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Les Fabacées : généralités

samedi 3 mai 2008, par Allerdata


Les Fabacées regroupent plus de 20000 espèces différentes, parmi lesquelles des plantes de toute première importance sur le plan allergologique, comme l’arachide, le soja, le lupin, les lentilles, …

Les usages des Fabacées sont multiples :

  • pâtures et fourrages : trèfles, sainfoins, mélilots, luzerne, ...
  • alimentation animale : protéagineux et tourteaux : pois, féverole, lupin, soja,…
  • nutrition humaine : soja et tous les féculents que sont les pois, les haricots, les lentilles et qui représentent une source essentielle de protéines dans beaucoup de pays
  • huiles alimentaires : arachide, soja
  • produits dérivés : à usage traditionnel ou diététique (ex. tofu) ou techno-alimentaire (farines, extrudés, isolats, …)
  • production de gommes : guar, adragante, caroube
  • sans compter l’arachide grillée (ou bouillie dans certains pays)

Sur le plan botanique, l’arachide se distingue par la croissance sous-terraine de ses gousses et par son caractère nettement oléagineux au sein des graines de Fabacées.

Le soja est, de ce point de vue, dans une position intermédiaire : ni une graine protéagineuse comme les pois, haricots, etc.. ni un oléagineux comme l’arachide.

Il n’est pas exclu que cette distinction révèle une composante non encore bien étudiée, à savoir l’influence des lipides sur l’allergénicité des protéines au sein d’un produit allergisant .

En ce qui concerne les Fabacées, on a une gradation qui pourrait concorder avec cette hypothèse, l’allergénicité augmentant depuis les pois, haricots, etc.. jusqu’à l’arachide en passant par le soja.

De nombreuses graines oléagineuses provenant d’autres familles botaniques montrent une allergénicité prononcée se révélant d’autant mieux que la graine elle-même est consommée : par exemple le sésame ou la noix, par opposition au colza, au tournesol, etc.. dont on consomme plutôt un dérivé pratiquement dénué de protéines, l’huile.

Les dénominations anglo-saxonnes des Fabacées peuvent générer des confusions : le terme de « legumes », par exemple qui désigne en fait les Légumineuses, c’est-à-dire les Fabacées en général.

Le terme de « pulses » se rapporte aux graines protéagineuses entrant dans l’alimentation comme les pois, les haricots, etc…

Par ailleurs, un grand nombre de graines comportent « bean » ou « pea » ou « gram » dans leur nom. Les équivalences entre anglais et français ne sont pas constantes et, là où « bean » représente une notion de graine en général, on peut trouver « pois », « fève », « haricot », etc.. en français. En dehors des Fabacées on a aussi des équivalences inconstantes comme « bean » pour les grains (café, ricin) ou les fèves (cacao).


Le tableau ci-dessous liste un certain nombre de graines de Fabacées avec leurs dénominations latines, françaises et anglo-saxonnes. Il ne saurait être exhaustif du fait de la multitude des appellations locales que l’usage ancien et populaire de ces graines a généré. En Inde, par exemple, on dénomme de nombreuses graines de Fabacées avec le mot « dal » en suffixe .

EspèceFrançaisAnglaisavec « bean »avec « pea »
Arachis hypogea arachide, cacahuète peanut
Cajanus cajan pois d’Angola, ambrevade red gram pigeon pea
Canavalia ensiformis jack bean
Canavalia gladiata pois sabre sword bean
Ceratonia siliqua caroube carob algarroba bean
Cicer arietinum pois chiche chickpea
Dipteryx odorata fève de Tonka tonka bean
Glycine max soja soya bean, soybean
Lablab purpureus dolique d’Egypte, pois nourrice hyacinth bean
Lathyrus sativus gesse commune vetchling indian pea, grass pea
Lens culinaris lentille lentil
Lupinus spp. lupin lupine
Macrotyloma uniflorum haricot de kulthi horse gram kulthi bean
Medicago sativa luzerne alfalfa
Phaseolus coccineus haricot d’Espagne runner bean
Phaseolus lunatus haricot du Cap / pois de sept ans lima bean
Phaseolus vulgaris haricot, haricot vert, flageolet,
haricot pinto, coco rose		 kidney bean, snap bean, green bean, string bean, butter bean, flageolet bean, French bean, navy bean, pinto bean
Phaseolus vulgaris (nanus group)
Physostigma venenosum fève de calabar calabar bean
Pisum sativum pois, petit pois, pois cassé green pea, garden pea, split pea
Pisum sativum arvense pois fourrager field pea
Pisum sativum macrocarpon pois mange-tout snow pea, snap pea
Psophocarpus tetragonolobus pois ailé / pois asperge winged bean
Tamarindus indica tamarin tamarind
Trigonella foenum-graecum fenugrec, trigonelle fenugreek
Vicia ervilia ers, lentille ers ervil, lentil vetch, bitter vetch
Vicia faba fève broad bean, fava bean
Vicia sativa vesce cultivée common vetch
Vigna aconitifolia haricot papillon moth bean
Vigna angularis haricot anguleux adzuki bean
Vigna mungo haricot mung(o) blackgram urad bean
Vigna radiata haricot mung(o) à grain vert/jaune green/golden gram mung bean
Vigna umbellata haricot (grain dee) riz rice bean
Vigna unguiculata cylindrica dolique des vaches, dolique cajun catjang bean cowpea
Vigna unguiculata sesquipedalis dolique asperge, haricot kilomètre yardlong bean, asparagus bean
Vigna unguiculata unguiculata dolique à œil noir, niébé black-eye pea, common cowpea

Les protéines IgE-réactives des Fabacées

Les graines de Fabacées sont souvent très riches en protéines, ce qui justifie leur importance alimentaire. Parmi ces protéines on trouve principalement des protéines dites de stockage, c’est-à-dire accumulées en vue de la germination et de la croissance de la plantule.

Ces protéines de stockage sont, notamment, des vicilines et des légumines. Bien que ces 2 sortes de protéines soient rangées dans la super-famille des cupines (cf. Les protéines des graines), il ne faut pas confondre vicilines et légumines car ces protéines ne croisent pas les unes avec les autres.

Aussi, le terme de « cupines » employé par certains peut être trompeur pour désigner tel ou tel allergène.

On trouve aussi des viclines et des légumines dans des graines d’autres familles botaniques : le sésame, les noix, les noisettes, etc..

Des protéines plus petites, sédimentant en « 2S » au lieu de 7S ou 11S comme les vicilines et légumines, sont trouvées dans les graines de Fabacées. Elles ont un rôle souvent mixte, à la fois de stockage et de défense .

On rencontre des équivalents de 2S albumines dans des graines d’autres familles botaniques : le sésame, le tournesol, la moutarde, la noix de cajou, etc…

Il n’est pas surprenant de trouver des LTP (lipid transfer proteins) dans les graines de Fabacées, ces protéines étant rencontrées dans la plupart des espèces végétales.

En revanche, la présence de protéines PR-10, Bet v 1-like, dans plusieurs graines de Fabacées (arachide, soja, etc..) crée une opportunité supplémentaire de réactivité croisée.

En plus des associations immunologiques et/ou cliniques entre graines de Fabacées, on peut donc observer des tableaux associant Fabacées et bouleau, Fabacées et fruits des Rosacées, ou encore Fabacées, bouleau et Rosacées.

On comprendra que la dissection des sensibilisations responsables de la réactivité finalement présente chez le patient soit parfois difficile à opérer.

Ces différents cadres étiologiques sont discutés ailleurs (cf. Fabacées réactions croisées)

Fabacées : monographies

Sont envisagés séparément :

Sont décrits ci-après :

Dans l’ensemble, ces graines génèrent assez rarement des réactions sévères en France : sur 900 déclarations au Réseau d’Allergo-Vigilance en mai 2010, 8 concernaient les lentilles, 2 les pois, 1 la fève, 1 le haricot vert et 1 le fenugrec. Ces chiffres sont à comparer avec ceux du soja (23 cas), du lupin (33) et, bien sûr, de l’arachide (111).

Haricots

L’espèce Phaseolus vulgaris comporte de multiples variétés et sous-espèces donnant une égale multitude de produits : haricots blancs, rouges, flageolets, cocos, haricots verts, mange-tout, etc..


Malgré leur large consommation, ces différents haricots semblent ne générer que rarement des réactions allergiques dans de nombreux pays d’Europe et en Amérique du nord.

On connaît plutôt des cas d’allergie par voie respiratoire : haricots verts crus ou vapeurs de cuisson de haricots blancs .


Ce n’est pas que les graines de haricot soient dépourvues de protéines appartenant à des familles d’allergènes : on trouve bien une PR-10, une profiline , une chitinase de classe 1 , une thaumatine-like , ainsi que la phaséoline qui est une viciline glycosylée présente en grande quantité dans les graines.


L’absence d’allergie alimentaire associée peut se comprendre dans le cas du haricot vert par une fragilité (ou une dilution ?) des allergènes. Ainsi Sanchez Monge a noté la disparition de l’IgE-réactivité de la chitinase après cuisson .

Malgré tout, des blots réalisés avec les vapeurs de cuisson étaient positifs au moins pour une bande (38 kD) , faisant penser qu’un allergène de haricot, au moins, est thermostable.


Compte tenu de cette thermolabilité partielle des allergènes de haricot, il a été suggéré d’utiliser pour les tests cutanés des extraits cuits plutôt que crus, comme le sont les extraits commerciaux : la spécificité des tests était alors améliorée .


Les recherches en technologie alimentaire ne cessant d’inventer de nouveaux procédés, les protéines de haricot pourraient venir allonger la liste des protéines de blé, de soja, de lupin, etc.., utilisées dans des préparations industrielles.

Un des buts de ces ajouts dans les aliments manufacturés étant d’obtenir des réseaux moléculaires (gels, élasticité, cohésion, etc..), le recours à des « colles » comme la transglutaminase est envisagé .
Le résultat sur le plan immunologique de ces transformations moléculaires est en grande partie inconnu , mais, s’agissant du haricot, une étude a montré une baisse de la digestibilité de la phaséoline traitée par la transglutaminase .

Lentilles



Les lentilles représentent une cause non négligeable d’allergie alimentaire dans certains pays : ainsi, en Espagne, elles sont la cause de 10% des allergies alimentaires de l’enfant.

Ailleurs en Europe, les cas d’allergie alimentaire aux lentilles sont rares. La responsabilité des habitudes alimentaires est souvent avancée pour expliquer ces écarts car, contrairement à d’autres légumineuses, l’IgE-réactivité des lentilles n’est pas affectée par la cuisson .

On a d’ailleurs décrit des réactions au contact des vapeurs de cuisson .


On connaît quelques allergènes :

  • Len c 1 est une viciline glycosylée, moyennement homologue avec Ara h 1 (50% d’identité) et positive chez la plupart des patients (Espagne) . Cet allergène pourrait être responsables des réactivités croisées avec l’arachide et le pois chiche
  • Len c 2 est une protéine de 66kD, homologue d’une « SBP65 » trouvée dans le pois. Ces 2 protéines ont la particularité d’être biotinylées naturellement. Il n’a pas été étudié si cette présence de biotine pouvait interférer dans certains tests in vitro utilisant un complexe de détection à base de streptavidine (ex. peroxydase-streptavidine) : on pourrait alors avoir une liaison directe à Len c 2 sans passer par la chaîne réactionnelle normale allergène – IgE du patient – anti-IgE biotinylée.

