from Hevea brasiliensis. Type I latex hypersensitivity is observed in certain occupational and other high-risk groups with frequent exposure to NRL products. This includes health care workers (HCWs), workers in the latex industry, children with spina bifida, and atopic individuals . OBJECTIVES: Early reliable diagnosis and avoidance are required for better patient care. Standardized reagents are not presently available for in vitro and in vivo testing and treatment of patients with latex allergy. However, a number of allergens have been isolated and characterized from Hevea latex and NRL products. Currently, a total of 11 major and minor allergens are designated by the International Allergen Nomenclature Committee. This article reviews the structural and functional characteristics of these latex allergenic proteins . RESULTS: NRL-allergenic proteins include those involved in the biosynthesis of polyisoprene and coagulation of latex rubber elongation factor, small rubber particle protein, prohevein, and patatin. Pathogenesis-related proteins include beta-1,3-glucanases, chitinases, and hevamine; and the structural proteins include microhelix protein complex, proline-rich protein, profilins, enolases, and manganese superoxide dismutase. Recombinant allergens demonstrated skin test reactivity in patients with latex allergy. The minimal level of skin test reactivity was about 70 pg/mL for NRL and 1 ng/mL for recombinant allergens. The use of selected recombinant latex allergens (Hev b 5, Hev b 6, and Hev b 7) in skin prick tests identified 93% of allergic individuals, mainly health care workers . CONCLUSIONS: Recombinant latex allergens are clinically reactive and can be produced in a standardized manner, which could potentially provide safe and sensitive reagents for the diagnosis and treatment of type I latex allergy.
[81] - Posch A, Chen Z, Wheeler CH, Dunn MJ, Raulf-Heimsoth M, Baur X. Characterization and identification of latex allergens by two-dimensional electrophoresis and protein microsequencing. J Allergy Clin Immunol 1997;99:385-395
Proteins of natural rubber latex cause IgE-mediated sensitization in 3% to 18% of health care workers and in up to 50% of patients with spina bifida. OBJECTIVE: This study was aimed at the generation of a comprehensive latex protein database by two-dimensional electrophoresis (2-DE). METHODS: Proteins extracted from fresh Hevea brasiliensis latex were separated by 2-DE. IgE-reactive proteins were analyzed by immunoblotting with sera of health care workers with latex allergy. Protein microsequencing and monoclonal antibodies were used to identify the latex allergens. RESULTS: The latex C-serum 2-DE map was very complex and exhibited about 200 distinct polypeptides. The proteins eluted from the latex particles consisted primarily of two groups of acidic proteins located in the 8 to 14 kd and 22 to 24 kd areas of the 2-DE map. Major IgE-reactivity was detected with C-serum proteins in the 56, 45, 30, 20, 14, and <6.5 kd areas of the immunoblots. The 8 to 14 kd particle proteins exhibited distinct IgE reactivity, whereas the 22 to 24 kd proteins were not stained. Seven of the soluble IgE-reactive protein spots showed high homology with enolase, superoxide dismutase, triosephosphate isomerase, proteasome subunit, and chitinase and represent previously undescribed latex allergens; whereas nine protein spots corresponded to known latex allergens, namely prohevein, hevein, prohevein C-domain, and hevamine. As identified by monoclonal antibodies, the IgE-reactive latex particle proteins mainly represent the allergenic rubber elongation factor. CONCLUSIONS: Two-dimensional electrophoresis, followed by immunoblotting and protein microsequencing, can rapidly identify a large number of IgE-binding latex proteins. The 2-DE latex maps generated will provide valuable information for the development of strategies to isolate the relevant latex allergens. Because the novel latex allergens are common plant enzymes, they may also act as cross-reacting proteins in various foods.
[82] - Hemery ML, Verdier R, Daban P, Sellier N, Dujols P, Demoly P. Sensibilisation aux gants en latex poudrés: une prévalence élevée chez le personnel hospitalier. Presse Med 2005;34:1363-1369
INTRODUCTION: The prevalence of latex allergies in industrial countries has skyrocketed since the 1980s. Between 2.6 and 22% of hospital workers are diagnosed with latex allergy, which has been recognized as an occupational hazard in France since 1997. AIM: To assess the prevalence of latex allergy among Montpellier University Hospital Center staff . METHODS: From 1999 to 2002, we interviewed and conducted skin-prick tests on 537 hospital staff members from different departments and with different degrees of exposure to airborne latex allergens . RESULTS: Symptoms while using natural-rubber latex gloves (most often contact dermatitis) were reported by 88 (16.4%) staff members. Overall work-related allergic symptoms included rhinitis, reported by 65 (12.1%), contact urticaria by 28 (5.2%), and bronchial symptoms by 16 (1.1%). In all, sensitization to latex was identified in 7.1% of our staff, and this percentage was higher (11.3%) in units where latex gloves were used more often. Sensitization was associated with high latex exposure and atopy . CONCLUSION: This high rate of latex sensitization and the risk factors are similar to those already published. Based on this study, we have eliminated powdered latex gloves, as have many other hospitals.
[83] - Lavaud F, Prévost A, Cossart C, Guérin L, Bernard J, Kochman S. Allergy to latex, avocado pear, and banana: evidence for a 30 kd antigen in immunoblotting. J Allergy Clin Immunol 1995;95:557-564
Allergens of natural latex, latex gloves, avocado pear, and banana extracts were investigated by an immunoblotting technique in sera of patients experiencing associated latex and fruit allergies. Extracts were separated by sodium dodecyl-sulfate-polyacrylamide gel electrophoresis and electroblotted onto nitrocellulose. After incubation with patients' sera, IgE antibodies were revealed by a goat anti-human IgE alkaline-phosphatase conjugate. Seventeen serum samples from patients with well-documented latex allergy were studied. Among these patients, 10 demonstrated an allergy to avocado pear sometimes associated with banana. In sera from patients with latex and fruit allergy, prominent IgE binding was revealed at about 30 kd with latex and fruit extracts. Serum controls remained negative. Cross-inhibition of immunoblotting confirmed that this main allergen is linked to a common epitope present in latex and fruits. This must be related to clinical findings and previous observations of cross-reactivity.
[84] - O'Riordain G, Radauer C, Hoffmann-Sommergruber K, Adhami F, Peterbauer CK, Blanco C, et al. Cloning and molecular characterization of the Hevea brasiliensis allergen Hev b 11, a class I chitinase. Clin Exp Allergy 2002;32:455-462
In the last 10 years type-I allergy against proteins from Hevea brasiliensis latex has become an acknowledged medical issue. Fruit-allergic patients represent one risk group for developing latex allergy. Class I chitinases have been identified from chestnut, avocado and banana as relevant allergens. The chitin binding (hevein) domain from these class I chitinases has been postulated to bear the important IgE binding epitopes. Objective : To clone the cDNA of an allergenic latex class I chitinase, to express the recombinant protein and to determine its IgE cross-reactivity with hevein (Hev b 6.02). Method s : A full-length cDNA coding for a class I chitinase has been isolated from Hevea latex RNA by reverse transcription followed by PCR. The chitinase encoding sequence has been subcloned into the pMAL expression vector and expressed in E. coli as a fusion protein to maltose binding protein. The highly enriched recombinant protein fraction has been tested for its IgE binding capacity in immunoblots and ELISA. Furthermore, the pathogenesis-related function of the recombinant protein was tested in a fungal growth inhibition assay. Result s : The Hevea brasiliensis latex chitinase, designated Hev b 11, displays 70% identity to the endochitinase from avocado and its hevein-domain 58% to hevein (Hev b 6.02). The recombinant Hev b 11-maltose binding protein is recognized by latex- and fruit-allergic patients with IgE binding in both, ELISA and immunoblots. Pre-incubation of sera with rHev b 11-maltose binding protein showed an overall 16% inhibition of subsequent binding to rHev b 6.02-maltose binding protein on solid phase. The growth of F. oxysporum was inhibited in a dose dependent manner by addition of rHev b 11-maltose binding protein to the culture. Conclusions : Hev b 11, a class I chitinase, is another allergen from Hevea latex with a chitin binding domain and displays a different IgE binding capacity compared with hevein.
[85] - Wagner S, Breiteneder H. Hevea brasiliensis Latex Allergens: Current Panel and Clinical Relevance. Int Arch Allergy Immunol 2005;136:90-97
Latex allergy has been studied in detail in Europe and the US over the past two decades, resulting in specific guidelines that succeeded in reducing its incidence in high-risk populations within the medical field. How these developments have affected high-risk populations outside the health care scenario is an important unanswered question. In addition, a second wave of latex allergy may occur in nations that are striving to attain higher economic and technologic standards, including population-dense countries such as China. Therefore, the application of Hevea allergens in novel diagnostic assays and the development of specific latex immunotherapy will provide new opportunities for latex allergy research. In this review, we summarize current knowledge on the immunological properties of the 13 officially accepted Hevea brasiliensis latex allergens.
[86] - Rihs HP, Dumont B, Rozynek P, Lundberg M, Cremer R, Brüning T, et al. Molecular cloning, purification, and IgE-binding of a recombinant class I chitinase from Hevea brasiliensis leaves (rHev b 11.0102). Allergy 2003;58:246-251
BACKGROUND: Class I chitinase in natural rubber latex (NRL) has been assumed to be an important allergen, especially concerning its cross-reactivity with fruits like avocado and banana . OBJECTIVES: The present study aimed to produce a recombinant latex class I chitinase from Hevea brasiliensis leaves and to study its immunoglobulin (Ig)E-binding reactivity . METHODS: A class I chitinase-specific complementary DNA from H. brasiliensis leaves was synthesized, subcloned, sequenced and overexpressed in fusion with the maltose-binding protein (MBP) in Escherichia coli. The IgE-binding reactivity of this protein was studied by the Pharmacia CAP System and by immunoblot experiments using sera from latex-allergic patients . RESULTS: The rHev b 11.0102 was found to have a length of 295 amino acid residues and contains an N-terminal hevein-like domain with a 56% homology to hevein. Analysis by the CAP method revealed the presence of rHev b 11.0102-specific IgE antibodies in 17 of 58 sera (29%) of IgE-mediated latex-allergic subjects tested. Immunoblot analysis of the MBP-rHev b 11.0102 fusion protein and the MBP carrier protein as a negative control confirmed the IgE-reactivity of rHev b 11.0102 . CONCLUSION: Due to its IgE-reactivity rHev b 11.0102 represents an allergen of intermediate prevalence in NRL. Its property to cross-react with certain fruits makes it an important supplement in the diagnostic panel of recombinant NRL allergens.
[87] - Beezhold DH, Hickey VL, Kostyal DA, Puhl H, Zuidmeer L, van Ree R, et al. Lipid transfer protein from Hevea brasiliensis (Hev b 12), a cross-reactive latex protein. Ann Allergy Asthma Immunol 2003;90:439-445
BACKGROUND: Latex-allergic individuals experience clinical cross-reactivity to a large number of fruits and vegetables. Much of the cross-reactivity can be attributed to Hev b 6, but evidence indicates that additional cross-reactive allergens may be present. A common pan-allergen, which has not previously been identified in latex, but may contribute to this cross-reactivity is lipid transfer protein (LTP). We sought to determine whether Hevea brasiliensis produces LTP and whether it would bind immunoglobulin E from latex-allergic patients. METHODS: LTP was identified in H. brasiliensis RNA by polymerase chain reaction using degenerate primers. The entire cDNA was obtained by polymerase chain reaction using rapid amplification of cDNA ends reactions. The complete coding sequence for LTP was determined and produced as a recombinant protein using the glutathione S-transferase and pET32 expression systems. Immunoblot analysis of sera from latex-allergic patients was used to determine whether patients recognize LTP as an allergen. RESULTS: We identified a 662-basepair cDNA with a 351-basepair open reading frame that encodes for a 116-amino acid protein. The protein has significant homology to the family of nonspecific LTPs. We expressed the protein as a mature LTP of 92 amino acids with a predicted isoelectric point of 10.8 and molecular weight of 9.3 kDa. Immunoblots demonstrated specific immunoglobulin E for LTP in the sera of 9 of 37 (24%) latex-allergic individuals. CONCLUSIONS: We describe the initial identification of rLTP in H. brasiliensis that may be important as a cross-reactive pan-allergen (Hev b 12).
[88] - Wagner S, Breiteneder H. Hevea brasiliensis Latex Allergens: Current Panel and Clinical Relevance. Int Arch Allergy Immunol 2005;136:90-97
Latex allergy has been studied in detail in Europe and the US over the past two decades, resulting in specific guidelines that succeeded in reducing its incidence in high-risk populations within the medical field. How these developments have affected high-risk populations outside the health care scenario is an important unanswered question. In addition, a second wave of latex allergy may occur in nations that are striving to attain higher economic and technologic standards, including population-dense countries such as China. Therefore, the application of Hevea allergens in novel diagnostic assays and the development of specific latex immunotherapy will provide new opportunities for latex allergy research. In this review, we summarize current knowledge on the immunological properties of the 13 officially accepted Hevea brasiliensis latex allergens.
[89] - Rihs HP, Rueff F, Lundberg M, Rozynek P, Garcia-Selles J, Barber D, et al. Specific IgE-Binding of rHev b 12 Is Restricted to Fruit-Allergic Patients. AAAAI 60th Annual Meeting, San Francisco, 19-23 March 2004, Poster n°205
Rationale Lipid transfer proteins (LTP) have been identified as relevant allergens. Their role in latex allergy is unclear. This study aimed to produce a recombinant latex-LTP (rHev b 12) to examine its IgE-binding properties. Methods A recombinant generated Maltose-Binding-Protein (MBP)-rHev b 12 fusion protein was coupled to ImmunoCAP for the determination of specific IgE. 48 sera of atopic patients (14 from Spain, 34 from Germany) with fruit-allergy were examined. 25 of these sera displayed positive latex-specific IgE values. Results rHev b 12-specific IgE was measured in four Spanish peach-allergic patients (CAP-values: 0.88-2.27 kU/L), whereby one had an additional latex-sensitization. Two sera from cherry-allergic German patients with latex-sensitization displayed also rHev b 12-specific IgE antibodies (0.68 and 0.96 kU/L). All other sera and the MBP controls revealed negative results (<0.35 kU/L). Conclusions Latex-LTP (rHev b 12)-specific IgE-binding in peach- and cherry-allergic patients seem to be a result of partially common IgE-binding epitopes with the LTPs of peach (homology to Pru p 3: 65%) and cherry (homology to Pru av 3: 61%). Although rHev b 12 seems to have minor relevance as a latex allergen in Central Europe, its significance as a cross-reactive allergen in Mediterranean countries like Spain has to be kept in mind.
[90] - Palosuo T, Lehto M, Kotovuori A, Kalkkinen N, Blanco C, Poza P, et al. Latex allergy: low prevalence of immunoglobulin E to highly purified proteins Hev b 2 and Hev b 13. Clin Exp Allergy 2007;37:1502-1511
BACKGROUND: Hevea brasiliensis (Hev b) 2 and Hev b 13 have recently been identified as major latex allergens by detecting specific IgE antibodies in >50% of sera from Hev b latex-allergic individuals . OBJECTIVE: We assessed the prevalence rates for sensitization to extensively purified latex allergens in patients from three diverse geographical areas . METHODS: Native Hev b 2, Hev b 5, Hev b 6.01 and Hev b 13 were purified by non-denaturating chromatography and were used in ELISAs to assess sera from 215 latex-allergic patients and 172 atopic non-sensitized controls from Finland, Spain and the United States to detect allergen-specific IgE antibodies . RESULTS: Unexpectedly, even highly purified Hev b 13 contained epitope(s) to which Hev b 6-specific human IgE antibodies bound effectively. Further purification, however, reduced the prevalence of IgE antibody reactivity to low levels: 15%, 5% and 11% for Hev b 2, and 18%, 30% and 27% for Hev b 13 among latex-allergic Finnish, Spanish and American patients, respectively. Interestingly, Finnish patients had a lower prevalence of Hev b 5-specific IgE antibody (28%) as compared with Spanish (49%) and American (71%) patients. The prevalence of Hev b 6.01-specific IgE reactivity was uniformly >50% in all three populations . CONCLUSION: Neither Hev b 2 nor Hev b 13 appear to be major latex allergens when evaluated in serological assays using highly purified allergens. The reason(s) for the observed differences in published sensitization rates in various geographic regions requires further study. The purity of the allergen preparations has a marked impact on the accuracy of latex-specific IgE antibody detection in epidemiological studies and in the serological diagnosis of latex allergy.
[91] - Bernstein DI, Biagini RE, Karnanni R, Hamilton R, Murphy K, Bernstein C, et al. In vivo sensitization to purified Hevea brasiliensis proteins in health care workers sensitized to natural rubber latex. J Allergy Clin Immunol 2003;111:610-616
BACKGROUND: Thirteen proteins of natural rubber latex (Hevea brasiliensis) known to bind human IgE have been isolated and characterized as Hev b allergens. However, the in vivo importance of native Hev b allergens has not been defined in health care workers (HCWs) with natural rubber latex (NRL) allergy . OBJECTIVES: The principal aim of this study was to identify the major in vivo Hev b allergens in HCWs with NRL allergy confirmed by percutaneous sensitivity to nonammoniated latex (NAL) . METHODS: Skin prick testing was performed with 7 (native) proteins purified from NAL (Hev b 1, 2, 3, 4, 6.01, 7.01, and a newly described Hev b 13) and recombinant Hev b 5 in 62 HCWs with histories of NRL allergy (group 1) confirmed by percutaneous reactivity to NAL and in 49 atopic HCWs without NRL allergy (group 2). Serial 10-fold concentrations of Hev b proteins (5 x 10(-5) microg/mL to 50 microg/mL) were tested; serum samples of subjects were assayed for serum specific IgE by immunoassays . RESULTS: Hev b 2, Hev b 5, Hev b 6.01, and Hev b 13 produced skin reactions in more than 60% of group 1 subjects, with Hev b 1, 3, 4, and 7.01 eliciting reactions in less than 50%. Only 1 of 49 group 2 workers reacted to a single Hev b antigen (Hev b 13). Specificity of 7 Hev b allergens was 100% and 98% for Hev b 13 in identifying workers with confirmed NRL allergy. Specific IgE by AlaSTAT and CAP immunoassays was elevated in 40 of 60 (67%) and 33 of 62 (53%) of NAL-reactive workers and produced false-positive test results in 4 of 49 (8%) and 3 of 48 (6%) group 2 subjects, respectively . CONCLUSION: Hev b 2, 5, and 6.01 are major in vivo allergens and Hev b 13 is a new major in vivo allergen among HCWs with allergy to NRL.
[92] - Jekel PA, Hartmann BH, Beintema JJ. The primary structure of hevamine, an enzyme with lysozyme/chitinase activity from Hevea brasiliensis latex. Eur J Biochem 1991;200:123-130
The primary structure of hevamine, an enzyme with lysozyme/chitinase activity from Hevea brasiliensis latex, has been determined predominantly with conventional non-automatic methods. The positions of three disulfide bridges have been determined. The sequence has about 60% identity with that of a chitinase from cucumber and 95% with the N-terminal sequence of the lysozyme/chitinase of Parthenocissus quinquefolia. The half-cystine residues in hevein and cucumber chitinase are located at identical positions. Hevamine is a basic protein from the lutoids (vacuoles) of rubber latex and may have a role in plugging the latex vessels and cessation of latex flow. The differences in cellular location, charge properties and sequence between hevamine and cucumber chitinase are similar to those between class I and class II chitinases from tobacco and other plant species.
[93] - Posch A, Chen Z, Wheeler CH, Dunn MJ, Raulf-Heimsoth M, Baur X. Characterization and identification of latex allergens by two-dimensional electrophoresis and protein microsequencing. J Allergy Clin Immunol 1997;99:385-395
Proteins of natural rubber latex cause IgE-mediated sensitization in 3% to 18% of health care workers and in up to 50% of patients with spina bifida. OBJECTIVE: This study was aimed at the generation of a comprehensive latex protein database by two-dimensional electrophoresis (2-DE). METHODS: Proteins extracted from fresh Hevea brasiliensis latex were separated by 2-DE. IgE-reactive proteins were analyzed by immunoblotting with sera of health care workers with latex allergy. Protein microsequencing and monoclonal antibodies were used to identify the latex allergens. RESULTS: The latex C-serum 2-DE map was very complex and exhibited about 200 distinct polypeptides. The proteins eluted from the latex particles consisted primarily of two groups of acidic proteins located in the 8 to 14 kd and 22 to 24 kd areas of the 2-DE map. Major IgE-reactivity was detected with C-serum proteins in the 56, 45, 30, 20, 14, and <6.5 kd areas of the immunoblots. The 8 to 14 kd particle proteins exhibited distinct IgE reactivity, whereas the 22 to 24 kd proteins were not stained. Seven of the soluble IgE-reactive protein spots showed high homology with enolase, superoxide dismutase, triosephosphate isomerase, proteasome subunit, and chitinase and represent previously undescribed latex allergens; whereas nine protein spots corresponded to known latex allergens, namely prohevein, hevein, prohevein C-domain, and hevamine. As identified by monoclonal antibodies, the IgE-reactive latex particle proteins mainly represent the allergenic rubber elongation factor. CONCLUSIONS: Two-dimensional electrophoresis, followed by immunoblotting and protein microsequencing, can rapidly identify a large number of IgE-binding latex proteins. The 2-DE latex maps generated will provide valuable information for the development of strategies to isolate the relevant latex allergens. Because the novel latex allergens are common plant enzymes, they may also act as cross-reacting proteins in various foods.
[94] - Alenius H, Kalkkinen N, Lukka M, Reunala T, Turjanmaa K, Makinen-Kiljunen S, et al. Prohevein from the rubber tree (Hevea brasiliensis) is a major latex allergen. Clin Exp Allergy 1995;25:659-665
There is general agreement that proteins eluting from different natural rubber latex products can cause immediate type hypersensitivity reactions in latex-allergic patients. However, there is as yet no consensus as to what are the most important allergens in natural rubber latex. OBJECTIVE: We wanted to purify and characterize at the primary structure level three natural latex proteins, suggested to represent significant allergens. METHODS: Proteins were purified from ultracentrifuged bottom fraction of natural rubber latex using high performance liquid chromatography gel filtration and reversed phase chromatography. Purified proteins were subjected to tryptic cleavage, peptide separation and amino acid sequencing. Immunoblotting was used to demonstrate IgE antibodies to the purified proteins in sera from latex-allergic patients. RESULTS: A 20 kDa protein was identified by amino acid sequencing as prohevein, a major protein in the rubber tree Hevea brasiliensis, and a 30 kDa natural rubber latex protein as hevamine, another essential rubber tree protein. A third, previously undescribed natural rubber latex protein, showed high homology to several plant endo-1,3-beta-glucosidases. In immunoblotting, the purified prohevein bound IgE antibodies from 24/29 (83%) sera of latex-allergic patients including positive results in 4/6 latex-allergic children with spina bifida or other congenital anomalies. The purified prohevein elicited positive skin-prick test reactions in all six latex-allergic patients showing IgE to prohevein. The purified 36 kDa protein bound IgE from 6/29 (21%) latex-allergic sera, and the purified hevamine from only 1/29 patient sera. CONCLUSION: The observed high frequency of IgE antibodies to prohevein suggests that this protein is a major natural rubber latex allergen.
[95] - Baur X, Chen Z, Rozynek P, Duser M, Raulf-Heimsoth M. Cross-reacting IgE antibodies recognizing latex allergens, including Hev b 1, as well as papain. Allergy 1995;50:604-609
The cross-reactivity of IgE antibodies recognizing epitopes of latex allergens and papain was studied in sera of 36 latex-exposed subjects and 22 papain workers. Eight out of 24 latex-sensitized persons also showed a positive reaction to papain in the CAP assay (mostly of low or moderate degree). On the other hand, six out of the 12 sensitized papain workers also revealed IgE binding to latex allergen(s). Reciprocal inhibition experiments confirmed that groups monosensitized to one of the two allergens can be separated from a group showing partial or nearly complete immunologic cross-reactivity. Papain inhibited IgE binding by 20-33% in these subjects, whereas IgE binding to papain was strongly blocked in most cases. Comparison between the primary sequences of Hev b 1, a major latex allergen, and papain suggests that the cross-reactivity may be due to several identical trimers and tetramers.
[96] - Rozynek P, Posch A, Baur X. Cloning, expression and characterization of the major latex allergen prohevein. Clin Exp Allergy 1998;28:1418-1426
BACKGROUND: About 70-80% of latex allergic health care workers are sensitized to prohevein (Hev b 6.01), a 20 kDa cysteine-rich chitin-binding protein of Hevea latex. OBJECTIVE: This study reports on the bacterial cloning, expression and immunochemical characterization of rHev b 6.01. METHODS: Prohevein was expressed in the periplasmatic space of Escherichia coli as maltose binding protein (MBP) fusion protein and purified to homogeneity after factor Xa cleavage. The IgE binding capacity of both rHev b 6.01 and prohevein isolated from fresh Hevea latex was compared by immunoblotting experiments using sera of latex-allergic patients. The diagnostic value of rHev b 6.01 was analysed by enzyme allergosorbent test (EAST). RESULTS: Two different cDNA clones of rHev b 6.01 were established. The deduced amino acid sequence of both clones revealed two and three amino acid differences in the C-terminal domain of prohevein compared with the original database entry. Purified rHev b 6.01 bound with high affinity to chitin as its natural counterpart isolated from natural latex. In IgE-immunblotting using sera of affected subjects binding intensity to both proteins was comparable indicating a very high antigenic similarity. The diagnostic value of MBP-prohevein was tested in EAST using sera of 33 latex-allergic subjects. The in vitro test showed high sensitivity and specificity and proved the diagnostic value of uncleaved MBP-prohevein. CONCLUSIONS: The production of recombinant latex key allergens with defined quality like prohevein is a straightforward strategy for the development of standardized in vitro test systems.
[97] - Posch A, Chen Z, Wheeler CH, Dunn MJ, Raulf-Heimsoth M, Baur X. Characterization and identification of latex allergens by two-dimensional electrophoresis and protein microsequencing. J Allergy Clin Immunol 1997;99:385-395
Proteins of natural rubber latex cause IgE-mediated sensitization in 3% to 18% of health care workers and in up to 50% of patients with spina bifida. OBJECTIVE: This study was aimed at the generation of a comprehensive latex protein database by two-dimensional electrophoresis (2-DE). METHODS: Proteins extracted from fresh Hevea brasiliensis latex were separated by 2-DE. IgE-reactive proteins were analyzed by immunoblotting with sera of health care workers with latex allergy. Protein microsequencing and monoclonal antibodies were used to identify the latex allergens. RESULTS: The latex C-serum 2-DE map was very complex and exhibited about 200 distinct polypeptides. The proteins eluted from the latex particles consisted primarily of two groups of acidic proteins located in the 8 to 14 kd and 22 to 24 kd areas of the 2-DE map. Major IgE-reactivity was detected with C-serum proteins in the 56, 45, 30, 20, 14, and <6.5 kd areas of the immunoblots. The 8 to 14 kd particle proteins exhibited distinct IgE reactivity, whereas the 22 to 24 kd proteins were not stained. Seven of the soluble IgE-reactive protein spots showed high homology with enolase, superoxide dismutase, triosephosphate isomerase, proteasome subunit, and chitinase and represent previously undescribed latex allergens; whereas nine protein spots corresponded to known latex allergens, namely prohevein, hevein, prohevein C-domain, and hevamine. As identified by monoclonal antibodies, the IgE-reactive latex particle proteins mainly represent the allergenic rubber elongation factor. CONCLUSIONS: Two-dimensional electrophoresis, followed by immunoblotting and protein microsequencing, can rapidly identify a large number of IgE-binding latex proteins. The 2-DE latex maps generated will provide valuable information for the development of strategies to isolate the relevant latex allergens. Because the novel latex allergens are common plant enzymes, they may also act as cross-reacting proteins in various foods.
[98] - Mengumpun K, Tayapiwatana C, Hamilton RG, Sangsupawanich P, Wititsuwannakul R. Hydrophobic allergens from the bottom fraction membrane of Hevea brasiliensis. Asian Pac J Allergy Immunol 2008;26:129-136
Several proteins of rubber latex have been recognized as allergens causing immediate hypersensitivity in humans. In this study, a bottom fraction membrane (BFM) protein preparation from Hevea brasiliensis trees grown in southern Thailand was used to detect specific IgE in four groups of serum samples. The first group included 170 samples of latex glove factory workers (LGWs); group 2 consisted of the sera of 35 health care workers (HCWs) who were repeatedly exposed to powdered latex gloves; groups 3 and 4 were 31 positive and 22 negative sera, respectively, obtained from Johns Hopkins University School of Medicine, Baltimore, USA, tested for IgE to latex allergen. It was found that 56/170 (33%), 5/35 (14%), 11/31 (35.5%) and 1/22 (4.5%) samples of the LGWs, HCWs, CAP+ and CAP- groups had significant IgE to the BFM proteins, respectively. However, of all subjects only one subject of group 1 had experienced allergic morbidity consisting of eczema, conjunctivitis and asthma. The IgE of this subject bound to a 55 kDa component in the rubber latex BFM preparation. Thus, this protein may be regarded as a novel, although minor, latex allergen. Further investigation is needed to characterize the component and to pinpoint its allergenic role.
[99] - Yip E, Cacioli P. The manufacture of gloves from natural rubber latex. J Allergy Clin Immunol 2002;110(2 part 2):S3-S14
Gloves that will provide a barrier of protection from infectious organisms are an essential feature of medical practice for the protection of both patients and medical personnel. Natural rubber latex has consistently been the most satisfactory raw material for the manufacture of gloves. Certain latex proteins, carried over into the finished product by inadequate manufacturing processes, may pose a risk of provoking allergic reactions in some patients and medical workers. As with any allergy, the risk depends on the route of exposure and dose. Hence, the method of manufacture, including the means used to coat gloves to make donning easy, can influence the eventual exposure of sensitive people to latex allergens. In this article, we describe the several processes in use and their effects on latex protein content.
[100] - Sutherland MF, Drew A, Rolland JM, Slater JE, Suphioglu C, O'Hehir RE. Specific monoclonal antibodies and human immunoglobulin E show that Hev b 5 is an abundant allergen in high protein powdered latex gloves. Clin Exp Allergy 2002;32:583-589
BackgroundHev b 5 is a major latex allergen recognized predominantly by latexallergic health care workers (HCWs). Recombinant Hev b 5 (rHev b 5) was previously expressed as a fusion protein with maltose binding protein (MBP), itself an immunogenic molecule; therefore nonfusion rHev b 5 is desirable. Moreover, standardized immunological assays for the detection of Hev b 5 are currently lacking and may have important implications for both allergen avoidance and diagnosis in latex allergy. ObjectivesTo generate and use Hev b 5specific mAbs to determine the relative abundance of Hev b 5 in different latex extracts, correlating this with the IgE reactivity of latexallergic HCWs and to produce nonfusion rHev b 5. MethodsFor the production of mAbs, mice were immunized with rHev b 5/MBP fusion protein and mAbs selected with rHev b 5/MBP but not MBP reactivity. The mAb reactivity was compared with polyclonal IgE from latexallergic HCWs using direct and inhibition ELISA and immunoblot assays. Recombinant Hev b 5 was expressed and purified in the pPROEXHTa bacterial expression system. ResultsFour Hev b 5specific mAbs were produced. Immunoblotting and ELISA using the mAbs indicate abundant Hev b 5 in high protein powdered latex glove extracts as compared with crude latex sap extracts. High quality surgical gloves with no detectable protein have no detectable Hev b 5. Inhibition ELISAs using serum IgE from latexallergic HCWs and Hev b 5specific mAbs gave strong correlation. Nonfusion recombinant Hev b 5 was successfully expressed and purified, showing reactivity with both the Hev b 5specific mAbs and serum IgE of latexallergic HCWs. ConclusionHev b 5specific mAbs and human IgE from latexallergic HCWs demonstrate the greater content of Hev b 5 in high protein powdered glove extracts. This may explain the observed higher frequency of sensitization to this allergen in HCWs.
[101] - Marchetti-Deschmann M, Allmaier G. Allergenic compounds on the inner and outer surfaces of natural latex gloves: MALDI mass spectrometry and imaging of proteinous allergens. J Mass Spectrom 2009;44:61-70
Natural latex gloves are the cause of a severe health problem to an increasing number of healthcare workers or patients due to the presence of protein allergens as Hevein or Rubber Elongation Factor (REF). One of the most challenging problems is the in situ localization of theses allergens in, e.g. gloves, to estimate the allergenic potential of the latex material. A sample preparation protocol applying a binary matrix-assisted laser desorption/ionization(MALDI) matrix containing alpha-cyano-4-hydroxy cinnamic acid (CHCA) and 2,5-dihydroxy benzoic acid (DHB) on trifluoro acetic acid (TFA) etched latex glove surfaces allowed the direct determination (exact molecular weight) of Hevein, REF and a truncated form of REF (tREF) within nine different brands of natural latex gloves by means of MALDI-TOF-MS in the linear mode. MALDI mass spectrometry demonstrated that Hevein, tREF and REF were present on the inner surfaces (in direct contact with the skin) of many, but not all, investigated gloves without any prior extraction procedure. Additionally, different isoforms of the allergen Hevein were detected (exhibiting ragged C-termini). tREF and REF could always be detected beside each other, but were not observed on every latex glove sample, which contained Hevein. It was also demonstrated that there is a significant difference in terms of proteins and polymers between inner and outer surfaces of gloves, which helps to explain the different allergenic potential of these.MALDI imaging allowed for the first time the unambiguous localization of all three allergens in parallel and showed that Hevein was present on 36% of the investigated area of a latex glove with a certain localization, whereupon, tREF and REF were only found on 25% of the investigated material.
