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La pêche

vendredi 2 avril 2010, par Allerdata

La pêche est un fruit des Rosacées.

A ce titre, elle fait partie du syndrome associant une pollinose au bouleau avec la pomme et d’autres fruits des Rosacées.

Mais la pêche surtout joue un rôle particulier dans les pays du pourtour méditerranéen où elle représente fréquemment le chef de file des aliments entrant dans le cadre d’un « syndrome LTP » et où elle génère volontiers des réactions plus sévères que celles rencontrées dans le syndrome bouleau-Rosacées.

Ces différences régionales sont nettes. Par exemple, en France, la survenue d’une réaction sévère à la pêche est rare :5 cas pour la pêche et 2 cas pour la nectarine ont été rapportés par le Réseau d’Allergo-Vigilance parmi 900 déclarations (mai 2010).

On pourra se rapporter au chapitre consacré aux Rosacées pour une approche détaillée des diverses composantes d’une réactivité à la pêche, et notamment :

fréquences selon les pays et le recrutement des patients
prévalences de positivités aux allergènes de pêche
démarche diagnostique face à une suspicion d’allergie à la pêche.

Dans un cadre professionnel il n’est pas rare, sur les zones de production, d’observer des urticaires de contact. On a rapporté aussi des cas d’asthme aux feuilles de pêchers, avec réactivité pour les LTP . Dans une observation le patient a développé peu à peu un syndrome oral à la pêche.

Peu de données spécifiques concernant la nectarine : elles sont présentées à part (cf. la nectarine).

Les allergènes de la pêche

Pru p 1

Pru p 1 est une protéine PR-10, présente à des concentration similaires dans la pulpe (0,26 µg/g) et dans la peau de pêche (0,6µg/g) .

Pru p 1 possède 59 % d’identité séquentielle avec Bet v 1. L’homologie avec la PR-10 de cerise, Pru av 1, est beaucoup plus forte (98 %). Et des sujets positifs pour Pru av 1 sont régulièrement trouvés positifs in vitro pour Pru p 1 .

Avec un tel taux d’homologie entre ces 2 allergènes d’une même famille on pourrait s’attendre à une très fréquente allergie simultanée pour la pêche et pour la cerise dans des pays comme l’Allemagne ou la Suisse. En fait une dissociation est fréquente dont l’origine est mal cernée car ces fruits sont consommés de façon analogue.

Pru p 2

La pêche possède une protéine thaumatine-like IgE-réactive (une "TLP"). Un travail a montré que ce type de protéine était exprimé dans le fruit, notamment à l’approche de la maturité . Les auteurs de ce travail ont dénommé cette thaumatine-like « Pru p 2 », à l’instar des Mal d 2 (pomme) ou Pru av 2 (cerise). Comme Pru p 3, cette protéine se trouve en concentrations plus élevées dans la peau de la pêche .

Récemment, il a été montré qu’environ 50% de patients allergiques à la pêche présentaient un TC positif pour Pru p 2 . Mais l’impact clinique réel de Pru p 2 reste difficile à apprécier car ces patients, Espagnols, étaient majoritairement réactifs à Pru p 3 : la symptomatologie était-elle dès lors le résultat de la seule sensibilisation à la LTP Pru p 3 ? Ou était-elle majorée par la réactivité supplémentaire à la TLP Pru p 2 ? Il est impossible de le discerner, faute de patients mono-positifs pour Pru p 2.

Pru p 3

Pru p 3 est le LTP de la pêche. L’importance prise par cet allergène et par le syndrome LTP dans l’allergologie moléculaire est remarquable pour une protéine dont l’allergénicité n’a été démontrée que récemment (en 1999) .

On a d’abord pensé que cette LTP était absente dans la pulpe . En fait, Pru p 3 est trouvé en concentration beaucoup plus élevée dans la peau de pêche comparativement à la pulpe .