Pois

Les différentes variétés de pois (petit pois, pois cassés, pois gourmands, etc..) appartiennent à une même espèce de Fabacée, Pisum sativum.



Les cas publiés d’allergie aux pois sont rares. Peut-être est-ce en relation avec la thermolabilité de certains des allergènes : dans une étude d’enfants Danois aucun des 10 TPO positifs avec le pois cru n’était positif avec le pois cuit .


Il est probable que cela traduit une réactivité liée au bouleau chez ces enfants et une PR-10 a été repérée dans le pois au moins avec un anticorps anti-Bet v 1, faute de preuve directe d’IgE-réactivité .

Mais d’autres protéines sont IgE-réactives et le peu de réactions cliniques avec le pois mériterait de trouver une explication à travers des travaux complémentaires.

On connaît une profiline, une viciline (Pis s 1) et une conviciline (Pis s 2), cette dernière appartenant à une catégorie proche des vicilines. Toutes ces protéines ont montré leur capacité à se lier aux IgE des patients.


Le pois est porteur de réactivité CCD également .

Pois chiche

Les études concernant le pois chiche ont principalement été conduites dans des pays où la consommation de cette Fabacée est importante (ex. Inde, Espagne).



Si les cas d’allergie alimentaires y sont fréquents, avec volontiers une allergie à d’autres légumineuses , on a aussi décrit des cas d’asthme par inhalation de vapeurs de cuisson de pois chiches . De fait, l’IgE-réactivité du pois chiche résiste bien à la cuisson (ébullition) .


On connaît très mal les protéines IgE-réactives du pois chiche :

  • une viciline est probable car Len c 1 (lentille) est capable d’inhiber la réactivité pour un extrait de pois chiche
  • une lectine pourrait être IgE-réactive  : cette protéine de 20-23 kD est parfois présentée comme une 2S albumine mais, si cela est vrai en vitesse de sédimentation, cette protéine n’est pas un homologue des 2S classiques comme Ara h 2 (arachide) ou Ber e 1 (noix du Brésil). Elle est en fait affiliée aux protéines « PA2 » qui possèdent une activité lectine de type hémopexine. On en trouve également dans le pois, le haricot mungo et la vesce commune, par exemple .


Une « allergie croisée » entre pois chiche et latex a été invoquée chez un enfant atteint de spina bifida et présentant un syndrome oral avec les pois chiches .

En fait, la preuve d’une réactivité croisée n’a pas été apportée dans ce travail, le pois chiche s’avérant ne pouvoir inhiber significativement le latex in vitro.

Pousses de soja

Contrairement aux apparences, les pousses de soja ne proviennent pas du vrai soja (Glycine max) mais d’une autre Fabacée, Vigna radiata.

A noter qu’il est possible de produire des « pousses » avec d’autres Fabacées, comme le vrai soja ou la luzerne, et que leur allergénicité est très probable .

D’ailleurs l’espèce Vigna mungo, très proche de Vigna radiata et consommée non germée en Inde (« urad dal »), est à l’origine de réactions allergiques .

Les anglo-saxons parlent de « mungbean seedlings », ce qui est plus conforme au nom de la graine qui, germée, donne les pousses de soja : le haricot mungo.


La réactivité aux pousses de soja est très souvent associée à une allergie au vrai soja car l’allergène principal de Vigna radiata, Vig r 1, est proche de Gly m 4, la PR-10 trouvée dans le soja (74% d’identité).

En fait, ces deux protéines ont une IgE-réactivité induite par une même sensibilisation, à savoir la pollinose au bouleau. Cela survient alors que l’homologie entre Vig r 1 et Bet v 1 est somme toute modérée (43%), ce qui rappelle les limites de la prédiction bio-informatique des réactivités croisées.

Vig r 1 n’est pas la seule protéine IgE-réactive dans les pousses de soja, même si sa concentration augmente beaucoup au moment de la germination.

On peut s’attendre à trouver des protéines similaires à celles d’autres graines de Fabacées. On a identifié une profiline et Mittag remarque une bande de 35kD positive chez la plupart des patients .

On a caractérisé aussi une LTP et une viciline sans que la preuve d’une IgE-réactivité n’ait été étudiée jusqu’à présent pour ces protéines.


Vigna radiata contient, comme d’autres Fabacées, des facteurs anti-nutritionnels qui sont inactivés par la chaleur et qui justifient donc une cuisson avant consommation (lectines, inhibiteurs de trypsine,..).

Bien que les protéines PR-10 soient réputées ne pas résister à la chaleur, les conditions environnantes dans le produit chauffé doivent jouer un rôle car, s’agissant des pousses de soja, une étude a montré que plus de la moitié des patients positifs en TC avant cuisson le restaient après cuisson .

Fabacées diverses

Fenugrec

Le fenugrec est utilisé comme épice (ex. curry). Il est à l’origine de rares observations d’allergie alimentaire où la responsabilité d’une sensibilisation à l’arachide est évoquée .

Le fenugrec a aussi provoqué des réactions par inhalation ou large contact cutané .

Fève et vesces

Ces graines proviennent de plantes du genre Vicia.

La fève, Vicia faba, a très rarement fait l’objet d’observations publiées d’allergie alimentaire. Dans un cas il s’agissait de farine de fève en tant qu’ingrédient dans du pain industriel . Dans un autre cas, la patiente avait présenté une réaction anaphylactique après ingestion de fèves fraîches et bouillies . Un asthme professionnel a été décrit aussi (cf. Bessot ).

Normalement les autres vesces sont des graines données aux animaux (Vicia sativa, Vicia ervilla).

Gesses

Les gesses sont également des plantes destinées aux animaux. La gesse commune, Lathyrus sativus, contient un agent neurotoxique qui peut aboutir à un trouble nommé lathyrisme.

Il a été observé des cas de rhinite/asthme professionnel avec la farine de gesse commune dans un contexte très particulier : la fabrique de lames de parquets .