[102] - Chen Z, Cremer R, Posch A, Raulf-Heimsoth M, Rihs HP, Baur X. On the allergenicity of Hev b 1 among health care workers and patients with spina bifida allergic to natural rubber latex. J Allergy Clin Immunol 1997;100:684-693
BACKGROUND: Recent studies have caused much controversy about the prevalence of IgE antibodies to Hev b 1 among health care workers (HCWs) and patients with spina bifida (SB) who are allergic to latex. This investigation was carried out to verify the results reported. METHOD: Serum samples from 140 patients with SB as well as from 105 HCWs allergic to latex were tested by enzyme allergosorbest test (EAST) and EAST-inhibition assay to evaluate the rate and degree of sensitization to highly purified Hev b 1. RESULTS: Eighty-one percent of patients with SB who were allergic to latex had IgE antibodies against Hev b 1. The prevalence of anti-Hev b 1 antibodies among HCWs allergic to latex was 52.3%. In 15 of 33 serum samples from patients with SB that were randomly tested, the IgE binding to commercial latex allergens could be completely inhibited by Hev b 1; in only six cases was the maximum inhibition of IgE binding to latex by Hev b 1 less than 50%. Testing two monoclonal anti-Hev b 1 antibodies with extracts of five brands of latex gloves revealed a predominant presence of Hev b 1 protein as a monomer or its aggregates. Molecular analysis of human leukocyte antigen-D region genes DRB and DQB1 suggested no statistically significant correlation between the human leukocyte antigen alleles tested and IgE responsiveness to Hev b 1. CONCLUSIONS: Our results indicate that Hev b 1 not only makes significant contributions to the IgE binding to latex, but it is also the unique sensitizer in about 45% of patients with SB who are allergic to latex
[103] - Chardin H, Chen Z, Raulf-Heimsoth M, Mayer C, Senechal H, Desvaux FX, et al. Identification of Hev b 1 in natural latex mattresses. Int Arch Allergy Immunol 2000;121:211-214
BACKGROUND: Type 1 hypersensitivity to natural rubber latex proteins is a well-recognized health problem. Recent data have shown that allergens can be extracted from natural latex mattresses. As Hev b 1 (rubber elongation factor) and Hev b 6.02 (hevein) were described as major allergens, the present work was carried out to evaluate their presence in latex mattresses as well as in latex gloves. METHODS: Extracted proteins from latex mattresses and gloves were separated by SDS-PAGE or two-dimensional gel electrophoresis, transferred onto nitrocellulose and detected with monoclonal antibodies specific for Hev b 1 and Hev b 6.02. RESULTS: The results showed that various forms of Hev b 1, as well as degradation products of Hev b 1 were detected in latex mattresses and gloves, whereas Hev b 6.02 was not detected either in mattresses or in gloves. In a standardized latex extract, Hev b 1 and Hev b 6.01 (prohevein) were identified by the monoclonal antibodies. CONCLUSION: The fact that only Hev b 1 was detected by im munoblot in latex articles indicates that Hev b 1 may be the last protein to be washed out of latex products and that the Hev b 1 content may be used as a criterion for the estimation of the allergenicity of the latex products.
[104] - La Grutta S, Mistrello G, Varin E, Pajno GB, Passalacqua G. Comparison of ammoniated and nonammoniated extracts in children with latex allergy. Allergy 2003;58:814-818
BACKGROUND: The use of ammoniated or nonammoniated latex extracts for the diagnosis of latex allergy is still a matter of debate. The aim of our study was to compare the characteristics of the two types of extracts by immunoblotting and RAST techniques in children with ascertained latex allergy . METHODS: Ammoniated (AL) and nonammoniated latex (NAL) extracts were prepared and blotted on SDS-PAGE to resolve their components. Also a solid phase for RAST assays was prepared with the two extracts. The sera from 18 children (mean age 11.4 years, range 6-15 years), with ascertained latex allergy (clinical history, skin test, CAP-RAST and provocation) were used for the experiments . RESULTS: The NAL extract is resolved in many bands (5-100 kDa), whereas AL showed only few components, likely Hev b 4, 6 and 7. IgE reactivity against AL was observed only in 5/18 patients, whereas 12/18 were positive with NAL. The blotting profile against NAL was complex and the IgE recognition pattern involved different bands . CONCLUSION: The extract obtained from NAL is able to detect specific IgE against a greater number of allergenic determinants, and therefore a greater diagnostic accuracy can be expected.
[105] - Beezhold DH, Hickey VL, Slater JE, Sussman GL. Human IgE-binding epitopes of the latex allergen Hev b 5. J Allergy Clin Immunol 1999;103:1166-1172
BACKGROUND: Hev b 5 is an acidic protein (isoelectric point, 3.5) rich in glutamic acid with 9 repeated amino acid (AA) sequences of XEEX or XEEEX. Although its function in Hevea brasiliensis is unknown, Hev b 5 has been identified as a major latex allergen. Immunoblot inhibition studies suggest Hev b 5 exists as multiple isoforms or contains a common epitope found in several other proteins. OBJECTIVE: The purpose of this study was to further characterize Hev b 5 and to identify linear IgE-binding epitopes. METHODS: Octapeptides spanning the entire Hev b 5 protein were synthesized on a derivatized cellulose membrane. The membrane was reacted with sera pooled from health care workers allergic to latex or rabbits immunized with latex proteins. B-cell epitopes were identified by subsequent incubations with the appropriate secondary antibodies and detected by using chemifluorescence. RESULTS: Sera from patients allergic to latex recognized 6 IgE-binding regions located throughout the molecule. Two epitopes (2 and 4) had the common AA sequence of KTEEP. Epitopes 3 and 5 had a similar AA sequence of EEXXA, where X was P, T, or K. Epitopes 1 and 6 appeared to be unrelated to the other epitopes. Database analysis could not identify other proteins with similar sequences. Neither of the XEEEX sequences bound IgE. Control sera failed to react to any peptides. CONCLUSIONS: Hev b 5 exists as multiple isoforms, but only small amounts are present in the nonammoniated latex preparations, such as those used for diagnostic tests, and this may help to explain the relatively poor sensitivity of some in vitro tests.
[106] - Yeang HY, Cheong KF, Sunderasan E, Hamzah S, Chew NP, Hamid S, et al. The 14.6 kd rubber elongation factor (Hev b 1) and 24 kd (Hev b 3) rubber particle proteins are recognized by IgE from patients with spina bifida and latex allergy. J Allergy Clin Immunol 1996;98:628-639
Two major water-insoluble proteins are located on the surface of rubber particles in Hevea brasiliensis latex. A 14.6 kd protein (Hev b 1), found mainly on large rubber particles (> 350 mm in diameter), and a 24 kd protein (Hev b 3), found mainly on small rubber particles (average diameter, 70 nm), are recognized by IgE from patients with spina bifida and latex allergy. Although Hev b 1 (also called the rubber elongation factor [REF]) has previously been reported as a major latex allergen, this conclusion has been disputed on the basis of results from other studies. The allergenicity of Hev b 1 is verified in this study by testing the recombinant protein generated from its gene. Because allergenicity is confined to patients with spina bifida and not observed in adults sensitive to latex, it is not a major latex allergen. The identification of Hev b 3 as another allergen originating from rubber particles is confirmed by immunogold labeling and electron microscopy. Observations with the monoclonal antibody USM/RC2 developed against Hev b 3 show that the protein has a tendency to fragment into several polypeptides of lower molecular weight (from 24 kd to about 5 kd) when stored at -20 degrees C. There is also indication of protein aggregation from the appearance of proteins with molecular weights greater than 24 kd. Fragmentation of Hev b 3 is induced immediately on he addition of latex B-serum, which is normally compartmentalized in the lutoids in fresh latex. In the preparation of ammoniated latex (used for the manufacture of latex products), the lutoids are ruptured, and the released B-serum reacts with Hev b 3 on the rubber particles to give rise to an array of low molecular weight polypeptides that are allergenic to patients with spina bifida.
[107] - Banerjee B, Kanitpong K, Fink JN, Zussman M, Sussman GL, Kelly KJ, et al. Unique and shared IgE epitopes of Hev b 1 and Hev b 3 in latex allergy. Mol Immunol 2000;37:789-798
Of the several latex proteins cloned and expressed, the rubber elongation factor, Hev b 1, and the closely related Hev b 3, represent two major allergens associated with latex allergy. Although both allergens demonstrated IgE binding with sera from latex allergic patients, it was not known whether these two molecules shared any epitopes. Hence, in the present study using health care workers (HCW) and spina bifida (SB) patients with latex allergy, we investigated the IgE binding epitopes in Hev b 1 and Hev b 3. Recombinant Hev b 1 and Hev b 3 were expressed in a prokaryotic expression system, while overlapping decapeptides of Hev b 1 and Hev b 3 were synthesized on derivatized cellulose membrane. Eight IgE binding epitopes for Hev b 1 and eleven for Hev b 3 were identified using sera from latex allergic patients with SB. On further analysis of synthetic peptides encompassing these epitopes, similar IgE antibody reactivity was demonstrated with three Hev b 1 epitopes b1E3, b1E5, b1E6 and two Hev b 3 epitopes: b3E10 and b3E 11. For Hev b 1, a unique IgE binding epitope was identified in the region of amino acid residues 16-25. In competitive ELISA, peptides bIE2 and bIE4 together inhibited 58% of IgE binding of Hev b 1, while b3E5 showed 22% inhibition in the IgE binding of Hev b 3. The results of the present study suggest that the understanding of linear and conformational IgE epitopes in the major latex allergens may provide better insight into the structure-function relationship of the allergens, and may lead to the development of better patient care and management strategies in latex allergy.
[108] - Broekaert W, Lee HI, Kush A, Chua NH, Raikhel N. Wound-induced accumulation of mRNA containing a hevein sequence in laticifers of rubber tree (Hevea brasiliensis). Proc Natl Acad Sci U S A 1990;87:7633-7637
Hevein is a chitin-binding protein that is present in laticifers of the rubber tree (Hevea brasiliensis). A cDNA clone (HEV1) encoding hevein was isolated by using the polymerase chain reaction with mixed oligonucleotides corresponding to two regions of hevein as primers and a Hevea latex cDNA library as a template. HEV1 is 1018 base pairs long and includes an open reading frame of 204 amino acids. The deduced amino acid sequence contains a putative signal sequence of 17 amino acid residues followed by a 187-amino acid polypeptide. This polypeptide has two striking features. The amino-terminal region (43 amino acids) is identical to hevein and shows homology to several chitin-binding proteins and to the amino termini of wound-inducible proteins in potato and poplar. The carboxyl-terminal portion of the polypeptide (144 amino acids) is 74-79% homologous to the carboxyl-terminal region of wound-inducible genes of potato. Wounding, as well as application of the plant hormones abscisic acid and ethylene, resulted in accumulation of hevein transcripts in leaves, stems, and latex but not in roots.
[109] - Kujala V, Alenius H, Palosuo T, Karvonen J, Pfäffli P, Reijula K. Extractable latex allergens in airborne glove powder and in cut glove pieces. Clin Exp Allergy 2002;32:1077-1081
Background: Quantification of natural rubber latex (NRL) allergens of NRL glove extracts has been reported in several studies. Similarly, immunoassay studies reporting the level of NRL aeroallergens in air samples have been published. When studying the NRL allergens of gloves, however, little attention has been focused on identifying the relationship between extractable NRL allergens of medical gloves and NRL aeroallergens in indoor air. Objective: In an experimental study we analysed NRL aeroallergens of medical gloves in joint relation to total airborne dust concentration and NRL allergen concentration in gloves. Methods: NRL aeroallergen level was measured using a chamber setting with 18 lots of powdered medical gloves. In each setting 10 pairs of powdered NRL gloves were swinging in an unventilated chamber (9m3). Air samples were collected using airflow through Millipore filters (pore size 0.8µm). The filters were weighed before and after the experiment, and total airborne dust concentration in the chamber was calculated. The filter samples were then extracted and the NRL allergen level measured by IgE ELISA-inhibition assay. Furthermore, cut NRL gloves were extracted and analysed by the same method. Finally, levels of two major NRL allergens, Hev b1 and Hev b 6.02, were measured in three selected NRL glove brands. Results: The NRL aeroallergen level in the chamber air ranged from <0.9 to 2.9 allergen units (AU)/m3. The total airborne dust concentration in the chamber air remained low with all lots of gloves measured (range <20 to 80µg/m3). The NRL allergen level in cut glove extracts varied over 100-fold (<10 to 1050 AU/mL). Statistically significant correlation between aeroallergen concentration and airborne dust (r=0.8, P=0.0015) concentration was found. Moreover, significant correlation between aeroallergen levels and allergen content of cut glove pieces was observed (r=0.59, P<0.05). Hev b 1 levels varied from 9 to 25ng/mL and the levels of Hev b 6.02 from 1720 to 14460ng/mL in the glove extracts. In the extracts from airborne dust samples, Hev b 6.02 content varied from 61 to 183ng/m3, whereas Hev b 1 levels were very low (0.4 to 3ng/m3). Conclusion: An elevated NRL aeroallergen level is rather related to a high level of airborne glove powder than to a high concentration of extractable NRL allergen in medical gloves.
[110] - Baur X. Are we closer to developing threshold limit values for allergens in the workplace ? Ann Allergy Asthma Immunol 2003;90(5 Suppl. 2):11-18
OBJECTIVE: To define threshold limit values and legally binding occupational exposure limits. DATA SOURCES: Review of suitable literature. STUDY SELECTION: Studies based on detailed descriptions and/or measurements of airborne allergenic dust, total allergens, or even key allergens were selected. RESULTS: Prevalences of IgE-mediated sensitization and occupational asthma are related to the aeroallergen load in workplaces. Data to set threshold limit values for flour, latex, a-amylase, and isocyanates are already sufficiently available. CONCLUSIONS: To optimize primary prevention in workplaces,health-based occupational exposure limits should be set for major occupational allergens.
[111] - Nettis E, Colanardi MC, Ferrannini A. Type I latex allergy in health care workers with latex-induced contact urticaria syndrome: a follow-up study. Allergy 2004;59:718-723
BACKGROUND: Latex allergy is an important occupational health issue among health care workers (HCWs). Secondary prevention in sensitized/allergic individuals involves avoiding exposure to NRL products. AIMS OF THE STUDY: The aim of this follow-up study is to determine the long-term health consequences in HCWs with type I latex allergy with latex-related contact urticaria syndrome, of providing appropriate information and practical avoidance education . METHODS: Seventeen HCWs with latex-induced contact urticaria syndrome, as ascertained by the glove use test, were investigated. Initial and follow-up visits included: a detailed questionnaire, skin prick test (SPT) with glove eluates and with commercial latex extract, SPT with common inhalant and food extracts, serum specific immunoglobulin (Ig)E to latex and some foods and the glove use test . RESULTS: On re-examination, 11 (64.7%) subjects showed positive SPTs to extemporaneous extract and 10 (58.8%) patients showed a positive SPT to commercial extract. Of the nine patients with detectable levels of serum latex specific IgE at first evaluation, four (44.4%) became negative and four were assigned to at least one class lower. Only one (11.1%) employee had higher radioallergosorbent test values than those at the latex allergy diagnosis. At follow-up, the 17 individuals had positive latex challenge results, although the duration of exposure causing a reaction increased . CONCLUSION: Our study shows that preventive measures are sufficient to induce a reduction of sensitization. Continued avoidance is needed to prevent re-sensitization or adverse reactions on re-exposure.
[112] - Turjanmaa K, Kanto M, Kautiainen H, Reunala T, Palosuo T. Long-term outcome of 160 adult patients with natural rubber latex allergy. J Allergy Clin Immunol 2002;110(2 suppl.):S70-S74
Allergy to natural rubber latex is a major occupational problem in the health care sector and a problem even in other occupations in which protective gloves are used. There is little information available about the long-term outcome at work in large patient populations. To study the occupational outcome when all gloves in the working environment were changed either to low-allergen latex or non-latex gloves, in 1995 to 1996 we re-examined 160 of 174 adult subjects who were diagnosed with natural rubber latex allergy between 1982 and 1994 after a median of 3 years (range, 0.5 to 24 years) after the diagnosis. The outcome in daily work or as a patient in health care was investigated with the use of a questionnaire. Special attention was paid to the assessment of the occurrence of hand eczema. Glove selection policy as an intervention in the treatment of natural rubber latex allergy is described. Of 71 health care workers and 89 non-health care workers, 72% and 83% were atopic; 54% and 65% had hand eczema at the time of diagnosis, and 89% and 19% had work-related allergy to natural rubber latex, respectively. On re-examination, none of the health care workers had changed work because of natural rubber latex allergy, and only 38% had hand eczema (significant decrease). Ninety-eight percent of the non-health care workers, of which 58% had hand eczema, continued with their previous jobs. The use of low-allergen latex or non-latex gloves throughout the health care sector seems to be an adequate step for health care workers who have natural rubber latex allergy; non-health care workers get along with personal avoidance of latex gloves if they are not working directly with natural rubber latex-containing materials in production.
[113] - Ruëff F, Schöpf P, Putz K, Przybilla B. Effect of reduced exposure on natural rubber latex sensitization in health care workers. Ann Allergy Asthma Immunol 2004;92:530-537
BACKGROUND: Knowledge about the long-term course of allergy or sensitization to natural rubber latex (NRL) is insufficient . OBJECTIVE: To investigate the long-term effect of preventive measures on sensitization variables in health care workers who had been diagnosed as having NRL allergy (NRLA) or NRL sensitization (NRLS) without clinical symptoms . METHODS: Repeated follow-up investigations, skin prick tests, and NRL specific IgE serum antibodies were performed in 88 health care workers-33 with NRLA and 55 with NRLS. All workers had been instructed to avoid NRL exposure. At the workplace, powder-free NRL gloves for all other employees were gradually introduced. Re-evaluations were done at 14 +/- 3.7 (N = 86) and 38 +/- 4.0 (N = 78) months after the first examination . RESULTS: At the last follow-up, a loss of skin prick test reactivity to NRL was observed in 1 of 29 subjects with NRLA (3.4%) and 16 of 35 with NRLS (45.7%) with previous skin test reactions (P < .001). Among those subjects who demonstrated a kU/L level (CAP class) equal to or greater than class I to NRL at the initial examination, NRL-specific IgE was absent at the last follow-up in 8 (32.0%) of 25 subjects with NRLA and 14 (38.9%) of 36 with NRLS. At the final examination, we could no longer demonstrate sensitization to NRL by any method in 24 (27.3%) of 88 health care workers. Complete loss of NRL sensitization was less frequent in subjects with NRLA than in those with NRLS (1 of 33 or 3.0% vs 23 of 55 or 41.8%; P < .001) . CONCLUSIONS: Implementation of simple preventive measures lowers markers of sensitization to NRL quickly in many health care workers with NRLA or NRLS.
[115] - Sunderasan E, Chow KS, Ward MA, Yeang HY. Hev b 4 Heavy Peptide Identified as Latex Cyanogenic Glucosidase. AAAAI 58th Annual Meeting, New York, 1-6 March, 2002, Poster n°1031
The latex protein Hev b 4 comprises a triplet of allergenic peptides of molecular weight 50 to 57 kDa. The three peptides share a common characteristic in remaining soluble only in the presence of high ionic concentration (e.g. 0.2M NaCl and higher). Dilution or dialysis induces precipitation of all three peptides that might have related biochemical functions in the latex, and that might be components of a protein complex. Hev b 4 has been shown to be allergenic by IgE-binding assays and in skin prick tests. The heaviest component of the Hev b 4 triplet, a 57 kDa peptide, was isolated from the latex B-serum by dialysis-induced protein precipitation and Sephacryl gel filtration. Further purification was carried out by preparative SDS-polyacrylamide gel electrophoresis. Following trypsin digestion, the 57 kDa peptide was scanned using a nano-electrospray tandem mass spectrometry (ESI-MS) and amino acid sequencing was carried out by MS/MS. Partial amino acid sequences of the protein matched published sequences of several plant glucosidases, including those of cyanogenic glucosidases. Enzymatic assays confirmed that the protein possessed glucosidase activity, the enzyme activity being further demonstrated by isozyme staining of the protein after electrophoretic separation on native polyacrylamide gels. Enzymic activity was also observed when linamarin was used as the specific enzymic substrate, indicating that the protein was a cyanogenic glucosidase (linamarase) as suggested by the partial amino acid sequences. A partial cDNA sequence with homology to plant glucosidases was obtained by reverse transcriptionˆPCR using sense and antisense strand degenerate primers. The 5´/3´RACE is being employed to generate the complete cDNA of H. brasiliensis latex glucosidase.
[116] - Chen Z, Cremer R, Posch A, Raulf-Heimsoth M, Rihs HP, Baur X. On the allergenicity of Hev b 1 among health care workers and patients with spina bifida allergic to natural rubber latex. J Allergy Clin Immunol 1997;100:684-693
BACKGROUND: Recent studies have caused much controversy about the prevalence of IgE antibodies to Hev b 1 among health care workers (HCWs) and patients with spina bifida (SB) who are allergic to latex. This investigation was carried out to verify the results reported. METHOD: Serum samples from 140 patients with SB as well as from 105 HCWs allergic to latex were tested by enzyme allergosorbest test (EAST) and EAST-inhibition assay to evaluate the rate and degree of sensitization to highly purified Hev b 1. RESULTS: Eighty-one percent of patients with SB who were allergic to latex had IgE antibodies against Hev b 1. The prevalence of anti-Hev b 1 antibodies among HCWs allergic to latex was 52.3%. In 15 of 33 serum samples from patients with SB that were randomly tested, the IgE binding to commercial latex allergens could be completely inhibited by Hev b 1; in only six cases was the maximum inhibition of IgE binding to latex by Hev b 1 less than 50%. Testing two monoclonal anti-Hev b 1 antibodies with extracts of five brands of latex gloves revealed a predominant presence of Hev b 1 protein as a monomer or its aggregates. Molecular analysis of human leukocyte antigen-D region genes DRB and DQB1 suggested no statistically significant correlation between the human leukocyte antigen alleles tested and IgE responsiveness to Hev b 1. CONCLUSIONS: Our results indicate that Hev b 1 not only makes significant contributions to the IgE binding to latex, but it is also the unique sensitizer in about 45% of patients with SB who are allergic to latex
[117] - Chardin H, Chen Z, Raulf-Heimsoth M, Mayer C, Senechal H, Desvaux FX, et al. Identification of Hev b 1 in natural latex mattresses. Int Arch Allergy Immunol 2000;121:211-214
BACKGROUND: Type 1 hypersensitivity to natural rubber latex proteins is a well-recognized health problem. Recent data have shown that allergens can be extracted from natural latex mattresses. As Hev b 1 (rubber elongation factor) and Hev b 6.02 (hevein) were described as major allergens, the present work was carried out to evaluate their presence in latex mattresses as well as in latex gloves. METHODS: Extracted proteins from latex mattresses and gloves were separated by SDS-PAGE or two-dimensional gel electrophoresis, transferred onto nitrocellulose and detected with monoclonal antibodies specific for Hev b 1 and Hev b 6.02. RESULTS: The results showed that various forms of Hev b 1, as well as degradation products of Hev b 1 were detected in latex mattresses and gloves, whereas Hev b 6.02 was not detected either in mattresses or in gloves. In a standardized latex extract, Hev b 1 and Hev b 6.01 (prohevein) were identified by the monoclonal antibodies. CONCLUSION: The fact that only Hev b 1 was detected by im munoblot in latex articles indicates that Hev b 1 may be the last protein to be washed out of latex products and that the Hev b 1 content may be used as a criterion for the estimation of the allergenicity of the latex products.
[118] - Tomazic VJ, Withrow TJ, Hamilton RG. Characterization of the allergen(s) in latex protein extracts. J Allergy Clin Immunol 1995;96:635-642
BACKGROUND: Immediate hypersensitivity to latex, induced by natural latex proteins remaining on the finished products, may lead to severe anaphylactic reactions. METHODS: We investigated the distribution of latex proteins by molecular weight and identified the specific allergenic molecules. Proteins extracted from various latex products were compared with those extracted from raw latex sap, both ammoniated and nonammoniated. RESULTS: Variations in the levels of extractable protein, as well as in the number of molecules and the molecular weight distribution, were observed especially among finished latex products. To identify allergenic (i.e., IgE-binding) molecules, we performed immunoblots with the sera from latex-sensitive persons. The results indicated that antigenic molecule profiles differed among the products and also between the finished products and the raw material. In addition, specificities of the anti-latex IgE antibodies varied among the sensitized persons. CONCLUSIONS: It appeared that persons with the same history of sensitization had similar patterns of antigenic specificities. If the history of exposure, as well as genetic predisposition and medical history of the patient, plays a significant role in the specific IgE response, it may be difficult to select a "standard" antigen and a "standard" antiserum for the evaluation of the latex sensitivity and allergenicity.
[119] - Vallier P, Balland S, Harf R, Valenta R, Deviller P. Identification of profilin as an IgE-binding component in latex from Hevea brasiliensis: clinical implications. Clin Exp Allergy 1995;25:332-339
Considering the high occurrence of profilin as an allergen in many plant species, the assumption was made that profilin might be an allergen in Hevea brasiliensis, a member of the latex producing Euphorbiaceae family Using IgE-binding inhibition by purified profilins we demonstrated that profilin is an IgE-binding component in the cytosolic fraction of natural latex and, to a lower extent, in the rubber fraction. Thirty-five out of 36 sera containing IgE to ragweed-profilin reacted with profilin from latex, indicating structural homologies between profilins from latex and ragweed. A large percentage (59%) of these sera were found to be positive in CAP latex assay. The preincubation of these sera with purified ragweed profilin greatly inhibited the CAP latex. Because profilin is also present in banana extract, it is likely to be involved in cross-sensitivity to banana and latex. In a group of 19 individuals allergic to latex only two had anti-profilin IgE antibodies. Profilin was barely detectable on glove extract immunoblots, whereas some sera from patients allergic to latex reacted with a 15 kDa allergen which was not profilin. Consequently, IgE antibodies to latex-profilin is a questionable factor for sensitization of occupationally-exposed patients; however, sensitization to profilin should be taken into account when interpreting the results of latex IgE antibody assays.
[121] - Muguerza J, Capo C, Porri F, Jacob JL, Mege JL, Vervloet D. Latex allergy: allergen identification in Hevea brasiliensis fractions by immunoblotting. Clin Exp Allergy 1996;26:1177-1181
BACKGROUND: Latex is the cause of several clinical symptoms of allergy, but the identification of allergens is not completely known. OBJECTIVE: The aim of this report was to study the immunoreactivity of purified stable latex fractions from Hevea braziliensis. METHODS: We purified the cytoplasm of Hevea braziliensis and obtained three fractions: latex particles (LP), lutoids (L) and cytosolic serum (CS). Using Western blot, specific IgE directed to latex allergens was found in 80 patients with latex allergy. RESULTS: Five major groups of allergens migrating as 14, 25, 29, 37-45 and 50 kDa were recognized. They were unequally distributed with the latex fractions: 37-45 kDa proteins were essentially recognized in CS and LP, whereas 14 and 29 kDa proteins were mainly labelled in the L fraction. As a control, aqueous glove extracts exhibited a more restricted pattern of reactivity, because only 14 and 29 kDa proteins were recognized by patient sera. The pattern of reactivity was not correlated specifically with IgE levels, but sera from patients suffering from spina bifida reacted specifically with the minor protein of 25 kDa located in LP. CONCLUSIONS: The present results show that latex allergic patients recognize several allergens which are differently distributed in subcellular fractions extracted from H. braziliensis and aqueous GE. The L fraction and GE were enriched in low molecular weight proteins and apparently contained the same allergens.
[122] - Fuchs T, Spitzauer S, Vente C, Hevler J, Kapiotis S, Rumpold H, et al. Natural latex, grass pollen, and weed pollen share IgE epitopes. J Allergy Clin Immunol 1997;100:356-364
Because of the frequent use of natural latex products, IgE-mediated reactions to latex proteins represent an important health threat in industrialized countries. Although several latex allergens have been characterized and IgE cross-reactivities with allergens present in plant-derived food have been described, limited information is available regarding the presence of common IgE-binding components in latex and plant pollen. METHODS: By using serum IgE from 56 individuals with latex allergy, the IgE-binding components in ammoniated latex milk and latex glove extracts were characterized by immunoblotting. The presence of cross-reactive IgE-binding components in the different latex extracts, extracts from mugwort, ragweed, timothy grass pollen, and recombinant birch pollen allergens (Bet v 1 and Bet v 2 [birch profilin]) was studied by immunoblot inhibitions and quantitative competition experiments. The involvement of carbohydrates in the constitution of cross-reactive IgE epitopes was studied by periodate treatment of extracts. RESULTS: Although sera from certain individuals with latex allergy showed IgE reactivity with protein bands of different molecular weights in Western-blotted latex milk and glove extracts, both extracts contained common IgE epitopes. Although preincubation with recombinant Bet v 1 and Bet v 2 did not significantly inhibit IgE binding to latex proteins, weed and, in particular, timothy grass pollen extract strongly inhibited IgE binding to latex allergens. The cross-reactive IgE epitopes were sensitive to periodate treatment. CONCLUSIONS: Mugwort, ragweed, and timothy grass pollen share IgE epitopes with glycoprotein latex allergens. The presence of common epitopes might in part explain clinical symptoms in patients allergic to pollen on contact with latex.
[123] - Yeang HY, Arif SA, Raulf-Heimsoth M, Loke YH, Sander I, Sulong SH, et al. Hev b 5 and Hev b 13 as allergen markers to estimate the allergenic potency of latex gloves. J Allergy Clin Immunol 2004;114:593-598
BACKGROUND: Sensitization to natural rubber latex has been linked to proteins from medical latex gloves. Various assays to estimate the amount of residual allergenic proteins extractable from latex gloves to assess their potential exposure hazard have inherent weaknesses. OBJECTIVE: This investigation was aimed at developing 2-site immunoenzymetric assays and identifying appropriate protein markers to assess the allergenic potential of latex gloves. METHODS: The presence of 6 latex allergens-Hev b 1, 2, 3, 5, 6, and 13-was measured in a cross-section of commercial latex medical gloves by using monoclonal and polyclonal antibody-based 2-site immunoenzymetric assays. The overall allergenic potential of these gloves was assessed by IgE-inhibition assay. Stepwise multiple regression analyses were performed to identify marker allergens that best explained the variation in latex glove allergenicity. RESULTS: All 6 latex allergens were detected in at least some of the glove samples. Hev b 5 and Hev b 13 were identif ied as the marker allergens that combined best to explain the variation in the glove allergenicity. The significant multiple correlation (R=0.855) between these 2 markers and glove allergenic potency forms the basis of an assay to gauge latex glove allergenicity. CONCLUSION: The overall allergenic potential of latex gloves can be estimated by using Hev b 5 and Hev b 13 as indicator allergens. The correlation between glove allergenicity and the level of these allergens was maintained for low-protein gloves ( < 200 microg/g). This estimation of glove allergenicity was superior to that obtained by using total protein readings
[124] - Palosuo T, Reinikka-Railo H, Kautiainen H, Alenius H, Kalkkinen N, Kulomaa M, et al. Latex allergy: the sum quantity of four major allergens shows the allergenic potential of medical gloves. Allergy 2007;62:781-786
Background: Assessment of allergenic potential of medical devices made of natural rubber latex (NRL) requires the measurement of concentrations of specific allergenic proteins or polypeptides eluting from rubber. METHODS: Four NRL allergens (Hev b 1, 3, 5, and 6.02) were quantified in all medical glove brands marketed in Finland in 1999, 2001, and 2003 (n = 208) by a capture enzyme immunoassay. The results were compared with those obtained from previous nationwide market surveys, using a skin prick test-validated human IgE-based ELISA-inhibition method. RESULTS: A high overall correlation (r = 0.87, 95% CI 0.83-0.90) emerged between the sum values of the four allergens(mug/g glove) and IgE-ELISA inhibition (allergen units, AU/ml, 1 : 5 diluted glove extract). The sum of four allergens when set at 0.15 mug/g discriminated low allergenic ( < 10 AU/ml) from moderate- to high-allergenic (>=10 AU/ml) gloves at a sensitivity of 0.93 (95% CI 0.85-0.98) and specificity of 0.90 (95% CI 0.83-0.94). When the sum was below the detection limit (0.03 mug/g) all gloves belonged to the previously defined low-allergen category. CONCLUSIONS: By comparing the sum concentration of four selected NRL allergens with results obtained in human IgE-ELISA inhibition, it was possible set a cut-off level (0.15 mug/g) below which virtually all gloves contain low or insignificant amounts of allergens, and can be considered as low allergenic. At different cut-off-points, one could calculate the likelihood of a given glove to belong to the previously defined low, moderate or high allergen categories
[125] - Barbara J, Santais MC, Levy DA, Ruff F, Leynadier F. Immunoadjuvant properties of glove cornstarch powder in latex-induced hypersensitivity. Clin Exp Allergy 2003;33:106-112
Cornstarch powder present in medical gloves plays an important role in latex-induced hypersensitivity as allergen carrier either, by the inhalation route, by skin contact or by direct contact with mucous membranes. ObjectiveOur objective was to test the hypothesis that cornstarch could act as an immunoadjuvant in immediate type-I latex-induced hypersensitivity. MethodsGuinea-pigs were sensitized by intraperitoneal route with two different antigens (latex proteins and ovalbumin) with or without cornstarch powder. Airway responsiveness after specific bronchial provocation was evaluated and specific IgG and IgG1 levels were determined by enzyme-linked immunosorbant assay (ELISA). Controls were treated with cornstarch powder or saline alone. ResultsAnimals sensitized with latex proteins (n = 7 in each group) showed significant bronchoconstriction (P < 0.03) and higher anti-latex antibody levels than the controls (P < 0.005). Guinea-pigs sensitized with latex-contaminated cornstarch had higher levels of specific antibodies than those sensitized with latex alone (P < 0.05). Animals sensitized to latex mixed with cornstarch showed higher bronchospasm than those treated with latex alone (P < 0.003). Animals sensitized to ovalbumin mixed with cornstarch also showed higher antibody and bronchoconstriction levels (P < 0.05) than those immunized with ovalbumin alone but antibody titres were significantly lower than those of the animals treated with ovalbumin and Freund's complete adjuvant (P < 0.01; n = 5 in each group). ConclusionOur findings show that cornstarch powder increases antigen-induced bronchoconstriction and antibody production. This role of immunoadjuvant is not antigen-specific. The cornstarch powder used as donning agent in latex gloves is an allergen carrier and it can enhance latex-induced hypersensitivity. .
[126] - Sutherland MF, Drew A, Rolland JM, Slater JE, Suphioglu C, O'Hehir RE. Specific monoclonal antibodies and human immunoglobulin E show that Hev b 5 is an abundant allergen in high protein powdered latex gloves. Clin Exp Allergy 2002;32:583-589
BackgroundHev b 5 is a major latex allergen recognized predominantly by latexallergic health care workers (HCWs). Recombinant Hev b 5 (rHev b 5) was previously expressed as a fusion protein with maltose binding protein (MBP), itself an immunogenic molecule; therefore nonfusion rHev b 5 is desirable. Moreover, standardized immunological assays for the detection of Hev b 5 are currently lacking and may have important implications for both allergen avoidance and diagnosis in latex allergy. ObjectivesTo generate and use Hev b 5specific mAbs to determine the relative abundance of Hev b 5 in different latex extracts, correlating this with the IgE reactivity of latexallergic HCWs and to produce nonfusion rHev b 5. MethodsFor the production of mAbs, mice were immunized with rHev b 5/MBP fusion protein and mAbs selected with rHev b 5/MBP but not MBP reactivity. The mAb reactivity was compared with polyclonal IgE from latexallergic HCWs using direct and inhibition ELISA and immunoblot assays. Recombinant Hev b 5 was expressed and purified in the pPROEXHTa bacterial expression system. ResultsFour Hev b 5specific mAbs were produced. Immunoblotting and ELISA using the mAbs indicate abundant Hev b 5 in high protein powdered latex glove extracts as compared with crude latex sap extracts. High quality surgical gloves with no detectable protein have no detectable Hev b 5. Inhibition ELISAs using serum IgE from latexallergic HCWs and Hev b 5specific mAbs gave strong correlation. Nonfusion recombinant Hev b 5 was successfully expressed and purified, showing reactivity with both the Hev b 5specific mAbs and serum IgE of latexallergic HCWs. ConclusionHev b 5specific mAbs and human IgE from latexallergic HCWs demonstrate the greater content of Hev b 5 in high protein powdered glove extracts. This may explain the observed higher frequency of sensitization to this allergen in HCWs.