Ahrazem a trouvé, par exemple, 133 µg/g dans la peau contre 0,6 µg/g dans la pulpe . Mieux, la peau de pêche non lavée contient plus de LTP que la peau lavée, le duvet recouvrant la peau étant particulièrement riche en Pru p 3 .

Les épitopes de Pru p 3 ont été étudiés :

dans le cas des épitopes T, la zone correspondant aux acides aminés 65 à 80 est importante
elle correspond plus ou moins à un épitope B (AA 70-79), ce dernier semblant avec la zone 35-46 concentrer une grande partie de l’IgE-réactivité vis à vis de Pru p 3 . On ne peut, cependant, utiliser fidèlement ces peptides épitopiques pour différencier, chez un patient donné, un syndrome oral et une réaction possiblement systémique.

Pru p 4

Pru p 4 est une profiline ayant 77 % d’identité avec Bet v 2 et 93 % avec Pru av 4 (cerise).

Autres protéines IgE-réactives

D’autres protéines IgE-réactives ont été suggérées : une 45 kDa et une isoflavone réductase .

On trouve dans Allergome des protéines ayant des noms de type IUIS (de Pru p 5 à Pru p 11) et dont le référencement est pour le moins mystérieux.

Pêche et autres allergies alimentaires liées à des LTP

La prévalence d’une histoire clinique positive pour la pêche est parfois très élevée sur la base d’un recrutement pour une autre allergie alimentaire dans les pays méditerranéens :

laitue : 7 cas sur 8
raisin : 11 cas sur 37
cerise : dans 90 % des cas

Malgré tout, dans de nombreux cas, la pêche semble jouer un rôle initiateur (ou amplificateur) dans le syndrome alimentaire du aux LTP.

A noter qu’en Grêce ce rôle est apparemment tenu par le raisin plutôt que la pêche .

Voir aussi Les Rosacées

Pêche, LTP et pollens

Pêche et armoise

Le pollen d’armoise contient une LTP (Art v 3). Une pollinose à l’armoise pourrait être à l’origine d’une sensibilisation croisée pour la pêche par des LTP.

La question a été étudiée en Espagne et en Italie du nord, mais pas dans des régions d’Europe à climat continental.

En Espagne la positivité pour la pêche et pour la LTP de pêche Pru p 3 est nettement plus fréquente en cas de pollinose à l’armoise associée à d’autres pollinoses, qu’en cas de mono-pollinose armoise (60 % vs 22 %), bien que la positivité pour Art v 3 en TC soit similaire d’un groupe à l’autre (73% vs 67%) .

L’armoise seule semble donc peu impliquée dans la réactivité pour la pêche : c’est plus le niveau d’atopie des sujets qui pourrait déterminer cette réactivité, les sujets les plus atopiques étant à la fois multi-polliniques et allergiques à la pêche.

Ce constat est soutenu par les résultats obtenus à Milan .

Parmi des sujets allergiques à la pêche et montrant un TC positif pour l’armoise, on ne trouve des mono-réactivités pour Pru p 3 que chez ceux qui ne sont pas cliniquement polliniques pour l’armoise.
Inversement, on ne trouve aucun sujet Pru p 3 positif parmi des polliniques à l’armoise sans allergie à la pêche.
De plus, les tests d’inhibitions réciproques armoise/pêche révèlent une prédominance de la pêche sur l’armoise (ou l’olivier) s’agissant des bandes LTP (9 kDa) . Cela est en faveur d’une sensibilisation à la pêche (Pru p 3) induisant une réactivité à Art v 3 (armoise), plutôt que l’inverse.

Pour autant, certains auteurs retiennent la possibilité d’un effet direct d’Art v 3 :

certains patients sont positifs pour Art v 3 sans l’être pour Pru p 3, bien qu’allergiques à la pêche (1 cas/9 dans l’étude de Lombardero )
et les patients positifs pour Pru p 3 présentent un plus large éventail de réactions à des aliments divers s’ils sont également positifs pour Art v 3 .