[1] - Malandain H. Non-peptide components of proteins and allergenicity: should we generalize the hapten concept? EAACI 22th Congress, Paris, 7-11 June, 2003, Poster n°285
Background: Why are some proteins allergens is still an unresolved question. Besides proteolytic and/or pro-inflammatory activities, recent data have shown that major allergens like Bet v 1 or Der p 2 bind lipid ligands. Objective : To check if such biochemical and/or structural features are a frequent finding among proteins known as allergens. Methods: Published literature and protein databases were studied. Airborne allergens were chosen because they directly reach immune system in native form, eg. not affected by digestion. Results: We found a wide array of allergens displaying a lipophilic binding structure, such as a groove or a tunnel : lipid transfer proteins, lipocalins, Bet v 1 family, Bla g 2, mite groups 2, 13, and 14, pollen 2 EF-hand Ca binding proteins, ... Noticeably, most of these allergens were not glycosylated. Conclusion: These observations suggest that protein-bound lipid moieties could play a role in conjunction with the peptide backbone for initiating a Th-2 response. One might hypothesize that surface receptors of surveillance immune cells, like Toll receptors, are programmed to detect non-self proteins as a whole : lipid + peptide, carbohydrate + peptide, and even metal ion + peptide. This hypothesis could also match the hapten concept of a self structure bearing a non-self ligand. Globally, such a mechanism would be similar to innate defense systems adopted by many eukaryotes, like hypersensitive reactions observed in plants in response to elicitins.
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[2] - Ayuso R, Vereda A, Han Y, Bardina L, Sampson HA. IgE-binding Profiles of Legume-allergic Children to Dal Proteins. J Allergy Clin Immunol 2008;121:S242
RATIONALE: Legumes are a frequent cause of allergic reactions in the pediatric population. The Indian term dal includes a variety of lentils and other legumes (toor dal, mung dal, urad dal, chana dal). Since such legumes are also frequently consumed in western countries, their potential allergenicity requires evaluation. METHODS: Extracts were prepared from yellow toor dal (Cajanus cajan), mung dal (Vigna radiata), urad dal (Vigna mungo), chana dal (Cicer aretinum), lentil (Lens culinaris) soybean (Glycine max) and peanut (Arachis hypogea). Serum from a pediatric patient with prior reactions to toor dal and other legumes but who tolerated urad dal, and from 4 legume-allergic children with elevated IgE to legumes (40-100 kUA/L; ImmunoCAP) were used for immunoblot and ELISA studies. RESULTS: IgE binding profiles of the different dal crops showed IgEbinding proteins of 98, 62, 49, 38, 28, 20 and 14 kD. Important inhibition of IgE reactivity to most legumes was obtained with lentil, chana dal, and toor dal. Less inhibition was observed with mung dal or urad dal suggesting non-crossreactive IgE-binding proteins. ELISA-inhibition of toor dal was highest with European lentil (90%), chana dal (85%) and peanut (82%) and lowest with urad dal (55%). Immunoblot inhibition showed cross-reactive proteins at 62, 49, 17 and 14kD among lentil, toor and urad dal. A 38 kD protein was lentil- and toor dal-specific. CONCLUSIONS: High cross-reactivity among chana dal, lentil and toor dal, and limited crossreactivity with mung dal and urad dal may have clinical implications for recommendations of food avoidance in legumeallergic children.
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[3] - 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.
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[4] - García González MC, González Mendiola MR, Prieto Montaño P, Muñoz Martín T, Martín C, de la Hoz Caballer B. Asthma and rhinoconjunctivitis induced by potato and green bean aeroallergens. EAACI 21th Congress, Naples, 1-5 June, 2002, Poster n°280
Background: Vegetables can cause allergic symptoms after being ingested. Less frecuently their allergens act as inhalants inducing rhinoconjunctivitis and asthmatic attacks. The most affected patients are housewives whose problems usually start at the age of 30-40 years. They often have a previous history of pollinic rhinoconjunctivitis or asthma and cooked vegetables do not cause them any symptom. Method: We describe the cases of two pollinic women whith a history of rhinoconjunctivitis and asthma due to vegetable aeroallergens. The first one referred these symptoms when peeling potatoes, and the second one when peeling green beans. Both could eat the vegetable implicated if it had been cooked before. Several test were performed including skin prick test ( with raw and boiled potatoe/green bean and with some extracts prepared in our laboratory), specific IgE antibodies, rub-test, bronchial provocation, challenge test and SDS-PAGE-immunoblotting. Results: First patient: The result of the skin prick test with raw potatoe was positive, but negative with boiled potatoe. The specific Ig E was 5 KU/l for peel and 9 KU/l for pulp. Rub test with the potatoes on the patient's forearm elicited wheals and pruritus whithin 5 minutes. Challenge test was positive (rhinoconjunctivitis), and SDS-PAGE-immunoblotting showed several peptidic bands of 100,47,38.5,33.5,29 and 26KDa. Second patient: The result of the skin prick test with raw green bean was positive, but negative with boiled green bean. The specific Ig E was 13 KU/l . Bronchial provocation was positive and immunoblotting showed peptidic bands of 78,72,50.4 and 17.3 KDa. Conclusion: We present two cases in which vegetable aeroallergens cause symptoms of rhinoconjunctivitis and asthma in two patients who however are able to eat the vegetable involved without any problem. Most of the few patients described presenting this syndrome present previous pollinic symptoms and get worse in the pollinic season when peeling the implicated vegetables. These facts lead us to think about a common allergen between pollens and the vegetables implicated. According to the results of the immunoblotting peptidic bands, it seems to be a high molecular weight protein. The allergen is still unknown.
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[5] - Daroca P, Crespo JF, Reano M, James JM, Lopez-Rubio A, Rodriguez J. Asthma and rhinitis induced by exposure to raw green beans and chards. Ann Allergy Asthma Immunol 2000;85:215-218
Abstract BACKGROUND: Although the vast majority of IgE-mediated allergic reactions to foods occurs through ingestion, a few cases of unexpected allergic reactions to foods may occur through the exposure to airborne food allergen particles. METHODS: Case reports. Skin prick tests and serum-specific IgE (CAP-FEIA) were used to identify specific IgE antibodies. Bronchial provocation tests were performed to determine the clinical relevance of inhaled exposure to raw and cooked green beans and raw chards. After demonstrating specific reactivity to them, SDS- PAGE and immunoblotting of raw and cooked green beans were carried out to identify relevant antigens. RESULTS: Three women developed bronchial asthma and rhinitis after exposure to raw green beans, and one of them also when exposed to raw chards. All women tolerated ingestion of green beans. Patients reported multiple episodes while handling these vegetables for cooking activities. Allergy to green beans and chards was demonstrated by skin testing and serum-specific Ig E. Bronchial challenge test with these allergens showed positive responses to raw, but not cooked, green beans and chards. Oral food challenges with green beans (raw and cooked) and chards were negative in all patients. In order to further characterize the allergenic components of these extracts, SDS-PAGE and electroblotting studies were also performed. Immunoblots of raw and cooked green beans extract showed two IgE- binding bands with apparent molecular weights of 41.1 and 70.6 kD. Interestingly, a 47-kD IgE-binding protein was detected only in raw green bean extracts. CONCLUSIONS: We report three patients who developed asthma and rhinitis caused by exposure to raw, but not to cooked, green beans and chards in a non-occupational environment. Only minor differences of IgE reactivity between nitrocellulose-blotted raw and boiled green bean extract were found
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[6] - Martinez Alonso J, Callejo Melgosa A, Fuentes Gonzalo MJ, Martin Garcia C. Angioedema induced by inhalation of vapours from cooked white bean in a child. Allergol Immunopathol (Madr) 2005;33:228-230
Background: There are few references of allergic reaction to beans in childhood. We report the case of a seven years old boy who suffered from angioedema associated to inhalation of vapours from cooked white bean. Methods: skin prick tests (SPT) were performed by prick-by-prick with cooked white bean and legumes. It was also determined total IgE and specific IgE antibodies to bean and legumes with the use of the CAP enzymo-immunoassay. Subsequently, a oral challenge test was carried out with white bean. RESULTS: The prick-by-prick with white bean was positive in our case, and negative in ten controls patients. Specific IgE in patient serum, assayed by CAP was positive for white bean and green bean. The patient developed angioedema after ingestion cooked white bean. CONCLUSION: we demonstrated a type I hypersensitivity to white bean in a seven years old child by SPT, specific IgE antibodies and challenge test.
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[7] - Guillen G, Lopez-Sanchez LM, Roman-Roque CS, Sanchez F, Villanueva MA. Biochemical characterization of profilin from seeds of Phaseolus vulgaris. Plant Cell Physiol 2001;42:54-62
The isoform composition of the 14.4 kDa profilin polypeptide was analyzed in seeds, leaves, flowers, roots and root-nodules from Phaseolus vulgaris L. Isoforms of pIs approximately 4.4-5 were present in all the tissues analyzed. The biochemical features of the protein present in seed tissue were determined. Seed profilin bound to Phenyl-Sepharose under low salt conditions which suggested a hydrophobic interaction; however, it was not associated with microsomal membranes nor it partitioned as a hydrophobic protein in Triton X-114. Fractions eluting from poly-L-proline or Phenyl-Sepharose columns contained well detectable amounts of profilin but no actin, suggesting that most of the protein was not present as profilactin in the seed. However, seed profilin appeared to be in some kind of complex since several molecular weight species were observed on native gels. In addition, profilin was found preferentially in the embryo axis and light microscopic immunolocalization showed a cytoplasmic distribution in this tissue.
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[8] - Sanchez-Monge R, Blanco C, Perales AD, Collada C, Carrillo T, Aragoncillo C, et al. Class I chitinases, the panallergens responsible for the latex-fruit syndrome, are induced by ethylene treatment and inactivated by heating. J Allergy Clin Immunol 2000;106:190-195
Class I chitinases have been identified as the major panallergens in fruits associated with the latex-fruit syndrome, such as avocado, banana, and chestnut. However, other plant foods containing these enzymes have not been related to this syndrome. OBJECTIVE: We sought out class I chitinases in the green bean, a legume that is known to express chitinases but is not associated with latex allergy, and examined whether the content or allergenic activity of chitinases can be modified by physical or chemical treatments. METHODS: IgE-binding proteins in untreated bean samples, as well as in ethylene- and heat-treated samples, were detected by using a pool of sera from patients with latex-fruit allergy. Putative allergens were purified by cation-exchange chromatography and characterized by N-terminal sequencing, enzymatic activity assays, immunodetection with sera and antichitinase antibodies, and immunoblot inhibition tests. Skin prick tests with untreated and heated purified allergens were also carried out. RESULTS: An IgE-binding protein of 32 kd that was also recognized by antichitinase antibodies was detected in green bean extracts. This reactive component was strongly induced by ethylene treatment. The protein, designated PvChI, was identified as a class I chitinase closely related to the major avocado allergen Prs a 1. Immunoblot inhibition assays demonstrated cross-reactivity between both allergens. Purified PvChI induced positive skin prick test responses in 7 of 8 patients with latex-fruit allergy. Heat treatment of both Prs a 1 and PvChI produced a full loss of their allergenic capacities both in vitro and in vivo. No IgE-binding component was detected in the white mature bean in which the main isolated 32-kd protein corresponded to a nonreactive phytohemagglutinin. CONCLUSIONS: Ethylene treatment induces the expression of plant class I chitinases. The allergenic activity of plant class I chitinases seems to be lost by heating. This fact could explain why plant foods containing these putative allergens that are consumed after cooking are not usually associated with the latex-fruit syndrome.
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[9] - Ye XY, Ng TB. First chromatographic isolation of an antifungal thaumatin-like protein from French bean legumes and demonstration of its antifungal activity. Biochem Biophys Res Commun 1999;263:130-134