[127] - Mitakakis TZ, Tovey ER, Yates DH, Toelle BG, Johnson A, Sutherland MF, et al. Particulate masks and non-powdered gloves reduce latex allergen inhaled by healthcare workers. Clin Exp Allergy 2002;32:1166-1169
BACKGROUND: Although allergy to latex is a well-characterized phenomenon, some hospitals continue to provide staff with powdered latex gloves as an option to low- or non-powdered gloves. OBJECTIVE: We aimed to measure the extent to which inhalation of latex particles could be reduced by the use of protective masks or by replacing powdered latex gloves with non-powdered latex gloves. METHODS: Twenty healthcare workers in a hospital setting wore nasal air samplers (NAS) and Institute of Occupational Medicine (IOM) samplers for four 20-min periods. Subjects wore powdered gloves, non-powdered gloves and no gloves during three sampling periods, and in the fourth, subjects applied an aerosol barrier face-mask or a particulate face-mask (N95) while wearing powdered gloves. All samples were stained for particles bearing Hev b 5 allergen by the Halogen assay. RESULTS: All subjects inhaled Hev b 5 bearing particles in all sampling periods. IOM samplers collected particles at 70% of the rate of NAS. The number of partic les inhaled while wearing powdered gloves was 23.8-fold higher than when not wearing gloves and 9.7-fold higher than when wearing non-powdered latex gloves (P < 0.0001). Wearing an aerosol barrier mask did not significantly reduce the number of particles inhaled (P = 0.108), while use of particulate masks significantly reduced the number of particles inhaled by 17.4-fold (P = 0.003). CONCLUSIONS: Use of non-powdered gloves is the most effective method of reducing occupational aeroallergen exposure to latex arising from gloves. However, secondary protection using particulate masks is a valid alternative, and may be helpful for preventing respiratory sensitization
[128] - Poulos LM, O'Meara TJ, Hamilton RG, Tovey ER. Inhaled latex allergen (Hev b 1). J Allergy Clin Immunol 2002;109:701-706
Background: IgE-mediated responses to natural rubber latex allergens have become a major health problem among recognized risk groups. Objective: The purpose of this investigation was to measure the amounts of Hevea brasiliensis latex allergen (Hev b 1) inhaled and deposited on surfaces when latex or vinyl gloves were worn and compare the results with the conventional measures (breathing zone samplers) of occupational exposure. Methods: Hev b 1 exposure was measured by nasal sampling and breathing zone sampling. Latex allergen exposure was generated by having each subject don a pair of powdered latex examination gloves and continuing his or her normal daily activity for 30 minutes. By means of adhesive tape, surface dust samples were collected from the surfaces of gloves, the subject's hands, and work areas. Sampling was performed with subjects wearing no gloves, subjects wearing powdered vinyl gloves, subjects wearing powdered latex gloves, and nearby colleagues wearing latex gloves. All samples were assayed through use of the HALOgen assay (Inhalix, Sydney, Australia) with a Hev b 1ˆspecific mAb. Particles transporting latex allergen were identified by a surrounding immunostain halo, and these were quantified and reported as total numbers of particles inhaled, airborne, or found on surface areas evaluated. Results: Study subjects inhaled 26 times more allergen when powdered latex gloves were worn than under the „no glove‰ and powdered vinyl glove conditions. During the same period, Hev b 1 particle levels measured in the ambient air through use of the breathing zone sampler increased by 24-fold. The median numbers of particles carrying Hev b 1 allergen per square centimeter on the surface of the hands after the wearing of latex and vinyl gloves were 1964 and 5, respectively. Latex allergen was physically associated both with cornstarch granules and with larger dust particles having a darker, more irregular appearance. Conclusion: In a laboratory where gloves are worn for protection, the use of latex gloves resulted in a 26-fold increase in inhaled latex allergen over background levels measured while vinyl gloves were worn as controls. Low levels of latex exposure also occurred when vinyl gloves or no gloves were worn; the reasons for this are under investigation.
[129] - Tomazic-Jezic VJ, Sanchez BA. Allergen Profiles of Natural Rubber Latex (NRL) Proteins on Gloves and Glove Powders. J Long Term Eff Med Implants 2005;15:115-124
The contributing role of glove powder in sensitization to natural rubber latex (NRL) proteins has been well documented in laboratory studies and through clinical evaluations. However, the quantitative relationship of the respiratory and topical exposures in the sensitization process remains unknown because the relative levels of protein on the glove powders in relation to the total levels of protein on NRL gloves have not been determined. In NRL allergens-Hev b 1, Hev b 3, Hev b 5, and Hev b 6.02-on randomly selected surgical and examination NRL gloves. We also examined the binding pattern of the four allergens to several glove powders that showed a different affinity to NRL proteins. The level of powder-bound protein was determined by the ELISA Inhibition Assay (ASTM D6499 standard method). Two cross-linked corn starch powders, one sample of cooking corn starch and one oat starch sample, were exposed to ammoniated (AL) or nonammoniated (NAL) raw NRL protein extracts. The levels of individual allergens were determined using the NRL allergen kit. In the NRL glove extracts we observed a wide range in the total allergen levels and a great diversity in the proportion of the four allergens. On the other hand, the evaluated starches had similar ratios of four individual allergens, regardless of the differences in their total allergen levels. The exposure of starches to NRL proteins with different allergen profiles did not affect the allergen ratio. All samples demonstrated a selective affinity for binding Hev b 1 and Hev b 5 allergens and a lesser affinity for the Hev b 6.02 allergen. Allergen Hev b 6.02 made up about 60% of the total allergen in the NAL extract, but only 12-30% of Hev b 6.02 was bound to starches. In contrast, there was only 3-7% of Hev b 1 allergen in the NAL extract, but powders had 35-45% of Hev b 1. These findings indicate that allergenic properties of NRL gloves and respective glove powders may be different.
[130] - Marchetti-Deschmann M, Allmaier G. Allergenic compounds on the inner and outer surfaces of natural latex gloves: MALDI mass spectrometry and imaging of proteinous allergens. J Mass Spectrom 2009;44:61-70
Natural latex gloves are the cause of a severe health problem to an increasing number of healthcare workers or patients due to the presence of protein allergens as Hevein or Rubber Elongation Factor (REF). One of the most challenging problems is the in situ localization of theses allergens in, e.g. gloves, to estimate the allergenic potential of the latex material. A sample preparation protocol applying a binary matrix-assisted laser desorption/ionization(MALDI) matrix containing alpha-cyano-4-hydroxy cinnamic acid (CHCA) and 2,5-dihydroxy benzoic acid (DHB) on trifluoro acetic acid (TFA) etched latex glove surfaces allowed the direct determination (exact molecular weight) of Hevein, REF and a truncated form of REF (tREF) within nine different brands of natural latex gloves by means of MALDI-TOF-MS in the linear mode. MALDI mass spectrometry demonstrated that Hevein, tREF and REF were present on the inner surfaces (in direct contact with the skin) of many, but not all, investigated gloves without any prior extraction procedure. Additionally, different isoforms of the allergen Hevein were detected (exhibiting ragged C-termini). tREF and REF could always be detected beside each other, but were not observed on every latex glove sample, which contained Hevein. It was also demonstrated that there is a significant difference in terms of proteins and polymers between inner and outer surfaces of gloves, which helps to explain the different allergenic potential of these.MALDI imaging allowed for the first time the unambiguous localization of all three allergens in parallel and showed that Hevein was present on 36% of the investigated area of a latex glove with a certain localization, whereupon, tREF and REF were only found on 25% of the investigated material.
[131] - Peixinho C, Tavares-Ratado P, Tomás MR, Taborda-Barata L, Tomaz CT. Latex allergy: new insights to explain different sensitization profiles in different risk groups. Br J Dermatol 2008;159:132-136
BACKGROUND: Differences in latex allergen sensitization profiles have been described between children subjected to repetitive surgical interventions and health care workers (HCW). 'Major' allergens for patients with spina bifida are Hev b 1, 3 and 7, while for HCW, 'major' allergens are Hev b 2, 5, 6.01 and 13. The reason for these differential sensitization profiles is currently unknown. OBJECTIVES: To investigate latex allergen profiles on internal and external surfaces of natural rubber latex gloves. METHODS: Eighty-two samples of commonly used surgical gloves (41 glove brands) were used for analysis. Specific allergen levels of Hev b 1, 3, 5 and 6.02 on both surfaces of the gloves were quantified using an enzyme immunometric assay, a FITkit (FIT Biotech, Tampere, Finland). RESULTS: Differences in allergen levels were observed between internal and external surfaces of all glove types. Concentrations of Hev b 1 and Hev b 3 were significantly higher on external surfaces, while internal surfaces had higher allergen levels of Hev b 5 and Hev b 6.02. Analysis of surgical and examination gloves, powdered and nonpowdered gloves also showed that the content of Hev b 5 and Hev b 6.02 was significantly higher on internal surfaces while that of Hev b 1 and Hev b 3 was higher on external surfaces. CONCLUSIONS: Our study showed different allergen profiles on internal and external surfaces of natural rubber latex gloves. These results may suggest a relationship between latex allergen localization and sensitization routes in different risk groups.
[132] - Yeang HY, Ward MA, Zamri AS, Dennis MS, Light DR. Amino acid sequence similarity of Hev b 3 to two previously reported 27- and 23-kDa latex proteins allergenic to spina bifida patients. Allergy 1998;53:513-519
Separate studies have reported spina bifida patients to be especially allergic to proteins of 27 and 23 kDa found in the serum of centrifuged natural rubber latex. An insoluble latex protein located on the surface of small rubber particles, Hev b 3, has similarly been found to be allergenic to spina bifida patients. In this study, internal amino acid sequences of Hev b 3 showed similarity to the published sequences for the 27- and 23-kDa latex proteins. The latter allergens are hence identified as Hev b 3. Determination of the molecular weight of Hev b 3 revealed various species of 22-23 kDa. The consistent gaps of about 266 Da observed between various forms of the intact protein suggest that the protein undergoes post-translational modification. To determine whether Hev b 3 also occurs in a soluble form in the latex serum, its presence in molecular-filtered serum was checked by ELISA and Western blot. The results showed Hev b 3 to be largely absent in the C-serum from fresh latex. The protein is therefore in soluble in its native state. However, a small amount of the solubilized protein was detected in ammonia-stabilized latex (commonly used in the manufacture of latex products)
[133] - Yeang HY, Hamilton RG, Bernstein DI, Arif SAM, Chow KS, Loke YH, et al. Allergen concentration in natural rubber latex. Clin Exp Allergy 2006;36:1078-1086
BACKGROUND: Hevea brasiliensis latex serum is commonly used as the in vivo and in vitro reference antigen for latex allergy diagnosis as it contains the full complement of latex allergens . OBJECTIVE: This study quantifies the concentrations of the significant allergens in latex serum and examines its suitability as an antigen source in latex allergy diagnosis and immunotherapy . METHODS: The serum phase was extracted from centrifuged latex that was repeatedly freeze-thawed or glycerinated. Quantitation of latex allergens was performed by two-site immunoenzymetric assays. The abundance of RNA transcripts of the latex allergens was estimated from the number of their clones in an Expressed Sequence Tags library . RESULTS: The latex allergens, Hev b 1, 2, 3, 4, 5, 6, 7 and 13, were detected in freeze-thawed and glycerinated latex serum at levels ranging from 75 (Hev b 6) to 0.06 nmol/mg total proteins (Hev b 4). Hev b 6 content in the latex was up to a thousand times higher than the other seven latex allergens, depending on source and/or preparation procedure. Allergen concentration was reflected in the abundance of mRNA transcripts. When used as the antigen, latex serum may bias the outcome of latex allergy diagnostic tests towards sensitization to Hev b 6. Tests that make use of latex serum may fail to detect latex-specific IgE reactivity in subjects who are sensitized only to allergens that are present at low concentrations . CONCLUSION: Latex allergy diagnostics and immunotherapy that use whole latex serum as the antigen source may not be optimal because of the marked imbalance of its constituent allergens.
[135] - Palosuo T, Lehto M, Kotovuori A, Kalkkinen N, Blanco C, Poza P, et al. Latex allergy: low prevalence of immunoglobulin E to highly purified proteins Hev b 2 and Hev b 13. Clin Exp Allergy 2007;37:1502-1511
BACKGROUND: Hevea brasiliensis (Hev b) 2 and Hev b 13 have recently been identified as major latex allergens by detecting specific IgE antibodies in >50% of sera from Hev b latex-allergic individuals . OBJECTIVE: We assessed the prevalence rates for sensitization to extensively purified latex allergens in patients from three diverse geographical areas . METHODS: Native Hev b 2, Hev b 5, Hev b 6.01 and Hev b 13 were purified by non-denaturating chromatography and were used in ELISAs to assess sera from 215 latex-allergic patients and 172 atopic non-sensitized controls from Finland, Spain and the United States to detect allergen-specific IgE antibodies . RESULTS: Unexpectedly, even highly purified Hev b 13 contained epitope(s) to which Hev b 6-specific human IgE antibodies bound effectively. Further purification, however, reduced the prevalence of IgE antibody reactivity to low levels: 15%, 5% and 11% for Hev b 2, and 18%, 30% and 27% for Hev b 13 among latex-allergic Finnish, Spanish and American patients, respectively. Interestingly, Finnish patients had a lower prevalence of Hev b 5-specific IgE antibody (28%) as compared with Spanish (49%) and American (71%) patients. The prevalence of Hev b 6.01-specific IgE reactivity was uniformly >50% in all three populations . CONCLUSION: Neither Hev b 2 nor Hev b 13 appear to be major latex allergens when evaluated in serological assays using highly purified allergens. The reason(s) for the observed differences in published sensitization rates in various geographic regions requires further study. The purity of the allergen preparations has a marked impact on the accuracy of latex-specific IgE antibody detection in epidemiological studies and in the serological diagnosis of latex allergy.
[136] - Palosuo T, Lehto M, Kotovuori A, Kalkkinen N, Blanco C, Poza P, et al. Latex allergy: low prevalence of immunoglobulin E to highly purified proteins Hev b 2 and Hev b 13. Clin Exp Allergy 2007;37:1502-1511
BACKGROUND: Hevea brasiliensis (Hev b) 2 and Hev b 13 have recently been identified as major latex allergens by detecting specific IgE antibodies in >50% of sera from Hev b latex-allergic individuals . OBJECTIVE: We assessed the prevalence rates for sensitization to extensively purified latex allergens in patients from three diverse geographical areas . METHODS: Native Hev b 2, Hev b 5, Hev b 6.01 and Hev b 13 were purified by non-denaturating chromatography and were used in ELISAs to assess sera from 215 latex-allergic patients and 172 atopic non-sensitized controls from Finland, Spain and the United States to detect allergen-specific IgE antibodies . RESULTS: Unexpectedly, even highly purified Hev b 13 contained epitope(s) to which Hev b 6-specific human IgE antibodies bound effectively. Further purification, however, reduced the prevalence of IgE antibody reactivity to low levels: 15%, 5% and 11% for Hev b 2, and 18%, 30% and 27% for Hev b 13 among latex-allergic Finnish, Spanish and American patients, respectively. Interestingly, Finnish patients had a lower prevalence of Hev b 5-specific IgE antibody (28%) as compared with Spanish (49%) and American (71%) patients. The prevalence of Hev b 6.01-specific IgE reactivity was uniformly >50% in all three populations . CONCLUSION: Neither Hev b 2 nor Hev b 13 appear to be major latex allergens when evaluated in serological assays using highly purified allergens. The reason(s) for the observed differences in published sensitization rates in various geographic regions requires further study. The purity of the allergen preparations has a marked impact on the accuracy of latex-specific IgE antibody detection in epidemiological studies and in the serological diagnosis of latex allergy.
[137] - Raulf-Heimsoth M, Rihs HP, Rozynek P, Cremer R, Gaspar A, Pires G, et al. Quantitative analysis of immunoglobulin E reactivity profiles in patients allergic or sensitized to natural rubber latex (Hevea brasiliensis). Clin Exp Allergy 2007;37:1657-1667
BACKGROUND: Characterized native and recombinant Hevea brasiliensis (rHev b) natural rubber latex (NRL) allergens are available to assess patient allergen sensitization profiles . OBJECTIVE: Quantification of individual IgE responses to the spectrum of documented NRL allergens and evaluation of cross-reactive carbohydrate determinants (CCDs) for more definitive diagnosis . METHODS: Sera of 104 healthcare workers (HCW; 51 German, 21 Portuguese, 32 American), 31 spina bifida patients (SB; 11 German, 20 Portuguese) and 10 Portuguese with multiple surgeries (MS) were analysed for allergen-specific IgE antibody (sIgE) to NRL, single Hev b allergens and CCDs with ImmunoCAP technology . RESULTS: In all patient groups rHev b 5-sIgE concentrations were the most pronounced. Hev b 2, 5, 6.01 and 13 were identified as the major allergens in HCW and combined with Hev b 1 and Hev b 3 in SB. In MS Hev b 1 displayed an intermediate relevance. Different sIgE antibody levels to native Hevea brasiliensis (nHev b) 2 and rHev b 6.01 allowed discrimination of SB with clinical relevant latex allergy vs. those with latex sensitization. Sensitization profiles of German, Portuguese and American patients were equivalent. rHev b 5, 6.01 and nHev b 13 combined detected 100% of the latex-allergic HCW and 80.1% of the SB. Only 8.3% of the sera showed sIgE response to CCDs . CONCLUSIONS: Hev b 1, 2, 5, 6.01 and 13 were identified as the major Hev b allergens and they should be present in standardized latex extracts and in vitro allergosorbents. CCDs are only of minor relevance in patients with clinical relevant latex allergy. Component-resolved diagnostic analyses for latex allergy set the stage for an allergen-directed immunotherapy strategy.
[138] - Peixinho C, Tavares P, Tomaz MR, Taborda-Barata L, Tomaz C. Differential expression of allergens on the internal and external surfaces of latex surgical gloves. Allergol Immunopathol (Madr) 2006;34:206-211
BACKGROUND: Differences in latex allergen sensitization profiles have been described between children undergoing repeated surgical interventions and health care workers. The purpose of this study was to determine whether such sensitization profiles are associated with differences in the expression of latex allergen between the internal and external surfaces of surgical gloves. METHODS: Extracts were obtained from whole surgical gloves as well as from their external and internal surfaces. The extracts were centrifuged, filtered, concentrated, dialyzed and lyophilized. The protein profile of the extracts was analyzed using hydrophobic interaction chromatography (HIC) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Immunoblotting was performed using sera from two patients with confirmed latex allergy. Latex recombinant allergen-specific IgE in these two patients was determined using a fluorescence enzyme immunoassay (FEIA) method. Latex allergen quantification was determined on both glove surfaces using an ELISA method. RESULTS: HIC and SDS-PAGE showed qualitative and quantitative differences in proteins between the internal and external glove surfaces, with the former being much richer in proteins. Immunoblotting of glove extracts using sera from two latex-allergic health workers showed differences between glove surface extracts. ELISA quantification of latex allergens demonstrated that the internal glove surface had high amounts of Hev b 5 and Hev b 6.02 whereas the external surface showed Hev b 1, Hev b 3, and Hev b 6.02. CONCLUSIONS: Our results reveal substantial differences in the composition of latex allergen profiles between the internal and external surfaces of surgical latex gloves, which may suggest a relationship between latex allergen localization and sensitization routes in different risk groups.
[139] - Poulos LM, O'Meara TJ, Hamilton RG, Tovey ER. Inhaled latex allergen (Hev b 1). J Allergy Clin Immunol 2002;109:701-706
Background: IgE-mediated responses to natural rubber latex allergens have become a major health problem among recognized risk groups. Objective: The purpose of this investigation was to measure the amounts of Hevea brasiliensis latex allergen (Hev b 1) inhaled and deposited on surfaces when latex or vinyl gloves were worn and compare the results with the conventional measures (breathing zone samplers) of occupational exposure. Methods: Hev b 1 exposure was measured by nasal sampling and breathing zone sampling. Latex allergen exposure was generated by having each subject don a pair of powdered latex examination gloves and continuing his or her normal daily activity for 30 minutes. By means of adhesive tape, surface dust samples were collected from the surfaces of gloves, the subject's hands, and work areas. Sampling was performed with subjects wearing no gloves, subjects wearing powdered vinyl gloves, subjects wearing powdered latex gloves, and nearby colleagues wearing latex gloves. All samples were assayed through use of the HALOgen assay (Inhalix, Sydney, Australia) with a Hev b 1ˆspecific mAb. Particles transporting latex allergen were identified by a surrounding immunostain halo, and these were quantified and reported as total numbers of particles inhaled, airborne, or found on surface areas evaluated. Results: Study subjects inhaled 26 times more allergen when powdered latex gloves were worn than under the „no glove‰ and powdered vinyl glove conditions. During the same period, Hev b 1 particle levels measured in the ambient air through use of the breathing zone sampler increased by 24-fold. The median numbers of particles carrying Hev b 1 allergen per square centimeter on the surface of the hands after the wearing of latex and vinyl gloves were 1964 and 5, respectively. Latex allergen was physically associated both with cornstarch granules and with larger dust particles having a darker, more irregular appearance. Conclusion: In a laboratory where gloves are worn for protection, the use of latex gloves resulted in a 26-fold increase in inhaled latex allergen over background levels measured while vinyl gloves were worn as controls. Low levels of latex exposure also occurred when vinyl gloves or no gloves were worn; the reasons for this are under investigation.
[140] - Tomazic-Jezic VJ, Sanchez BA. Allergen Profiles of Natural Rubber Latex (NRL) Proteins on Gloves and Glove Powders. J Long Term Eff Med Implants 2005;15:115-124
The contributing role of glove powder in sensitization to natural rubber latex (NRL) proteins has been well documented in laboratory studies and through clinical evaluations. However, the quantitative relationship of the respiratory and topical exposures in the sensitization process remains unknown because the relative levels of protein on the glove powders in relation to the total levels of protein on NRL gloves have not been determined. In NRL allergens-Hev b 1, Hev b 3, Hev b 5, and Hev b 6.02-on randomly selected surgical and examination NRL gloves. We also examined the binding pattern of the four allergens to several glove powders that showed a different affinity to NRL proteins. The level of powder-bound protein was determined by the ELISA Inhibition Assay (ASTM D6499 standard method). Two cross-linked corn starch powders, one sample of cooking corn starch and one oat starch sample, were exposed to ammoniated (AL) or nonammoniated (NAL) raw NRL protein extracts. The levels of individual allergens were determined using the NRL allergen kit. In the NRL glove extracts we observed a wide range in the total allergen levels and a great diversity in the proportion of the four allergens. On the other hand, the evaluated starches had similar ratios of four individual allergens, regardless of the differences in their total allergen levels. The exposure of starches to NRL proteins with different allergen profiles did not affect the allergen ratio. All samples demonstrated a selective affinity for binding Hev b 1 and Hev b 5 allergens and a lesser affinity for the Hev b 6.02 allergen. Allergen Hev b 6.02 made up about 60% of the total allergen in the NAL extract, but only 12-30% of Hev b 6.02 was bound to starches. In contrast, there was only 3-7% of Hev b 1 allergen in the NAL extract, but powders had 35-45% of Hev b 1. These findings indicate that allergenic properties of NRL gloves and respective glove powders may be different.
[141] - Kujala V, Alenius H, Palosuo T, Karvonen J, Pfäffli P, Reijula K. Extractable latex allergens in airborne glove powder and in cut glove pieces. Clin Exp Allergy 2002;32:1077-1081
Background: Quantification of natural rubber latex (NRL) allergens of NRL glove extracts has been reported in several studies. Similarly, immunoassay studies reporting the level of NRL aeroallergens in air samples have been published. When studying the NRL allergens of gloves, however, little attention has been focused on identifying the relationship between extractable NRL allergens of medical gloves and NRL aeroallergens in indoor air. Objective: In an experimental study we analysed NRL aeroallergens of medical gloves in joint relation to total airborne dust concentration and NRL allergen concentration in gloves. Methods: NRL aeroallergen level was measured using a chamber setting with 18 lots of powdered medical gloves. In each setting 10 pairs of powdered NRL gloves were swinging in an unventilated chamber (9m3). Air samples were collected using airflow through Millipore filters (pore size 0.8µm). The filters were weighed before and after the experiment, and total airborne dust concentration in the chamber was calculated. The filter samples were then extracted and the NRL allergen level measured by IgE ELISA-inhibition assay. Furthermore, cut NRL gloves were extracted and analysed by the same method. Finally, levels of two major NRL allergens, Hev b1 and Hev b 6.02, were measured in three selected NRL glove brands. Results: The NRL aeroallergen level in the chamber air ranged from <0.9 to 2.9 allergen units (AU)/m3. The total airborne dust concentration in the chamber air remained low with all lots of gloves measured (range <20 to 80µg/m3). The NRL allergen level in cut glove extracts varied over 100-fold (<10 to 1050 AU/mL). Statistically significant correlation between aeroallergen concentration and airborne dust (r=0.8, P=0.0015) concentration was found. Moreover, significant correlation between aeroallergen levels and allergen content of cut glove pieces was observed (r=0.59, P<0.05). Hev b 1 levels varied from 9 to 25ng/mL and the levels of Hev b 6.02 from 1720 to 14460ng/mL in the glove extracts. In the extracts from airborne dust samples, Hev b 6.02 content varied from 61 to 183ng/m3, whereas Hev b 1 levels were very low (0.4 to 3ng/m3). Conclusion: An elevated NRL aeroallergen level is rather related to a high level of airborne glove powder than to a high concentration of extractable NRL allergen in medical gloves.
[142] - Moneret-Vautrin DA, Kanny G, Guenard L, Beaudouin E, Aublet-Cuvelier A. Intérêt de l'utilisation associée de trois matériels de latex pour prick-tests, dans le diagnostic de l'allergie au latex. Allerg Immunol (Paris) 2000;32:298-302
The diagnosis of latex allergy is mainly based on prick-tests. In order to improve the efficiency of the diagnosis, a comparative study of three latex materials is carried out in 64 controls and 29 patients allergic to latex = Stallergenes and Allerbio extracts, prepared from crude natural latex, and an ammoniated emulsion of rubber latex (AEL). No adverse reactions are recorded. The specificity is 100% for both extracts, 85% for AEL. Sensitivity is respectively 68%, 63%, 72%. The range of values of the wheal is significantly narrower for Stallergenes extract. 46% of allergic patients are reactive to the three materials. The combined use of three materials reaches an 80% efficiency similar to that of Rast Cap System and the addition of three prick-tests and Rast obtains an efficiency of 93.1%. Using several latex materials could increase the efficiency of the diagnosis, inasmuch as the extracts are generated from natural latex, whereas patients are sensitized to manufactured products originating from ammoniated latex.
[145] - Sanz ML, Garcia-Avilés MC, Tabar AI, Anda M, Garcia BE, Barber D, et al. Basophil Activation Test and specific IgE measurements using a panel of recombinant natural rubber latex allergens to determine the latex allergen sensitization profile in children. Pediatr Allergy Immunol 2006;17:148-156
There are no documented studies that describe natural rubber latex (NRL) sensitization in children with a history of surgical intervention but without any congenital malformation (urogenital anomalies, spina bifida, etc.), although some authors have studied NRL allergy in children without a history of surgical intervention. The aim of this work was to evaluate the sensitization profile to single NRL allergens in children without spina bifida and without repeated surgical interventions, by using different recombinant and natural latex allergens in two analytical techniques: specific serum immunoglobulin E (IgE) quantification and flow cytometry determination of activated basophils expressing CD63, after stimulating cells from patients with NRL allergens. A total of 23 patients and 10 healthy children were selected. Conjunctival and in-use NRL provocation tests were carried out, as well as specific IgE determination in all patients' and controls' sera with the recombinant NRL allergens: rHev b 1, rHev b 2, rHev b 3, rHev b 5, rHev b 6.01, rHev b 6.02, rHev b 8, rHev b 9 and rHev b 11 and with NRL (k82) using appropriate ImmunoCAPs. The Basophil Activation Test (BAT) was performed with whole latex extract and with the recombinant allergens rHev b 5 and rHev b 6.01, as well as with the natural allergen Hev b 6.02. The sensitivity and the specificity of NRL-specific IgE (k82) were 100%. Positive IgE responses to rHev b 5 were found in sera of 10 children, to rHev b 6.01 in 16 and for rHev b 6.02 in 15 children's sera. Specific IgE to rHev b 8 was found in four sera of the children. We only found significant differences in sensitization to rHev b 5 in children with two or more surgical interventions compared with the non-intervened group or those with only one intervention. Specific IgE in sera of children with latex-fruit syndrome recognized rHev b 6.02, but not to rHev b 11. The patients sensitized to Hev b 8, Hev b 9 and/or Hev b 11 were atopic. The four patients presenting a positive response to the NRL profilin Hev b 8 were allergic to pollen. The BAT against whole NRL extract was positive in 22 of 23 children; against rHev b 5 in 14 of the patients studied; against rHev b 6.01 in seven cases and against nHev b 6.02 in 19 children. In all the control subjects, the results using this technique were negative. If combined rHev b 5, rHev b 6.01 and nHev b 6.02 together, BAT could detect 20 of the 23 children with latex allergy. The combined use of ImmunoCAP with all the recombinant NRL allergens and BAT with rHev b 5, rHev b 6.01 and nHev b 6.02, enabled the identification of NRL allergy in 22 of 23 patients. There is a positive and significant correlation between sensitization to Hev b 5 and the number of interventions. BAT and allergen-specific IgE determination could be used as first-line in vitro diagnostic tests in patients with NRL allergy.
[146] - Erdmann SM, Moll-Slodowy S, Sauer I, Merk HF. CD63 expression on basophils as a tool for the diagnosis of latex allergy: sensitivity and specificity. EAACI 22th Congress, Paris, 7-11 June, 2003, Poster n°1305
Background: Basophil activation is associated with the expression of CD63. Since allergens can induce basophil activation by cross linking specific IgE, increased CD63 expression has been proposed as a novel in vitro test for immediate type allergy. Objective: We compared the CD63 based basophil activation test (BAT) in the diagnosis of latex allergy with skin prick tests (SPT) and measurement of allergen specific IgE. Methods: 20 patients with a most suggestive history of latex allergy (contact urticaria, angioedema, dyspnea, anaphylaxis) and 10 controls (5 atopics and 5 non atopics) were studied. SPT were performed with a commercially available latex allergen extract (ALK-Abello). Allergen specific IgE was determined by flow cytometry upon double staining with anti-IgE/anti-CD63 mAb. Results: SPT with latex allergen extract showed a sensitivity of 100 % and a specificity of 90 %. Sensitivity of allergen specific IgE and the BAT was 75 % vs. 90 % with corresponding specificities of 90 % vs. 80 %. The cut-off for a positive BAT was 10 % CD63+ basophils. Moreover, there was a positive correlation between IgE reactivity and the number of CD63+ basophils. Conclusions: Quantitation of basophil activation by CD63 expression is a valuable new in vitro method for diagnosis of immediate type allergy to latex. The CD63 based BAT may supplement routine diagnostic tests such as SPT or allergen-specific IgE in the future.
[147] - Sanz ML, Gamboa PM, García-Avilés C, Vila L, Diéguez I, Antépara I, et al. Flow-Cytometric Cellular Allergen Stimulation Test in Latex Allergy. Int Arch Allergy Immunol 2003;130:33-39
Background: The use of flow-cytometric basophil activation to different allergens has been recommended in recent years. In this study, we analyzed the diagnostic reliability of the flow-cytometric allergen stimulation test (FAST) after latex-specific stimulation in vitro. The diagnostic reliability of the technique was assessed as well as its correlation with other in vitro diagnostic parameters. METHODS: 43 patients allergic to latex with a positive history and skin test participated in the study. Thirty subjects (20 of them exposed to latex) with a negative history, skin tests and serum-specific IgE determination to latex were used as controls. In FAST the percentage of basophils that express CD63 as an activation marker after in vitro stimulation with allergen (latex) is determined by flow cytometry, following double labelling with the monoclonal antibodies anti-CD63-PE and anti-IgE FITC. RESULTS: Intraclass correlation coefficient in FAST with latex was 0.995 (p < 0.0001), which demonstrates the excellent reproducibility of this technique. Taking a cutoff point of 10% by means of ROC curves, FAST yields a sensitivity of 93% and a specificity of 100%. The FAST positive predictive value in latex allergy was 100% and the negative predictive value was 99.9%. We found a positive and significant correlation between FAST and specific IgE (CAP) with the histamine release test and specific sulphidoleukotriene production [cellular allergen stimulation test (CAST); p < 0.05]. CONCLUSIONS: FAST is a highly reliable technique (93% sensitivity and 100% specificity) in the in vitro diagnosis of IgE-mediated latex allergy.
[148] - Bernardini R, Pucci N, Azzari C, Novembre E, De Martino M, Milani M. Sensitivity and specificity of different skin prick tests with latex extracts in pediatric patients with suspected natural rubber latex allergy – A cohort study. Pediatr Allergy Immunol 2008;19:315-318
Skin prick tests (SPT) with latex extracts are commonly used in the diagnostic approach of natural rubber latex (NRL) allergy. Non-ammoniated latex extracts are commonly utilized for this purpose. There is only one commercially available ammoniated latex extract for both diagnostic and therapeutic purposes. To our knowledge, no direct clinical comparisons have been made so far comparing sensitivity and specificity of different latex extracts in patients with clinically confirmed latex allergy. The objective of this study was to evaluate sensitivity and specificity of non-ammoniated or ammoniated SPT of latex extracts in children with a clinical history suggestive of NRL allergy. Forty-two subjects (age 4-15 yr), referred to our Allergy Clinic Center, with suspected clinical signs of NRL allergy were evaluated. Patients were selected according to the following inclusion criteria: pediatric patients of age above 4 yr with a suspected clinical history of urticaria, rhinoconjunctivitis and/or asthma due to sensitization to NRL. A definitive NRL allergy diagnosis was made on the identification of latex-specific IgE (>0.7 kU/l), positive glove use test and a highly indicative clinical history. NRL allergy was confirmed in 26 (62%) subjects. SPT for latex were performed with three different SPT solutions containing non-ammoniated NRL extracts (product A: Lofarma SpA, Milan, Italy and product B: Stallergenes, Milan, Italy) or an ammoniated extract (product C: ALK-Abellò, Madrid, Spain). A positive result was considered as the appearance of a wheal with a diameter >3 mm. In patients with confirmed NRL allergy, wheal mean (s.d.) diameters were 7.3 (2.1) with product C, and 5.7 (2.3) and 4.4 (1.8) with products B and A respectively (p = 0.004 C vs. B and A). Mean wheal diameters were significantly higher for product C in comparison with product B and A (p < 0.04). Sensitivity according to the different products tested was 65% (product A); 81% (product B) and 96% (product C) (p = 0.0084; C vs. A). No difference between the three products was observed regarding diagnostic specificity (94%, 88% and 94%, product A, B and C, respectively). Our study shows that diagnostic sensitivity of different latex extract for SPT could differ. In this particular clinical setting (i.e., children with NRL allergy) ammoniated latex extract shows a higher sensitivity in comparison with non-ammoniated products. This aspect should be considered in the diagnostic approach in subjects with suspected NRL allergy. In children with clinical confirmed latex allergy, sensitivity and specificity of different commercially available skin prick test could vary. Ammoniated latex extract has shown a higher sensibility in comparison with non-ammoniated products.