Au total, on pourrait faire l’hypothèse que la LTP d’armoise joue un rôle parfois additionnel

soit en renforçant l’exposition globale du patient aux LTP,
soit par l’existence d’une « image épitopique » légèrement différente (et complémentaire) de celle de Pru p 3.

Pêche et olivier

Le pollen d’olivier, comme celui d’armoise, contient une LTP (Ole e 7). Mais, comme pour l’armoise, la pollinose à l’olivier

ne semble pas générer en soi une réactivité à la pêche
peut jouer un rôle additionnel : on trouve plus de réactions sévères à la pêche chez les polliniques à l’olivier qui, de plus, sont positifs pour Ole e 7 .

Par contre, l’olivier, ainsi que les graminées, participe à une réactivité profilinique qui peut, dans certains cas, être cliniquement relevante pour certains aliments, dont la pêche.

On pourra se reporter au texte sur les oléacées pour la discussion portant sur les liens entre olivier et aliments végétaux contenant des LTP.

Pêche et platane

Le pollen de platane contient également une LTP (Pla a 3). Comme pour Ole e 7, Pla a 3 ne semble pas se comporter en promoteur unique d’une réactivité aux LTP alimentaires.

On pourra se reporter au texte sur le Platane pour une discussion détaillée de la relation platane-LTP-aliments.

Pêche et pollens hors LTP

Pêche et cyprès

Quelques cas d’allergie à la pêche chez des polliniques aux Cupressacées ont été observés dans le midi de la France .

Les résultats in vitro (blots, inhibitions) ne permettent pas pour l’instant de conclure à une allergie croisée.

Pêche et bouleau

Dans les régions concernées par la survenue d’un syndrome bouleau-pomme, un nombre non négligeable de sujets rapportent aussi des réactions cliniques pour la pêche (29 % , 33 % , 64 % ).

L’association bouleau-pêche a pour origine une réactivité croisée entre protéines PR-10 : Bet v 1 et Pru p 1.

Pêche et profilines

En l’absence de bouleaux, la réactivité pour la pêche peut provenir d’une sensibilisation aux LTP (Pru p 3 pour la pêche) ou aux profilines (Pru p 4).

Dans la mesure où la grande majorité des sujets allergiques à la pêche sont polliniques, il faut s’attendre à trouver des résultats positifs pour des profilines conjointement à une réactivité LTP.

L’essentiel des résultats relatifs à pêche et profiline provient d’études espagnoles.

Malgré l’absence de bouleaux, rBet v 2 s’avère souvent positif par réactivité croisée avec les profilines des pollens locaux (graminées, olivier, diverses herbacées).

Cela contraste avec les résultats pour rBet v 1 :

aucun rBet v 1 positif parmi des allergiques à la pêche multipolliniques . Un seul sur 12 dans une autre étude où pourtant la pollinose positivait les tests cutanés pour le bouleau (12/16).
Aucun rBet v 1 positif mais 9 rBet v 2 (profiline) positifs parmi 10 polliniques avec syndrome oral dont 5 sont TC positifs pour la pêche .
7 % de rBet v 1 positifs contre 32 % pour rBet v 2 chez des allergiques à la pêche
51 % de rBet v 2 positifs parmi un groupe d’allergiques à la pêche ayant débuté leur allergie après l’âge de 15 ans
44 % de rBet v 2 positifs dans une autre cohorte Madrilène
100 % de rBet v 2 positifs dans une série de patients Basques avec syndrome oral pour la pêche .

Pêche et latex

La pêche est-elle un fruit à classer parmi les aliments du syndrome "latex-fruits" ?

De rares cas de réaction croisée latex-pêche
e ont été rapportés, les allergènes en jeu n’étant, par ailleurs, pas concordants .