A protein, with a molecular weight of 20 kDa, and an N-terminal sequence analogous to those of thaumatin-like proteins (TLPs) and thaumatins, was first isolated from the legume of the French bean Phaseolus vulgaris cv Kentucky wonder using a simple procedure involving affinity and ion exchange chromatography. The protein was adsorbed on both CM-Sepharose and Affi-gel Blue Gel. It was the first leguminous TLP-like protein demonstrated to exert antifungal activity against Fusarium oxysporum, Pleurotus ostreatus, and Coprinus comatus but not against Rhizoctonia solani
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[10] - García González MC, González Mendiola MR, Prieto Montaño P, Muñoz Martín T, Martín C, de la Hoz Caballer B. Asthma and rhinoconjunctivitis induced by potato and green bean aeroallergens. EAACI 21th Congress, Naples, 1-5 June, 2002, Poster n°280
Background: Vegetables can cause allergic symptoms after being ingested. Less frecuently their allergens act as inhalants inducing rhinoconjunctivitis and asthmatic attacks. The most affected patients are housewives whose problems usually start at the age of 30-40 years. They often have a previous history of pollinic rhinoconjunctivitis or asthma and cooked vegetables do not cause them any symptom. Method: We describe the cases of two pollinic women whith a history of rhinoconjunctivitis and asthma due to vegetable aeroallergens. The first one referred these symptoms when peeling potatoes, and the second one when peeling green beans. Both could eat the vegetable implicated if it had been cooked before. Several test were performed including skin prick test ( with raw and boiled potatoe/green bean and with some extracts prepared in our laboratory), specific IgE antibodies, rub-test, bronchial provocation, challenge test and SDS-PAGE-immunoblotting. Results: First patient: The result of the skin prick test with raw potatoe was positive, but negative with boiled potatoe. The specific Ig E was 5 KU/l for peel and 9 KU/l for pulp. Rub test with the potatoes on the patient's forearm elicited wheals and pruritus whithin 5 minutes. Challenge test was positive (rhinoconjunctivitis), and SDS-PAGE-immunoblotting showed several peptidic bands of 100,47,38.5,33.5,29 and 26KDa. Second patient: The result of the skin prick test with raw green bean was positive, but negative with boiled green bean. The specific Ig E was 13 KU/l . Bronchial provocation was positive and immunoblotting showed peptidic bands of 78,72,50.4 and 17.3 KDa. Conclusion: We present two cases in which vegetable aeroallergens cause symptoms of rhinoconjunctivitis and asthma in two patients who however are able to eat the vegetable involved without any problem. Most of the few patients described presenting this syndrome present previous pollinic symptoms and get worse in the pollinic season when peeling the implicated vegetables. These facts lead us to think about a common allergen between pollens and the vegetables implicated. According to the results of the immunoblotting peptidic bands, it seems to be a high molecular weight protein. The allergen is still unknown.
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[11] - Daroca P, Crespo JF, Reano M, James JM, Lopez-Rubio A, Rodriguez J. Asthma and rhinitis induced by exposure to raw green beans and chards. Ann Allergy Asthma Immunol 2000;85:215-218
Abstract BACKGROUND: Although the vast majority of IgE-mediated allergic reactions to foods occurs through ingestion, a few cases of unexpected allergic reactions to foods may occur through the exposure to airborne food allergen particles. METHODS: Case reports. Skin prick tests and serum-specific IgE (CAP-FEIA) were used to identify specific IgE antibodies. Bronchial provocation tests were performed to determine the clinical relevance of inhaled exposure to raw and cooked green beans and raw chards. After demonstrating specific reactivity to them, SDS- PAGE and immunoblotting of raw and cooked green beans were carried out to identify relevant antigens. RESULTS: Three women developed bronchial asthma and rhinitis after exposure to raw green beans, and one of them also when exposed to raw chards. All women tolerated ingestion of green beans. Patients reported multiple episodes while handling these vegetables for cooking activities. Allergy to green beans and chards was demonstrated by skin testing and serum-specific Ig E. Bronchial challenge test with these allergens showed positive responses to raw, but not cooked, green beans and chards. Oral food challenges with green beans (raw and cooked) and chards were negative in all patients. In order to further characterize the allergenic components of these extracts, SDS-PAGE and electroblotting studies were also performed. Immunoblots of raw and cooked green beans extract showed two IgE- binding bands with apparent molecular weights of 41.1 and 70.6 kD. Interestingly, a 47-kD IgE-binding protein was detected only in raw green bean extracts. CONCLUSIONS: We report three patients who developed asthma and rhinitis caused by exposure to raw, but not to cooked, green beans and chards in a non-occupational environment. Only minor differences of IgE reactivity between nitrocellulose-blotted raw and boiled green bean extract were found
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[12] - Sanchez-Monge R, Blanco C, Perales AD, Collada C, Carrillo T, Aragoncillo C, et al. Class I chitinases, the panallergens responsible for the latex-fruit syndrome, are induced by ethylene treatment and inactivated by heating. J Allergy Clin Immunol 2000;106:190-195
Class I chitinases have been identified as the major panallergens in fruits associated with the latex-fruit syndrome, such as avocado, banana, and chestnut. However, other plant foods containing these enzymes have not been related to this syndrome. OBJECTIVE: We sought out class I chitinases in the green bean, a legume that is known to express chitinases but is not associated with latex allergy, and examined whether the content or allergenic activity of chitinases can be modified by physical or chemical treatments. METHODS: IgE-binding proteins in untreated bean samples, as well as in ethylene- and heat-treated samples, were detected by using a pool of sera from patients with latex-fruit allergy. Putative allergens were purified by cation-exchange chromatography and characterized by N-terminal sequencing, enzymatic activity assays, immunodetection with sera and antichitinase antibodies, and immunoblot inhibition tests. Skin prick tests with untreated and heated purified allergens were also carried out. RESULTS: An IgE-binding protein of 32 kd that was also recognized by antichitinase antibodies was detected in green bean extracts. This reactive component was strongly induced by ethylene treatment. The protein, designated PvChI, was identified as a class I chitinase closely related to the major avocado allergen Prs a 1. Immunoblot inhibition assays demonstrated cross-reactivity between both allergens. Purified PvChI induced positive skin prick test responses in 7 of 8 patients with latex-fruit allergy. Heat treatment of both Prs a 1 and PvChI produced a full loss of their allergenic capacities both in vitro and in vivo. No IgE-binding component was detected in the white mature bean in which the main isolated 32-kd protein corresponded to a nonreactive phytohemagglutinin. CONCLUSIONS: Ethylene treatment induces the expression of plant class I chitinases. The allergenic activity of plant class I chitinases seems to be lost by heating. This fact could explain why plant foods containing these putative allergens that are consumed after cooking are not usually associated with the latex-fruit syndrome.
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[13] - García González MC, de la Hoz Caballer B, Cerecedo I, Carnés J, Fernández-Caldas E. Collection and identification of aerosolised proteins during the boiling process of vegetables and legumes. EAACI 23th Congress, Amsterdam, 12-16 June, 2004, Poster n°964
Background: Although most food allergic reactions are caused by ingestion, IgE mediated respiratory reactions caused by the inhalation of vapours from boiling vegetables and legumes have been described. In these cases, IgE-mediated mechanisms have been suggested, but the responsible allergens have not been fully characterised. The aims of this study were to develop a method to collect allergens from vegetable vapours and to investigate if these proteins are capable of binding specific IgE. Material and Methods: Three types of extracts (A, B and C) were prepared with potato, carrot, Swiss chard and green beans. Extracts A (raw), extracted in PBS 0.01 M (1:2 w/v); Extracts B (boiled), cooked in PBS 0.01 M (1:2 w/v) for 30 minutes; and Extract C (lyophilised vapours), collected during the boiling process. The vapours were passed through a refrigeration column and the drops collected and freeze-dried. Raw and boiled extracts were extracted for 4 hours, dialyzed against bidistilled water in a 3.5 kDa cut-off dialysis membrane, frozen and freeze-dried. Antigenic profile of the extracts was evaluated by SDS-PAGE and analysed by scanning densitometry. Results: Several bands were visualised in the raw extracts in the molecular weight range of 9 to 90 KDa in the 4 analysed raw extracts. The protein profile of the boiled extracts showed low molecular weights bands, which could correspond to denatured proteins from the raw extracts. The total yield of vapour proteins obtained in each extract was 0,64% in potato, 0,48% in carrot, 0,4% in Swiss chard and 0,352% in green bean. Vapours extracts (Extracts C) showed several prominent bands; 1 band in potatoes (35 kDa), 2 bands in carrot vapours (60 and a 66 kDa), 2 bands in the Swiss chard vapours (62 and 71 kDa) and 1 band (38 kDa) in green bean vapours. Almost all these molecules had a corresponding band in their respective raw extracts. Faint IgE binding was observed to green bean vapours. Conclusions: We have developed a reproducible method to collect vapours during the boiling process of foods. The results suggest that proteins are present in vapour drops aerosolised during the boiling process. More studies are necessary to elucidate if these proteins are capable of inducing respiratory symptoms and to what extent these proteins are denatured and capable of binding specific IgE.
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[14] - Mateo Borrega MB, Sánchez Fernández C, Losada Cosmes E, de la Hoz Caballer B, Sánchez Cano M. Skin tests with pulses: a comparison of the results with natural foodstuffs and extracts, and effects of heating. Alergol Immunol Clin 2001;16:271-278
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[15] - Dube M, Schäfer C, Neidhart S, Carle R. Texturisation and modification of vegetable proteins for food applications using microbial transglutaminase. Eur Food Res Technol 2007;225:287-299
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[16] - Malandain H. Transglutaminases : a meeting point for wheat allergy, celiac disease, and food safety. Eur Ann Allergy Clin Immunol 2005;37:397-403
Wheat is the staple cereal in many countries and its uses in manufactured foods are ever growing due to the technological qualities of gluten proteins. Transglutaminases (TG) are ubiquitous enzymes with many functions. They are able to transform proteins by deamidation and/or transamidation. This last reaction can cross-link proteins together. Intestinal tissue TG has been shown to play an important role in two kinds of immune reactions to wheat: celiac disease and wheat-dependent exercise-induced anaphylaxis. In addition, new epitopes have been suspected in cases of anaphylaxis to wheat isolates, a food ingredient consisting mainly of deamidated gluten proteins. As a microbial TG is included in many food technological processes, its safe use should be checked. This assessment must cover not only the safety of the TG itself but also that of the deamidated/cross-linked proteins generated by this enzyme. This article aims at discussing the possible consequences of using TG in food industry in the light of today knowledge about immune reactions to wheat.
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[17] - Mariniello L, Giosafatto CV, Pierro PD, Sorrentino A, Porta R. Synthesis and Resistance to in Vitro Proteolysis of Transglutaminase Cross-Linked Phaseolin, the Major Storage Protein from Phaseolus vulgaris. J Agric Food Chem 2007;55:4717-4721
The ability of phaseolin to act as an acyl donor and acceptor substrate of transglutaminase was studied by using an enzyme isolated from Streptoverticillium mobarense. Phaseolin, a trimeric storage protein from Phaseolus vulgaris L., was shown to possess both glutamine and lysine residues reactive for the enzyme. The extent of transglutaminase-catalyzed cross-linking has been studied in function of both incubation time and enzyme concentration. Native- and SDS-PAGE demonstrated that phaseolin is intra- and intermolecularly cross-linked by transglutaminase and gives rise to different polymers as well as to modified forms of the protein having a similar molecular weight but lower Stokes radius if compared to unmodified phaseolin. Cross-linked phaseolin was found to be more resistant to proteolytic cleavage than the unmodified counterpart, as demonstrated by in vitro trypsin and pepsin digestion experiments. This behavior could suggest novel possible uses of the transglutaminase-modified phaseolin. Keywords: Enzymatic modification; phaseolin; proteolysis resistance; transglutaminase.