[150] - Hamilton RG, Rossi CE, Yeang HY, Bernstein DI, Biagini R. Latex-Specific IgE Assay Sensitivity Enhanced Using Hev b 5 Enriched Latex Allergosorbent. AAAAI 59th Annual Meeting, Denver, 7-12 March, 2003, Poster n°424
RATIONALE: All clinically-available FDA-cleared serological assays for latex-specific IgE display less than ideal diagnostic sensitivity as a result of allergenic epitopes from multiple Hevb allergens that are either missing or denatured during the allergosorbent manufacturing process (JACI 2002;109:S259, abstract 790). We have studied the extent of performance enhancement in IgE anti-latex detection using a recombinant Hevb5-enriched allergosorbent. METHODS: Serum from 68 latex allergic (clinical history [Hx] positive, Greer investigational skin test [G-ST] positive) and 48 non-latex allergic (Hx negative, G-ST negative) were analyzed in the Pharmacia CAP System using the standard latex (K82) and Hevb5-enriched latex (K85-S) allergosorbents. RESULTS: Based on this cohort of subjects, the diagnostic sensitivity of the CAP System increased significantly (p<0.001) from 51.5% to 61.8% with the use of recombinant Hevb5 supplemented latex on the allergosorbent. Diagnostic specificity remained essentially unchanged [93.8% (K82) vs 91.7% (K82-S)]. There was also a >2-fold increase in quantitative kIU/L levels of IgE antibody detected in sera from 37% of the Hx/G-ST positive subjects, with one third of these representing a seroconversion from negative-(K82) to positive-(K82-S). CONCLUSIONS: The enrichment of the latex CAP allergosorbent with recombinant Hevb5 results in modest improvements in diagnostic sensitivity and qualitative latex-specific IgE antibody detection, with no appreciable loss in diagnostic specificity. Even so, the Pharmacia CAP System still fails to detect latex-specific IgE antibody in up to 38% of latex Hx and G-ST positive subjects. Supplementation of native latex allergens with other recombinant allergens should be a future manufacturing goal.
[151] - Pecquet C, Rouquette AM, Rajoely B, Rollet S, Mouillard S, Leynadier F. Contribution of the new K82 recombi + (Pharmacia) in the diagnostic of latex allergy. EAACI 22th Congress, Paris, 7-11 June, 2003, Poster n°1032
Aim of the study: To evaluate the recombinant hevb5 antigen enhancement of the k82 test (k82 recombi +, Pharmacia) on sera previously negative with the old k82. Method: A blind analysis of 57 sera that gave negative results from January 2001 to November 2002, has been done with the k82 recombi+ (UniCAP 100). Sera were collected from 52 women and five men aged from 17 to 70 (mean : 37,46 + 5,66) including 33 atopic and 34 latex-exposed patients (29 occupational and 5 non- occupational expositions). Medical check-ups were performed for these patients because of suggestive latex immediate hypersensitivity (25 times), food allergy to kiwi or figus or banana (4), local or general per-anesthetic reaction (6), delayed reaction due to the use of gloves (5), one chronic urticaria exploration (5), or for less suggestive symptoms (12). Skin prick tests performed with latex extracts from Laboratoire des Stallergènes (Antony, France) and from Allerbio (Varenne en Argonne, France) were positive (9 times) with both commercial extracts, positive for only one extract (8), non interpretable (dermographism: 10, drug interference or not tested: 2), negative (28). Results: Determination of specific IgE antibodies to k82 recombi + were > 0,35 kUA/l for 6 sera (10%) including 4 (7%) results between 0,99 and 6,72 kUA/l. Three of these 6 patients had an occupationnal exposition, 4 had suggestive symptoms, 3 had positive skin tests with both extracts, 2 positive skin tests with only one extract and non interpretable in one case. Conclusion: In this population, enhancement of the k82 test with recombinant hev b5 allows a gain of 7% additional positive serological specific IgE by the UniCAP Pharmacia method, for latex-exposed patients, suspected of latex allergy.
[152] - Jaby-Ardelean D, Traube C, Cailliez M, Chabane H. In vitro detection of specific IgE to latex: comparison between the old K82 and the new K82 'recombinant improved' (UniCAP Pharmacia). EAACI 22th Congress, Paris, 7-11 June, 2003, Poster n°1033
Latex allergy has become a significant problem in the two last decades, among health care personnel and patients with a history of repeated surgery. The diagnosis is based on clinical history and skin prick tests, completed with specific IgE tests (k82) whose sensibility needs to be improved. Aim of the study: We compared performances of the old k82 test for latex-specific IgE antibodies measurement, with those of the new k82 "spiked" with recombinant protein of hev B5. Material and methods: We analyzed 40 sera, 24 from patients with positive skin tests to latex (Stallergènes, Antony), and 16 with negative skin tests. We measured specific IgE antibodies to latex with the UniCAP system (Pharmacia, Uppsala), both with the old and the new ImmunoCAP k82. Results: Among the 24 patients with positive skin tests, 19 had detectable specific IgE antibodies to latex (> 0.35 KUA/L) with the old k82, versus 21 with the new k82 Latex. 14/19 results were higher with the new k82 than with the old k82. Only 1/16 patients with negative skin tests had a slightly positive value with both k82 tests. Conclusion: The new in vitro test K82 latex is more sensitive for the detection of specific IgE antibodies, with the same specificity.
[153] - Bienvenu F, Benoit Y, Robert O, Bouvier M, Pain V, Mezzarobba C, et al. Usefulness of recombinant allergens in the diagnosis of latex allergy. 16th European Congress of Immunology, Paris, September 6-9, 2006, poster n° 1975
Aim of the study: To investigate the contribution of recombinant allergens in the diagnosis and follow- up of latex allergy as previously done for birch and timothy pollens. Method: Selection of 61 sera with specific IgE against latex (k82), from adults who presented: peranaesthetic anaphylactic shocks (28), occupational allergies (28) and food allergies (5). Each serum was tested, using the ImmunoCAP 250 system, for specific IgE against each of the 9 latex recombinant allergens (rHev b 1, rHev b 2, rHev b 3, rHev b 5, rHev b 6.01, rHev b 6.02, rHev b 8, rHev b 9, rHev b 11) and bromelain. Results: rHev b 6.01, 6.02, rHev b2 et rHev b 5 were the most frequently positive allergens (52%, 49%, 48% and 43% respectively). rHev b 6.02 was strongly correlated with rHev b 6.01; the same is true for rHev b 2 with rHev b 6.01/02. rHev b 9 is slightly positive in only 2 patients/61. Positive results for rHev b 1 and/or rHev b 3 were observed only in the anaphylactic shock group (9/28). Two patients, with discrepant results between negative prick-test (PT) and recent evocative clinical symptoms of latex allergy had an isolated positive rHev b 5. In this series, 11 patients positive for k82 (range: 0,51 to 6.2 kU/L) had only a positive test for rHev b 8 which was always associated with a negative clinical history for latex together with a negative PT; these patients presented with pollinosis. In the 8/61 patients with negative results for all recombinants, the positivity of k82 was related to the presence of anti-CCD IgE (Carbohydrate Cross-reactive Determinants) detected by the bromelain test. Conclusion: These preliminary results provide interesting information which has to be confirmed in a larger population. The utilisation of three recombinants rHev b 5, rHev b 6.01 and rHev b 8 represents a new useful tool to confirm the clinical diagnosis of latex allergy and to understand cross-reactions. rHev b 1 and rHev b 3 are helpful in case of surgical procedures.
[154] - Raulf-Heimsoth ME, Yeang HY, Sander I, Rozynek P, Arif SAM, Fleischer C, et al. Is ENSP (Hev b 13) the Missing Latex Allergen to Fill the Gap in the Repertoire of Isolated Allergens for the Determination of Sensitization Profiles ? AAAAI 59th Annual Meeting, Denver, 7-12 March, 2003, Poster n°96
RATIONALE: Recombinant latex allergens (rHev b 1, rHev b 3, rHev b 5, rHev b 6.01, rHev b 8, rHev b 9, rHev b 10, rHev b 11) are useful tools to determine an individual latex allergen sensitization profile. Although more than 88% of all sera obtained from latex-allergic health care workers (HCW) responded to one or more of these allergens, six out of 53 sera were negative to all 8 recombinant latex allergens. Therefore, we carried out to study further latex allergens. METHODS: Sera of latex-allergic HCW (n=53) and spina bifida patients [SB]; n=11) were tested for their IgE response to the native latex protein that is a homologue of the early nodule specific protein (ENSP) (prepared at the RRIM) by EAST. This new allergen, now designated as Hev b 13 (IUIS), was isolated from the latex B-serum. RESULTS: 86.8% of the HCW sera and 75% of the SB sera were positive to ENSP. In addition, all 6 HCW sera which were negative to the recombinant latex allergens had specific IgE to ENSP (0.36-2.63 kU/L). Furthermore, we found a good correlation between specific IgE to rHev b 6.01 and ENSP. Inhibition-studies revealed a 45-95% inhibition in 5 out of 7 cases of ENSP response by using rHev b 6.01. CONCLUSION: ENSP is an allergen showing high prevalence among HCW and SB with good correlation of specific IgE-responses to rHev b 6.01. It will be necessary to confirm these data by using recombinant ENSP when this will become available.
[155] - Bernstein DI, Biagini RE, Karnanni R, Hamilton R, Murphy K, Bernstein C, et al. In vivo sensitization to purified Hevea brasiliensis proteins in health care workers sensitized to natural rubber latex. J Allergy Clin Immunol 2003;111:610-616
BACKGROUND: Thirteen proteins of natural rubber latex (Hevea brasiliensis) known to bind human IgE have been isolated and characterized as Hev b allergens. However, the in vivo importance of native Hev b allergens has not been defined in health care workers (HCWs) with natural rubber latex (NRL) allergy . OBJECTIVES: The principal aim of this study was to identify the major in vivo Hev b allergens in HCWs with NRL allergy confirmed by percutaneous sensitivity to nonammoniated latex (NAL) . METHODS: Skin prick testing was performed with 7 (native) proteins purified from NAL (Hev b 1, 2, 3, 4, 6.01, 7.01, and a newly described Hev b 13) and recombinant Hev b 5 in 62 HCWs with histories of NRL allergy (group 1) confirmed by percutaneous reactivity to NAL and in 49 atopic HCWs without NRL allergy (group 2). Serial 10-fold concentrations of Hev b proteins (5 x 10(-5) microg/mL to 50 microg/mL) were tested; serum samples of subjects were assayed for serum specific IgE by immunoassays . RESULTS: Hev b 2, Hev b 5, Hev b 6.01, and Hev b 13 produced skin reactions in more than 60% of group 1 subjects, with Hev b 1, 3, 4, and 7.01 eliciting reactions in less than 50%. Only 1 of 49 group 2 workers reacted to a single Hev b antigen (Hev b 13). Specificity of 7 Hev b allergens was 100% and 98% for Hev b 13 in identifying workers with confirmed NRL allergy. Specific IgE by AlaSTAT and CAP immunoassays was elevated in 40 of 60 (67%) and 33 of 62 (53%) of NAL-reactive workers and produced false-positive test results in 4 of 49 (8%) and 3 of 48 (6%) group 2 subjects, respectively . CONCLUSION: Hev b 2, 5, and 6.01 are major in vivo allergens and Hev b 13 is a new major in vivo allergen among HCWs with allergy to NRL.
[156] - Jappe U, Raulf-Heimsoth M, Hoffmann M, Burow G, Hübsch-Müller C, Enk A. In vitro hymenoptera venom allergy diagnosis: improved by screening for cross-reactive carbohydrate determinants and reciprocal inhibition. Allergy 2006;61:1220-1229
BACKGROUND: Immunoglobulin (Ig) E-double positivity for honeybee (HB) and yellow jacket (YJ) venom causes diagnostic difficulties concerning therapeutical strategies. The aim of this study was to clarify the cause and relation of the cross-reactivity in patients with insect venom allergy . METHODS: For this purpose, 147 patients with suspected stinging insect allergy and CAP-FEIA-double positivity were investigated for specific sIgE to additional cross-reactive carbohydrate determinant (CCD)-containing allergens: timothy grass pollen, rape pollen, natural rubber latex (NRL), bromelain, and horseradish peroxidase (HRP). Sera with sIgE to NRL were further investigated with the commercially available recombinant latex allergens. Reciprocal inhibition assays with both venoms and HRP were performed . RESULTS: About 36 of 147 (24.5%) patients had sIgE to both venoms only. However, 111 of 147 (75.5%) additionally reacted to CCD-carrying allergens. 89 of 111 CCD-reactive sera had NRL-sIgE. In cases where inhibition experiments were performed, the NRL-sIgE binding was completely abolished in the presence of HRP. Only nine of 61 sera were positive for at least one recombinant latex allergen; all of them were negative in history and NRL-skin prick test. In 43 sera containing sIgE to CCD, HRP inhibition revealed unequivocal results: In 28 of 43 (65%) an HRP-inhibition >70% of sIgE to one venom occurred, pointing out the relevant venom. In three of 43 sIgE proved to be entirely CCD-specific . CONCLUSIONS: Our data indicate that in cases of IgE positivity to both insect venoms supplementary screening tests with at least one CCD-containing allergen should be performed; HRP being a suitable tool for this test. In addition, subsequent reciprocal inhibition is an essential diagnostic method to specify cross-reacting sIgE results.
[157] - Rihs HP, Rueff F, Lundberg M, Rozynek P, Garcia-Selles J, Barber D, et al. Specific IgE-Binding of rHev b 12 Is Restricted to Fruit-Allergic Patients. AAAAI 60th Annual Meeting, San Francisco, 19-23 March 2004, Poster n°205
Rationale Lipid transfer proteins (LTP) have been identified as relevant allergens. Their role in latex allergy is unclear. This study aimed to produce a recombinant latex-LTP (rHev b 12) to examine its IgE-binding properties. Methods A recombinant generated Maltose-Binding-Protein (MBP)-rHev b 12 fusion protein was coupled to ImmunoCAP for the determination of specific IgE. 48 sera of atopic patients (14 from Spain, 34 from Germany) with fruit-allergy were examined. 25 of these sera displayed positive latex-specific IgE values. Results rHev b 12-specific IgE was measured in four Spanish peach-allergic patients (CAP-values: 0.88-2.27 kU/L), whereby one had an additional latex-sensitization. Two sera from cherry-allergic German patients with latex-sensitization displayed also rHev b 12-specific IgE antibodies (0.68 and 0.96 kU/L). All other sera and the MBP controls revealed negative results (<0.35 kU/L). Conclusions Latex-LTP (rHev b 12)-specific IgE-binding in peach- and cherry-allergic patients seem to be a result of partially common IgE-binding epitopes with the LTPs of peach (homology to Pru p 3: 65%) and cherry (homology to Pru av 3: 61%). Although rHev b 12 seems to have minor relevance as a latex allergen in Central Europe, its significance as a cross-reactive allergen in Mediterranean countries like Spain has to be kept in mind.
[158] - Rihs HP, Chen Z, Schumacher S, Rozynek P, Cremer R, Lundberg M, et al. Recombinant Hev b 1: large-scale production and immunological characterization. Clin Exp Allergy 2000;30:1285-1292
BACKGROUND: Hev b 1 represents one of the most important allergens in Hevea brasiliensis latex. It is difficult to get an appropriate amount of native Hev b 1 (nHev b 1) for research purposes. OBJECTIVE: The aim of this study was to produce sufficient amounts of Hev b 1 by recombinant methods to prove its suitability for latex allergy diagnostics. METHODS: We isolated total RNA of Hevea brasiliensis leaves and synthesized cDNA by RT PCR. Recombinant Hev b 1 (rHev b 1) as well as three fragments (amino acid residues 29-137, 48-137, 78-137) were subcloned and expressed as fusion proteins with Maltose-binding protein (MBP) in Escherichia coli. The MBP-rHev b 1 fusion protein was examined by RAST with the CAP method, histamine release test and immunoblots with human sera from spina bifida patients as well as from health care workers with latex allergy and monoclonal antibodies. RESULTS: Histamine release test and immunoblots revealed the high allergenicity of the MBP-rHev b 1 construct. By the CAP method, 54 out of 58 serum samples (93%) from latex-sensitized spina bifida patients previously showing immunoglobulin (Ig) E to nHev b 1 exhibited IgE-binding to rHev b 1. Among 71 latex-allergic health care workers tested, 16 (22.5%) had IgE antibodies to rHev b 1. The analysis of the fusion proteins carrying rHev b 1 fragments revealed that the loss of the N-terminal 28 amino acid residues did not affect IgE-binding. In contrast, the lack of the first 47 amino acid residues led to decreased IgE-binding reactivity in two out of four sera tested, whereas the absence of the N-terminal 77 residues abolished IgE-binding in these two sera. CONCLUSION: The MBP-rHev b 1 fusion protein exhibits a corresponding IgE-binding reactivity to nHev b 1 and may therefore substitute natural Hev b 1 for both in vitro diagnostics and research purposes.
[159] - Rozynek P, Posch A, Baur X. Cloning, expression and characterization of the major latex allergen prohevein. Clin Exp Allergy 1998;28:1418-1426
BACKGROUND: About 70-80% of latex allergic health care workers are sensitized to prohevein (Hev b 6.01), a 20 kDa cysteine-rich chitin-binding protein of Hevea latex. OBJECTIVE: This study reports on the bacterial cloning, expression and immunochemical characterization of rHev b 6.01. METHODS: Prohevein was expressed in the periplasmatic space of Escherichia coli as maltose binding protein (MBP) fusion protein and purified to homogeneity after factor Xa cleavage. The IgE binding capacity of both rHev b 6.01 and prohevein isolated from fresh Hevea latex was compared by immunoblotting experiments using sera of latex-allergic patients. The diagnostic value of rHev b 6.01 was analysed by enzyme allergosorbent test (EAST). RESULTS: Two different cDNA clones of rHev b 6.01 were established. The deduced amino acid sequence of both clones revealed two and three amino acid differences in the C-terminal domain of prohevein compared with the original database entry. Purified rHev b 6.01 bound with high affinity to chitin as its natural counterpart isolated from natural latex. In IgE-immunblotting using sera of affected subjects binding intensity to both proteins was comparable indicating a very high antigenic similarity. The diagnostic value of MBP-prohevein was tested in EAST using sera of 33 latex-allergic subjects. The in vitro test showed high sensitivity and specificity and proved the diagnostic value of uncleaved MBP-prohevein. CONCLUSIONS: The production of recombinant latex key allergens with defined quality like prohevein is a straightforward strategy for the development of standardized in vitro test systems.
[160] - Bienvenu F, Benoit Y, Robert O, Bouvier M, Pain V, Mezzarobba C, et al. Usefulness of recombinant allergens in the diagnosis of latex allergy. 16th European Congress of Immunology, Paris, September 6-9, 2006, poster n° 1975
Aim of the study: To investigate the contribution of recombinant allergens in the diagnosis and follow- up of latex allergy as previously done for birch and timothy pollens. Method: Selection of 61 sera with specific IgE against latex (k82), from adults who presented: peranaesthetic anaphylactic shocks (28), occupational allergies (28) and food allergies (5). Each serum was tested, using the ImmunoCAP 250 system, for specific IgE against each of the 9 latex recombinant allergens (rHev b 1, rHev b 2, rHev b 3, rHev b 5, rHev b 6.01, rHev b 6.02, rHev b 8, rHev b 9, rHev b 11) and bromelain. Results: rHev b 6.01, 6.02, rHev b2 et rHev b 5 were the most frequently positive allergens (52%, 49%, 48% and 43% respectively). rHev b 6.02 was strongly correlated with rHev b 6.01; the same is true for rHev b 2 with rHev b 6.01/02. rHev b 9 is slightly positive in only 2 patients/61. Positive results for rHev b 1 and/or rHev b 3 were observed only in the anaphylactic shock group (9/28). Two patients, with discrepant results between negative prick-test (PT) and recent evocative clinical symptoms of latex allergy had an isolated positive rHev b 5. In this series, 11 patients positive for k82 (range: 0,51 to 6.2 kU/L) had only a positive test for rHev b 8 which was always associated with a negative clinical history for latex together with a negative PT; these patients presented with pollinosis. In the 8/61 patients with negative results for all recombinants, the positivity of k82 was related to the presence of anti-CCD IgE (Carbohydrate Cross-reactive Determinants) detected by the bromelain test. Conclusion: These preliminary results provide interesting information which has to be confirmed in a larger population. The utilisation of three recombinants rHev b 5, rHev b 6.01 and rHev b 8 represents a new useful tool to confirm the clinical diagnosis of latex allergy and to understand cross-reactions. rHev b 1 and rHev b 3 are helpful in case of surgical procedures.
[161] - Bernstein DI, Biagini RE, Karnanni R, Hamilton R, Murphy K, Bernstein C, et al. In vivo sensitization to purified Hevea brasiliensis proteins in health care workers sensitized to natural rubber latex. J Allergy Clin Immunol 2003;111:610-616
BACKGROUND: Thirteen proteins of natural rubber latex (Hevea brasiliensis) known to bind human IgE have been isolated and characterized as Hev b allergens. However, the in vivo importance of native Hev b allergens has not been defined in health care workers (HCWs) with natural rubber latex (NRL) allergy . OBJECTIVES: The principal aim of this study was to identify the major in vivo Hev b allergens in HCWs with NRL allergy confirmed by percutaneous sensitivity to nonammoniated latex (NAL) . METHODS: Skin prick testing was performed with 7 (native) proteins purified from NAL (Hev b 1, 2, 3, 4, 6.01, 7.01, and a newly described Hev b 13) and recombinant Hev b 5 in 62 HCWs with histories of NRL allergy (group 1) confirmed by percutaneous reactivity to NAL and in 49 atopic HCWs without NRL allergy (group 2). Serial 10-fold concentrations of Hev b proteins (5 x 10(-5) microg/mL to 50 microg/mL) were tested; serum samples of subjects were assayed for serum specific IgE by immunoassays . RESULTS: Hev b 2, Hev b 5, Hev b 6.01, and Hev b 13 produced skin reactions in more than 60% of group 1 subjects, with Hev b 1, 3, 4, and 7.01 eliciting reactions in less than 50%. Only 1 of 49 group 2 workers reacted to a single Hev b antigen (Hev b 13). Specificity of 7 Hev b allergens was 100% and 98% for Hev b 13 in identifying workers with confirmed NRL allergy. Specific IgE by AlaSTAT and CAP immunoassays was elevated in 40 of 60 (67%) and 33 of 62 (53%) of NAL-reactive workers and produced false-positive test results in 4 of 49 (8%) and 3 of 48 (6%) group 2 subjects, respectively . CONCLUSION: Hev b 2, 5, and 6.01 are major in vivo allergens and Hev b 13 is a new major in vivo allergen among HCWs with allergy to NRL.
[162] - Malandain H, Giroux F, Cano Y. The influence of carbohydrate structures present in common allergen sources on specific IgE results. Eur Ann Allergy Clin Immunol 2007;39:216-220
BACKGROUND: Cross-reactive carbohydrate determinants (CCD) are well known interferants in specific IgE assays (sIgE). Glyco-epitopes are not restricted to CCD and extracts used to prepare in vitro tests contain many other glycoproteins able to bind glycan-specific IgE. The overall amounts of IgE-bindable glycan structures in allergen sources are unknown . OBJECTIVE: We aimed at quantifying the influence of N-glycan structures on IgE reactivity to commonly tested allergen sources . METHODS: IgE reactivity to 51 allergen extracts, one purified natural allergen and 10 recombinant allergens was measured on Phadia UniCAP system using 2 sera demonstrating significant levels of glycan-related IgE reactivity. Immobilized bromelain and horseradish peroxidase (HRP) were used to capture N-glycan-specific IgE from these sera. Residual IgE reactivity was measured for 42 allergen sources and 4 recombinant/purified allergens . RESULTS: An obviously excessive number of positive CAP-results were obtained with both sera, especially for plant-based allergen sources. Capture of glycan-specific IgE led to a decrease of serum IgE ractivity, variable among allergen sources and between sera. Among others, peanut results were proven largely interfered by the presence of glycan-specific IgE. Unexpectedly some allergen sources showed a slight influence of glycan-related reactivity, such as cockroach, mosquito, mussel, shrimp and domestic mites . CONCLUSION: In patients sensitized to pollens or to Hymenoptera venoms sIgE results should be interpreted with caution. One cannot substract the result of a glyco-reporter test (bromelain and/or HRP) in order to compute glycan-free slgE results for common allergen sources like peanuts. As long as the demonstration of a significant role for glycan structures in clinical allergic reactions is lacking, a simple pre-treatment able to discard glycan-specific IgE from serum would be useful to improve accuracy of in vitro diagnostic tests.
[163] - Bienvenu F, Benoit Y, Robert O, Bouvier M, Pain V, Mezzarobba C, et al. Usefulness of recombinant allergens in the diagnosis of latex allergy. 16th European Congress of Immunology, Paris, September 6-9, 2006, poster n° 1975
Aim of the study: To investigate the contribution of recombinant allergens in the diagnosis and follow- up of latex allergy as previously done for birch and timothy pollens. Method: Selection of 61 sera with specific IgE against latex (k82), from adults who presented: peranaesthetic anaphylactic shocks (28), occupational allergies (28) and food allergies (5). Each serum was tested, using the ImmunoCAP 250 system, for specific IgE against each of the 9 latex recombinant allergens (rHev b 1, rHev b 2, rHev b 3, rHev b 5, rHev b 6.01, rHev b 6.02, rHev b 8, rHev b 9, rHev b 11) and bromelain. Results: rHev b 6.01, 6.02, rHev b2 et rHev b 5 were the most frequently positive allergens (52%, 49%, 48% and 43% respectively). rHev b 6.02 was strongly correlated with rHev b 6.01; the same is true for rHev b 2 with rHev b 6.01/02. rHev b 9 is slightly positive in only 2 patients/61. Positive results for rHev b 1 and/or rHev b 3 were observed only in the anaphylactic shock group (9/28). Two patients, with discrepant results between negative prick-test (PT) and recent evocative clinical symptoms of latex allergy had an isolated positive rHev b 5. In this series, 11 patients positive for k82 (range: 0,51 to 6.2 kU/L) had only a positive test for rHev b 8 which was always associated with a negative clinical history for latex together with a negative PT; these patients presented with pollinosis. In the 8/61 patients with negative results for all recombinants, the positivity of k82 was related to the presence of anti-CCD IgE (Carbohydrate Cross-reactive Determinants) detected by the bromelain test. Conclusion: These preliminary results provide interesting information which has to be confirmed in a larger population. The utilisation of three recombinants rHev b 5, rHev b 6.01 and rHev b 8 represents a new useful tool to confirm the clinical diagnosis of latex allergy and to understand cross-reactions. rHev b 1 and rHev b 3 are helpful in case of surgical procedures.
[164] - Mari A, Scala E, D’Ambrosio C, Breiteneder H, Wagner S. Latex Allergy within a Cohort of Not-at-Risk Subjects with Respiratory Symptoms: Prevalence of Latex Sensitization and Assessment of Diagnostic Tools. Int Arch Allergy Immunol 2007;143:135-143
BACKGROUND: Immunoglobluin E (IgE)-mediated hypersensitivity to natural rubber latex (NRL) is a major problem in allergy practice. Currently, the use of skin prick tests (SPTs) with latex extracts and specific IgE detection for the diagnosis of NRL allergy in suspected patients is directed to identification of risk factors. Many cases of NRL allergy remain undiagnosed due to misreporting of symptoms by the patients or lack of proper questions asked by the physician. MATERIALS AND METHODS: A total of 6,126 subjects referred for respiratory symptoms underwent SPTs with NRL. Positive subjects were resurveyed for exposure to NRL, and specific IgE for NRL extracts and recombinant molecules was determined. Immunoblots of NRL extracts were performed to identify IgE patterns . RESULTS: Forty-six of 3,930 sensitized subjects had a positive SPT with NRL, displaying a prevalence of NRL sensitization of 0.75% for the general and 1.2% for the sensitized population. Eleven out of 46 (23.9%) subjects could be defined as NRL asymptomatic, whereas 35 (76.1%) developed symptoms upon exposure to NRL. Specific IgE to NRL was detected for 22 (75.86%) of 29 tested sera. Seventeen out of 22 (77%) sera displayed specific IgE to recombinant allergens with most reactions to Hev b 5, Hev b 6.01 and Hev b 6.02. Immunoblots of NRL extract fractions with patients' sera showed heterogeneous patterns . CONCLUSIONS: SPTs with NRL extract should be routinely performed in patients with respiratory symptoms. Hev b 5, Hev b 6.01 and Hev b 6.02 are the most important allergens, but further characterization of NRL extracts is needed to identify novel allergens and to clarify the role of crossreactive carbohydrate determinants.
[165] - Bernstein DI, Biagini RE, Karnanni R, Hamilton R, Murphy K, Bernstein C, et al. In vivo sensitization to purified Hevea brasiliensis proteins in health care workers sensitized to natural rubber latex. J Allergy Clin Immunol 2003;111:610-616
BACKGROUND: Thirteen proteins of natural rubber latex (Hevea brasiliensis) known to bind human IgE have been isolated and characterized as Hev b allergens. However, the in vivo importance of native Hev b allergens has not been defined in health care workers (HCWs) with natural rubber latex (NRL) allergy . OBJECTIVES: The principal aim of this study was to identify the major in vivo Hev b allergens in HCWs with NRL allergy confirmed by percutaneous sensitivity to nonammoniated latex (NAL) . METHODS: Skin prick testing was performed with 7 (native) proteins purified from NAL (Hev b 1, 2, 3, 4, 6.01, 7.01, and a newly described Hev b 13) and recombinant Hev b 5 in 62 HCWs with histories of NRL allergy (group 1) confirmed by percutaneous reactivity to NAL and in 49 atopic HCWs without NRL allergy (group 2). Serial 10-fold concentrations of Hev b proteins (5 x 10(-5) microg/mL to 50 microg/mL) were tested; serum samples of subjects were assayed for serum specific IgE by immunoassays . RESULTS: Hev b 2, Hev b 5, Hev b 6.01, and Hev b 13 produced skin reactions in more than 60% of group 1 subjects, with Hev b 1, 3, 4, and 7.01 eliciting reactions in less than 50%. Only 1 of 49 group 2 workers reacted to a single Hev b antigen (Hev b 13). Specificity of 7 Hev b allergens was 100% and 98% for Hev b 13 in identifying workers with confirmed NRL allergy. Specific IgE by AlaSTAT and CAP immunoassays was elevated in 40 of 60 (67%) and 33 of 62 (53%) of NAL-reactive workers and produced false-positive test results in 4 of 49 (8%) and 3 of 48 (6%) group 2 subjects, respectively . CONCLUSION: Hev b 2, 5, and 6.01 are major in vivo allergens and Hev b 13 is a new major in vivo allergen among HCWs with allergy to NRL.
[167] - Raulf-Heimsoth M, Rihs HP, Rozynek P, Cremer R, Gaspar A, Pires G, et al. Quantitative analysis of immunoglobulin E reactivity profiles in patients allergic or sensitized to natural rubber latex (Hevea brasiliensis). Clin Exp Allergy 2007;37:1657-1667
BACKGROUND: Characterized native and recombinant Hevea brasiliensis (rHev b) natural rubber latex (NRL) allergens are available to assess patient allergen sensitization profiles . OBJECTIVE: Quantification of individual IgE responses to the spectrum of documented NRL allergens and evaluation of cross-reactive carbohydrate determinants (CCDs) for more definitive diagnosis . METHODS: Sera of 104 healthcare workers (HCW; 51 German, 21 Portuguese, 32 American), 31 spina bifida patients (SB; 11 German, 20 Portuguese) and 10 Portuguese with multiple surgeries (MS) were analysed for allergen-specific IgE antibody (sIgE) to NRL, single Hev b allergens and CCDs with ImmunoCAP technology . RESULTS: In all patient groups rHev b 5-sIgE concentrations were the most pronounced. Hev b 2, 5, 6.01 and 13 were identified as the major allergens in HCW and combined with Hev b 1 and Hev b 3 in SB. In MS Hev b 1 displayed an intermediate relevance. Different sIgE antibody levels to native Hevea brasiliensis (nHev b) 2 and rHev b 6.01 allowed discrimination of SB with clinical relevant latex allergy vs. those with latex sensitization. Sensitization profiles of German, Portuguese and American patients were equivalent. rHev b 5, 6.01 and nHev b 13 combined detected 100% of the latex-allergic HCW and 80.1% of the SB. Only 8.3% of the sera showed sIgE response to CCDs . CONCLUSIONS: Hev b 1, 2, 5, 6.01 and 13 were identified as the major Hev b allergens and they should be present in standardized latex extracts and in vitro allergosorbents. CCDs are only of minor relevance in patients with clinical relevant latex allergy. Component-resolved diagnostic analyses for latex allergy set the stage for an allergen-directed immunotherapy strategy.
[168] - Radauer C, Hafner C, Ganglberger E, Wagner B, Scheiner O, Breiteneder H. Profilin-specific immunoglobulin E is capable of eliciting clinical symptoms of latex allergy. EAACI 21th Congress, Naples, 1-5 June, 2002, Communication n°31
Patients allergic to Hevea brasiliensis latex often show allergic symptoms to various fruits such as banana, avocado, chestnut, or kiwi. Many of these patients also exhibit pollen allergy. The cross-reactivity between latex and fruits has been attributed to the major latex allergens Hev b 2 and Hev b 6. Profilins are important cross-reactive pollen and food allergens, while their role in latex allergy is a matter of debate, since it is unclear if sensitization to profilin can elicit allergic reactions to latex or if the observed cross-reactivity of latex profilin (Hev b 8) is merely an in vitro phenomenon. In order to clarify this issue, we selected patients visiting an allergy clinic according to the presence of birch pollen profilin-specific IgE as determined by Pharmacia CAP or IgE immunoblot. Seventeen of these patients were subjected to a latex glove provocation test, which resulted in six positive outcomes. Sera of these six patients were tested in IgE immunoblots of latex B and C extracts as well as of purified recombinant Hev b 8 and Hev b 1, an allergen present in latex C extract with a molecular mass similar to profilin. Three sera recognized no proteins in latex B extract, a single 14 kDa band in latex C extract, and recombinant Hev b 8, but not recombinant Hev b 1. IgE binding to the 14 kDa band in latex C extract was completely abolished by preincubation with recombinant Hev b 8. From these data we conclude that monosensitization to profilin can elicit allergic symptoms to latex and the risk for profilinallergic patients to develop latex allergy should not be neglected.