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[18] - Ibanez Sandin D, Martinez San Ireneo M, Maranon Lizana F, Fernández-Caldas E, Alonso Lebrero E, Laso Borrego MT. Specific IgE determinations to crude and boiled lentil (Lens culinaris) extracts in lentil-sensitive children and controls. Allergy 1999;54:1209-1214
The aims of this study were to evaluate the allergenicity of boiled and crude lentil extracts and to compare specific IgE binding in tolerant and nontolerant lentil-allergic children. METHODS: Thirty-eight children were studied and divided into three groups. Group I comprised 24 children with a positive open oral challenge, or a convincing history of anaphylaxis after the ingestion of lentils; group II comprised nine children with a history of allergic reactions in the past, but currently tolerant of lentils; and group III comprised five children allergic to other legumes, but always tolerant of the ingestion of lentils. Specific IgE determinations and ELISA inhibitions were performed with the crude and boiled lentil extracts. The allergenic profile of both extracts was evaluated by SDS-PAGE and immunoblot. RESULTS: Mean specific IgE levels in group I were significantly higher than in groups II and III. The heating process caused a significant decrease in specific IgE binding. However, IgE-inhibition studies showed that the boiled lentil extract had a greater inhibitory capacity than the crude extract. Immunoblots revealed no important differences in IgE-binding patterns between the two extracts. Multiple allergens were detected in a wide range of molecular masses. CONCLUSIONS: Boiled lentil extracts maintain strong allergenicity. Patients who have developed tolerance of lentil ingestion have lower specific IgE levels than symptomatic patients.
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[19] - Martinez San Ireneo M, Ibanez Sandin MD, Fernández-Caldas E, Marañón F, Munoz Martinez MC, Laso Borrego MT. The diagnostic value of crude or boiled extracts to identify tolerant versus nontolerant lentil-sensitive children. Ann Allergy Asthma Immunol 2001;86:686-690
OBJECTIVE: The aim of this study was to compare two types of lentil extracts for use in skin prick tests for the diagnosis of lentil clinical allergy . METHODS: Thirty-six patients with a history of allergic reactions after the ingestion of lentils were skin tested with two types of lentil extracts at 0.05, 0.5, 5, and 10 mg/mL. Both extracts were extracted at 40 degrees C and afterward, one of them was boiled for 15 minutes. Thirty-three of these patients underwent oral challenges with lentils and three had a convincing recent history of lentil anaphylaxis . RESULTS: Twenty patients had a positive oral challenge; 13 were negative. Skin prick tests performed with the boiled extract at 0.5 and 5 mg/mL were positive in 96% and 100% of patients with positive food challenge, and in 31% and 85% of those with negative food challenge, respectively; positive skin test results were similar in both groups using the crude extract. Mean wheal sizes using the boiled extract at 0.5, 5, and 10 mg/mL were significantly greater in patients with a positive oral challenge than in those with a negative one (4.9, 6.8, and 7.4 mm versus 1.9, 3.5, and 5.1 mm, respectively; P < 0.05) These mean values were not statistically different using the crude extract . CONCLUSIONS: These data suggest that lentil extracts for the diagnosis of lentil hypersensitivity should be heated, since boiled extracts, used at a concentration of 0.5 or 5 mg/mL, best identify clinically sensitive individuals.
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[20] - Sanchez-Monge R, Pascual CY, Diaz-Perales A, Fernandez-Crespo J, Martin-Esteban M, Salcedo G. Isolation and characterization of relevant allergens from boiled lentils. J Allergy Clin Immunol 2000;106:955-961
BACKGROUND: Lentils seem to be the most common legume implicated in pediatric allergic patients in the Mediterranean area. However, no lentil allergen has been isolated and characterized. OBJECTIVE: We sought to purify and characterize relevant IgE-binding proteins from boiled lentil extracts. METHODS: IgE-binding proteins from crude and boiled lentil extracts were detected with a pool of sera from patients with lentil allergy. Allergens were isolated by gel-filtration chromatography followed by cation- and anion-exchange chromatography or by reverse-phase HPLC. Their characterization included N-terminal amino acid sequencing, complex asparagine-linked glycan detection, specific IgE immunodetection with 22 individual sera from allergic patients, and immunoblot and CAP inhibition assays. RESULTS: Heat treatment of lentils produced substantial changes in the SDS-PAGE patterns of whole extracts, mainly a strong increase of 12- to 16-kd bands and a decrease of 25- to 45-kd components. Major IgE-binding proteins from the boiled lentil extract were located in the 12- to 16-kd and 45- to 70-kd ranges. Two allergens of 16 kd, proteins L1 and L2, and another one of 12 kd, protein L3, were purified. N-terminal sequencing indicated that all 3 were related and allowed their identification as gamma-vicilin subunits. Protein L1 was recognized by 68% of the individual sera tested and inhibited 64% of the IgE binding by commercial lentil CAPs. A second type of allergen of 66 kd, named protein H, was also isolated and identified as a seed-specific biotinylated protein. Protein H reacted with 41% of the individual sera and produced 45% inhibition in CAP inhibition assays. CONCLUSIONS: Two different types of allergens have been identified in boiled lentils. Those of 12 to 16 kd, called Len c 1, correspond to gamma-vicilin subunits, and those of 66 kd, designated Len c 2, correspond to seed-specific biotinylated protein. Homology with proteins from other legume species can explain potential cross-reactions among these foods.
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[21] - Martin JA, Compaired JA, de la Hoz B, Quirce S, Alonso MD, Igea JM, et al. Bronchial asthma induced by chick pea and lentil. Allergy 1992;47:185-187
Allergic reactions to legumes through inhalation have rarely been described. We report the case of a 20-year-old man who experienced asthmatic attacks when exposed to the steam from cooking either chick pea or lentil. Type I hypersensitivity to the antigens in these legumes was demonstrated by means of immediate skin reactivity, histamine release tests, RAST and RAST inhibition. Specific bronchial challenges with the heated (75 degrees for 30 min) extracts of chick pea and lentil elicited isolated immediate responses.
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[22] - López-Torrejón G, Salcedo G, Martín-Esteban M, Díaz-Perales A, Pascual CY, Sánchez-Monge R. Len c 1, a major allergen and vicilin from lentil seeds: Protein isolation and cDNA cloning. J Allergy Clin Immunol 2003;112:1208-1215
BACKGROUND: Lentils are among the main plant foods causing allergic reactions in pediatric patients in the Mediterranean area and in many Asian communities. However, very few reports have been devoted to identifying lentil allergens. Seed storage proteins of the vicilin family have been characterized as major allergens in several seed legumes and tree nuts . OBJECTIVE: We sought to evaluate the role of lentil vicilins as food allergens . METHODS: A serum pool and individual sera from 22 patients with lentil allergy were used in different IgE-binding assays. Mature lentil vicilin was isolated by means of cation-exchange chromatography, followed by reverse-phase HPLC, and characterized by means of N-terminal amino acid sequencing, matrix-assisted laser desorption/ionization mass spectrometry (MALDI) analysis, complex asparagine-linked glycan detection, specific IgE immunodetection with individual sera, and ELISA inhibition assays. Complete cDNAs encoding lentil vicilin variants were isolated by means of PCR with primers based on the amino acid sequence of the allergen . RESULTS: A major IgE-binding component of approximately 50 kd was detected in lentil extracts. This component was isolated and characterized, showing a single N-terminal amino acid sequence homologous to those of legume vicilins and a broad peak (maximum at 48613 d) in MALDI analysis. The purified allergen was recognized by 77% (17/22) of the individual sera from patients with lentil allergy and reached up to 65% inhibition of the IgE binding to the crude lentil extract. The allergen showed 3 isoforms varying in their degree of N-glycosylation. Two cDNA clones encoding different allergen variants were isolated. The amino acid sequences deduced from both clones (415 and 418 residues; 47.4 and 47.8 kd) showed greater than 50% identity with major peanut (Ara h 1) and soybean (conglutinin subunits) allergens belonging to the vicilin family. Furthermore, these sequences included those of the previously characterized lentil allergen Len c 1.02 (108 amino acid residues of the C-terminal domain) and those of a novel lentil IgE-binding protein of 26 kd . CONCLUSION: The mature 48-kd lentil vicilin, designated Len c 1.01, is a major allergen. Two of its processing fragments, corresponding to subunits of 12 to 16 kd (previously named Len c 1) and 26 kd, are also relevant lentil IgE-binding proteins. The sequence homology of Len c 1.01 to those of major allergens from peanut, soybean, walnut, and cashew can help to investigate potential cross-reactions among these plant foods.
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[23] - Pereira MJ, Pascual CY, Belver MT, Diaz Pena JM, Sanchez Monge R, Gomez Palacios A, et al. Lentil allergen and cross-reactivity with chickpeas, peanuts and soy. EAACI 21th Congress, Naples, 1-5 June, 2002, Poster n°231
Background: Legumes are among the most common antigens causing allergenic reaction in children. Lentils seem to be the most common legume implicated in pediatric allergic patients in the Mediterranean area. Legumes had demonstrated great degree of immunological cross-reactivity. Homology with protein from other legumes species can explain potential cross-reactions among these foods. Objetive: The aim of this study is to assess the posible cross-reactivity between peanut, soy, lentil and chickpea, and the role of the two allergens from lentils in the cross-sensitization. Methods: We included 22 patients with histories of adverse reactions to lentils. All of them had positive specific IgE antibodies measured by CAP-FEIA to the four legumes. Individual immunoblot patterns for each legume were performed. CAP-inhibition with the four heated legumes in liquid phase was assessed. CAP-inhibition with Len c1 and Len c2 in liquid phase to the four legumes in solid phase, also was performed. Results: The CAP-FEIA inhibition was performed to assess the amount of inhibition by boiled lentils, boiled chickpeas, roasted peanuts and boiled soybeans in liquid phase to commercial lentils in CAP system as solid phase. The CAP-inhibition with Len c1 and Len c2 in liquid phase to the four legumes in solid phase (with doses of 100 ngr, 1 ngr, 10 pgr and 0.1 pgr/ml) are described at maximal dose of 100 ngr. Len c1 achieve a higher CAP- inhibition than Len c2 in every legume but soy in wich both only inhibited 30%. We performed as control, an auto-inhibition of lentils with boiled lentils at the same concentration that the two allergens: Len c1 and Len c2. Len c1 inhibit more than 50% of specific binding in lentils and peanuts at 100 ngr/ml. Len c2 also inhibit lentil and peanut in a very similar amount, but minor than Len c1. Both lentils allergens release histamine from an allergic patients basophiles. Conclusions: The allergenicity in legumes is mostly related to allergens from their seed storage proteins. Vicilins from this group of proteins could be an important common allergen in legumes clinical allergy and may be it could be used as indicator in diagnosis of this type of sensitization.
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[24] - Pascual CY, Pastor SS, De Miguel S, Sanchez GG, Crespo JF, Salcedo G, et al. Cross-reactivity to legumes in mediterranean pediatric patients. J Allergy Clin Immunol 2000;105(1 part 2):S133
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[25] - Bjerremann Jensen L, Andersen M, Skov P, Poulsen L, Bindslev-Jensen C. Investigating the common allergic reactivity within the legume botanical family using skin prick test (SPT), specific immunoglobulin E (IgE) and histamine release (HR). EAACI 25th Congress, Vienna, 10-14 June, 2006, Poster n°578
Background: The aim of the study was to investigate possible common allergic reactions between peanut and different foods from the legume family. Methods: The legumes investigated were peanut, soybean, lupine, and fresh as well as blanched green pea. Patients were included on the basis of confirmed food allergy to peanut according to EAACI guidelines. SPTs were performed by prick-prick with legumes using as cutoff a wheal with a diameter >3 mm. IgE were determined using the CAP system (Pharmacia, Sweden) with a cutoff of 0.35 kUA/L. HR was performed by direct stimulation of patient basophils with legume extracts and the released histamine was measured spectrofluorometrically (RefLab ApS, Denmark). Results: 72 peanut allergic patients were included. The sensitivity for SPT, IgE and HR was 92-97% for peanut. Testing the other legumes with SPT and HR produced most positives for lupine (45%, 80%) and fresh pea (41%, 91%) followed by soybean (28%, 19%) and blanched pea (16%, 5%), whereas soybean produced most positives in IgE (58%) followed by pea (48%) and lupine (47%). Pairwise comparison between the three diagnostic tests in general gave the highest concordance between SPT and HR. A subpopulation of the patients was challenged with the legumes. The most common reported symptom for reactions related to the various foods was the oral allergy syndrome (OAS). 22 patients were challenged with soybean, and of these 2 were positive. 1 of 6 challenges was positive with lupine whereas 0 of 12 was positive with fresh pea. This suggests that clinical coreactivity between peanut and other legumes is substantially lower than indicated by diagnostic tests. Conclusion: Patients with peanut allergy might in a few cases be clinically allergic to other members of the legume family. However, application of SPT, IgE or HR tests results in an overestimation of the positive reactions.
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[26] - Barkholt V, Jorgensen PB, Sorensen D, Bahrenscheer J, Haikara A, Lemola E, et al. Protein modification by fermentation: effect of fermentation on the potential allergenicity of pea. Allergy 1998;53(46 Suppl.):106-108
The effect of fermentation on components of potential significance for the allergenicity of pea was analyzed. Pea flour was fermented with three lactic acid bacteria, Pediococcus pentosaceus, Lactococcus raffinolactis, and Lactobacillus plantarum, and two fungi, Rhizopus microsporus, var. oligosporus and Geotrichum candidum. Residual antigenicity against antipea antibodies was reduced to 10% by the three lactic acid bacteria and R. microsporus. Reactions to anti-pea profilin and anti-Bet v 1 were still detectable after fermentation. The contents of lectin and pea protease inhibitor were not reduced by the microorganisms.
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[27] - Garcia-Casado G, Sanchez-Monge R, Chrispeels MJ, Armentia A, Salcedo G, Gomez L. Role of complex asparagine-linked glycans in the allergenicity of plant glycoproteins. Glycobiology 1996;6:471-477
Many plant proteins, particularly those found in foods and pollen, are known to act as sensitizing agents in humans upon repeated exposure. Among the cereal flour proteins involved in asthmatic reactions, those members of the alpha-amylase inhibitor family which are glycosylated, polypeptides, BMAI-1, BTAI-CMb*, and WTAI-CM16* are particularly reactive both in vivo and in vitro. We show here that these major glycoprotein allergens carry a single asparagine-linked complex glycan that contains both beta 1-->2 xylose and alpha 1-->3 fucose. Evidence is presented that the xylosyl residue and, to a lesser extent, the fucosyl residue are key IgE-binding epitopes and largely responsible for the allergenicity of these and unrelated proteins from plants and insects. Our results suggest that the involvement of xylose- and fucose-containing complex glycans in allergenic responses may have been underestimated previously these glycans provide a structural basis to help explain the cross-reactivities often observed between pollen, vegetable food, and insect allergens.
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[28] - van Ree R, Aalberse RC. Pollen-vegetable food crossreactivity: serological and clinical relevance of crossreactive IgE. J Clin Immunoassay 1993;16:124-130
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[29] - Patil SP, Niphadkar PV, Bapat MM. Chickpea: a major food allergen in the Indian subcontinent and its clinical and immunochemical correlation. Ann Allergy Asthma Immunol 2001;87:140-145
BACKGROUND: The food allergy pattern of a country is influenced by the foods most commonly consumed. In India, the majority of the population consumes a vegetarian diet made up of pulse (legumes), cereals, and vegetables. In contrast to many western countries, chickpea preparations are consumed in large quantities in India. This study reports for the first time chickpea hypersensitivity reactions diagnosed with in vivo and in vitro tests. METHODS: One thousand four hundred patients visiting allergy clinics were randomly selected for the study. Those patients reporting an allergic reaction on every occasion after eating chickpea were considered history-positive. Modified prick tests were performed with chickpea and other members of the legume family on all these patients. The claims of the history-positive patients were verified with double-blind, placebo-controlled food challenges (DBPCFCs). Proteins in chickpea extracts were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred on nitrocellulose paper. Serum specimens from history-positive patients were analyzed by immunoblot and ELISA. To verify the IgE specificity, an immunoblot inhibition assay was also performed. RESULTS: Of the 1,400 patients screened, 142 patients were history-positive to some food and 59 of these implicated chickpeas. Forty-one patients were skin test-positive and 31 were DBPCFC-positive for chickpea. The predominant symptoms after chickpea ingestion were respiratory. The ELISA results did not correlate well with the DBPCFC results; however, the skin test results correlated with DBPCFC in 75% of patients. Immunoblot analysis showed that 70, 64, 35, and 26 kD proteins were major allergens. CONCLUSIONS: Chickpea is an important source of allergen that can cause IgE-mediated hypersensitivity reactions ranging from rhinitis to anaphylaxis
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[30] - Martin JA, Compaired JA, de la Hoz B, Quirce S, Alonso MD, Igea JM, et al. Bronchial asthma induced by chick pea and lentil. Allergy 1992;47:185-187
Allergic reactions to legumes through inhalation have rarely been described. We report the case of a 20-year-old man who experienced asthmatic attacks when exposed to the steam from cooking either chick pea or lentil. Type I hypersensitivity to the antigens in these legumes was demonstrated by means of immediate skin reactivity, histamine release tests, RAST and RAST inhibition. Specific bronchial challenges with the heated (75 degrees for 30 min) extracts of chick pea and lentil elicited isolated immediate responses.
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[31] - Martinez San Ireneo M, Ibanez Sandin MD, Fernanadez-Caldas E, Maranon Lizana F, Rosales Fletes MJ, Laso Borrego MT. Specific IgE levels to Cicer arietinum (Chickpea) in tolerant and nontolerant children: evaluation of boiled and raw extracts. Int Arch Allergy Immunol 2000;121:137-143
The chick pea, Cicer arietinum, is a legume commonly consumed in Spain and other Mediterranean countries. The sera of 29 children (mean age: 8.4 years) with a current or past history of allergic reactions after ingestion of chick pea, and positive skin tests to this legume, were used to study the allergenic composition of raw and boiled chick pea extracts. The patient population was divided into 2 groups: group 1 consisted of 19 patients with clinical sensitivity confirmed by either positive oral challenges or a convincing recent history of anaphylaxis after eating chick peas, and group 2 consisted of 10 patients with clinical sensitivity in the past, but tolerant at the time of blood extraction. Six atopic children, not allergic to legumes, were included as controls. Specific IgE to the raw and boiled extracts was measured by ELISA. The allergenic composition of both extracts was analyzed by SDS-PAGE and immunoblots. There were no significant differences between specific IgE levels to the raw and boiled extracts (p = 0.23). The mean levels in group 1 were significantly higher than in group 2 and controls (p = 0.0001). Multiple IgE binding proteins/peptides were detected in both extracts in the molecular weight range of 10-106 kD. Only nontolerant patients recognized a similar number of bands in both extracts. Chick pea extracts contain a majority of heat-stable allergens, which could be responsible for the clinical sensitivity to chick pea. Patients with a current clinical allergy to chick pea have statistically higher specific IgE levels than tolerant patients and controls.
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[32] - Sánchez-Monge R, Valdés I, López-Torrejón G, Pascual C, Martin-Esteban M, Salcedo G. A new lectin family involved in paediatric chickpea (Cicer arietinum) allergy. EAACI 25th Congress, Vienna, 10-14 June, 2006, Poster n°588
Background: Legumes (mainly lentil, chickpea and green pea) are the fourth cause of food allergy among Spanish children. Vicilins and convicilins have been identified as major allergens in lentil and pea but no chickpea allergen has been identified to date. Methods: A pool or individual sera from 16 allergic patients, with clinical history of chickpea allergy and positive SPT to chickpea and lentil, were used in IgE immunodetection of a crude chickpea protein extract. The relationship of chickpea IgE binding proteins with lentil allergenic vicilins was studied by immunodetection with anti-Len c 1 rabbit polyclonal antibodies and ELISA inhibition assays. The chickpea protein extract was fractionated by anion exchange FPLC and reverse phase HPLC. N- terminal amino acid sequencing of IgE binding proteins was attempted. Results: The N-terminal amino acid sequence of two of the IgE binding proteins (23 and 44 kDa and 40% prevalence) is identical and show a high degree of sequence identity with a previously described pea albumin and chickpea lectin (67 and 87% respectively). Additionally some of the IgE binding proteins were recognized by the anti-vicilin antibodies, and lentil allergenic vicilin Len c 1 inhibited up to 60% of the IgE binding to a chickpea protein extract in ELISA assay. Conclusion: Two novel proteins of a lectin family, not related with peanut allergenic agglutinin, were described as chickpea IgE binding proteins implicated in Spanish paediatric chickpea allergy. Furthermore, some of the chickpea IgE binding proteins ( 45-55 kDa and 70 kDa) could be tentatively assigned to the vicilin family.
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[33] - Vioque J, Clemente A, Sanchez-Vioque R, Pedroche J, Bautista J, Millan F. Comparative study of chickpea and pea PA2 albumins. J Agric Food Chem 1999;47:3609-3613
PA2 albumins from 5 different cv. of kabuli-type chick peas and from one pea cv. were purified and characterized. Selective extraction of albumins was followed by 2 gel filtration purification steps. In all chick pea cv., PA2 was found to be made up of one type of protein, while pea PA2 was made up of 2 isoforms as observed by nondenaturating PAGE. PA2 has, so far, no defined function in peas, while in chick peas, a similar protein to PA2 has been described as a lectin. Results confirmed that PA2 from peas and chick peas agglutinates papainized human erythrocytes, and that chick pea and pea PA2 albumins are allergens for sensitive individuals
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[34] - Vioque J, Sanchez-Vioque R, Clemente A, Pedroche J, Bautista J, Millan F. Purification and partial characterization of chickpea 2S albumin. J Agric Food Chem 1999;47:1405-1409
A chickpea 2S albumin has been purified by solubilization in 60% methanol and ion-exchange chromatography. Under denaturing conditions it is composed of two peptides of 10 and 12 kDa. Native molecular mass determined by gel filtration chromatography is 20 kDa. Amino acid composition shows that it is rich in sulfur amino acids, mainly cysteine with 4.6% of the total. On the other hand, it has antinutritional characteristics of being allergenic for chickpea-sensitive individuals and inhibitory against porcine chymotrypsin with a lesser degree toward trypsin. The results of interest from a nutritional point of view are discussed.
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[35] - Qureshi IA, Sethi DK, Salunke DM. Purification, identification and preliminary crystallographic studies of an allergenic protein from Lathyrus sativus. Acta Crystallogr Sect F Struct Biol Cryst Commun 2006;63:869-872
A 24 kDa protein was purified from the seeds of Lathyrus sativus by ammonium sulfate fractionation and ion-exchange chromatography. The N-terminal amino-acid sequence showed significant homology with the 2S albumin class of seed storage proteins. The protein showed 85% sequence homology with the seed albumin of Pisum sativum within the 40 N-terminal residues. Crystals were obtained by the hanging-drop vapour-diffusion method. The crystals belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 43.5, b = 82.7, c = 153.4 A.
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[36] - Branco Ferreira M, Pedro E, Meneses Santos J, Pereira dos Santos MC, Palma Carlos ML, Bartolome B, et al. Latex and chickpea (Cicer arietinum) allergy: first description of a new association. Eur Ann Allergy Clin Immunol 2004;36:366-371