[169] - Bienvenu F, Benoit Y, Robert O, Bouvier M, Pain V, Mezzarobba C, et al. Usefulness of recombinant allergens in the diagnosis of latex allergy. 16th European Congress of Immunology, Paris, September 6-9, 2006, poster n° 1975
Aim of the study: To investigate the contribution of recombinant allergens in the diagnosis and follow- up of latex allergy as previously done for birch and timothy pollens. Method: Selection of 61 sera with specific IgE against latex (k82), from adults who presented: peranaesthetic anaphylactic shocks (28), occupational allergies (28) and food allergies (5). Each serum was tested, using the ImmunoCAP 250 system, for specific IgE against each of the 9 latex recombinant allergens (rHev b 1, rHev b 2, rHev b 3, rHev b 5, rHev b 6.01, rHev b 6.02, rHev b 8, rHev b 9, rHev b 11) and bromelain. Results: rHev b 6.01, 6.02, rHev b2 et rHev b 5 were the most frequently positive allergens (52%, 49%, 48% and 43% respectively). rHev b 6.02 was strongly correlated with rHev b 6.01; the same is true for rHev b 2 with rHev b 6.01/02. rHev b 9 is slightly positive in only 2 patients/61. Positive results for rHev b 1 and/or rHev b 3 were observed only in the anaphylactic shock group (9/28). Two patients, with discrepant results between negative prick-test (PT) and recent evocative clinical symptoms of latex allergy had an isolated positive rHev b 5. In this series, 11 patients positive for k82 (range: 0,51 to 6.2 kU/L) had only a positive test for rHev b 8 which was always associated with a negative clinical history for latex together with a negative PT; these patients presented with pollinosis. In the 8/61 patients with negative results for all recombinants, the positivity of k82 was related to the presence of anti-CCD IgE (Carbohydrate Cross-reactive Determinants) detected by the bromelain test. Conclusion: These preliminary results provide interesting information which has to be confirmed in a larger population. The utilisation of three recombinants rHev b 5, rHev b 6.01 and rHev b 8 represents a new useful tool to confirm the clinical diagnosis of latex allergy and to understand cross-reactions. rHev b 1 and rHev b 3 are helpful in case of surgical procedures.
[172] - Quercia O, Stefanini GF, Scardovi A, Asero R. Patients monosensitised to Hev b 8 (Hevea brasiliensis latex profilin) may safely undergo major surgery in a normal (non-latex safe) environment. Eur Ann Allergy Clin Immunol 2009;41:112-116
BACKGROUND: Natural rubber latex allergy is a condition at high risk of anaphylaxis during surgery. However, latex contains several protein allergens and not all of them may show the same clinical relevance. OBJECTIVE: To investigate the clinical relevance of Hey b 8, the natural rubber latex profilin. METHODS: Seven patients without a clinical history of latex allergy but identified as being latex hypersensitive by positive SPT (3/7) and or positive latex-specific IgE during routine pre-surgery allergy investigations were studied. All patients were monosensitized to Hev b 8 (Hevea brasiliensis latex profilin) as shown by the detection of specific IgE to recombinant latex allergen components. Ten subjects with a history of latex allergy (urticaria, asthma, and/or rhinitis), sensitised to latex allergens other than profilin were enrolled as controls. Both patients and controls underwent a latex glove-wearing test; in case of a negative test, patients underwent surgery in a normal surgical setting. RESULTS: All 7 patients scored negative on latex glove wearing test and underwent major surgery (orthopaedic, Caesarean section, pilonidal sinus, vascular, tonsillectomy, uterine revision, and uretral surgery) in a normal (non-latex safe) surgical setting without any consequence. In contrast, 9/10 (90%) controls showed a positive latex glove-wearing test (p < 0.01). CONCLUSION: Latex profilin is either clinically irrelevant or is no longer present in latex products. This study highlights the importance of a component-resolved diagnosis of latex sensitisation as a tool to get a more precise assessment of the risk and to reduce the costs of healthcare.
[176] - Raulf-Heimsoth M, Rihs HP, Rozynek P, Cremer R, Gaspar A, Pires G, et al. Quantitative analysis of immunoglobulin E reactivity profiles in patients allergic or sensitized to natural rubber latex (Hevea brasiliensis). Clin Exp Allergy 2007;37:1657-1667
BACKGROUND: Characterized native and recombinant Hevea brasiliensis (rHev b) natural rubber latex (NRL) allergens are available to assess patient allergen sensitization profiles . OBJECTIVE: Quantification of individual IgE responses to the spectrum of documented NRL allergens and evaluation of cross-reactive carbohydrate determinants (CCDs) for more definitive diagnosis . METHODS: Sera of 104 healthcare workers (HCW; 51 German, 21 Portuguese, 32 American), 31 spina bifida patients (SB; 11 German, 20 Portuguese) and 10 Portuguese with multiple surgeries (MS) were analysed for allergen-specific IgE antibody (sIgE) to NRL, single Hev b allergens and CCDs with ImmunoCAP technology . RESULTS: In all patient groups rHev b 5-sIgE concentrations were the most pronounced. Hev b 2, 5, 6.01 and 13 were identified as the major allergens in HCW and combined with Hev b 1 and Hev b 3 in SB. In MS Hev b 1 displayed an intermediate relevance. Different sIgE antibody levels to native Hevea brasiliensis (nHev b) 2 and rHev b 6.01 allowed discrimination of SB with clinical relevant latex allergy vs. those with latex sensitization. Sensitization profiles of German, Portuguese and American patients were equivalent. rHev b 5, 6.01 and nHev b 13 combined detected 100% of the latex-allergic HCW and 80.1% of the SB. Only 8.3% of the sera showed sIgE response to CCDs . CONCLUSIONS: Hev b 1, 2, 5, 6.01 and 13 were identified as the major Hev b allergens and they should be present in standardized latex extracts and in vitro allergosorbents. CCDs are only of minor relevance in patients with clinical relevant latex allergy. Component-resolved diagnostic analyses for latex allergy set the stage for an allergen-directed immunotherapy strategy.
[177] - Bienvenu F, Benoit Y, Robert O, Bouvier M, Pain V, Mezzarobba C, et al. Usefulness of recombinant allergens in the diagnosis of latex allergy. 16th European Congress of Immunology, Paris, September 6-9, 2006, poster n° 1975
Aim of the study: To investigate the contribution of recombinant allergens in the diagnosis and follow- up of latex allergy as previously done for birch and timothy pollens. Method: Selection of 61 sera with specific IgE against latex (k82), from adults who presented: peranaesthetic anaphylactic shocks (28), occupational allergies (28) and food allergies (5). Each serum was tested, using the ImmunoCAP 250 system, for specific IgE against each of the 9 latex recombinant allergens (rHev b 1, rHev b 2, rHev b 3, rHev b 5, rHev b 6.01, rHev b 6.02, rHev b 8, rHev b 9, rHev b 11) and bromelain. Results: rHev b 6.01, 6.02, rHev b2 et rHev b 5 were the most frequently positive allergens (52%, 49%, 48% and 43% respectively). rHev b 6.02 was strongly correlated with rHev b 6.01; the same is true for rHev b 2 with rHev b 6.01/02. rHev b 9 is slightly positive in only 2 patients/61. Positive results for rHev b 1 and/or rHev b 3 were observed only in the anaphylactic shock group (9/28). Two patients, with discrepant results between negative prick-test (PT) and recent evocative clinical symptoms of latex allergy had an isolated positive rHev b 5. In this series, 11 patients positive for k82 (range: 0,51 to 6.2 kU/L) had only a positive test for rHev b 8 which was always associated with a negative clinical history for latex together with a negative PT; these patients presented with pollinosis. In the 8/61 patients with negative results for all recombinants, the positivity of k82 was related to the presence of anti-CCD IgE (Carbohydrate Cross-reactive Determinants) detected by the bromelain test. Conclusion: These preliminary results provide interesting information which has to be confirmed in a larger population. The utilisation of three recombinants rHev b 5, rHev b 6.01 and rHev b 8 represents a new useful tool to confirm the clinical diagnosis of latex allergy and to understand cross-reactions. rHev b 1 and rHev b 3 are helpful in case of surgical procedures.
[178] - Beezhold DH, Hickey VL, Slater JE, Sussman GL. Human IgE-binding epitopes of the latex allergen Hev b 5. J Allergy Clin Immunol 1999;103:1166-1172
BACKGROUND: Hev b 5 is an acidic protein (isoelectric point, 3.5) rich in glutamic acid with 9 repeated amino acid (AA) sequences of XEEX or XEEEX. Although its function in Hevea brasiliensis is unknown, Hev b 5 has been identified as a major latex allergen. Immunoblot inhibition studies suggest Hev b 5 exists as multiple isoforms or contains a common epitope found in several other proteins. OBJECTIVE: The purpose of this study was to further characterize Hev b 5 and to identify linear IgE-binding epitopes. METHODS: Octapeptides spanning the entire Hev b 5 protein were synthesized on a derivatized cellulose membrane. The membrane was reacted with sera pooled from health care workers allergic to latex or rabbits immunized with latex proteins. B-cell epitopes were identified by subsequent incubations with the appropriate secondary antibodies and detected by using chemifluorescence. RESULTS: Sera from patients allergic to latex recognized 6 IgE-binding regions located throughout the molecule. Two epitopes (2 and 4) had the common AA sequence of KTEEP. Epitopes 3 and 5 had a similar AA sequence of EEXXA, where X was P, T, or K. Epitopes 1 and 6 appeared to be unrelated to the other epitopes. Database analysis could not identify other proteins with similar sequences. Neither of the XEEEX sequences bound IgE. Control sera failed to react to any peptides. CONCLUSIONS: Hev b 5 exists as multiple isoforms, but only small amounts are present in the nonammoniated latex preparations, such as those used for diagnostic tests, and this may help to explain the relatively poor sensitivity of some in vitro tests.
[179] - Hamilton RG, Rossi CE, Yeang HY, Bernstein DI, Biagini R. Latex-Specific IgE Assay Sensitivity Enhanced Using Hev b 5 Enriched Latex Allergosorbent. AAAAI 59th Annual Meeting, Denver, 7-12 March, 2003, Poster n°424
RATIONALE: All clinically-available FDA-cleared serological assays for latex-specific IgE display less than ideal diagnostic sensitivity as a result of allergenic epitopes from multiple Hevb allergens that are either missing or denatured during the allergosorbent manufacturing process (JACI 2002;109:S259, abstract 790). We have studied the extent of performance enhancement in IgE anti-latex detection using a recombinant Hevb5-enriched allergosorbent. METHODS: Serum from 68 latex allergic (clinical history [Hx] positive, Greer investigational skin test [G-ST] positive) and 48 non-latex allergic (Hx negative, G-ST negative) were analyzed in the Pharmacia CAP System using the standard latex (K82) and Hevb5-enriched latex (K85-S) allergosorbents. RESULTS: Based on this cohort of subjects, the diagnostic sensitivity of the CAP System increased significantly (p<0.001) from 51.5% to 61.8% with the use of recombinant Hevb5 supplemented latex on the allergosorbent. Diagnostic specificity remained essentially unchanged [93.8% (K82) vs 91.7% (K82-S)]. There was also a >2-fold increase in quantitative kIU/L levels of IgE antibody detected in sera from 37% of the Hx/G-ST positive subjects, with one third of these representing a seroconversion from negative-(K82) to positive-(K82-S). CONCLUSIONS: The enrichment of the latex CAP allergosorbent with recombinant Hevb5 results in modest improvements in diagnostic sensitivity and qualitative latex-specific IgE antibody detection, with no appreciable loss in diagnostic specificity. Even so, the Pharmacia CAP System still fails to detect latex-specific IgE antibody in up to 38% of latex Hx and G-ST positive subjects. Supplementation of native latex allergens with other recombinant allergens should be a future manufacturing goal.
[180] - Pecquet C, Rouquette AM, Rajoely B, Rollet S, Mouillard S, Leynadier F. Contribution of the new K82 recombi + (Pharmacia) in the diagnostic of latex allergy. EAACI 22th Congress, Paris, 7-11 June, 2003, Poster n°1032
Aim of the study: To evaluate the recombinant hevb5 antigen enhancement of the k82 test (k82 recombi +, Pharmacia) on sera previously negative with the old k82. Method: A blind analysis of 57 sera that gave negative results from January 2001 to November 2002, has been done with the k82 recombi+ (UniCAP 100). Sera were collected from 52 women and five men aged from 17 to 70 (mean : 37,46 + 5,66) including 33 atopic and 34 latex-exposed patients (29 occupational and 5 non- occupational expositions). Medical check-ups were performed for these patients because of suggestive latex immediate hypersensitivity (25 times), food allergy to kiwi or figus or banana (4), local or general per-anesthetic reaction (6), delayed reaction due to the use of gloves (5), one chronic urticaria exploration (5), or for less suggestive symptoms (12). Skin prick tests performed with latex extracts from Laboratoire des Stallergènes (Antony, France) and from Allerbio (Varenne en Argonne, France) were positive (9 times) with both commercial extracts, positive for only one extract (8), non interpretable (dermographism: 10, drug interference or not tested: 2), negative (28). Results: Determination of specific IgE antibodies to k82 recombi + were > 0,35 kUA/l for 6 sera (10%) including 4 (7%) results between 0,99 and 6,72 kUA/l. Three of these 6 patients had an occupationnal exposition, 4 had suggestive symptoms, 3 had positive skin tests with both extracts, 2 positive skin tests with only one extract and non interpretable in one case. Conclusion: In this population, enhancement of the k82 test with recombinant hev b5 allows a gain of 7% additional positive serological specific IgE by the UniCAP Pharmacia method, for latex-exposed patients, suspected of latex allergy.
[181] - Jaby-Ardelean D, Traube C, Cailliez M, Chabane H. In vitro detection of specific IgE to latex: comparison between the old K82 and the new K82 'recombinant improved' (UniCAP Pharmacia). EAACI 22th Congress, Paris, 7-11 June, 2003, Poster n°1033
Latex allergy has become a significant problem in the two last decades, among health care personnel and patients with a history of repeated surgery. The diagnosis is based on clinical history and skin prick tests, completed with specific IgE tests (k82) whose sensibility needs to be improved. Aim of the study: We compared performances of the old k82 test for latex-specific IgE antibodies measurement, with those of the new k82 "spiked" with recombinant protein of hev B5. Material and methods: We analyzed 40 sera, 24 from patients with positive skin tests to latex (Stallergènes, Antony), and 16 with negative skin tests. We measured specific IgE antibodies to latex with the UniCAP system (Pharmacia, Uppsala), both with the old and the new ImmunoCAP k82. Results: Among the 24 patients with positive skin tests, 19 had detectable specific IgE antibodies to latex (> 0.35 KUA/L) with the old k82, versus 21 with the new k82 Latex. 14/19 results were higher with the new k82 than with the old k82. Only 1/16 patients with negative skin tests had a slightly positive value with both k82 tests. Conclusion: The new in vitro test K82 latex is more sensitive for the detection of specific IgE antibodies, with the same specificity.
[183] - Smith AM, Amin HS, Biagini RE, Hamilton RG, Arif SAM, Yeang HY, et al. Percutaneous reactivity to natural rubber latex proteins persists in health-care workers following avoidance of natural rubber latex. Clin Exp Allergy 2007;37:1349-1356
BACKGROUND: Long-term avoidance of natural rubber latex [Hevea brasiliensis (Hev b)] is currently recommended for health-care workers (HCWs) with established natural rubber latex (NRL) allergy. Percutaneous sensitivity to eight Hev b NRL allergens was evaluated in HCWs in 2000. To date, no studies have evaluated the longitudinal effects of NRL avoidance on percutaneous sensitivity to NRL allergens . OBJECTIVE: The aims of this study were to evaluate changes in percutaneous reactivity to non-ammoniated latex (NAL) and NRL allergens in HCWs 5 years after a recommendation to avoid NRL and to evaluate factors that predict the persistence of in vivo sensitivity to NAL and NRL allergens . METHODS: Skin prick testing was performed with NAL, seven NRL allergens (Hev b 1, 2, 3, 4, 6.01, 7.01, and 13), and recombinant Hev b 5 (rHev b 5) in 34 HCWs who were initially evaluated in 2000 for occupationally related NRL allergy. Serial 10-fold dilutions of NAL and NRL allergens were employed in skin testing. Sera from the HCWs were assayed for latex and enhanced latex (rHev b 5-enriched allergosorbent)-specific IgE antibodies using the ImmunoCAP assay . RESULTS: The prevalence of work-related symptoms significantly decreased between 2000 and 2005 with avoidance of NRL (P<0.05). A >/=100-fold reduction in percutaneous sensitivity to Hev b 2 and Hev b 7 was less likely in those with prior history of systemic reactions to NRL (P=0.0053), reported history of reaction to cross-reactive foods (P=0.014), continued local reactions to NRL gloves (P<0.0001), or high NRL glove exposure since the initial study (P=0.0075). The diagnostic sensitivity and specificity of the latex-specific IgE serology was 54% and 87.5%, respectively, in comparison with NAL skin tests. The addition of rHev b 5 to the ImmunoCAP (enhanced latex) allergosorbent altered the diagnostic sensitivity and specificity of the ImmunoCAP to 77% and 75%, respectively . CONCLUSION: While symptoms may resolve quickly with NRL avoidance therapy, detectable IgE indicating continued sensitization remains beyond 5 years, and thus continued avoidance of NRL should be recommended.
[184] - Yagami T, Haishima Y, Tsuchiya T, Tomitaka-Yagami A, Kano H, Matsunaga K. Proteomic Analysis of Putative Latex Allergens. Int Arch Allergy Immunol 2004;135:3-11
BACKGROUND: Extensive analysis of allergenic proteins is generally time-consuming and labor-intensive. Accordingly, a rapid and easy procedure for allergen identification is required. As sequence information on proteins and genes is accumulated in databases, it is becoming easier to identify a candidate protein using proteomic strategies, i.e. two-dimensional gel electrophoresis, site-specific fragmentation, mass spectrometry and then database search. In this study, we evaluated the usefulness of a proteomic strategy for identifying putative allergens through its application to latex proteins . METHODS: Latex proteins were separated with two-dimensional gel electrophoresis, and putative allergens were visualized by IgE immunoblotting using pooled serum from latex-sensitive patients. The IgE-interactive proteins were cut out from the negatively stained two-dimensional gel and subjected to in-gel digestion by trypsin. Then the resulting peptides were analyzed with mass spectrometry. Based on the mass spectrometric data we obtained, the allergen candidates were assigned by a database search . RESULTS: Five previously reported allergens and five new allergen candidates were identified with the proteomic approach without isolating the individual proteins. Less than 1 mg of crude latex protein was sufficient for the entire protocol. Because plural proteins can be processed in parallel, analysis of about 50 IgE-interactive proteins was accomplished within 1 week . CONCLUSIONS: Analysis of putative allergens with proteomic strategies (allergenomics) is a promising avenue for rapid and exhaustive research. The high resolving power of two-dimensional gel electrophoresis is superior to conventional gel electrophoresis. Moreover, the notable sensitivity and speed of mass spectrometry have pronounced advantages over the N-terminal sequencing that has generally been used for protein identification.
[185] - Yagami T, Haishima Y, Tsuchiya T, Tomitaka-Yagami A, Kano H, Matsunaga K. Proteomic Analysis of Putative Latex Allergens. Int Arch Allergy Immunol 2004;135:3-11
BACKGROUND: Extensive analysis of allergenic proteins is generally time-consuming and labor-intensive. Accordingly, a rapid and easy procedure for allergen identification is required. As sequence information on proteins and genes is accumulated in databases, it is becoming easier to identify a candidate protein using proteomic strategies, i.e. two-dimensional gel electrophoresis, site-specific fragmentation, mass spectrometry and then database search. In this study, we evaluated the usefulness of a proteomic strategy for identifying putative allergens through its application to latex proteins . METHODS: Latex proteins were separated with two-dimensional gel electrophoresis, and putative allergens were visualized by IgE immunoblotting using pooled serum from latex-sensitive patients. The IgE-interactive proteins were cut out from the negatively stained two-dimensional gel and subjected to in-gel digestion by trypsin. Then the resulting peptides were analyzed with mass spectrometry. Based on the mass spectrometric data we obtained, the allergen candidates were assigned by a database search . RESULTS: Five previously reported allergens and five new allergen candidates were identified with the proteomic approach without isolating the individual proteins. Less than 1 mg of crude latex protein was sufficient for the entire protocol. Because plural proteins can be processed in parallel, analysis of about 50 IgE-interactive proteins was accomplished within 1 week . CONCLUSIONS: Analysis of putative allergens with proteomic strategies (allergenomics) is a promising avenue for rapid and exhaustive research. The high resolving power of two-dimensional gel electrophoresis is superior to conventional gel electrophoresis. Moreover, the notable sensitivity and speed of mass spectrometry have pronounced advantages over the N-terminal sequencing that has generally been used for protein identification.
[186] - Posch A, Chen Z, Wheeler CH, Dunn MJ, Raulf-Heimsoth M, Baur X. Characterization and identification of latex allergens by two-dimensional electrophoresis and protein microsequencing. J Allergy Clin Immunol 1997;99:385-395
Proteins of natural rubber latex cause IgE-mediated sensitization in 3% to 18% of health care workers and in up to 50% of patients with spina bifida. OBJECTIVE: This study was aimed at the generation of a comprehensive latex protein database by two-dimensional electrophoresis (2-DE). METHODS: Proteins extracted from fresh Hevea brasiliensis latex were separated by 2-DE. IgE-reactive proteins were analyzed by immunoblotting with sera of health care workers with latex allergy. Protein microsequencing and monoclonal antibodies were used to identify the latex allergens. RESULTS: The latex C-serum 2-DE map was very complex and exhibited about 200 distinct polypeptides. The proteins eluted from the latex particles consisted primarily of two groups of acidic proteins located in the 8 to 14 kd and 22 to 24 kd areas of the 2-DE map. Major IgE-reactivity was detected with C-serum proteins in the 56, 45, 30, 20, 14, and <6.5 kd areas of the immunoblots. The 8 to 14 kd particle proteins exhibited distinct IgE reactivity, whereas the 22 to 24 kd proteins were not stained. Seven of the soluble IgE-reactive protein spots showed high homology with enolase, superoxide dismutase, triosephosphate isomerase, proteasome subunit, and chitinase and represent previously undescribed latex allergens; whereas nine protein spots corresponded to known latex allergens, namely prohevein, hevein, prohevein C-domain, and hevamine. As identified by monoclonal antibodies, the IgE-reactive latex particle proteins mainly represent the allergenic rubber elongation factor. CONCLUSIONS: Two-dimensional electrophoresis, followed by immunoblotting and protein microsequencing, can rapidly identify a large number of IgE-binding latex proteins. The 2-DE latex maps generated will provide valuable information for the development of strategies to isolate the relevant latex allergens. Because the novel latex allergens are common plant enzymes, they may also act as cross-reacting proteins in various foods.
[187] - Lee MF, Hwang GY, Chen YH, Lin HC, Wu CH. Molecular cloning of Indian jujube (Zizyphus mauritiana) allergen Ziz m 1 with sequence similarity to plant class III chitinases. Mol Immunol 2006;43:1144-1151
Indian jujube (Zizyphus mauritiana) is a sweet fruit that is abundantly cultivated in Taiwan. We have previously identified 42 and 30kDa allergens that are cross-reactive with latex allergen from crude Indian jujube extract. This study aimed to clone the 30kDa Ziz m 1 Z. mauritiana allergen. The Ziz m 1 encoding cDNA was isolated from a ZAPII cDNA library constructed from Z. mauritiana mRNA, sequenced and expressed in Pichia pastoris. The protein predicted from the cDNA sequence has 330 amino acids, the first 25 of which constituted a putative signal peptide. The deduced molecular mass of the mature protein is 33.86kDa, while its isoelectric point is estimated at 4.36. The recombinant Ziz m 1 showed chitinase activity, possessed IgE binding capacity, and had IgE cross-reactivity with the latex allergen. Moreover, anti-recombinant Ziz m 1 antibody-based ELISA was able to detect commercial skin testing latex reagent, laboratory prepared latex and Indian jujube extracts. Recombinant Ziz m 1 showed 87.5% skin reactivities on eight latex- and Indian jujube-sensitive subjects. Although no sequence similarity was found to other known allergens, Ziz m 1 was found to have amino acid sequence identity (39-45.3%) to many plant chitinases including chitinase (45.2%) of Hevea brasiliensis (hevamine), class III chitinases of Vigna angularis (45.3%), Capsicum annuum (44.7%) and Oryza sativa (41.2%). A conserved domain search revealed that Ziz m 1 belongs to the family 18 glycosyl hydrolases. The recombinant allergen may therefore be of value for diagnosis and therapeutic purposes, and the further characterization of Indian jujube allergen may help to elucidate the mechanism underlying latex-fruit syndrome.
[188] - Ganglberger E, Radauer C, Wagner S, O'Riordain G, Beezhold DH, Brehler R, et al. Hev b 8, the Hevea brasiliensis latex profilin, is a cross-reactive allergen of latex, plant foods and pollen. Int Arch Allergy Immunol 2001;125:216-227
BACKGROUND: Plant profilins are important pan-allergens. They are responsible for a significant percentage of pollen-related allergies. Limited information is available about their involvement in the latex-fruit syndrome and the cross-reactivities between latex and pollen. We aimed to clone and express the Hevea brasiliensis latex profilin to investigate its allergological significance and serological cross-reactivities to profilins from plant foods and pollens . METHODS: A DNA complementary to messenger RNA (cDNA) coding for the Hevea latex profilin, Hev b 8, was amplified by polymerase chain reaction from latex RNA. Recombinant (r)Hev b 8 was produced in Escherichia coli and used to screen sera from 50 latex- allergic health care workers (HCWs) with well-documented histories of food and pollen allergy and 34 latex-allergic spina bifida (SB) patients. The cross-reactivity of natural Hev b 8 and rHev b 8 with other plant profilins was determined by ELISA inhibition assays. A three-dimensional homology model of Hev b 8 was constructed based on known profilin structures . RESULTS: The cDNA of Hev b 8 encoded a protein of 131 amino acids with a predicted molecular mass of 14 kD. Twelve of the 50 HCWs and 2 of the 34 SB patients were sensitized to Hev b 8. All Hev b 8-sensitized patients showed allergic symptoms to pollen or plant foods. Cross-reactivities between profilins of latex, pollen and plant food were illustrated by their ability to inhibit IgE binding to rHev b 8. Homology modeling of Hev b 8 yielded a structure highly similar to Bet v 2, the birch pollen profilin, with the most distinct differences located at the N-terminus . CONCLUSIONS: We conclude that primary sensitization to latex profilin in the majority of cases takes place via pollen or food profilins. Additionally, pollinosis and food-allergic patients with profilin-specific IgE can be at risk of developing latex allergy
[189] - Beezhold DH, Hickey VL, Kostyal DA, Puhl H, Zuidmeer L, van Ree R, et al. Lipid transfer protein from Hevea brasiliensis (Hev b 12), a cross-reactive latex protein. Ann Allergy Asthma Immunol 2003;90:439-445
BACKGROUND: Latex-allergic individuals experience clinical cross-reactivity to a large number of fruits and vegetables. Much of the cross-reactivity can be attributed to Hev b 6, but evidence indicates that additional cross-reactive allergens may be present. A common pan-allergen, which has not previously been identified in latex, but may contribute to this cross-reactivity is lipid transfer protein (LTP). We sought to determine whether Hevea brasiliensis produces LTP and whether it would bind immunoglobulin E from latex-allergic patients. METHODS: LTP was identified in H. brasiliensis RNA by polymerase chain reaction using degenerate primers. The entire cDNA was obtained by polymerase chain reaction using rapid amplification of cDNA ends reactions. The complete coding sequence for LTP was determined and produced as a recombinant protein using the glutathione S-transferase and pET32 expression systems. Immunoblot analysis of sera from latex-allergic patients was used to determine whether patients recognize LTP as an allergen. RESULTS: We identified a 662-basepair cDNA with a 351-basepair open reading frame that encodes for a 116-amino acid protein. The protein has significant homology to the family of nonspecific LTPs. We expressed the protein as a mature LTP of 92 amino acids with a predicted isoelectric point of 10.8 and molecular weight of 9.3 kDa. Immunoblots demonstrated specific immunoglobulin E for LTP in the sera of 9 of 37 (24%) latex-allergic individuals. CONCLUSIONS: We describe the initial identification of rLTP in H. brasiliensis that may be important as a cross-reactive pan-allergen (Hev b 12).
[190] - Rihs HP, Rueff F, Lundberg M, Rozynek P, Garcia-Selles J, Barber D, et al. Specific IgE-Binding of rHev b 12 Is Restricted to Fruit-Allergic Patients. AAAAI 60th Annual Meeting, San Francisco, 19-23 March 2004, Poster n°205
Rationale Lipid transfer proteins (LTP) have been identified as relevant allergens. Their role in latex allergy is unclear. This study aimed to produce a recombinant latex-LTP (rHev b 12) to examine its IgE-binding properties. Methods A recombinant generated Maltose-Binding-Protein (MBP)-rHev b 12 fusion protein was coupled to ImmunoCAP for the determination of specific IgE. 48 sera of atopic patients (14 from Spain, 34 from Germany) with fruit-allergy were examined. 25 of these sera displayed positive latex-specific IgE values. Results rHev b 12-specific IgE was measured in four Spanish peach-allergic patients (CAP-values: 0.88-2.27 kU/L), whereby one had an additional latex-sensitization. Two sera from cherry-allergic German patients with latex-sensitization displayed also rHev b 12-specific IgE antibodies (0.68 and 0.96 kU/L). All other sera and the MBP controls revealed negative results (<0.35 kU/L). Conclusions Latex-LTP (rHev b 12)-specific IgE-binding in peach- and cherry-allergic patients seem to be a result of partially common IgE-binding epitopes with the LTPs of peach (homology to Pru p 3: 65%) and cherry (homology to Pru av 3: 61%). Although rHev b 12 seems to have minor relevance as a latex allergen in Central Europe, its significance as a cross-reactive allergen in Mediterranean countries like Spain has to be kept in mind.
[191] - Rihs HP, Rueff F, Lundberg M, Rozynek P, Barber D, Scheurer S, et al. Relevance of the recombinant lipid transfer protein of Hevea brasiliensis: IgE-binding reactivity in fruit-allergic adults. Ann Allergy Asthma Immunol 2006;97:643-649
BACKGROUND: Lipid transfer proteins (LTPs) are relevant allergens in certain plants. The role of the LTP of Hevea brasiliensis in the latex-fruit syndrome is widely unknown. OBJECTIVE: To study IgE reactivity with recombinant Hevea LTP in sera of fruit-allergic adults with and without natural rubber latex (NRL) allergy. METHODS: An LTP-specific complementary DNA of H brasiliensis leaves was amplified, subcloned into the pMAL expression system, and analyzed. The recombinant protein was coupled to ImmunoCAP, and the IgE-binding properties were studied in sera of 10 NRL-allergic patients without symptoms to fruit and 48 atopic patients with fruit allergy. Eleven of these 48 patients were also allergic to NRL, 14 displayed sensitization to NRL without symptoms on NRL exposure so far, and 23 had neither symptoms nor IgE antibodies to NRL. RESULTS: After expression in Escherichia coli, a soluble maltose-binding protein-rHev b 12 fusion protein was isolated and coupled to ImmunoCAP to determine rHev b 12 specific IgE reactivity. rHev b 12 specific IgE binding was found in 3 fruit-allergic patients with NRL sensitization (0.68, 0.88, and 0.96 kU/L) and in 3 fruit-allergic patients without NRL sensitization (1.58, 2.25, and 2.27 kU/L). The remaining 52 serum samples and all maltose-binding protein control test results were negative (< 0.35 kU/L). CONCLUSIONS: In these patients, rHev b 12 specific IgE reactivity seems to result from common cross-reactive epitopes with some of the fruit LTPs tested and underscores only an involvement in co-recognition. No clinical relevance of IgE binding to the LTP of H brasiliensis in association with NRL allergy was detected.
[192] - Majamaa H, Seppälä U, Palosuo T, Turjanmaa K, Kalkkinen N, Reunala T. Positive skin and oral challenge responses to potato and occurrence of IgE antibodies to patatin (Sol t 1) in infants with atopic dermatitis. Pediatr Allergy Immunol 2001;12:283-288
The clinical significance and molecular specificity of hypersensitivity reactions to raw and cooked potatoes remain ambiguous. We therefore investigated the clinical hypersensitivity to raw and cooked potato in infants suspected to have potato allergy and compared the findings with the occurrence of immunoglobulin E (IgE) antibodies to patatin (Sol t 1), characterized as the primary allergen of potato. Twelve infants (10 to 24 months of age) suffering from atopic dermatitis (AD) and suspected to have adverse reactions to potato, were examined. As a skin exposure test we used rubbing with both raw and cooked potato, and used open oral challenge with cooked potato for 7 days. A special eczema scoring system (SCORAD) was used to assess the severity of symptoms and signs of AD. Skin-prick tests (SPTs) were performed with raw potato and natural Sol t 1, and serological studies included measurement of total serum IgE and IgE antibodies to Sol t 1, and potato radioallergosorbent testing (RAST). The skin-rubbing test with raw potato was positive in seven (58%) and the oral challenge positive in eight (67%) infants. One infant presented with an immediate reaction and seven with a delayed reaction, i.e. exacerbation of AD, after oral challenge responses to cooked potato. Nine (75%) infants had IgE antibodies to Sol t 1 in enzyme-linked immunosorbent assay (ELISA), and SPT to natural Sol t 1 was positive in six (50%) potato-allergic infants. In conclusion, we observed positive challenge responses to both raw and cooked potato in food-allergic atopic infants. The presence of IgE antibodies and concomitant positive SPTs to the heat-stable potato allergen, Sol t 1, suggest that cooked potato can be an allergenic food for infants suffering from AD
[193] - Majamaa H, Seppälä U, Palosuo T, Reunala T, Kalkkinen N, Turjanmaa K. Hypersensitivity to Raw Potatoes in Adults. AAAAI 58th Annual Meeting, New York, 1-6 March, 2002, Poster n°659
Previous studies have shown that hypersensitivity reaction between raw and cooked potatoes (Solanum tuberosum) are substantially different. Young infants and children suffering from food allergies may develop allergy to cooked potatoes showing specific IgE to heat-stabile potato allergens, Sola t 1-4. In general, however, hypersensitivity reactions to raw potatoes are associated with atopic adults frequently showing positive skin prick test (SPT) to birch (Betula verrucosa) pollen. Immunological basis for this phenomenon has considered to result from IgE-mediated cross-reactivity between heat-labile pollen and potato allergens. Allergic reactions to raw potatoes are usually immediate, appearing within minutes after peeling and/or handling of them. The clinical spectrum includes symptoms ranging from contact urticaria and itching of hands to non-cutaneous symptoms such as rhinitis and wheezing. In the present study 9 adults (ranging from 15 to 46 years) showing clinical history of hypersensitivity against raw potatoes were examined. Total and potato specific IgE was measured by RAST (Pharmacia, Sweden) and all the patients were SPT with purified potato allergens, Sola t 1-4, raw potato, birch pollen (ALK-Abello, Denmark), and natural rubber latex (NRL) (Stallergénes SPT reagent, France). Skin prick test to raw potato and birch pollen was positive in all the 9 patients but only 1 patient showed positive response to NRL reagent. SPT with purified potato allergen Sola t 1 was positive in 2/9 patients whereas SPT with Sola t 2-4 remained negative. Interestigly, potato RAST was positive (0.7-3.9 kU/L) in 5/9 and only 6 patients had increased total IgE levels (217-10772 kU/L). The present study with adult patients showed that clinical reactivity in handling of raw potatoes correlates well to positive SPT to raw potato and birch pollen. However, SPT results with purified heat-stabile food allergens of potato suggest that Sola t 1-4 do not play a major role in allergic reactions to raw potato. In conclusion, the heat-labile potato allergen(s), allegedly causing the clinical cross-reactivity for potato and birch pollen allergic patients, remain(s) to be identified.