In this paper we describe the existence of cross-reactivity between allergens from latex and chickpea, a food from the Leguminosae family, which is common in the Mediterranean diet. We present the case report of a spina bifida boy with a clinical relevant food allergy to chickpea (oral syndrome + dysphonia), developing after the appearance of latex allergy symptoms (lip angioedema + intraoperative anaphylaxis). Specific IgE to latex and chickpea was demonstrated by skin prick tests, measurement of patient's serum specific IgE and IgE-immunoblotting. Cross-reactivity was studied by means of EAST-inhibition and western blotting-inhibition. A strong inhibition was observed in several IgE-binding bands when latex extract was used in solid phase and patient serum was preincubated with chickpea extract (chickpea extract as inhibitor phase). As far as we know, this is the first report of cross-reactivity between latex and chickpea, a food which should therefore be added to the extensive list of latex cross-reactive foods.
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[37] - Bjerremann Jensen L, Torp AM, Pedersen MH, Skov PS, Poulsen LK, Bindslev-Jensen C, et al. Legume sprouts might pose a risk for peanut allergic patients. Allergy 2007;62(suppl. 83):105-106
Background Peanut allergic patients are often advised to avoid all legumes in their diet. Studies have shown that despite an observed in vitro cross-reactivity this does not necessarily reflect a clinically relevant reaction. This might be due to degradation of allergens during cooking. A relatively new source of protein in especially vegetarian meals is legume sprouts, which are often consumed raw and peanut allergic patients have reported symptoms upon ingestion. In this study we report that sera from peanut allergic patients cross-react to extracts of legume sprouts. Methods Crude extracts of seeds and hypocotyls (part of embryo located below cotyledon attachment) / epicotyls (shoot of embryo above cotyledon) from peanut, soybean, pea, lupine, mung, alfalfa, broad bean and azuki were prepared. Stripped human blood basophils were passively sensitized with sera from 10 peanut allergic patients or a healthy control. The cells were challenged with the legume extracts in 6 concentrations. The released histamine was measured spectrofluorometrically (RefLab). The protein concentration resulting in 20% histamine release (HR) was used as a measure for biologic activity. Enzyme-allergosorbent test (EAST) was performed with MaxisorpTM microtiterplates coated with legume extracts (2µg/ml) and developed by HRP-conjugated anti-human-IgE. Direct EAST was performed with 3 patient sera and inhibition experiments using plates coated with peanut were performed with 8 patient sera. Cutoff was 15% inhibition. Results Histamine release (HR) experiments indicated an extensive co-reactivity between peanut and the legumes. Most sensitive was lupine, followed by soybean and pea. Surprisingly, hypocotyls/epicotyls from pea, lupine, mung, alfalfa, broad bean, and azuki were more sensitive than the corresponding seeds. Only peanut gave OD>0.1 in direct EAST. EAST inhibition experiments showed that lupine seeds could inhibit peanut reaction with up to 63%, soybean seeds up to 56%, and pea seeds up to 60%. Lupine hypocotyl was the only sprout able to inhibit >15%. Interestingly, lower concentrations of lupine hypocotyl were necessary to induce inhibition compared to lupine seed. Conclusion Patient sera recognize legume seeds and sprouts in vitro. Interestingly, the sprout extracts were more potent than the seeds in HR. The symptoms reported by peanut allergic patients after legume sprout intake might therefore be caused by cross-reactivity.
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[38] - Kumari D, Kumar R, Sridhara S, Arora N, Gaur SN, Singh BP. Sensitization to blackgram in patients with bronchial asthma and rhinitis: clinical evaluation and characterization of allergens. Allergy 2006;61:104-110
BACKGROUND: Legumes are important causative agents of type I hypersensitivity in south Asia and Europe but such studies are lacking in Indian population. The present study investigates blackgram sensitization in asthma and rhinitis patients and identifies immunoglobulin E (IgE)-binding proteins . METHODS: Respiratory allergy patients were evaluated using standard questionnaire and skin prick tests (SPT) with common foods and aeroallergens. Blackgram-specific IgE level was estimated by enzyme-linked immunosorbent assay (ELISA) and sensitization was established by a double-blind, placebo-controlled food challenge (DBPCFC). The cross-reactivity of blackgram with other legumes was studied by immunobiochemical methods . RESULT: Of 816 patients, 35 gave history of blackgram hypersensitivity. From these, 16 patients were SPT positive and 14 showed elevated specific IgE (three times of negative control) to blackgram. DBPCFC established blackgram sensitivity in four of 14 patients. Immunoblotting with individual patient's sera recognized eight most prevalent allergens of 78, 56, 47, 43, 40, 30, 28 and 16 kDa. Roasted blackgram showed six major allergens whereas 47, 43 and 28 kDa proteins retained IgE reactivity upon boiling. Blackgram extract required 14 ng of self protein for 50% ELISA inhibition whereas roasted and boiled blackgram required 16 and 120 ng protein. ELISA and immunoblot inhibition show partial inhibition to blackgram proteins by lentil, limabean and pea . CONCLUSION: Blackgram induces IgE-mediated reactions in 1.7% of asthma and rhinitis patients and contains eight major IgE-binding components, of which six retained IgE reactivity after roasting. Blackgram shares allergenicity with lentil and limabean.
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[39] - Mittag D, Vieths S, Vogel L, Wagner-Loew D, Starke A, Hunziker P, et al. Birch pollen-related food allergy to legumes: identification and characterization of the Bet v 1 homologue in mungbean (Vigna radiata), Vig r 1. Clin Exp Allergy 2005;35:1049-1055
BACKGROUND: Recently allergic reactions to legumes mediated by Bet v 1-homologous food allergens were described for soy and peanut. In this study we assessed allergic reactions to another legume, to mungbean seedlings, and identified its Bet v 1-homologous allergen Vig r 1 . METHODS: Ten patients were selected who had a history of allergic reactions to mungbean seedlings and a respiratory allergy to birch pollen. The Bet v 1 homologue in mungbean seedlings, Vig r 1, was cloned by a PCR strategy, expressed in Escherichia coli, and purified by preparative SDS-PAGE. In all sera, specific IgE against birch pollen, Bet v 1, Bet v 2, Vig r 1, and the Bet v 1 homologues in soy (Gly m 4) and cherry (Pru av 1) was determined by CAP-FEIA. Cross-reactivity of specific IgE with Vig r 1, Bet v 1, Gly m 4, and Pru av 1 was assessed by immunoblot inhibition. Expression of Vig r 1 during development of mungbean seedlings and under wounding stress was analysed by immunoblotting. The Vig r 1 double band was analysed by matrix-assisted laser desorption/ionization time-of-flight and liquid chromatography/tandem mass spectrometry (LC/MS/MS) . RESULTS: All patients were sensitized to birch pollen and Bet v 1, 20% to Bet v 2, and 90% to Gly m 4. Seventy percent of the patients showed IgE binding to a double band at 15 kDa in mungbean extract that was inhibited after pre-incubation of sera with rBet v 1. PCR cloning revealed that the mungbean homologue of Bet v 1 had a molecular weight of 16.2 kDa, a calculated pI of 4.6% and 42.8% amino acid sequence identity with Bet v 1. MS analysis confirmed similarity of the double band with the deduced Vig r 1 sequence, but also indicated the existence of other Vig r 1 isoforms. ImmunoCAP analysis detected IgE against Vig r 1 in 80% of the sera. IgE binding to Vig r 1 was inhibited with Gly m 4 in six of six and with rPru av 1 in four of six patients. Vig r 1 expression occurred during development of seedlings and was increased by wounding stress . CONCLUSIONS: Food allergy to mungbean seedlings can be caused by primary sensitization to birch pollen and is mediated by Vig r 1 in the majority of the patients with birch pollen-related allergy to mungbean seedlings.
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[40] - Jung P, Sesztak-Greinecker G, Wantke F, Goetz M, Jarisch R, Hemmer W. Prevalence of cross-sensitisation to soy allergens in patients with birch pollen allergy and allergenicity of different soy products. EAACI 25th Congress, Vienna, 10-14 June, 2006, Poster n°1530
Background: Soybean Gly m 4, a member of the PR-10 protein family, has been recognized as a cross-reactive food allergen in birch pollen-related food hypersensitivity with the potential to elicit severe systemic reactions. We investigated the prevalence of cross-sensitisation to soybean and the allergenicity of various soy-based food items in patients with birch pollen allergy. Method: A commercial brand of soy milk was added to our routine skin prick test panels and tested in consecutive patients with suspect inhalant or food allergy. Consumption habits concerning soy products and eventual adverse reactions were recorded by questionnaire. Additional prick-to-prick testing with different soy products was done in selected patients. Results: Among 292 patients with a positive skin prick test to birch pollen, 72 (24.7%) reacted to soy milk but only 18 (6.2%) reacted to a commercial soy skin prick test. 34/97 (35%) of birch pollen-allergic patients reported to have knowingly consumed soy milk before with 11/34 (31%) of them having experienced side effects. Mild reactions to tofu and soybean sprouts were occasionally reported. Prick-to-prick testing with different soy products in 16 patients with a positive skin test to soy milk revealed positive reactions to raw and cooked soybean sprouts (94%/50%), raw and cooked tofu (87%/40%), soy dessert (86%), and soy joghurt (21%). No significant differences in skin test responses were seen between seven different brands of soy milk (protein content 3.0-3.7g/100ml). Skin tests remained positive even after boiling soy milk für 5, 10 and 30 minutes. Conclusions: Cross-sensitisation to soy is frequent among patients with birch pollen allergy and many soy-based foods retain considerable allergenicity. As soy products are becoming increasingly popular on the market, birch pollen-allergic patients may be at growing risk of experiencing allergic reactions to these products.
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[41] - Mittag D, Vieths S, Vogel L, Wagner-Loew D, Starke A, Hunziker P, et al. Birch pollen-related food allergy to legumes: identification and characterization of the Bet v 1 homologue in mungbean (Vigna radiata), Vig r 1. Clin Exp Allergy 2005;35:1049-1055
BACKGROUND: Recently allergic reactions to legumes mediated by Bet v 1-homologous food allergens were described for soy and peanut. In this study we assessed allergic reactions to another legume, to mungbean seedlings, and identified its Bet v 1-homologous allergen Vig r 1 . METHODS: Ten patients were selected who had a history of allergic reactions to mungbean seedlings and a respiratory allergy to birch pollen. The Bet v 1 homologue in mungbean seedlings, Vig r 1, was cloned by a PCR strategy, expressed in Escherichia coli, and purified by preparative SDS-PAGE. In all sera, specific IgE against birch pollen, Bet v 1, Bet v 2, Vig r 1, and the Bet v 1 homologues in soy (Gly m 4) and cherry (Pru av 1) was determined by CAP-FEIA. Cross-reactivity of specific IgE with Vig r 1, Bet v 1, Gly m 4, and Pru av 1 was assessed by immunoblot inhibition. Expression of Vig r 1 during development of mungbean seedlings and under wounding stress was analysed by immunoblotting. The Vig r 1 double band was analysed by matrix-assisted laser desorption/ionization time-of-flight and liquid chromatography/tandem mass spectrometry (LC/MS/MS) . RESULTS: All patients were sensitized to birch pollen and Bet v 1, 20% to Bet v 2, and 90% to Gly m 4. Seventy percent of the patients showed IgE binding to a double band at 15 kDa in mungbean extract that was inhibited after pre-incubation of sera with rBet v 1. PCR cloning revealed that the mungbean homologue of Bet v 1 had a molecular weight of 16.2 kDa, a calculated pI of 4.6% and 42.8% amino acid sequence identity with Bet v 1. MS analysis confirmed similarity of the double band with the deduced Vig r 1 sequence, but also indicated the existence of other Vig r 1 isoforms. ImmunoCAP analysis detected IgE against Vig r 1 in 80% of the sera. IgE binding to Vig r 1 was inhibited with Gly m 4 in six of six and with rPru av 1 in four of six patients. Vig r 1 expression occurred during development of seedlings and was increased by wounding stress . CONCLUSIONS: Food allergy to mungbean seedlings can be caused by primary sensitization to birch pollen and is mediated by Vig r 1 in the majority of the patients with birch pollen-related allergy to mungbean seedlings.
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[42] - Lin KF, Liu YN, Hsu ST, Samuel D, Cheng CS, Bonvin AM, et al. Characterization and structural analyses of nonspecific lipid transfer protein 1 from mung bean. Biochemistry 2005;44:5703-5712
Plant nonspecific lipid transfer proteins (nsLTPs) are thermal stable proteins that are capable of transferring lipid molecules between bilayers in vitro. This family of proteins, abundant in plants, is proposed to be involved in defense, pollination, and germination; the in vivo biological function remains, however, elusive. Here we report the purification and sequencing of an nsLTP1 from mung bean sprouts. We have also determined the solution structure of this nsLTP1, which represents the first 3D structure of the dicotyledonous nsLTP1 family. The global fold of mung bean nsLTP1 is similar to those of the monocotyledonous nsLTP1 structures and consists of four alpha-helices stabilized by four disulfide bonds. There are, however, some notable differences in the C-terminal tails and internal hydrophobic cavities. Circular dichroism and fluorescence spectroscopy were used to compare the thermodynamics and lipid transfer properties of mung bean nsLTP1 with those of rice nsLTP1. Docking of a lipid molecule into the solution structure of mung bean nsLTP1 reveals similar binding cavities and hydrophobic interactions as in rice nsLTP1, consistent with their comparable lipid transfer properties measured experimentally.
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[43] - Jung P, Sesztak-Greinecker G, Wantke F, Goetz M, Jarisch R, Hemmer W. Prevalence of cross-sensitisation to soy allergens in patients with birch pollen allergy and allergenicity of different soy products. EAACI 25th Congress, Vienna, 10-14 June, 2006, Poster n°1530
Background: Soybean Gly m 4, a member of the PR-10 protein family, has been recognized as a cross-reactive food allergen in birch pollen-related food hypersensitivity with the potential to elicit severe systemic reactions. We investigated the prevalence of cross-sensitisation to soybean and the allergenicity of various soy-based food items in patients with birch pollen allergy. Method: A commercial brand of soy milk was added to our routine skin prick test panels and tested in consecutive patients with suspect inhalant or food allergy. Consumption habits concerning soy products and eventual adverse reactions were recorded by questionnaire. Additional prick-to-prick testing with different soy products was done in selected patients. Results: Among 292 patients with a positive skin prick test to birch pollen, 72 (24.7%) reacted to soy milk but only 18 (6.2%) reacted to a commercial soy skin prick test. 34/97 (35%) of birch pollen-allergic patients reported to have knowingly consumed soy milk before with 11/34 (31%) of them having experienced side effects. Mild reactions to tofu and soybean sprouts were occasionally reported. Prick-to-prick testing with different soy products in 16 patients with a positive skin test to soy milk revealed positive reactions to raw and cooked soybean sprouts (94%/50%), raw and cooked tofu (87%/40%), soy dessert (86%), and soy joghurt (21%). No significant differences in skin test responses were seen between seven different brands of soy milk (protein content 3.0-3.7g/100ml). Skin tests remained positive even after boiling soy milk für 5, 10 and 30 minutes. Conclusions: Cross-sensitisation to soy is frequent among patients with birch pollen allergy and many soy-based foods retain considerable allergenicity. As soy products are becoming increasingly popular on the market, birch pollen-allergic patients may be at growing risk of experiencing allergic reactions to these products.
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[44] - Pecquet C, Leynadier F. Réaction allergique aux grains de fénugrec et de nigelle. Rev Fr Allergol Immunol Clin 1997;37:1160
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[45] - Patil SP, Niphadkar PV, Bapat MM. Allergy to fenugreek (Trigonella foenum graeccum). Ann Allergy Asthma Immunol 1997;78:297-300
BACKGROUND: Allergic reactions after consumption of spices are well-known. In Asia, fenugreek seeds are consumed as spices and also as medicines. Literature survey carried out does not reveal reports of allergic reactions to fenugreek. In our survey carried out on patients with food allergy, we found two cases of severe allergy to fenugreek. METHODS: We report here two cases of immediate allergy following ingestion, inhalation, and external application of fenugreek seed powder. In the first case, inhalation of the fenugreek seed powder resulted in rhinorrhea, wheezing, and fainting. The second case was of a patient with chronic asthma who developed numbness of head, facial angioedema, and wheezing after application of fenugreek paste to her scalp as a treatment for dandruff. Skin scratch test was performed with fenugreek and other members of the Leguminosae family as fenugreek also belongs to Leguminosae. Objective evidence of the reaction was obtained by conducting double-blind placebo-controlled challenges (DBPCFC). For detecting IgE binding by immunoblotting method, the proteins of the fenugreek extract were resolved using sodium dodecyl sulphate polyacrylamide gel electrophoresis. RESULTS: Skin scratch tests for the patients revealed strong sensitivity to fenugreek and chickpeas. None of the controls showed such response with fenugreek extract. During DBPCFC, both patients showed > 20% drop in peak flow rate after consumption of fenugreek and chickpea. Immunoblots demonstrated binding of specific IgE from the patients' sera with the protein from extracts between 20 kD to 70 kD bands. CONCLUSION: This case report has enlarged the list of food allergens with the addition of fenugreek.
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[46] - Nicolie B, Leduc V, Drouet M. Allergie alimentaire croisée entre les graines de fenugrec (Trigonella foenum-graecum) et d’arachide. Rev Fr Allergol Immunol Clin 2007;47:283
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[47] - Patil SP, Niphadkar PV, Bapat MM. Allergy to fenugreek (Trigonella foenum graeccum). Ann Allergy Asthma Immunol 1997;78:297-300
BACKGROUND: Allergic reactions after consumption of spices are well-known. In Asia, fenugreek seeds are consumed as spices and also as medicines. Literature survey carried out does not reveal reports of allergic reactions to fenugreek. In our survey carried out on patients with food allergy, we found two cases of severe allergy to fenugreek. METHODS: We report here two cases of immediate allergy following ingestion, inhalation, and external application of fenugreek seed powder. In the first case, inhalation of the fenugreek seed powder resulted in rhinorrhea, wheezing, and fainting. The second case was of a patient with chronic asthma who developed numbness of head, facial angioedema, and wheezing after application of fenugreek paste to her scalp as a treatment for dandruff. Skin scratch test was performed with fenugreek and other members of the Leguminosae family as fenugreek also belongs to Leguminosae. Objective evidence of the reaction was obtained by conducting double-blind placebo-controlled challenges (DBPCFC). For detecting IgE binding by immunoblotting method, the proteins of the fenugreek extract were resolved using sodium dodecyl sulphate polyacrylamide gel electrophoresis. RESULTS: Skin scratch tests for the patients revealed strong sensitivity to fenugreek and chickpeas. None of the controls showed such response with fenugreek extract. During DBPCFC, both patients showed > 20% drop in peak flow rate after consumption of fenugreek and chickpea. Immunoblots demonstrated binding of specific IgE from the patients' sera with the protein from extracts between 20 kD to 70 kD bands. CONCLUSION: This case report has enlarged the list of food allergens with the addition of fenugreek.
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[48] - Mur Gimeno P, Feo Brito F, Martin Iglesias A, Lombardero Vega M, Bautista Martinez P. Allergic reaction caused by a new hidden food, broad bean flour. Allergy 2007;62:1340-1341
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[49] - Campina Costa S, Neto M, Trindade M, Bartolome B, Campos Melo A, Pereira Santos MC. Anaphylaxis to fava bean (Vicia fava). Allergy 2007;62(suppl. 83):369
Background: Legume allergy is frequent in Mediterranean countries where their consumption is common and may cause life-threatening reactions in sensitized individuals. They contain multiple allergens with a significant degree of cross-reactivity among different species. It has been suggested that legume allergens are heat-stable and sensitive to the action of acids and enzymes. There is little information about allergy to other legumes besides peanut and soybean. Fava bean (Fb) belongs to the Papilionaceae family of the Fabales botanical order, and until now few data has been published on Fb allergy. We report the clinical case of a 34-year-old woman who suffered from anaphylactic reactions after fresh Fb ingestion. Methods: Skin prick test (SPT) was performed with extracts from common pollens and with Fb, beans, pea, chick-pea, lentil, peanut and soybean (Bial Aristegui®). Prick-prick test was performed to fresh and fronzen Fb, on its raw and boiled forms. Total IgE and specific IgE to these legumes were determined by UniCAP Pharmacia® method. Specific IgE to Fb was measured by enzyme allergo-sorbent test (EAST) and molecular mass of the IgE binding proteins was determined by SDS-PAGE IgE-Immunoblotting assay, (Bial Aristegui®). Glucose 6-phosphate dehydrogenase (G6PD) was measured. Basophil activation test (BAT), using IgE/CD63 double labelling was performed using serial dilution of Fb extract (Bial Aristegui®), and was evaluated by flow-citometry (FACSCALIBUR, Bectan Dickinson). Oral challenge (OC) with boiled frozen and fresh Fb was also carried out. Results: SPT, prick-prick test, specific IgE to Fb as well as to the other legumes tested were all negative. Immunoblotting to Fb did not reveal any IgE-binding band. Total serum IgE was 88,6 kU/L. G6PD was 8,16 UI/gHb. For a stimulation concentration of 2mg/ml Fb extract expression of CD63 above spontaneous expression was <5%, stimulation index <2. OC was positive with boiled fresh Fb however it was negative with boiled frozen Fb. Conclusion: we present a case of anaphylactic reaction after ingestion of fresh Fb. Neither SPT nor specific IgE were able to confirm an IgE mediated mechanism. OC was the unique useful test for the diagnosis of legume allergy in this case, the process of freezing could be responsible for altering the allergenicity of the allergens. The usefulness of BAT as a tool for investigating allergy to Fb is discussed.
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[50] - Bessot JC. Allergènes végétaux non polliniques. Rev Fr Allergol Immunol Clin 2003;43:40-52
L'inventaire des allergènes végétaux non polliniques s'est considérablement élargi au cours des 12 dernières années. Ces allergènes, initialement répertoriés dans l'environnement professionnel peuvent aussi être rencontrés dans l'environnement domestique. Certains d'entre eux se comportent à la fois comme des pneumallergènes ou des trophallergènes. Dans cette revue générale, seront envisagés la prévalence, les mécanismes, les aspects cliniques, la démarche diagnostique des allergies aux gommes végétales, aux graines, aux racines, aux feuilles et aux plantes d'intérieur. Le rôle des allergènes du latex, des bois, des farines, des enzymes végétales ne sera pas traité ici. Les gommes végétales (arabique, karaya, guar, psyllium...) provoquent des rhinites ou des asthmes IgE médiés, bien que leurs allergènes soient des polysaccharides. La colophane fait partie des 5 allergènes le plus souvent responsables d'asthmes professionnels en Grande-Bretagne. Parmi les graines, le rôle des graines provenant de céréales, du café, du ricin, du soja sera privilégié. Parmi les racines, certaines plantes médicinales (sanyak, bahna, salsepareille...) ont une importance croissante. Le henné, le tabac, le thé, le lycopode sont aussi des sources d'allergènes. Des allergies IgE dépendantes ont été décrites pour le ficus, mais aussi pour d'autres plantes d'appartement. Certaines étiologies, se limitant actuellement à un ou quelques cas publiés, peuvent annoncer des pathologies allergiques émergentes.
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[51] - Porcel S, Leon F, Valero AM, Calderin PM, Cuevas M, Cuesta EA. Occupational rhinitis and asthma by Lathyrus sativus flour: characterization of allergens. J Allergy Clin Immunol 2001;107:743-744
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[52] - Antón Gironés M, de la Hoz Caballer B, Muñoz Martín T, Cuevas Agustín M, Sánchez-Cano M. Occupational rhinoconjunctivitis and asthma by exposure to Lathyrus sativus flour. Allergol Immunopathol (Madr) 2005;33:326-328
We report the case of a 42-year-old non-smoking man, who had worked as a carpenter for 6 years and who reported a history of rhinorrhea, paroxysmal sneezing, nasocular pruritus, lacrimation, wheezing and dyspnea attacks while preparing a mixture to seal the junctures between wooden panels. Allergy study consisted of skin prick testing (SPT) to inhalants, foods and Lathyrus sativus flour (LSF) extract, specific bronchial provocation test with LSF extract, cytological analysis of sputum, specific IgE antibodies against LSF, and histamine releasing test with dilutions (1:5, 1:25, 1:125, 1:625) of LSF. The results demonstrated occupational rhinoconjunctivitis and asthma due to LSF exposure. We provide a review of published reports to date.
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