[194] - Wagner S, Wagner B, Niggemann B, Scheiner O, Breiteneder H. Sensitisation of mould allergic patients to latex allergens indicate a latex-mould syndrome. EAACI 22th Congress, Paris, 7-11 June, 2003, Poster n°113
Latex allergy is a well-defined medical problem within recognised risk groups such as health-care workers, children with spina bifida, or individuals with allergies to fruits. Cross-reactive allergens of Hevea latex and fruits are responsible for the high incidence of fruit allergy in subjects with latex allergy, resulting in the latex-fruit syndrome. Recently we have characterised the Hevea latex allergens Hev b 9 and Hev b 10 and demonstrated cross-reactivity of these allergens to homologous proteins from Cladosporium herbarum and Aspergillus fumigatus. Due to these findings we aimed to investigate whether mould allergic subjects show a higher incidence of sensitisation to the latex allergens Hev b 9 and Hev b 10 possibly resulting in a latex-mould syndrome. Therefore, sera of 139 subjects with positive clinical history and positive RAST to moulds were evaluated in ELISA experiments on recombinant (r)Hev b 9 and rHev b 10. Thirty two individuals (23%) displayed IgE antibodies against rHev b 9 and 17 individuals (12%) against rHev b 10. IgE reactivity to both allergens was observed for sera of 13 subjects, while sera of 19 subjects recognized only rHev b 9, and sera of 4 subjects only rHev b 10. In total, 36/139 (26%) subjects were found to be sensitised to Hev b 9 and/or Hev b 10. From our data we conclude that IgE reactivity to mould allergens could be a risk factor for sensitisation to Hevea latex. Subjects suffering from mould allergy represent a potential novel risk group for developing latex allergy.
[195] - Wagner S, Mari A, Simon-Nobbe B, Wagner B, Niggemann B, Breitenbach M, et al. Hev b 9, the Latex Enolase, a Necessary Reagent of a Mold Allergy Diagnostic Panel ? AAAAI 60th Annual Meeting, San Francisco, 19-23 March 2004, Poster n°1096
Rationale Enolases are important mold allergens. We investigated mold allergic patients' sensitization to the allergenic latex enolase Hev b 9 and its cross-reactivity with the allergenic mold enolase Alt a 5. Method s : Sera of 167 subjects with allergy to molds, 57 subjects with allergy to latex, and 19 patients with allergy to molds and latex˜characterized by positive clinical history and positive CAP˜were evaluated in ELISA experiments on recombinant (r)Hev b 9 or rAlt a 5, the enolase from Alternaria alternata. Result s : In the group of mold allergic subjects 45/167 displayed IgE to rAlt a 5, 35/167 to rHev b 9, and 16/167 to both allergens. Sera of 4/57 latex allergic patients contained IgE specific for rAlt a 5, 3/57 for rHev b 9, and 2/57 for both allergens. Sera of 2/19 of the mold and latex allergic patients displayed IgE to rAlt a 5 or rHev b 9, one recognizing both allergens. In inhibition studies with the 19 sera displaying IgE to rHev b 9 and rAlt a 5, IgE binding to both allergens with the homologous protein could be weakly inhibited for only one patient. Conclusions : Alt a 5 and Hev b 9 showed only very weak cross-reactivity. Nevertheless, the high number of mold allergic people sensitized to Hev b 9 indicates that cross-reactivity is induced by various enolases of other mold sources. Therefore, Hev b 9 should be part of an allergen panel for diagnosis of mold allergy.
[196] - Wagner S, Breiteneder H, Simon-Nobbe B, Susani M, Krebitz M, Niggemann B, et al. Hev b 9, an enolase and a new cross-reactive allergen from hevea latex and molds. Purification, characterization, cloning and expression. Eur J Biochem 2000;267:7006-7014
Natural rubber latex allergy is an IgE-mediated disease that is caused by proteins that elute from commercial latex products. A complementary DNA (cDNA) coding for Hev b 9, an enolase (2-phospho-D-glycerate hydrolyase) and allergen from latex of the rubber tree Hevea brasiliensis, was amplified by PCR. The PCR primers were designed according to conserved regions of enolases from plants. The obtained cDNA amplification product consisted of 1651 bp and encoded a protein of 445 amino-acid residues with a calculated molecular mass of 47.6 kDa. Sequence comparisons revealed high similarities of the Hevea latex enolase to mold enolases that have been identified as important allergens. In addition, the crucial amino-acid residues that participate in the formation of the catalytic site and the Mg2+ binding site of enolases were also conserved. Hevea latex enolase was produced as a recombinant protein in Escherichia coli with an N-terminal hexahistidyl tag, and purified by affinity chromatography. The yield amounted to 110 mg of purified Hev b 9 per litre of bacterial culture. The recombinant allergen bound IgE from latex, as well as mold-allergic patients, in immunoblot and ELISA experiments. The natural enolase was isolated from Hevea latex by (NH4)2SO4 precipitation and ion exchange chromatography. The natural and the recombinant (r)Hev b 9 showed equivalent enzymatic activity. Patients' IgE-antibodies preincubated with rHev b 9 lost their ability to bind to natural (n) Hev b 9, indicating the identity of the B-cell epitopes on both molecules. Cross-reactivity with two enolases from Cladosporium herbarum and Alternaria alternata was determined by inhibition of IgE-binding to these enolases by rHev b 9. Therefore, enolases may represent another class of highly conserved enzymes with allergenic potentials.
[197] - Sussman GL, Beezhold DH, Kurup VP. Allergens and natural rubber proteins. J Allergy Clin Immunol 2002;110(2 part 2):S33-S39
BACKGROUND: Allergy to natural rubber latex (NRL) results from exposure to proteins derived from Hevea brasiliensis. Type I latex hypersensitivity is observed in certain occupational and other high-risk groups with frequent exposure to NRL products. This includes health care workers (HCWs), workers in the latex industry, children with spina bifida, and atopic individuals . OBJECTIVES: Early reliable diagnosis and avoidance are required for better patient care. Standardized reagents are not presently available for in vitro and in vivo testing and treatment of patients with latex allergy. However, a number of allergens have been isolated and characterized from Hevea latex and NRL products. Currently, a total of 11 major and minor allergens are designated by the International Allergen Nomenclature Committee. This article reviews the structural and functional characteristics of these latex allergenic proteins . RESULTS: NRL-allergenic proteins include those involved in the biosynthesis of polyisoprene and coagulation of latex rubber elongation factor, small rubber particle protein, prohevein, and patatin. Pathogenesis-related proteins include beta-1,3-glucanases, chitinases, and hevamine; and the structural proteins include microhelix protein complex, proline-rich protein, profilins, enolases, and manganese superoxide dismutase. Recombinant allergens demonstrated skin test reactivity in patients with latex allergy. The minimal level of skin test reactivity was about 70 pg/mL for NRL and 1 ng/mL for recombinant allergens. The use of selected recombinant latex allergens (Hev b 5, Hev b 6, and Hev b 7) in skin prick tests identified 93% of allergic individuals, mainly health care workers . CONCLUSIONS: Recombinant latex allergens are clinically reactive and can be produced in a standardized manner, which could potentially provide safe and sensitive reagents for the diagnosis and treatment of type I latex allergy.
[198] - Randall A, Hillier A, Cole LK, Kwochka KW, Needham G, Wassom DL. Quantitation of house dust mites and house dust mite allergens in the microenvironment of dogs. Am J Vet Res 2003;64:1580-1588
OBJECTIVE: To quantitate the density of Dermatophagoides farinae and D pteronyssinus and concentrations of house dust mite (HDM) allergens (Der f 1, Der p 1, and Group 2 allergens) in the indoor microenvironment of dogs. SAMPLE POPULATION: 50 homes in Columbus, Ohio. PROCEDURES: In each home, samples of dust were collected from 3 locations in which dogs spent most time. Whenever possible, the species of mites collected was identified. Mite density (mites/g of dust) was assessed, and allergen concentrations were assayed by standardized ELISAs. Relative humidity and temperature in each home were monitored during a 5-day period. Characteristics of homes and sample sources were evaluated. RESULTS: Dust samples from all 50 homes contained > or = 1 HDM allergen; Der f 1 and Der p 1 were detected in 100 and 74% of homes, respectively. Fifteen homes had HDMs; compared with D pteronyssinus, D farinae was found more commonly (14/15 homes) and at a higher density. Basements, homes without central air-conditioning, and dog beds that were > or = 1 year old had high HDM allergen concentrations. Homes with > or = 2 microg of Der f 1 or Group 2 allergens/g of dust or > or = 100 mites/g of dust were significantly more likely to have a maximum relative humidity > or = 75%. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated the presence of HDMs and HDM allergens in the specific microenvironment of dogs in homes. Factors associated with high levels of exposure were identified, which may be associated with increased risk for sensitization and development of atopic diseases.
[199] - Wagner S, Wagner B, Niggemann B, Scheiner O, Breiteneder H. Sensitisation of mould allergic patients to latex allergens indicate a latex-mould syndrome. EAACI 22th Congress, Paris, 7-11 June, 2003, Poster n°113
Latex allergy is a well-defined medical problem within recognised risk groups such as health-care workers, children with spina bifida, or individuals with allergies to fruits. Cross-reactive allergens of Hevea latex and fruits are responsible for the high incidence of fruit allergy in subjects with latex allergy, resulting in the latex-fruit syndrome. Recently we have characterised the Hevea latex allergens Hev b 9 and Hev b 10 and demonstrated cross-reactivity of these allergens to homologous proteins from Cladosporium herbarum and Aspergillus fumigatus. Due to these findings we aimed to investigate whether mould allergic subjects show a higher incidence of sensitisation to the latex allergens Hev b 9 and Hev b 10 possibly resulting in a latex-mould syndrome. Therefore, sera of 139 subjects with positive clinical history and positive RAST to moulds were evaluated in ELISA experiments on recombinant (r)Hev b 9 and rHev b 10. Thirty two individuals (23%) displayed IgE antibodies against rHev b 9 and 17 individuals (12%) against rHev b 10. IgE reactivity to both allergens was observed for sera of 13 subjects, while sera of 19 subjects recognized only rHev b 9, and sera of 4 subjects only rHev b 10. In total, 36/139 (26%) subjects were found to be sensitised to Hev b 9 and/or Hev b 10. From our data we conclude that IgE reactivity to mould allergens could be a risk factor for sensitisation to Hevea latex. Subjects suffering from mould allergy represent a potential novel risk group for developing latex allergy.
[200] - Wagner S, Sowka S, Mayer C, Crameri R, Focke M, Kurup VP, et al. Identification of a Hevea brasiliensis latex manganese superoxide dismutase (Hev b 10) as a cross-reactive allergen. Int Arch Allergy Immunol 2001;125:120-127
BACKGROUND: Cross-reactive allergens play an increasingly important role in latex allergy in complicating both the diagnosis and time course of allergic symptoms. Manganese superoxide dismutase (MnSOD), a ubiquitous protein of prokaryotic and eukaryotic organisms, was described as a cross-reactive allergen in Aspergillus fumigatus. Little information is available on the importance of this pan-allergen in Hevea brasiliensis latex. The aim of this study was to clone and express MnSOD from H. brasiliensis latex, and to obtain the soluble and immunologically active recombinant allergen for diagnosis of latex allergy and to investigate possible cross-reactivities with the structurally related A. fumigatus and human MnSODs . METHODS: A complementary DNA coding for Hevea latex MnSOD was amplified by PCR. The recombinant protein was produced in Escherichia coli with an N-terminal hexahistidyl tag. Enzymatic activity of the recombinant protein was determined using an enzyme assay for SODs. IgE immunoblotting and IgE inhibition assays were performed to characterize the recombinant allergen and its cross-reactivity . RESULTS: A Hevea latex MnSOD consisting of 206 amino acid residues was cloned and expressed in E. coli. The allergen was designated Hev b 10. The recombinant protein was enzymatically active, indicating the correct folding of the protein. In immunoblots, latex- as well as A. fumigatus-allergic patients revealed IgE binding to recombinant (r)Hev b 10. Cross-reactivity to Asp f 6, the MnSOD from A. fumigatus, and human MnSOD was determined by inhibition of IgE binding to these MnSODs by rHev b 10 . CONCLUSIONS: Hev b 10 is a new cross-reactive allergen of H. brasiliensis which belongs to the 'latex-mold' group of latex allergens. Furthermore, it is a candidate for primary sensitization in patients allergic to the pan-allergen MnSOD.
[201] - Hemery ML, Verdier R, Daban P, Sellier N, Dujols P, Demoly P. Sensibilisation aux gants en latex poudrés: une prévalence élevée chez le personnel hospitalier. Presse Med 2005;34:1363-1369
INTRODUCTION: The prevalence of latex allergies in industrial countries has skyrocketed since the 1980s. Between 2.6 and 22% of hospital workers are diagnosed with latex allergy, which has been recognized as an occupational hazard in France since 1997. AIM: To assess the prevalence of latex allergy among Montpellier University Hospital Center staff . METHODS: From 1999 to 2002, we interviewed and conducted skin-prick tests on 537 hospital staff members from different departments and with different degrees of exposure to airborne latex allergens . RESULTS: Symptoms while using natural-rubber latex gloves (most often contact dermatitis) were reported by 88 (16.4%) staff members. Overall work-related allergic symptoms included rhinitis, reported by 65 (12.1%), contact urticaria by 28 (5.2%), and bronchial symptoms by 16 (1.1%). In all, sensitization to latex was identified in 7.1% of our staff, and this percentage was higher (11.3%) in units where latex gloves were used more often. Sensitization was associated with high latex exposure and atopy . CONCLUSION: This high rate of latex sensitization and the risk factors are similar to those already published. Based on this study, we have eliminated powdered latex gloves, as have many other hospitals.
[202] - Mari A, Scala E, D’Ambrosio C, Breiteneder H, Wagner S. Latex Allergy within a Cohort of Not-at-Risk Subjects with Respiratory Symptoms: Prevalence of Latex Sensitization and Assessment of Diagnostic Tools. Int Arch Allergy Immunol 2007;143:135-143
BACKGROUND: Immunoglobluin E (IgE)-mediated hypersensitivity to natural rubber latex (NRL) is a major problem in allergy practice. Currently, the use of skin prick tests (SPTs) with latex extracts and specific IgE detection for the diagnosis of NRL allergy in suspected patients is directed to identification of risk factors. Many cases of NRL allergy remain undiagnosed due to misreporting of symptoms by the patients or lack of proper questions asked by the physician. MATERIALS AND METHODS: A total of 6,126 subjects referred for respiratory symptoms underwent SPTs with NRL. Positive subjects were resurveyed for exposure to NRL, and specific IgE for NRL extracts and recombinant molecules was determined. Immunoblots of NRL extracts were performed to identify IgE patterns . RESULTS: Forty-six of 3,930 sensitized subjects had a positive SPT with NRL, displaying a prevalence of NRL sensitization of 0.75% for the general and 1.2% for the sensitized population. Eleven out of 46 (23.9%) subjects could be defined as NRL asymptomatic, whereas 35 (76.1%) developed symptoms upon exposure to NRL. Specific IgE to NRL was detected for 22 (75.86%) of 29 tested sera. Seventeen out of 22 (77%) sera displayed specific IgE to recombinant allergens with most reactions to Hev b 5, Hev b 6.01 and Hev b 6.02. Immunoblots of NRL extract fractions with patients' sera showed heterogeneous patterns . CONCLUSIONS: SPTs with NRL extract should be routinely performed in patients with respiratory symptoms. Hev b 5, Hev b 6.01 and Hev b 6.02 are the most important allergens, but further characterization of NRL extracts is needed to identify novel allergens and to clarify the role of crossreactive carbohydrate determinants.
[203] - O'Riordain G, Radauer C, Hoffmann-Sommergruber K, Adhami F, Peterbauer CK, Blanco C, et al. Cloning and molecular characterization of the Hevea brasiliensis allergen Hev b 11, a class I chitinase. Clin Exp Allergy 2002;32:455-462
In the last 10 years type-I allergy against proteins from Hevea brasiliensis latex has become an acknowledged medical issue. Fruit-allergic patients represent one risk group for developing latex allergy. Class I chitinases have been identified from chestnut, avocado and banana as relevant allergens. The chitin binding (hevein) domain from these class I chitinases has been postulated to bear the important IgE binding epitopes. Objective : To clone the cDNA of an allergenic latex class I chitinase, to express the recombinant protein and to determine its IgE cross-reactivity with hevein (Hev b 6.02). Method s : A full-length cDNA coding for a class I chitinase has been isolated from Hevea latex RNA by reverse transcription followed by PCR. The chitinase encoding sequence has been subcloned into the pMAL expression vector and expressed in E. coli as a fusion protein to maltose binding protein. The highly enriched recombinant protein fraction has been tested for its IgE binding capacity in immunoblots and ELISA. Furthermore, the pathogenesis-related function of the recombinant protein was tested in a fungal growth inhibition assay. Result s : The Hevea brasiliensis latex chitinase, designated Hev b 11, displays 70% identity to the endochitinase from avocado and its hevein-domain 58% to hevein (Hev b 6.02). The recombinant Hev b 11-maltose binding protein is recognized by latex- and fruit-allergic patients with IgE binding in both, ELISA and immunoblots. Pre-incubation of sera with rHev b 11-maltose binding protein showed an overall 16% inhibition of subsequent binding to rHev b 6.02-maltose binding protein on solid phase. The growth of F. oxysporum was inhibited in a dose dependent manner by addition of rHev b 11-maltose binding protein to the culture. Conclusions : Hev b 11, a class I chitinase, is another allergen from Hevea latex with a chitin binding domain and displays a different IgE binding capacity compared with hevein.
[204] - Rihs HP, Dumont B, Rozynek P, Lundberg M, Cremer R, Brüning T, et al. Molecular cloning, purification, and IgE-binding of a recombinant class I chitinase from Hevea brasiliensis leaves (rHev b 11.0102). Allergy 2003;58:246-251
BACKGROUND: Class I chitinase in natural rubber latex (NRL) has been assumed to be an important allergen, especially concerning its cross-reactivity with fruits like avocado and banana . OBJECTIVES: The present study aimed to produce a recombinant latex class I chitinase from Hevea brasiliensis leaves and to study its immunoglobulin (Ig)E-binding reactivity . METHODS: A class I chitinase-specific complementary DNA from H. brasiliensis leaves was synthesized, subcloned, sequenced and overexpressed in fusion with the maltose-binding protein (MBP) in Escherichia coli. The IgE-binding reactivity of this protein was studied by the Pharmacia CAP System and by immunoblot experiments using sera from latex-allergic patients . RESULTS: The rHev b 11.0102 was found to have a length of 295 amino acid residues and contains an N-terminal hevein-like domain with a 56% homology to hevein. Analysis by the CAP method revealed the presence of rHev b 11.0102-specific IgE antibodies in 17 of 58 sera (29%) of IgE-mediated latex-allergic subjects tested. Immunoblot analysis of the MBP-rHev b 11.0102 fusion protein and the MBP carrier protein as a negative control confirmed the IgE-reactivity of rHev b 11.0102 . CONCLUSION: Due to its IgE-reactivity rHev b 11.0102 represents an allergen of intermediate prevalence in NRL. Its property to cross-react with certain fruits makes it an important supplement in the diagnostic panel of recombinant NRL allergens.
[205] - O'Riordain G, Radauer C, Hoffmann-Sommergruber K, Adhami F, Peterbauer CK, Blanco C, et al. Cloning and molecular characterization of the Hevea brasiliensis allergen Hev b 11, a class I chitinase. Clin Exp Allergy 2002;32:455-462
In the last 10 years type-I allergy against proteins from Hevea brasiliensis latex has become an acknowledged medical issue. Fruit-allergic patients represent one risk group for developing latex allergy. Class I chitinases have been identified from chestnut, avocado and banana as relevant allergens. The chitin binding (hevein) domain from these class I chitinases has been postulated to bear the important IgE binding epitopes. Objective : To clone the cDNA of an allergenic latex class I chitinase, to express the recombinant protein and to determine its IgE cross-reactivity with hevein (Hev b 6.02). Method s : A full-length cDNA coding for a class I chitinase has been isolated from Hevea latex RNA by reverse transcription followed by PCR. The chitinase encoding sequence has been subcloned into the pMAL expression vector and expressed in E. coli as a fusion protein to maltose binding protein. The highly enriched recombinant protein fraction has been tested for its IgE binding capacity in immunoblots and ELISA. Furthermore, the pathogenesis-related function of the recombinant protein was tested in a fungal growth inhibition assay. Result s : The Hevea brasiliensis latex chitinase, designated Hev b 11, displays 70% identity to the endochitinase from avocado and its hevein-domain 58% to hevein (Hev b 6.02). The recombinant Hev b 11-maltose binding protein is recognized by latex- and fruit-allergic patients with IgE binding in both, ELISA and immunoblots. Pre-incubation of sera with rHev b 11-maltose binding protein showed an overall 16% inhibition of subsequent binding to rHev b 6.02-maltose binding protein on solid phase. The growth of F. oxysporum was inhibited in a dose dependent manner by addition of rHev b 11-maltose binding protein to the culture. Conclusions : Hev b 11, a class I chitinase, is another allergen from Hevea latex with a chitin binding domain and displays a different IgE binding capacity compared with hevein.
[206] - Raulf-Heimsoth M, Rihs HP, Rozynek P, Cremer R, Gaspar A, Pires G, et al. Quantitative analysis of immunoglobulin E reactivity profiles in patients allergic or sensitized to natural rubber latex (Hevea brasiliensis). Clin Exp Allergy 2007;37:1657-1667
BACKGROUND: Characterized native and recombinant Hevea brasiliensis (rHev b) natural rubber latex (NRL) allergens are available to assess patient allergen sensitization profiles . OBJECTIVE: Quantification of individual IgE responses to the spectrum of documented NRL allergens and evaluation of cross-reactive carbohydrate determinants (CCDs) for more definitive diagnosis . METHODS: Sera of 104 healthcare workers (HCW; 51 German, 21 Portuguese, 32 American), 31 spina bifida patients (SB; 11 German, 20 Portuguese) and 10 Portuguese with multiple surgeries (MS) were analysed for allergen-specific IgE antibody (sIgE) to NRL, single Hev b allergens and CCDs with ImmunoCAP technology . RESULTS: In all patient groups rHev b 5-sIgE concentrations were the most pronounced. Hev b 2, 5, 6.01 and 13 were identified as the major allergens in HCW and combined with Hev b 1 and Hev b 3 in SB. In MS Hev b 1 displayed an intermediate relevance. Different sIgE antibody levels to native Hevea brasiliensis (nHev b) 2 and rHev b 6.01 allowed discrimination of SB with clinical relevant latex allergy vs. those with latex sensitization. Sensitization profiles of German, Portuguese and American patients were equivalent. rHev b 5, 6.01 and nHev b 13 combined detected 100% of the latex-allergic HCW and 80.1% of the SB. Only 8.3% of the sera showed sIgE response to CCDs . CONCLUSIONS: Hev b 1, 2, 5, 6.01 and 13 were identified as the major Hev b allergens and they should be present in standardized latex extracts and in vitro allergosorbents. CCDs are only of minor relevance in patients with clinical relevant latex allergy. Component-resolved diagnostic analyses for latex allergy set the stage for an allergen-directed immunotherapy strategy.
[208] - Rihs HP, Dumont B, Rozynek P, Lundberg M, Cremer R, Brüning T, et al. Molecular cloning, purification, and IgE-binding of a recombinant class I chitinase from Hevea brasiliensis leaves (rHev b 11.0102). Allergy 2003;58:246-251
BACKGROUND: Class I chitinase in natural rubber latex (NRL) has been assumed to be an important allergen, especially concerning its cross-reactivity with fruits like avocado and banana . OBJECTIVES: The present study aimed to produce a recombinant latex class I chitinase from Hevea brasiliensis leaves and to study its immunoglobulin (Ig)E-binding reactivity . METHODS: A class I chitinase-specific complementary DNA from H. brasiliensis leaves was synthesized, subcloned, sequenced and overexpressed in fusion with the maltose-binding protein (MBP) in Escherichia coli. The IgE-binding reactivity of this protein was studied by the Pharmacia CAP System and by immunoblot experiments using sera from latex-allergic patients . RESULTS: The rHev b 11.0102 was found to have a length of 295 amino acid residues and contains an N-terminal hevein-like domain with a 56% homology to hevein. Analysis by the CAP method revealed the presence of rHev b 11.0102-specific IgE antibodies in 17 of 58 sera (29%) of IgE-mediated latex-allergic subjects tested. Immunoblot analysis of the MBP-rHev b 11.0102 fusion protein and the MBP carrier protein as a negative control confirmed the IgE-reactivity of rHev b 11.0102 . CONCLUSION: Due to its IgE-reactivity rHev b 11.0102 represents an allergen of intermediate prevalence in NRL. Its property to cross-react with certain fruits makes it an important supplement in the diagnostic panel of recombinant NRL allergens.
[209] - Raulf-Heimsoth M, Rihs HP, Rozynek P, Cremer R, Gaspar A, Pires G, et al. Quantitative analysis of immunoglobulin E reactivity profiles in patients allergic or sensitized to natural rubber latex (Hevea brasiliensis). Clin Exp Allergy 2007;37:1657-1667
BACKGROUND: Characterized native and recombinant Hevea brasiliensis (rHev b) natural rubber latex (NRL) allergens are available to assess patient allergen sensitization profiles . OBJECTIVE: Quantification of individual IgE responses to the spectrum of documented NRL allergens and evaluation of cross-reactive carbohydrate determinants (CCDs) for more definitive diagnosis . METHODS: Sera of 104 healthcare workers (HCW; 51 German, 21 Portuguese, 32 American), 31 spina bifida patients (SB; 11 German, 20 Portuguese) and 10 Portuguese with multiple surgeries (MS) were analysed for allergen-specific IgE antibody (sIgE) to NRL, single Hev b allergens and CCDs with ImmunoCAP technology . RESULTS: In all patient groups rHev b 5-sIgE concentrations were the most pronounced. Hev b 2, 5, 6.01 and 13 were identified as the major allergens in HCW and combined with Hev b 1 and Hev b 3 in SB. In MS Hev b 1 displayed an intermediate relevance. Different sIgE antibody levels to native Hevea brasiliensis (nHev b) 2 and rHev b 6.01 allowed discrimination of SB with clinical relevant latex allergy vs. those with latex sensitization. Sensitization profiles of German, Portuguese and American patients were equivalent. rHev b 5, 6.01 and nHev b 13 combined detected 100% of the latex-allergic HCW and 80.1% of the SB. Only 8.3% of the sera showed sIgE response to CCDs . CONCLUSIONS: Hev b 1, 2, 5, 6.01 and 13 were identified as the major Hev b allergens and they should be present in standardized latex extracts and in vitro allergosorbents. CCDs are only of minor relevance in patients with clinical relevant latex allergy. Component-resolved diagnostic analyses for latex allergy set the stage for an allergen-directed immunotherapy strategy.
[211] - Cremer R, Hoppe A, Kleine-Diepenbruck U, Blaker F. Longitudinal study on latex sensitization in children with spina bifida. Pediatr Allergy Immunol 1998;9:40-43
The course of latex sensitization is rarely documented, and only a few reports about the influence of prophylaxis in the occupational environment have been published concerning health care workers. Therefore we did a follow-up study in the high risk group of patients with spina bifida and evaluated the efficacy of latex prophylaxis. For this purpose we measured IgE antibodies (FEIA) against latex and inhalative allergens in the sera of 67 patients with spina bifida and reevaluated them 0.6 to 4.1 years later, having instructed the patients about prophylactic measures and established a latex-free environment for surgery of all spina bifida patients in our hospital. 37% of the patients did not develop latex antibodies during the follow-up period, 27% showed decreasing levels of antibodies (12% to non-detectable levels), 19% had an increase in latex sensitization (6% newly sensitized), and 9% showed no change in levels of latex antibodies. From our data it may be concluded that surgery without strict latex prophylaxis is the main cause of new sensitization and worsening of preexisting latex antibody levels. Mild sensitization can be reduced by prophylactic measurements to non-detectable antibody levels. With consistent prophylaxis, even symptomatic patients can be operated without risk of allergic complications or increasing antibodies.
[212] - Filon FL, Radman G. Latex allergy: a follow up study of 1040 healthcare workers. Occup Environ Med 2006;63:121-125
BACKGROUND: Natural rubber latex allergy can cause skin and respiratory symptoms The aim of this study was to evaluate the prevalence and incidence of latex related symptoms and sensitisation among a large group of healthcare workers in Trieste hospitals, followed for three years before and after the introduction of powder-free gloves with low latex release. METHODS: In the years 1997-99 the authors evaluated 1040 healthcare workers exposed to latex allergen for latex related symptoms and sensitisation by means of a questionnaire, a medical examination, skin prick tests, and IgE specific antibody assay. The second evaluation was carried out in the years 2000-02, subsequent to the changeover to a powder-free environment. RESULTS: Glove related symptoms were seen in 21.8% of the nurses (227), mostly consisting of mild dermatitis: 38 (3.6%) complaining of contact urticaria and 24 (2.3%) of asthma and/or rhinitis. These symptoms were significantly related to skin prick tests positive to latex (OR = 9.70; 95% CI 5.5 to 17) and to personal atopy (OR = 2.29; 95% CI 1.6 to 3.2). Follow up was completed in 960 subjects (92.3%): 19 new subjects (2.4%) complained of itching erythema when using gloves, but none was prick positive to latex. Symptoms significantly improved and in most cases disappeared (p<0.0001). CONCLUSIONS: Simple measures such as the avoidance of unnecessary glove use, the use of non-powdered latex gloves by all workers, and use of non-latex gloves by sensitised subjects can stop the progression of latex symptoms and can avoid new cases of sensitisation.
[214] - Ruëff F, Schöpf P, Putz K, Przybilla B. Effect of reduced exposure on natural rubber latex sensitization in health care workers. Ann Allergy Asthma Immunol 2004;92:530-537
BACKGROUND: Knowledge about the long-term course of allergy or sensitization to natural rubber latex (NRL) is insufficient . OBJECTIVE: To investigate the long-term effect of preventive measures on sensitization variables in health care workers who had been diagnosed as having NRL allergy (NRLA) or NRL sensitization (NRLS) without clinical symptoms . METHODS: Repeated follow-up investigations, skin prick tests, and NRL specific IgE serum antibodies were performed in 88 health care workers-33 with NRLA and 55 with NRLS. All workers had been instructed to avoid NRL exposure. At the workplace, powder-free NRL gloves for all other employees were gradually introduced. Re-evaluations were done at 14 +/- 3.7 (N = 86) and 38 +/- 4.0 (N = 78) months after the first examination . RESULTS: At the last follow-up, a loss of skin prick test reactivity to NRL was observed in 1 of 29 subjects with NRLA (3.4%) and 16 of 35 with NRLS (45.7%) with previous skin test reactions (P < .001). Among those subjects who demonstrated a kU/L level (CAP class) equal to or greater than class I to NRL at the initial examination, NRL-specific IgE was absent at the last follow-up in 8 (32.0%) of 25 subjects with NRLA and 14 (38.9%) of 36 with NRLS. At the final examination, we could no longer demonstrate sensitization to NRL by any method in 24 (27.3%) of 88 health care workers. Complete loss of NRL sensitization was less frequent in subjects with NRLA than in those with NRLS (1 of 33 or 3.0% vs 23 of 55 or 41.8%; P < .001) . CONCLUSIONS: Implementation of simple preventive measures lowers markers of sensitization to NRL quickly in many health care workers with NRLA or NRLS.
[215] - Turjanmaa K, Kanto M, Kautiainen H, Reunala T, Palosuo T. Long-term outcome of 160 adult patients with natural rubber latex allergy. J Allergy Clin Immunol 2002;110(2 suppl.):S70-S74
Allergy to natural rubber latex is a major occupational problem in the health care sector and a problem even in other occupations in which protective gloves are used. There is little information available about the long-term outcome at work in large patient populations. To study the occupational outcome when all gloves in the working environment were changed either to low-allergen latex or non-latex gloves, in 1995 to 1996 we re-examined 160 of 174 adult subjects who were diagnosed with natural rubber latex allergy between 1982 and 1994 after a median of 3 years (range, 0.5 to 24 years) after the diagnosis. The outcome in daily work or as a patient in health care was investigated with the use of a questionnaire. Special attention was paid to the assessment of the occurrence of hand eczema. Glove selection policy as an intervention in the treatment of natural rubber latex allergy is described. Of 71 health care workers and 89 non-health care workers, 72% and 83% were atopic; 54% and 65% had hand eczema at the time of diagnosis, and 89% and 19% had work-related allergy to natural rubber latex, respectively. On re-examination, none of the health care workers had changed work because of natural rubber latex allergy, and only 38% had hand eczema (significant decrease). Ninety-eight percent of the non-health care workers, of which 58% had hand eczema, continued with their previous jobs. The use of low-allergen latex or non-latex gloves throughout the health care sector seems to be an adequate step for health care workers who have natural rubber latex allergy; non-health care workers get along with personal avoidance of latex gloves if they are not working directly with natural rubber latex-containing materials in production.
[216] - Nettis E, Colanardi MC, Ferrannini A. Type I latex allergy in health care workers with latex-induced contact urticaria syndrome: a follow-up study. Allergy 2004;59:718-723
BACKGROUND: Latex allergy is an important occupational health issue among health care workers (HCWs). Secondary prevention in sensitized/allergic individuals involves avoiding exposure to NRL products. AIMS OF THE STUDY: The aim of this follow-up study is to determine the long-term health consequences in HCWs with type I latex allergy with latex-related contact urticaria syndrome, of providing appropriate information and practical avoidance education . METHODS: Seventeen HCWs with latex-induced contact urticaria syndrome, as ascertained by the glove use test, were investigated. Initial and follow-up visits included: a detailed questionnaire, skin prick test (SPT) with glove eluates and with commercial latex extract, SPT with common inhalant and food extracts, serum specific immunoglobulin (Ig)E to latex and some foods and the glove use test . RESULTS: On re-examination, 11 (64.7%) subjects showed positive SPTs to extemporaneous extract and 10 (58.8%) patients showed a positive SPT to commercial extract. Of the nine patients with detectable levels of serum latex specific IgE at first evaluation, four (44.4%) became negative and four were assigned to at least one class lower. Only one (11.1%) employee had higher radioallergosorbent test values than those at the latex allergy diagnosis. At follow-up, the 17 individuals had positive latex challenge results, although the duration of exposure causing a reaction increased . CONCLUSION: Our study shows that preventive measures are sufficient to induce a reduction of sensitization. Continued avoidance is needed to prevent re-sensitization or adverse reactions on re-exposure.
[217] - Smith AM, Amin HS, Biagini RE, Hamilton RG, Arif SAM, Yeang HY, et al. Percutaneous reactivity to natural rubber latex proteins persists in health-care workers following avoidance of natural rubber latex. Clin Exp Allergy 2007;37:1349-1356
BACKGROUND: Long-term avoidance of natural rubber latex [Hevea brasiliensis (Hev b)] is currently recommended for health-care workers (HCWs) with established natural rubber latex (NRL) allergy. Percutaneous sensitivity to eight Hev b NRL allergens was evaluated in HCWs in 2000. To date, no studies have evaluated the longitudinal effects of NRL avoidance on percutaneous sensitivity to NRL allergens . OBJECTIVE: The aims of this study were to evaluate changes in percutaneous reactivity to non-ammoniated latex (NAL) and NRL allergens in HCWs 5 years after a recommendation to avoid NRL and to evaluate factors that predict the persistence of in vivo sensitivity to NAL and NRL allergens . METHODS: Skin prick testing was performed with NAL, seven NRL allergens (Hev b 1, 2, 3, 4, 6.01, 7.01, and 13), and recombinant Hev b 5 (rHev b 5) in 34 HCWs who were initially evaluated in 2000 for occupationally related NRL allergy. Serial 10-fold dilutions of NAL and NRL allergens were employed in skin testing. Sera from the HCWs were assayed for latex and enhanced latex (rHev b 5-enriched allergosorbent)-specific IgE antibodies using the ImmunoCAP assay . RESULTS: The prevalence of work-related symptoms significantly decreased between 2000 and 2005 with avoidance of NRL (P<0.05). A >/=100-fold reduction in percutaneous sensitivity to Hev b 2 and Hev b 7 was less likely in those with prior history of systemic reactions to NRL (P=0.0053), reported history of reaction to cross-reactive foods (P=0.014), continued local reactions to NRL gloves (P<0.0001), or high NRL glove exposure since the initial study (P=0.0075). The diagnostic sensitivity and specificity of the latex-specific IgE serology was 54% and 87.5%, respectively, in comparison with NAL skin tests. The addition of rHev b 5 to the ImmunoCAP (enhanced latex) allergosorbent altered the diagnostic sensitivity and specificity of the ImmunoCAP to 77% and 75%, respectively . CONCLUSION: While symptoms may resolve quickly with NRL avoidance therapy, detectable IgE indicating continued sensitization remains beyond 5 years, and thus continued avoidance of NRL should be recommended.
[218] - Selman L, Bienvenu F, Benoit Y, Bouvier M, Robert O, Bérard F, et al. Les allergènes recombinants dans le diagnostic de l’allergie au latex. Rev Fr Allergol Immunol Clin 2007;47:267
Objectifs. ˆ _ optimiser le diagnostic de l‚allergie au latex par l‚utilisation des allerge`nes recombinants du latex (ARL) ; _ cibler les ARL d‚inte´re√t diagnostique. Me´thode.ˆ Population : 98 patients (74 femmes) sensibilise´s au latex (k82 : 0,10 >100 kU/L) re partis en deux groupes : allergie au latex confirme e (82 patients) ; sensibilisation biologique sans allergie clinique au latex (16 patients). De´termination des IgE spe´cifiques pour neuf ARL (rHev b1, b2, b3, b5, b6.01, b6.02, b8, b9, b11) et pour la brome´line (Immunocap 250, Phadia). Re´sultats. ˆ _ Population allergique au latex : rHev b6.01 (72 %), rHev b6.02 (66 %), rHev b2 (63 %), rHev b5 (60 %) sont les plus fre´quemment rencontre´s. Il est note´ une tre`s forte corre lation entre rHev b6.01 et 6.02 de me√me qu‚entre rHev b2 et 6.01/ 6.02. Le profil rHev b2, rHev b6.01 et rHev b6.02 est fortement corre le´ un test cutane´ (TC) positif (43 TC positifs sur 44 tests re´alise´s). rHev b1, rHev b3 et rHev b5 donnent des valeurs de kU/L plus leve´es chez les patients ayant pre´sente´ un choc au latex. Sur quatre patients avec une clinique vocatrice et un rHev b5 positif isole´, deux ont un TC ne´gatif ; _ Population sensibilise´e (k82 : 0,63 36 kU/L) mais non allergique : 14/16 patients ont un rHev b8 isole´ (1,68 >100 kU/L) et sont polliniques. Il faut noter que, parmi les patients allergiques non polliniques, rHev b8 n‚est jamais retrouve´. Deux patients, ne´gatifs pour tous les ARL et non polliniques, ont des IgE anti carbohydrates de´tecte´es par la brome´line. Conclusion. ˆ Les trois ARL, rHev b6.01, rHev b5 et rHev b8 apparaissent particulie` rement inte ressants : _ rHev b6.01 et rHev b5 pour confirmer le diagnostic d‚allergie au latex dans la majorite´ des cas ; _ rHev b8 pour comprendre les re´actions croise´es (k82 faussement positif) chez certains patients polliniques.
[219] - Sastre J, Raulf-Heimsoth M, Rihs HP, Fernandez-Nieto M, Barber D, Lombardero M, et al. IgE reactivity to latex allergens among sensitized healthcare workers before and after immunotherapy with latex. Allergy 2006;61:206-210
BACKGROUND: New IgE sensitizations to proteins in allergen extracts have been shown to occur during allergen-specific immunotherapy (IT) . METHODS: Twenty-four healthcare workers (HCWs) -- patients included in a latex IT study -- were analysed, 16 in active treatment and eight in placebo. Sera were obtained at baseline and after 6 months of IT and analysed with immunoblotting and CAP System with eight single recombinant latex allergens (rHev b 1, 3, 5, 6.01, 8, 9, 10, 11, and a mix of rHev b1, 5, 6.01 and 8) . RESULTS: After IT with latex, three patients in the active treatment group had new IgE sensitizations, one to Hev b 5, one to Hev b 11 and another to Hev b 6.01. No other significant variation in mean of specific IgE to latex or recombinant allergens were observed in patients who received placebo or active treatment. A significant (P = 0.012) negative correlation (-0.72) was observed between maximal tolerated dose and specific IgE to Hev b 6.01 at baseline. After IT, immunoblot analysis demonstrated a significant increase in IgE binding in a band of approximately 22 kDa (P = 0.032) that may correspond to Hev b 6.01. New or more intense bands appeared in seven patients of the active group, while in three subjects a reduction was observed . CONCLUSIONS: Hev b 6.01 seems to be the most relevant latex allergen in HCWs. New or more intense IgE binding to latex allergenic components occurs during latex immunotherapy. However, the levels of specific IgE against these new components are low and do not seem to have clinical relevance.
[220] - Spínola Santos A, Branco Ferreira M, Lopes Pregal A, Lopes da Silva S, Mendes A, Pedro E, et al. Safety of Sublingual Latex Immunotherapy. Are Specific IgE to Recombinant Latex Allergens a Risk Factor ? AAAAI 62nd Annual Meeting, Miami, 3-7 March 2006, Poster n°512
RATIONALE: Specific immunotherapy (IT) could be a valuable therapeutic tool for latex allergy. We studied the safety of sublingual latex rush IT and its relation with specific IgE to recombinant latex allergens METHODS: We studied nine patients (F=8, M=1;mean age=35.1 years) belonging to risk groups (multiple surgeries=2; occupationally exposed=7). Clinical manifestations were: anaphylaxis-1; asthma-5; urticaria and /or angioedema-5; rhinitis-6, hand eczema-1. We performed skin prick test with latex and serum measurements of specific IgE (sIgE)(ImmunoCAP, PharmaciaDiagnostics, Sweeden) to latex and its recombinant allergens (rHev b 1, rHev b 2, rHev b 3, rHev b 5, rHev b 6.01, rHev b 6.02, rHev b 8, rHev b 9, rHev b 11). All patients underwent rush sublingual IT with a standardized latex extract (SLIT-Latex, ALKAbelló, S.A., Spain) under medical surveillance, after giving informed consent RESULTS: In our series, latex sublingual IT was associated with adverse reactions in 55% of the patients, although these reactions were mild and easily controlled. In all patients with perioral or respiratory reactions, sIgE to rHev b 6.01 or to rHevb 6.02 were higher than sIgE to rHev b 5 On the other hand, in 3 of the 4 patients that tolerated rush IT without any adverse reactions, we observed the opposite pattern (sIgE to rHev b 5 higher than to rHev b 6) CONCLUSIONS: Sublingual rush latex IT is associated with same adverse reaction. In this series, the pattern of sIgE to rHev b 5/rHev b 6 seems to be associated with the occurrence of these adverse reactions Funding: PharmaciaDiagnostics
[221] - Sastre J, Raulf-Heimsoth M, Rihs HP, Fernandez-Nieto M, Barber D, Lombardero M, et al. IgE reactivity to latex allergens among sensitized healthcare workers before and after immunotherapy with latex. Allergy 2006;61:206-210
BACKGROUND: New IgE sensitizations to proteins in allergen extracts have been shown to occur during allergen-specific immunotherapy (IT) . METHODS: Twenty-four healthcare workers (HCWs) -- patients included in a latex IT study -- were analysed, 16 in active treatment and eight in placebo. Sera were obtained at baseline and after 6 months of IT and analysed with immunoblotting and CAP System with eight single recombinant latex allergens (rHev b 1, 3, 5, 6.01, 8, 9, 10, 11, and a mix of rHev b1, 5, 6.01 and 8) . RESULTS: After IT with latex, three patients in the active treatment group had new IgE sensitizations, one to Hev b 5, one to Hev b 11 and another to Hev b 6.01. No other significant variation in mean of specific IgE to latex or recombinant allergens were observed in patients who received placebo or active treatment. A significant (P = 0.012) negative correlation (-0.72) was observed between maximal tolerated dose and specific IgE to Hev b 6.01 at baseline. After IT, immunoblot analysis demonstrated a significant increase in IgE binding in a band of approximately 22 kDa (P = 0.032) that may correspond to Hev b 6.01. New or more intense bands appeared in seven patients of the active group, while in three subjects a reduction was observed . CONCLUSIONS: Hev b 6.01 seems to be the most relevant latex allergen in HCWs. New or more intense IgE binding to latex allergenic components occurs during latex immunotherapy. However, the levels of specific IgE against these new components are low and do not seem to have clinical relevance.
[222] - Spinola Santos A, Lopes da Silva S, Alves R, Pedro E, Pereira Santos C, Pereira Barbosa M. Sublingual Immunotherapy to Latex: Safety and Induction of New Sensitizations. J Allergy Clin Immunol 2008;121:S28
RATIONALE: Diverse data has been presented on short and long term safety of latex sublingual immunotherapy (latex-SLIT), namely frequency of adverse reactions (AR) occurrence and new sensitizations induction. METHODS: Clinical data were collected from SLIT registry of 22 patients (19F/3M; 35.5 6 7.5 years-old), including description of AR occurred during induction phase (4 days) Specific IgE (sIgE) to latex and recombinants (rHev b 1, 2, 3, 5, 6.01, 6.02, 8, 9, 11) were determined before SLIT (T0-22 patients) and repeated 1 year later (T1-9 patients), while maintaining SLIT. RESULTS: AR occurred in 12 patients (20 events), mostly systemic reactions (16/20), occurring within first 30 minutes after last dose (19/20). Medical treatment (anti-histamines, iv steroids and bronchodilator) was required in 15/20 reactions. Protocol schedule was adapted in 8 patients after AR and pre-medication was started in 6. There was no significant difference in mean sIgE to Hev b6.01 (T0) between patients with and without AR or a correlation between those levels and maximum tolerated dose. New sensitizations appeared in T1 to Hev b2 (2 patients) and Hev b3 (1). CONCLUSIONS: We verified the occurrence of systemic reactions in 54.5% of the patients, during latex-SLIT induction. We believe this rushprotocol should always be performed in hospital setting, under close clinical surveillance. In our population, sIgE to Hev b6.01 was not useful as a predictor of low tolerance to vaccine, as previously suggested. After one year of SLIT, 3 new sensitizations were detected to Hev b2 and Hev b3, but not to Hev b6.01.
[223] - Sanz M, García B, Tabar A, García-Avilés M, Salcedo G, Barber D, et al. Immunological changes after sublingual immunotherapy in latex-allergic children. EAACI 25th Congress, Vienna, 10-14 June, 2006, Poster n°815
The aim of the study was to assess the possible immunological changes expressed on the basophil activation marker CD63, on the specific IgE to latex and recombinant latex allergens; and on the IgG4 to latex during the treatment with antigen specific sublingual immunotherapy in children allergic to latex. Material: Twenty-three children diagnosed of latex allergy by anamnesis, positive skin tests to latex and positive specific IgE. Eighteen of them (9 males and 9 females, age 12 ± 4) received latex sublingual immunotherapy (SLIT-latex, ALK-Abelló, S.A). The other five children (3 males and 2 females, age 12 ± 5) who did not receive immunotherapy were the control group. METHODS: All the children underwent the following determinations before starting the administration of the immunotherapy (T0), six (T1) and 12 months (T2) after the treatment was started: - Basophil activation test (BAT) to latex and the following natural and recombinant latex allergens: rHev b 5, rHev b6.01, nHeb b6.02, according to the technique described (Pediatr Allergy Immunol in press). - Specific IgE to latex, rHev b 1, rHev b 2, rHev b 3, rHev b 5, rHev b 6.01, rHev b 6.02, rHev b 8, rHev b 11 (CAP Pharmacia). - Specific IgG4 to latex (CAP Pharmacia). RESULTS: The BAT shows a significant decrease with latex (p<0.001) as well as with rHev b 6.01 proteins (p<0.05) and nHev b 6.02 (p<0.01) in the first 6 months of treatment. Specific IgE suffers no significant changes in the course of immunotherapy for any of the recombinant proteins or for latex. IgG4 to latex significantly increases (p<0.05) twelve months after starting the treatment. No changes were observed in the untreated controls. CONCLUSIONS: SLIT to latex administered in children allergic to latex induces changes in the basophil activation to prohevein (rHev b6.01) and hevein (nHev b.02) which can be detected six months after starting the treatment, and in the IgG4 production to latex which can be detected twelve months after starting the treatment.
[224] - Yagami T, Osuna H, Kouno M, Haishima Y, Nakamura A, Ikezawa Z. Significance of Carbohydrate Epitopes in a Latex Allergen with beta-1,3-Glucanase Activity. Int Arch Allergy Immunol 2002;129:27-37
Background: One of the latex allergens, Hev b 2, has beta-1,3-glucanase activity. The entire sequence of this allergen is already known. There is one potential N-glycosylation site in this molecule (27Asn). Heterogeneous glycosylation of this Asn residue could be a source of the multiplicity of natural Hev b 2. Possible participation of the carbohydrate epitopes of latex beta-1,3-glucanase isoenzymes in their IgE-binding capacity and cross-reactivity was investigated in this study. Methods: beta-1,3-Glucanase isoenzymes were separated based on their affinities for concanavalin A. IgE-binding capacity and cross-reactivity were examined by immunoblotting and enzyme-linked immunosorbent assay (ELISA). Sequence heterogeneity among the isoenzymes was probed by peptide mass mapping after lysyl endopeptidase digestion. To clarify the relation to Hev b 2, N-terminal sequencing was performed on a fragmented peptide common to the separated isoenzymes. Results: Basic beta-1,3-glucanase was subdivided into two glycosylated isoenzymes (GI and GII) and one non-glycosylated isoenzyme (GIII). IgE antibodies in latex-positive sera chiefly recognized the glycosylated isoenzymes. Inhibition ELISA supported the significance of the carbohydrate epitopes for the IgE recognition and cross-reactivity. However, non-glycosylated GIII, as well as GI and GII, produced positive results in a skin prick test. The three beta-1,3-glucanase isoenzymes shared a partial sequence in common with Hev b 2. Conclusions: Our results suggest that the carbohydrate epitopes in Hev b 2 homologues are relevant to an in vitro diagnosis of latex allergy and the accompanying cross-reactivity. Carbohydrate epitopes do not necessarily provoke allergic symptoms. Therefore, the actual allergenicity of Hev b 2 and its homologues should be carefully evaluated not only by in vitro IgE tests but also by in vivo tests
[225] - Arif SAM, Hamilton RG, Yusof F, Chew NP, Loke YH, Nimkar S, et al. Isolation and characterization of the Early Nodule Specific Protein homologue (Hev b 13), an allergenic lipolytic esterase from Hevea brasiliensis latex. J Biol Chem 2004;279:23933-23941
Recurring reports of a highly allergenic 42-46-kDa protein in Hevea brasiliensis latex appeared to have been resolved with the discovery of a 43-kDa allergenic latex protein that was a homologue to patatin. However, the low to moderate prevalence of sensitization to the protein, designated Hev b 7, among latex-allergic patients could not adequately explain the frequent observations of the 42-46-kDa allergen. This led to the hypothesis that another, more allergenic protein of a similar molecular mass existed in Hevea latex. We report the isolation and purification of a 42.98-kDa latex glycoprotein showing homology to the early nodule-specific protein (ENSP) of the legumes Medicago sativa, Medicago truncatula, and Glycine max. The protein is allergenic, being recognized by immunoglobulin E (IgE) in sera from latex-allergic patients. The IgE epitope resides on the carbohydrate moiety of the protein, and the presence of a similar carbohydrate component on potato tuber patatin enables the latter to inhibit IgE binding to the ENSP homologue. The cDNA encoding the ENSP homologue was isolated by reverse transcription-PCR and cloned. The protein predicted from the cDNA sequence has 391 amino acids, the first 26 of which constitute a putative signal peptide. The deduced molecular mass of the mature protein is 40.40 kDa, while its isoelectric point is estimated at 5.0. The discrepancy between the predicted and observed molecular mass might be due to glycosylation, for which three N-sites on the protein are predicted. The purified protein showed lipase and esterase activities and may be involved in plant defense.
[226] - Yeang HY. Prevalence of latex may be vastly overestimated when determined by in vitro assays. Ann Allergy Asthma Immunol 2000;84:628-632
BACKGROUND: The prevalence of latex-specific IgE computed from the results of serologic assays is commonly thought to reflect, to a greater or lesser extent, the prevalence of latex allergy and its implied risk . OBJECTIVE: The study examines how imperfect test specificity of in vitro assays influences the precision of latex allergy prevalence that it estimates . METHODS: Various models encompassing a range of hypothetical test sensitivity and specificity values are investigated to gauge their influence on the estimate of latex allergy prevalence. The models examine these interactions in situations of high or low allergy prevalence . RESULTS: Serologic latex diagnostic assays with test specificity within the range of those of commercially available assays can greatly overestimate prevalence where the true prevalence is low (eg, of the order of one in 100 or one in 1,000). A formula to correct for errors in prevalence estimates arising from imperfect test sensitivity and specificity of an in vitro assay is presented . CONCLUSION: While serologic assays for latex IgE pose few hazards to the patient and are useful for confirming the diagnosis of latex allergy, the test results may vastly overestimate the true prevalence of latex allergy and its associated risks in situations where latex allergy is actually rare.
[227] - Merrett TG, Merrett J, Kekwick R. The prevalence of immunoglobulin E antibodies to the proteins of rubber (Hevea brasiliensis) latex and grass (Phleum pratense) pollen in sera of British blood donors. Clin Exp Allergy 1999;29:1572-1578
Although there have been many studies of the prevalence of latex allergy in populations deemed to be at risk, little is known of the potential allergic susceptibility to latex products prevailing in the general population. OBJECTIVE: To assess the possible prevalence of allergy to latex goods in a population of blood donors by measurement of specific antilatex immunoglobulin (Ig) E in blood, to relate this to prevalence of antigrass IgE in the blood donations, and to assess the prevalence of antibodies to grass, house dust mite and cat allergens in those donors having antilatex IgE antibodies. METHODS: Sera from two groups of donations obtained in the English West Midlands were assayed. A group of 2000 donations obtained in midwinter was assayed for antilatex and antigrass pollen IgE. A group of 5000 midsummer donations was assayed for total IgE, and antilatex IgE and the sera giving a positive reaction, assayed for antigrass pollen, antihouse dust mite and anticat IgE. The nature of the principal latex and grass pollen polypeptides reacting with IgE in the sera was assessed by immunoblotting. RESULTS: Anti-latex IgE was detected in approximately 4% of the winter and 7% of the summer donations. The prevalence of antigrass IgE in the winter donations was approximately 20% and amongst the latex-positive sera approximately 84% contained antigrass IgE. Of the summer donations of latex-positive sera, 96% contained antigrass, 48.6% antimite IgE and 34% anticat IgE. The prevalence of both antilatex and antigrass IgE was age and sex related. Inhibition studies indicated cross-reactivity of IgE with latex and grass pollen proteins. CONCLUSIONS: Whilst 4-7% of the population may have serum IgE reacting with latex, the levels are low compared with those reacting with the aeroallergens studied. The apparent cross-reactivity of some serum IgE with both latex and grass pollen taken with other evidence suggests that, in some individuals, allergy to latex may arise from an initial sensitization by grass pollen.
[228] - Bousquet J, Flahault A, Vandenplas O, Ameille J, Duron JJ, Pecquet C, et al. Natural rubber latex allergy among health care workers: A systematic review of the evidence. J Allergy Clin Immunol 2006;118:447-454
BACKGROUND: Natural rubber latex is a recognized allergen, but a recent meta-analysis failed to find any association between latex exposure and allergy in health care workers (HCWs) . OBJECTIVES: A meta-analysis was carried out under the auspices of the French National Regulatory Authority to assess the allergic risk induced by latex gloves in HCWs . METHODS: The risk of work-related exposure to latex for the development of latex allergy was assessed. Prevalence and incidence rates of latex sensitization or allergy were compared in HCWs and in the general population. Exposure-response relationships were assessed in HCWs . RESULTS: Latex allergy was found in 4.32% (range, 4.01% to 4.63%) of HCWs and in 1.37% (range, 0.43% to 2.31%) of the general population. Latex-positive skin prick test responses ranged from 2.1% to 3.7% in the general population and from 6.9% to 7.8% for the HCWs. HCWs exposed to latex showed an increased risk of hand dermatitis (odds ratio [OR], 2.46; 95% CI, 2.11-2.86), asthma or wheezing (OR, 1.55; 95% CI, 1.15-2.08), rhinoconjunctivitis (OR, 2.73; 95% CI, 1.97-3.81), and at least one generic symptom (OR, 1.27; 95% CI, 1.09-1.47). Sensitization to latex was significantly associated with asthma and rhinoconjunctivitis. By contrast, exposure to latex was not associated with a significantly increased risk of positive skin prick test responses to latex (OR, 1.47; 95% CI, 0.94-2.30) . CONCLUSION: HCWs have an increased risk of sensitization and allergic symptoms to latex. CLINICAL IMPLICATIONS: Prevention of latex allergy in HCWs is needed.
[231] - Hourihane JO, Allard JM, Wade AM, McEwan AI, Strobel S. Impact of repeated surgical procedures on the incidence and prevalence of latex allergy: a prospective study of 1263 children. J Pediatr 2002;140:479-482
Assessment with questionnaire, skin tests, and immunoglobulin E measurements showed 6 (0.5%) of 1263 children having elective surgery had latex allergy; 50 (4%) others were latex-sensitized. Any previous operation increased the odds of latex sensitization by 13 times; multiple operations had a less marked effect. All children having surgery may need primary prophylaxis for latex.
[232] - Blanco C, Carrillo T, Ortega N, Alvarez M, Dominguez C, Castillo R. Comparison of skin-prick test and specific serum IgE determination for the diagnosis of latex allergy. Clin Exp Allergy 1998;28:971-976
BACKGROUND: Latex IgE-mediated hypersensitivity has been recognized as an international health problem. However, there is poor information on the efficiency of the diagnostic methods available. OBJECTIVE: The purpose of this study was to specify the efficiency of several diagnostic methods for latex allergy. METHODS: We designed a prospective study involving 50 adult patients with latex allergy, as diagnosed by a suggestive clinical history and a positive skin-prick test (SPT) to a latex extract. One control group of 50 subjects paired for age, sex, total IgE levels and latex exposure, and another control group of 30 subjects with pollen allergy were used. A low-ammoniated natural-latex and several glove-latex extracts were elaborated. SPTs with these extracts, as well as with four different commercial-latex extracts were performed. Latex-specific serum IgE was determined by the CAP and the AlaSTAT methods. RESULTS: Diagnostic sensitivity was 98% for the natural-latex extract SPT, from 90% to 98% for the commercial-latex extract SPT, and from 64% to 96% for the glove-latex extract SPT. Diagnostic specificity of SPT was 100%, and no severe adverse reactions were observed during skin testing. With respect to the latex-specific serum IgE determinations, sensitivity was 86% for the CAP system and 84% for the AlaSTAT assay, and specificity was dependent on the population considered. CONCLUSION: SPT with natural latex extracts has shown a diagnostic efficiency close to 100%, significantly higher than that of latex-specific serum IgE determination.
[233] - Kochuyt AM, Van Hoeyveld EM, Stevens EA. Prevalence and clinical relevance of specific immunoglobulin E to pollen caused by sting-induced specific immunoglobulin E to cross-reacting carbohydrate determinants in Hymenoptera venoms. Clin Exp Allergy 2005;35:441-447
Summary Background Hymenoptera stings can induce specific IgE (sIgE) to carbohydrate determinants (CD) on venom glycoproteins that cross-react with CD in pollen. sIgE to such cross-reacting CD (CCD) are believed to have little or no biological activity and thus may cause misdiagnosis of pollen sensitization after a sting. Objective To determine the prevalence of multiple false positive CAP results to pollen because of sting induced anti-CCD sIgE in Hymenoptera venom (HV) allergic patients and to investigate the association of such anti-CCD sIgE with features of 'atopy'. Methods Skin prick tests (SPT) and CAP tests with grass, tree and weed pollen and with house dust mite (HDM) were carried out prospectively in 259 HV allergic patients and CAP tests with honeybee (HBV) and yellow jacket (YJV) venom were performed. Patients with negative pollen SPT associated with positive CAP tests to all three pollen groups were operationally defined as 'CCD positive'. We investigated in selected 'CCD positive' patients the presence of anti-CCD sIgE by CAP tests with bromelain and studied the identity of CD in HVs and pollen by mutual sIgE inhibition tests with CD from proteinase treated HBV (HBV-CD) and Lolium perenne (Lol-CD) extracts. Results sIgE to all three pollen groups without positive SPT or history was found in 16% of 259 patients. The presence of anti-CCD sIgE was substantiated by positive CAP tests with bromelain in 14/14 and by inhibition of all pollen CAP tests with HBV-CD in 8/9 and with Lol-CD in 2/2 patients. Double venom (DV) positive CAP tests were present in 93% of 'CCD positive' patients and were in some associated with DV skin test positivity and allergy. The prevalence of 'CCD positivity' was significantly higher among HBV (23%) than among YJV (11%) allergic patients, but was also unexpectedly high among those with DV allergy (47%). 'CCD positive' patients were younger, had a higher total IgE and more sIgE to HDM than 'CCD negative' patients. Conclusion We have shown that the risk in HV allergic patients for misdiagnosis of multivalent pollen sensitization is 16%, and we have confirmed that sting induced anti-pollen sIgE are directed to similar CD in venoms and pollen. We found evidence that the recognition of CCD might be related to the 'atopic' trait. Importantly, a positive bromelain CAP test does not exclude clinical reactivity to both venoms in 'CCD positive' HV allergic patients.
[234] - Jappe U, Raulf-Heimsoth M, Hoffmann M, Burow G, Hübsch-Müller C, Enk A. In vitro hymenoptera venom allergy diagnosis: improved by screening for cross-reactive carbohydrate determinants and reciprocal inhibition. Allergy 2006;61:1220-1229
BACKGROUND: Immunoglobulin (Ig) E-double positivity for honeybee (HB) and yellow jacket (YJ) venom causes diagnostic difficulties concerning therapeutical strategies. The aim of this study was to clarify the cause and relation of the cross-reactivity in patients with insect venom allergy . METHODS: For this purpose, 147 patients with suspected stinging insect allergy and CAP-FEIA-double positivity were investigated for specific sIgE to additional cross-reactive carbohydrate determinant (CCD)-containing allergens: timothy grass pollen, rape pollen, natural rubber latex (NRL), bromelain, and horseradish peroxidase (HRP). Sera with sIgE to NRL were further investigated with the commercially available recombinant latex allergens. Reciprocal inhibition assays with both venoms and HRP were performed . RESULTS: About 36 of 147 (24.5%) patients had sIgE to both venoms only. However, 111 of 147 (75.5%) additionally reacted to CCD-carrying allergens. 89 of 111 CCD-reactive sera had NRL-sIgE. In cases where inhibition experiments were performed, the NRL-sIgE binding was completely abolished in the presence of HRP. Only nine of 61 sera were positive for at least one recombinant latex allergen; all of them were negative in history and NRL-skin prick test. In 43 sera containing sIgE to CCD, HRP inhibition revealed unequivocal results: In 28 of 43 (65%) an HRP-inhibition >70% of sIgE to one venom occurred, pointing out the relevant venom. In three of 43 sIgE proved to be entirely CCD-specific . CONCLUSIONS: Our data indicate that in cases of IgE positivity to both insect venoms supplementary screening tests with at least one CCD-containing allergen should be performed; HRP being a suitable tool for this test. In addition, subsequent reciprocal inhibition is an essential diagnostic method to specify cross-reacting sIgE results.
[235] - Ghunaim N, Grönlund H, Kronqvist M, Grönneberg R, Söderström L, Ahlstedt S, et al. Antibody profiles and self-reported symptoms to pollen-related food allergens in grass pollen-allergic patients from northern Europe. Allergy 2005;60:185-191
BACKGROUND: Most studies on pollen-related food allergy have so far focused on the association of birch/weed pollen allergens and plant food allergy. The aim of this study was to elucidate the allergen spectrum among a group of grass pollen-allergic patients from northern Europe and to relate the results to clinical histories of pollen-related food allergy . METHODS: Fifty-eight grass pollen-allergic patients answered a questionnaire regarding allergy to foods. Blood samples were taken to test IgE-reactivity to a large panel of pollen allergens and pollen- and nonpollen-related food allergens using crude allergen extracts and recombinant and native allergens . RESULTS: Three different groups of grass pollen-allergic patients were identified according to their IgE antibody profile: a grass pollen group only (19%), a grass and tree pollen group (29%) and a grass, tree and compositae (pan-) pollen group (48%). No sensitization to Bet v 1 as well as almost no IgE to plant food was observed in the grass pollen group. In contrast, nearly all patients in the two tree-related groups had IgE to Bet v 1, which reflected the high frequency of adverse reactions to typical birch-related food in these groups. Only four patients belonging to the pan-pollen group displayed IgE to profilin Phl p 12/Bet v 2. Patients in the pan-pollen group reported significantly more symptoms to food allergens compared with patients in the two other groups. The most frequently reported symptom was the oral allergy syndrome . CONCLUSIONS: Sensitization to grass pollen alone is rare among grass pollen-allergic patients from northern Europe. The majority of patients are in addition sensitized to birch (Bet v 1), which seems to be closely related to their pollen-derived food allergy. The study highlights the advantage of using well-defined allergen molecules for the diagnosis of cross-reactivity between pollen and food allergens.
[236] - Coutinho V, Vidal C, Garrido M, Gude F, Lojo S, Linneberg A, et al. Interference of cross-reactive carbohydrates in the determination of specific IgE in alcohol drinkers and strategies to minimize it: the example of latex. Ann Allergy Asthma Immunol 2008;101:394-401
BACKGROUND: Cross-reactive carbohydrate determinants (CCDs) are N-glycans in plant and invertebrate proteins that interfere with specific IgE determinations. The prevalence of IgE to Man2XylFucGlcNAc2 (MUXF), the CCD from bromelain, may be increased in heavy drinkers. OBJECTIVE: To further investigate the relationship of alcohol consumption to CCD specific IgE. Latex was used as an example for investigating CCD interference with in vitro allergy testing and how to minimize the interference by using nonglycosylated recombinant allergens and inhibition assays. METHODS: We determined the levels of IgE to CCD markers (MUXF and ascorbate oxidase) and natural rubber latex in 270 adults without a history of latex allergy (73 abstainers or occasional drinkers, 76 light drinkers, 47 moderate drinkers, and 74 heavy drinkers). In cases with latex reactivity, we performed inhibition assays with MUXF and screened for IgE to a panel of recombinant latex allergens. Fourteen-day serologic follow-up was available for a subset of individuals. RESULTS: Moderate to heavy drinkers displayed an increased prevalence of IgE to CCD markers. The presence of CCD specific IgE was closely associated with latex IgE reactivity. Inhibition studies and the absence of reactivity to nonglycosylated recombinant latex allergens indicated CCD interference in latex IgE determinations. Serum levels of specific IgE decreased with alcohol abstention. CONCLUSIONS: In this population, alcohol consumption is associated with an increased prevalence of IgE reactivity to natural rubber latex due to CCD interference. The use of nonglycosylated recombinant allergens and inhibition assays may help to minimize CCD interference in populations in which IgE to CCDs is common.
[237] - Bridts CH, Ebo DG, Hagendorens MM, de Clerck LS, Stevens WJ. Cross-Reactive Carbohydrate Determinants (CCD): Mimickers of Allergy. AAAAI 59th Annual Meeting, Denver, 7-12 March, 2003, Poster n°379
RATIONALE: To investigate the prevalence of anti-CCD-IgE in healthy controls and patients allergic to pollen, house dust mite (HDM), pets, natural rubber latex (NRL) and hymenoptera venom. To study the contribution of anti-CCD-IgE as a cause of clinically irrelevant IgE for NRL and apple. METHODS: IgE antibodies were quantified by Immuno-CAP. Skin prick tests (SPT) were performed with HAL extracts, except for bromelain (500 µg/mL, Sigma-Aldrich). Patients were identified as anti-CCD-IgE positive if they had a negative SPT and positive IgE for bromelain. Sera containing IgE against apple or NRL were classified as true-positive or false-positive according to the presence or absence of an oral allergy syndrome (OAS) or NRL-induced anaphylaxis. RESULTS: No anti-CCD-IgE was found in controls (n=12), patients monosensitized to NRL (n=19), pets (n=7), HDM (n=28) or birch pollen (n=32). In contrast, anti-CCD-IgE was present in 21% (4/19) of patients with grass pollinosis, 25% (4/20) of patients with combined grass and birch pollinosis, in 4/8 of patients with combined grass, tree and weed pollinosis (p=0.03), and in 14% (4/28) of patients with venom anaphylaxis. False-positive NRL individuals had a higher prevalence of anti-CCD-IgE than NRL allergic patients (78% (18/23) versus 0% (0/19), p<0.001). In individuals with a false-positive IgE to apple, the frequency of anti-CCD-IgE was higher than in OAS patients (38% (15/40) versus 11% (3/27), p=0.03). CONCLUSIONS: Sensitization to CCD, caused by grass pollen or hymenoptera venom allergens, can mimic NRL and apple allergy. Patients monosensitized to NRL or birch pollen showed no anti-CCD-IgE.
[239] - Mari A, Scala E, D’Ambrosio C, Breiteneder H, Wagner S. Latex Allergy within a Cohort of Not-at-Risk Subjects with Respiratory Symptoms: Prevalence of Latex Sensitization and Assessment of Diagnostic Tools. Int Arch Allergy Immunol 2007;143:135-143
BACKGROUND: Immunoglobluin E (IgE)-mediated hypersensitivity to natural rubber latex (NRL) is a major problem in allergy practice. Currently, the use of skin prick tests (SPTs) with latex extracts and specific IgE detection for the diagnosis of NRL allergy in suspected patients is directed to identification of risk factors. Many cases of NRL allergy remain undiagnosed due to misreporting of symptoms by the patients or lack of proper questions asked by the physician. MATERIALS AND METHODS: A total of 6,126 subjects referred for respiratory symptoms underwent SPTs with NRL. Positive subjects were resurveyed for exposure to NRL, and specific IgE for NRL extracts and recombinant molecules was determined. Immunoblots of NRL extracts were performed to identify IgE patterns . RESULTS: Forty-six of 3,930 sensitized subjects had a positive SPT with NRL, displaying a prevalence of NRL sensitization of 0.75% for the general and 1.2% for the sensitized population. Eleven out of 46 (23.9%) subjects could be defined as NRL asymptomatic, whereas 35 (76.1%) developed symptoms upon exposure to NRL. Specific IgE to NRL was detected for 22 (75.86%) of 29 tested sera. Seventeen out of 22 (77%) sera displayed specific IgE to recombinant allergens with most reactions to Hev b 5, Hev b 6.01 and Hev b 6.02. Immunoblots of NRL extract fractions with patients' sera showed heterogeneous patterns . CONCLUSIONS: SPTs with NRL extract should be routinely performed in patients with respiratory symptoms. Hev b 5, Hev b 6.01 and Hev b 6.02 are the most important allergens, but further characterization of NRL extracts is needed to identify novel allergens and to clarify the role of crossreactive carbohydrate determinants.
[241] - Jappe U, Raulf-Heimsoth M, Hoffmann M, Burow G, Hübsch-Müller C, Enk A. In vitro hymenoptera venom allergy diagnosis: improved by screening for cross-reactive carbohydrate determinants and reciprocal inhibition. Allergy 2006;61:1220-1229
BACKGROUND: Immunoglobulin (Ig) E-double positivity for honeybee (HB) and yellow jacket (YJ) venom causes diagnostic difficulties concerning therapeutical strategies. The aim of this study was to clarify the cause and relation of the cross-reactivity in patients with insect venom allergy . METHODS: For this purpose, 147 patients with suspected stinging insect allergy and CAP-FEIA-double positivity were investigated for specific sIgE to additional cross-reactive carbohydrate determinant (CCD)-containing allergens: timothy grass pollen, rape pollen, natural rubber latex (NRL), bromelain, and horseradish peroxidase (HRP). Sera with sIgE to NRL were further investigated with the commercially available recombinant latex allergens. Reciprocal inhibition assays with both venoms and HRP were performed . RESULTS: About 36 of 147 (24.5%) patients had sIgE to both venoms only. However, 111 of 147 (75.5%) additionally reacted to CCD-carrying allergens. 89 of 111 CCD-reactive sera had NRL-sIgE. In cases where inhibition experiments were performed, the NRL-sIgE binding was completely abolished in the presence of HRP. Only nine of 61 sera were positive for at least one recombinant latex allergen; all of them were negative in history and NRL-skin prick test. In 43 sera containing sIgE to CCD, HRP inhibition revealed unequivocal results: In 28 of 43 (65%) an HRP-inhibition >70% of sIgE to one venom occurred, pointing out the relevant venom. In three of 43 sIgE proved to be entirely CCD-specific . CONCLUSIONS: Our data indicate that in cases of IgE positivity to both insect venoms supplementary screening tests with at least one CCD-containing allergen should be performed; HRP being a suitable tool for this test. In addition, subsequent reciprocal inhibition is an essential diagnostic method to specify cross-reacting sIgE results.
[242] - Malandain H, Giroux F, Cano Y. The influence of carbohydrate structures present in common allergen sources on specific IgE results. Eur Ann Allergy Clin Immunol 2007;39:216-220
BACKGROUND: Cross-reactive carbohydrate determinants (CCD) are well known interferants in specific IgE assays (sIgE). Glyco-epitopes are not restricted to CCD and extracts used to prepare in vitro tests contain many other glycoproteins able to bind glycan-specific IgE. The overall amounts of IgE-bindable glycan structures in allergen sources are unknown . OBJECTIVE: We aimed at quantifying the influence of N-glycan structures on IgE reactivity to commonly tested allergen sources . METHODS: IgE reactivity to 51 allergen extracts, one purified natural allergen and 10 recombinant allergens was measured on Phadia UniCAP system using 2 sera demonstrating significant levels of glycan-related IgE reactivity. Immobilized bromelain and horseradish peroxidase (HRP) were used to capture N-glycan-specific IgE from these sera. Residual IgE reactivity was measured for 42 allergen sources and 4 recombinant/purified allergens . RESULTS: An obviously excessive number of positive CAP-results were obtained with both sera, especially for plant-based allergen sources. Capture of glycan-specific IgE led to a decrease of serum IgE ractivity, variable among allergen sources and between sera. Among others, peanut results were proven largely interfered by the presence of glycan-specific IgE. Unexpectedly some allergen sources showed a slight influence of glycan-related reactivity, such as cockroach, mosquito, mussel, shrimp and domestic mites . CONCLUSION: In patients sensitized to pollens or to Hymenoptera venoms sIgE results should be interpreted with caution. One cannot substract the result of a glyco-reporter test (bromelain and/or HRP) in order to compute glycan-free slgE results for common allergen sources like peanuts. As long as the demonstration of a significant role for glycan structures in clinical allergic reactions is lacking, a simple pre-treatment able to discard glycan-specific IgE from serum would be useful to improve accuracy of in vitro diagnostic tests.
[243] - Ebo DG, Lechkar B, Schuerwegh AJ, Bridts CH, De Clerck LS, Stevens WJ. Validation of a two-color flow cytometric assay detecting in vitro basophil activation for the diagnosis of IgE-mediated natural rubber latex allergy. Allergy 2002;57:706-712
BACKGROUND: IgE-dependent triggering of basophils not only elicits the release of different mediators but also the up-regulation of certain markers, e.g. CD63, which can be detected by flow cytometry. We intended to investigate if flow cytometric analysis of basophil activation could be a valuable tool in the diagnosis of latex allergy, and to evaluate if the basophil activation test (BAT) could be helpful in determining the clinical significance of a positive latex IgE in individuals with negative history and negative latex skin test. Additionally we aimed to determine the role of cross-reactive carbohydrate determinants (CCDs) in causing positive latex IgE without apparent clinical significance . METHODS: Twelve healthy controls without a history of latex hypersensitivity with a negative latex IgE and skin test (group 1), 24 individuals without a history of latex hypersensitivity with a negative latex IgE and skin test but with other inhalant allergies (group 2), and 29 latex allergic patients with a compelling history of latex allergy with a positive latex IgE and prick test (group 3) were enrolled. The diagnostic performances of the BAT were further evaluated in 13 individuals with a history of latex allergy but with negative specific IgE and/or skin test (group 4). Twenty-four individuals with positive latex IgE without apparent clinical relevance, i.e. without history of latex hypersensitivity and negative latex skin tests, were also analyzed (group 5). The putative role of CCDs causing positive latex IgE results without apparent clinical significance was evaluated by quantification of IgE for bromelain . RESULTS: According, to the receiver operating characteristics(ROC)-generated threshold value of 17% between latex allergic patients and the pooled group of nonlatex allergic individuals, the sensitivity and specificity of the basophil activation test was 93.1% and 91.7%, respectively. In healthy controls, allergic patients without latex hypersensitivity and latex allergic patients the number of positive BATs was 0/12, 3/24 and 27/29, respectively. In the individuals with an evocative history of latex allergy but with negative latex IgE and/or skin test the BAT was positive in all 13 cases. Twenty of 24 individuals without apparent latex allergy but with positive latex IgE had a negative BAT. IgE for bromelain was positive in 1/19 sera from group 2, 1/24 sera from group 3, none of the 8 sera from group 4, but in 16/18 sera from group 5, respectively . CONCLUSION: Flow cytometric analysis of activated basophils seems a highly sensitive and specific tool for diagnosing latex allergy. In addition, the technique might help to determine the clinical relevance of positive IgE quantification in the absence of overt latex allergy. CCDs of natural rubber latex allergens were confirmed to mimic latex sensitization.
[244] - Malandain H, Giroux F, Cano Y. The influence of carbohydrate structures present in common allergen sources on specific IgE results. Eur Ann Allergy Clin Immunol 2007;39:216-220
BACKGROUND: Cross-reactive carbohydrate determinants (CCD) are well known interferants in specific IgE assays (sIgE). Glyco-epitopes are not restricted to CCD and extracts used to prepare in vitro tests contain many other glycoproteins able to bind glycan-specific IgE. The overall amounts of IgE-bindable glycan structures in allergen sources are unknown . OBJECTIVE: We aimed at quantifying the influence of N-glycan structures on IgE reactivity to commonly tested allergen sources . METHODS: IgE reactivity to 51 allergen extracts, one purified natural allergen and 10 recombinant allergens was measured on Phadia UniCAP system using 2 sera demonstrating significant levels of glycan-related IgE reactivity. Immobilized bromelain and horseradish peroxidase (HRP) were used to capture N-glycan-specific IgE from these sera. Residual IgE reactivity was measured for 42 allergen sources and 4 recombinant/purified allergens . RESULTS: An obviously excessive number of positive CAP-results were obtained with both sera, especially for plant-based allergen sources. Capture of glycan-specific IgE led to a decrease of serum IgE ractivity, variable among allergen sources and between sera. Among others, peanut results were proven largely interfered by the presence of glycan-specific IgE. Unexpectedly some allergen sources showed a slight influence of glycan-related reactivity, such as cockroach, mosquito, mussel, shrimp and domestic mites . CONCLUSION: In patients sensitized to pollens or to Hymenoptera venoms sIgE results should be interpreted with caution. One cannot substract the result of a glyco-reporter test (bromelain and/or HRP) in order to compute glycan-free slgE results for common allergen sources like peanuts. As long as the demonstration of a significant role for glycan structures in clinical allergic reactions is lacking, a simple pre-treatment able to discard glycan-specific IgE from serum would be useful to improve accuracy of in vitro diagnostic tests.
[245] - Hamilton RG, Cornish K. Absence of Cross-Reactivity Between Natural Rubber Latex From Production Lots of Hevea brasiliensis (Hev-b) and Parthenium argentatum (Guayule). J Allergy Clin Immunol 2005;115(2 suppl.):S111
RATIONALE: We investigated the potential allergenicity of production lots of Guayule latex using Hevea brasiliensis latex specific IgE antibodies to identify a non-allergenic alternative natural rubber latex for use by Hev-b latex allergic individuals METHODS: Two IgE anti-Hev-b serum pools [(53 children with spina bifida); (180 adult healthcare workers) with Hev-b latex allergy] were prepared and used in a Pharmacia CAP inhibition assay to investigate potential cross-reactivity of ammoniated and non-ammoniated Guayule latex and Guayule C-serum with non-ammoniated Hev-b C-serum (K82) Calibration was performed with 11 dilutions of FDA-E8 non-ammoniated Hev-b C-serum [(100,000 AU/ml = 38.9 mg/ml total protein) to 0.17 AU/ml]. Neoprene and two powdered Hev-b latex gloves were used as negative and positive controls, respectively RESULTS: The degree of inhibition produced in the latex CAP Inhibition Assay by all the ammoniated and non-ammoniated forms of Guayule latex and C-serum using both the adult and pediatric IgE anti-Hev-b latex serum pools was not significantly different from the neoprene negative control extract (< 1 AU/ml). This indicated an absence of any detectable cross-reactive allergenic protein (p = NS, student T-test) in the Guayule preparations. In contrast, the Redline and Triflex powdered latex examination glove positive controls produced 1,812 and 1,283,900 AU/ml of detectable allergen, respectively CONCLUSIONS: These results indicate that proteins present in production lots of Guayule latex are not cross-reactive with Hevea latex allergens This suggests that devices manufactured with Guayule latex as an alternative rubber source should be safe for use by Hev-b latex allergic individuals
[246] - Siler DJ, Cornish K, Hamilton RG. Absence of cross-reactivity of IgE antibodies from subjects allergic to Hevea brasiliensis latex with a new source of natural rubber latex from guayule (Parthenium argentatum). J Allergy Clin Immunol 1996;98:895-902
Natural rubber latex sensitization is caused by proteins from the latex of Hevea brasiliensis, the Brazilian rubber tree. Alternative non-cross-reactive latex rubber sources are needed for the manufacture of nonallergenic medical devices (e.g., catheters) that require elasticity, which is not currently provided by synthetic materials. METHODS: We have studied the ability of IgE antibodies from subjects allergic to Hevea latex and IgG antibodies from hyperimmunized mice to detect proteins derived from guayule (Parthenium argentatum), an alternative rubber source, using a direct binding and competitive inhibition RAST and Western blot analysis. RESULTS: IgE antibodies from 62 subjects allergic to Hevea latex (46 adults and 16 children with spina bifida) and from serum pools of adults allergic to Hevea latex (n = 183), pediatric patients (n = 101), and patients with spina bifida (n = 53), as well as IgG antibodies from hyperimmunized mice, were unable to detect any proteins in guayule by RAST or Western blot analysis. No competitive inhibition of IgE anti-Hevea binding to Hevea solid phase was detected by the preincubation of sera from subjects allergic to Hevea latex with soluble guayule latex before RAST analysis. CONCLUSIONS: These results demonstrate that proteins in guayule latex are not cross-reactive with Hevea latex allergens. Devices that require the elasticity provided by rubber may be manufactured with guayule latex as an alternative rubber source for patients with Hevea latex allergy.
[247] - Costa GE, Johnson JD, Hamilton RG. Cross-Reactivity studies of gutta-percha, gutta-balata, and natural rubber latex (Hevea brasiliensis). J Endod 2001;27:584-587
Gutta-percha and gutta-balata are derived from the Paliquium gutta and Mimusops globsa trees, respectively, that are in the same botanical family as the rubber tree Hevea brasiliensis. For this reason the potential for immunological cross-reactivity between the gutta-percha and gutta-balata used in endodontics and natural rubber latex (NRL) has been the subject of some controversy, because these products may be used in latex-allergic individuals. The objective of this study was to investigate the potential cross-reactivity between gutta-percha, gutta-balata, and NRL. Physiological extracts of seven commercially available gutta-percha products, raw gutta-percha, raw gutta-balata, and synthetic transpolyisoprene were each analyzed for cross-reactivity with NRL in a competitive radioallergosorbent test inhibition assay. No detectable cross-reactivity was observed with any of the raw or clinically used gutta-percha products. In contrast the raw gutta-balata released proteins that were cross-reactive with Hevea latex. We conclude that the absence of gutta-percha proteins that can react with Hevea latex-specific IgE antibody supports the minimal potential for commercially available gutta-percha to induce allergic symptoms in individuals sensitized to NRL. Because gutta-balata is sometimes added to commercial gutta-percha products caution should be exercised if products containing gutta-balata are used in endodontic care of latex-allergic individuals. Gutta-percha and gutta-balata are derived from the Paliquium gutta and Mimusops globsa trees, respectively, that are in the same botanical family as the rubber tree Hevea brasiliensis. For this reason the potential for immunological cross-reactivity between the gutta-percha and gutta-balata used in endodontics and natural rubber latex (NRL) has been the subject of some controversy, because these products may be used in latex-allergic individuals. The objective of this study was to investigate the potential cross-reactivity between gutta-percha, gutta-balata, and NRL. Physiological extracts of seven commercially available gutta-percha products, raw gutta-percha, raw gutta-balata, and synthetic transpolyisoprene were each analyzed for cross-reactivity with NRL in a competitive radioallergosorbent test inhibition assay. No detectable cross-reactivity was observed with any of the raw or clinically used gutta-percha products. In contrast the raw gutta-balata released proteins that were cross-reactive with Hevea latex. We conclude that the absence of gutta-percha proteins that can react with Hevea latex-specific IgE antibody supports the minimal potential for commercially available gutta-percha to induce allergic symptoms in individuals sensitized to NRL. Because gutta-balata is sometimes added to commercial gutta-percha products caution should be exercised if products containing gutta-balata are used in endodontic care of latex-allergic individuals
[249] - Hamann C, Rodgers PA, Alenius H, Halsey JF, Sullivan K. Cross-reactivity between gutta-percha and natural rubber latex: assumptions vs. reality. J Am Dent Assoc 2002;133:1357-1367
BACKGROUND: Immunological cross-reactivity between gutta-percha and natural rubber latex, or NRL, has not been demonstrated clearly despite recent concerns and several suspected cases reported in the literature. METHODS: The authors analyzed aqueous extracts of commercial gutta-percha points and raw gutta-percha samples for cross-reactivity to NRL by radioallergosorbent test, or RAST, inhibition; immunoblot inhibition; direct enzyme-linked immunosorbent assay, or ELISA; and ELISA inhibition using sera from NRL-allergic people as the source of anti-NRL immunoglobulin E, or IgE, antibodies. To confirm in vitro results, the authors conducted skin prick testing, or SPT, on a patient with type I NRL allergy using aqueous extracts from raw gutta-percha, ammoniated gutta-percha and gutta-percha points. RESULTS: Aqueous extracts from commercial gutta-percha points did not cross-react to NRL in RAST inhibition or immunoblot inhibition, ELISA or ELISA inhibition assays. However, three of 13 sera from subjects with type I NRL allergy exhibited IgE binding to raw gutta-percha extracts in direct ELISA. Moreover, in ELISA inhibition, the binding of IgE to raw gutta-percha extracts was inhibited in a dose-dependent manner by raw NRL and vice versa. SPT results from a subject with type I NRL allergy were positive for NRL and raw gutta-percha extracts but negative for gutta-percha point extracts. CONCLUSIONS: The authors found no detectable cross-reactivity between NRL and commercial gutta-percha points. However, their ELISA and SPT results demonstrated that some allergenic cross-reactivity exists between raw gutta-percha and raw NRL. CLINICAL IMPLICATIONS: Gutta-percha alone is not likely to induce symptoms in patients with type I NRL allergy. However, other materials used in obturating root canals may be irritating and potentially allergenic in patients with pre-existing allergies.
[251] - Kespohl S, Sander I, Merget R, Petersen A, Meyer HE, Gastaminza G, et al. Putative class I chitinase is a major allergen in obeche wood (Triplochiton scleroxylon). EAACI 23th Congress, Amsterdam, 12-16 June, 2004, Poster n°1152
Background: There is good evidence of an increased prevalence of allergic occupational asthma in wood dust exposed workers. Obeche (Triplochiton scleroxylon) is frequently used for sauna benches and modeling and its dust is known to be highly potent by means of inducing IgE-mediated sensitization. The aim of the study was to characterize major allergens in obeche wood sensitized persons and to elucidate cross-reactivity with IgE of latex-fruit allergic subjects. Method: Obeche allergens were detected in IgE Western blot studies with sera of subjects with work-related asthma and sensitization to obeche. The most abundant IgE binding protein was analyzed by N-terminal sequencing, mass spectrometric fingerprinting and sequencing of proteolytic peptides. IgE binding properties and cross-reactivity were further studied by ImmunoCAP and immunoblot inhibition experiments using sera of latex- and obeche-allergic patients. Results: In IgE binding studies a 38 kDa obeche protein was recognized by about 80% of the sera. Sequence analysis identified this protein as homologous to class I chitinases with a short N-terminal hevein-like domain followed by a catalytic protein region. Inhibition studies with wood allergic patients' sera on obeche as solid phase and latex hevein as inhibitor showed partial cross-reactivity; however, the isolated chitinase from obeche was the more effective inhibitor. Using latex sensitized patients' sera in inhibition experiments with obeche immunoblots, only in one out of four sera latex hevein inhibited IgE binding to the 38 kDa obeche protein completely while purified avocado chitinase revealed inhibition in three out of four cases. Thus, homology between the obeche class I chitinase and latex hevein is not the only reason for cross-reactivity of patients' IgE. Conclusion: Class I chitinases are already characterized as cross-reactive allergens in latex, fruit, and vegetables. As pathogenesis related (PR) proteins, they are obviously also expressed in woody plants like obeche and play an important role for allergic sensitization. Cross-reactivity between obeche wood and latex is only partially caused by homology to hevein, additional common epitopes on the catalytic domain of chitinases are assumed.
[252] - Venturini M, Gastaminza G, Kespohl S, Bernedo N, Garmendia M, Raulf-Heimsoth M, et al. Cross-reactivity between obeche wood (Triplochiton scleroxylon) and natural rubber latex. Allergy 2004;59:225-228
BACKGROUND: Several studies have shown that allergy to natural rubber latex is associated with cross reactivity to other plants. We have investigated a possible cross reactivity with a tree belonging to the Sterculiaceae family, Tryplochiton scleroxylon, in a patient allergic to obeche wood, that began to suffer symptoms of allergy in contact with latex . METHODS: Determinations of specific IgE by CAP to obeche, natural rubber latex (NRL) and recombinant allergens of latex were done. Immunoblotting was performed to study allergens detected by serum of our patient. CAP-inhibition methods were used to study cross-reactive between NRL and obeche wood . RESULTS: In obeche extract, 4 bands were found of an apparent molecular weight of 18, 38, 75 and 199 kDa. In NRL extract the allergens have a molecular weight around 18, 21, 32, 38, 60, 70 and 199 kDa. IgE to latex was almost completely inhibited (96%) by the obeche extract in CAP-inhibition studies. Using obeche extract as inhibitor, IgE binding to native hevein (nHevb 6.02) was reduced by 25% . CONCLUSIONS: The results obtained in CAP-inhibition demonstrate the existence of cross-reactivity between proteins in NRL and obeche wood. In our patient, a band of apparent molecular weight of 38 kDa could be the most important allergen.
[253] - Kespohl S, Sander I, Merget R, Petersen A, Meyer HE, Gastaminza G, et al. Putative class I chitinase is a major allergen in obeche wood (Triplochiton scleroxylon). EAACI 23th Congress, Amsterdam, 12-16 June, 2004, Poster n°1152
Background: There is good evidence of an increased prevalence of allergic occupational asthma in wood dust exposed workers. Obeche (Triplochiton scleroxylon) is frequently used for sauna benches and modeling and its dust is known to be highly potent by means of inducing IgE-mediated sensitization. The aim of the study was to characterize major allergens in obeche wood sensitized persons and to elucidate cross-reactivity with IgE of latex-fruit allergic subjects. Method: Obeche allergens were detected in IgE Western blot studies with sera of subjects with work-related asthma and sensitization to obeche. The most abundant IgE binding protein was analyzed by N-terminal sequencing, mass spectrometric fingerprinting and sequencing of proteolytic peptides. IgE binding properties and cross-reactivity were further studied by ImmunoCAP and immunoblot inhibition experiments using sera of latex- and obeche-allergic patients. Results: In IgE binding studies a 38 kDa obeche protein was recognized by about 80% of the sera. Sequence analysis identified this protein as homologous to class I chitinases with a short N-terminal hevein-like domain followed by a catalytic protein region. Inhibition studies with wood allergic patients' sera on obeche as solid phase and latex hevein as inhibitor showed partial cross-reactivity; however, the isolated chitinase from obeche was the more effective inhibitor. Using latex sensitized patients' sera in inhibition experiments with obeche immunoblots, only in one out of four sera latex hevein inhibited IgE binding to the 38 kDa obeche protein completely while purified avocado chitinase revealed inhibition in three out of four cases. Thus, homology between the obeche class I chitinase and latex hevein is not the only reason for cross-reactivity of patients' IgE. Conclusion: Class I chitinases are already characterized as cross-reactive allergens in latex, fruit, and vegetables. As pathogenesis related (PR) proteins, they are obviously also expressed in woody plants like obeche and play an important role for allergic sensitization. Cross-reactivity between obeche wood and latex is only partially caused by homology to hevein, additional common epitopes on the catalytic domain of chitinases are assumed.
[254] - Venturini M, Gastaminza G, Kespohl S, Bernedo N, Garmendia M, Raulf-Heimsoth M, et al. Cross-reactivity between obeche wood (Triplochiton scleroxylon) and natural rubber latex. Allergy 2004;59:225-228
BACKGROUND: Several studies have shown that allergy to natural rubber latex is associated with cross reactivity to other plants. We have investigated a possible cross reactivity with a tree belonging to the Sterculiaceae family, Tryplochiton scleroxylon, in a patient allergic to obeche wood, that began to suffer symptoms of allergy in contact with latex . METHODS: Determinations of specific IgE by CAP to obeche, natural rubber latex (NRL) and recombinant allergens of latex were done. Immunoblotting was performed to study allergens detected by serum of our patient. CAP-inhibition methods were used to study cross-reactive between NRL and obeche wood . RESULTS: In obeche extract, 4 bands were found of an apparent molecular weight of 18, 38, 75 and 199 kDa. In NRL extract the allergens have a molecular weight around 18, 21, 32, 38, 60, 70 and 199 kDa. IgE to latex was almost completely inhibited (96%) by the obeche extract in CAP-inhibition studies. Using obeche extract as inhibitor, IgE binding to native hevein (nHevb 6.02) was reduced by 25% . CONCLUSIONS: The results obtained in CAP-inhibition demonstrate the existence of cross-reactivity between proteins in NRL and obeche wood. In our patient, a band of apparent molecular weight of 38 kDa could be the most important allergen.
[255] - Vidal C, Gonzalez-Quintela A, Rodriguez V, Armisen M, Linares T, Fernandez-Caldas E. Anaphylaxis to Cyphomandra betacea Sendth (tamarillo) in an obeche wood (Triplochiton scleroxylon)--allergic patient. Ann Allergy Asthma Immunol 2006;96:870-873
BACKGROUND: Anaphylaxis after the first exposure to a food allergen is uncommon unless a cross-reaction is present. OBJECTIVE: To investigate a possible relationship between the fruit Cyphomandra betacea Sendth (commonly known as tamarillo) and the wood of Triplochiton scleroxylon (obeche) in a patient with allergic occupational bronchial asthma due to obeche wood who began to experience anaphylaxis episodes after eating tamarillo. METHODS: A 33-year-old carpenter exposed to obeche wood in his occupation was initially seen with rhinitis and bronchial asthma. The causal relationship of these symptoms to obeche wood exposure was investigated by means of peak flow monitoring and bronchial inhalation testing. Furthermore, the patient had 2 acute episodes of anaphylaxis a few minutes after eating salad containing tamarillo. He had never tasted tamarillo before. The allergologic study included skin prick tests, serum specific IgE determinations, bronchial challenges, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and immunoblotting. RESULTS: Results of skin prick tests with common aeroallergens were negative. Strong skin prick test responses were obtained with obeche and tamarillo. Results of bronchial challenge testing with obeche extract were positive. In tamarillo extract, a 28-kDa band appeared as the most relevant IgE-binding antigen. A similar band of 28 kDa happens to be frequently detected in obeche-allergic patients. CONCLUSION: To our knowledge, this is the first reported case of anaphylaxis to tamarillo presented in a patient allergic to obeche, which raises the question of a new cross-reactivity antigen
[257] - Piirilä P, Mäkinen-Kiljunen S, Kajosaari M, Kiistala R, Petman L, Haahtela T. Nonoccupational Sensitization to Indoor Plants. Allergy Clin Immunol Int 2006;18:106-113
Background: Indoor plants can cause imunoglobulin E (IgE)-mediated allergy. We studied sensitization to indoor plants and factors affecting sensitization in an allergy outpatient clinic. Methods/Data base: 205 atopic and 90 nonatopic patients were skin prick tested for sensitivity to 12 common indoor plants. Sera from 34 adult plant-positive patients were im-munospot tested. The occurrence of the plants in homes and associated respiratory symptoms were assessed by questionnaire. Results: Of the atopic patients, 39% had positive reactions to the plants; 54% to Ficus benjamina, 36% to Hibiscus rosa sinensis, 35% to Yucca aloifolia, 23% to Dracaena fragrans, and 21% to Asplenium nidus. Of the nonatopics, 2% had skin reactions to the plants. In immunospot studies, 62% of the plant-positive patients showed IgE specific to at least one plant; 44% to Ficus benjamina, 35% to Dieffenbachia picta, and 32% to Hibiscus rosa sinensis and Chamaedorea elegans. Forty percent of patients sensitized to Ficus benjamina and 33% of those sensitized to Dieffenbachia picta reported clinical symptoms. Conclusion: Sensitization to indoor plants, particularly to Ficus spp., is commonly detected by skin and immunospot tests in atopic patients. A minority (0-40% for various plants) of prick test-positive patients reported clinical symptoms. Clustering of pollen, vegetable food, and indoor plant sensitivities suggested crossreactivity of allergens.
[258] - Ranta PM, Dimitropoulou C, Ford J, Young P, Ownby DR. Identification and characterization of crossreactive Poinsettia (Euphorbia pulcherrima) and natural rubber latex (Hevea brasiliensis) allergens. ACAAI Annual Meeting, New Orleans, 7-12 Nov. 2003, Poster n° 19
Background: The poinsettia, Euphorbia pulcherrima, is a popular ornamental plant, especially around the Christmas holiday season. Natural rubber latex (NRL) is obtained from the Brazilian rubber tree, Hevea brasiliensis, and is in the same plant family as poinsettia, Euphorbiaceae. The specific cross-reactive allergenic protein(s) between poinsettia and NRL has not been described. OBJECTIVE: The objective of our study was to make a poinsettia extract and to investigate if poinsettia proteins cross-react with NRL allergens. METHODS: The Human Assurance Committee approved this study. Poinsettia extract was prepared from 2 poinsettia plants. Sera from subjects with sensitivity to NRL were included if they had anti-latex IgE values ?0.70 kIU/L and if they had a residual volume of ?1 ml. Latex sensitive sera were tested and included if the anti-poinsettia IgE value was ?0.35 kIU/L. A negative control consisted of pooled sera from 6 individuals who had anti-latex and anti-poinsettia IgE values of <0.35 kIU/L. SDS-PAGE was followed by immunoblotting with poinsettia extract and with NRL extract. Immunoblot inhibition was done with pooled serum with specific IgE to both NRL and poinsettia. Allergen binding was inhibited by poinsettia and NRL extracts. RESULTS: Of the latex sensitive sera, 40% (6/15) had poinsettia specific IgE. The group that was sensitive to both poinsettia and NRL had significantly fewer females than the control group (p=0.046), but there was no statistically significant difference in sex between the poinsettia/ NRL sensitive group and the NRL sensitive group (p=0.5) and the NRL sensitive group and the control group (p=0.26). There was no statistical significant differences in average age between the groups (p=0.29, p=0.16, p=0.20). The molecular weights of the two cross-reactive proteins were 18.24 kD and 24.64 kD. The table shows the extent of inhibition for 18.24 kD; „0‰ indicates no inhibition, „+‰ indicates some inhibition, „++‰ indicates considerable inhibition, „+++‰ indicates total inhibition. CONCLUSIONS: These cross-reactive proteins correspond to Hev b 6.01, a major NRL allergen, and Hev b 10, a minor NRL allergen, respectively. Individuals with latex allergy should be advised to avoid poinsettia plants, since exposure may result in a type I immediate hypersensitivity reaction.
[259] - Le Sellin J. Latex/tobacco : cross allergy. EAACI 22th Congress, Paris, 7-11 June, 2003, Poster n°882
A farmer's particular clinical case for a possibility of a cross allergy between latex and tobacco leave was found. Actually , this patient has been cured in 1990 about a latex glove allergy with the result of a positive biology and skin prick test. Later on , in 2002 , he showed a much severe contact urticaria all over his hands , forearms and his face : it was clearly due to cultivated tobacco (variety RO 201). Methods: Commercial latex skin prick test (SPT) , green tobacco leave SPT , dried tobacco leave SPT , STP using an in-house extract tobacco , latex CAP-RAST, tobacco CAP-RAST and tobacco CAP-RAST inhibited by commercial latex extract have been achieved. Results: SPT is posive for commercial latex . SPTs are positive for green tobacco leave and dried tobacco leave with the same response . SPT is sligthly positive for in- house extract tobacco . Both CAP-RAST are positive for latex ( 8,69 KUA/l ) and tobacco(Pharmacia allergen) ( 2,38 KUA /l ) . Tobacco CAP-RAST is inhibited by Commercial latex extract .This inhitibion is 60 % . Conclusions: Green tobacco sensibilization has been hnown for a long time , whereas dried tobacco is carried out . Both sensitization to latex and tobacco in relation with significant tobacco CAP-RAST inhibition clearly show a cross-allergy between latex and tobacco.
Imprimer la bibliographie