La pêche ne semble pas être un "fruit latex" car la prévalence des allergies à la pêche rapportées par les patients reste très faible parmi les sujets allergiques ou sensibilisés au latex (NB : les cas n’ont pas été confirmés par un TPO) :

1 cas parmi 17 allergiques à l’avocat (dont 10 au latex aussi)
aucun cas, malgré 20 % de TC positifs, chez des sujets non polliniques allergiques au latex
2 cas parmi 23 allergiques à la banane (avec 9 fois une allergie au latex) mais aucun de ces 2 cas n’est allergique au latex
3 cas parmi 28 allergiques au latex, contre 11 pour la châtaigne chez les mêmes patients
aucun cas parmi 35 allergiques aux fruits, malgré 6 cas/35 de TC positif pour le latex .
2 TC positifs pêche parmi 12 allergiques au latex, contre 13 parmi 21 sujets non latex et allergiques aux fruits .

Inversement, les cas d’allergie ou de positivité pour le latex parmi des sujets allergiques à la pêche sont rares :

10 TC positifs pour la pêche chez des sujets allergiques au melon mais aucun TC positif latex

Ces faibles prévalences sont en défaveur d’une association latex-pêche.

Pêche : effets de la chaleur et de la digestion

Pru p 1 ne résiste pas à la digestion gastrique mais récupère sa conformation spatiale initiale après chauffage à 95°C, pour autant que le pH du milieu ne soit pas acide . On peut remarquer, cependant, que cette apparente stabilité de la PR-10 Pru p 1 n’a pu être testée dans ce type d’expériences (technique de dichroïsme circulaire) à une température entraînant une ébullition de l’échantillon (100°C et au-delà, ex. confitures).

Sans surprise la LTP Pru p 3 résiste très bien à la digestion gastrique. Et après digestion duodénale des fragments subsistent qui sont toujours IgE-réactifs .

Facteurs agronomiques et techno-alimentaires

Une étude de différentes variétés de pêche n’a pas révélé de différences importantes pour les concentrations de Pru p 3 à maturité, selon les cultivars .

Pour d’autres auteurs, les cultivars d’origine américaine auraient des taux supérieurs à ceux de cultivars traditionnels espagnols .

L’influence du degré de maturité des fruits, de la quantité de lumière reçue par l’arbre et de la productivité de celui-ci a été étudiée :

l’expression des allergènes croît avec la maturité (sauf Pru p 4 et Pru p 3 dans la pulpe)
une plus forte illumination et un rendement plus faible tendent à limiter un peu l’expression des allergènes.

En fait, ces différences ne permettent pas de déclarer un cultivar significativement moins allergisant qu’un autre, ou un mode cultural moins allergisant.

Le stockage à 4 °C fait diminuer les taux de LTP pour certains cultivars. Le stockage à 20 °C ne change pas les taux de LTP .

Si ni le chauffage à 121°C pendant 30 min ni les protéases ne détruisent Pru p 3 , le pelage chimique des pêches et l’ultrafiltration du jus de pêche diminuent les taux de Pru p 3 .

Ceci est retrouvé cliniquement : certains sujets avec une allergie modérée à la pêche (syndrome oral) peuvent tolérer les pêches en boite ou le jus de pêche .

Cela pourrait résulter d’une modification partielle de la structure III de la LTP du fait du chauffage du produit, car Pru p 3 ne se refolde pas correctement à froid si le pH n’est pas acide .

Plus simplement, cette moindre allergénicité peut résulter du pelage des fruits, l’essentiel des LTP étant contenues dans la peau.

La question de la peau de la pêche, et notamment de l’allergénicité des fruits selon qu’ils sont consommés localement ou après être passés dans les circuits de la grande distribution, est abordée dans le texte des généralités sur les Rosacées : en bref, la sensibilisation aux LTP, plus fréquente en zone méditerranéenne, résulte-t-elle de la consommation de pêches venant directement des vergers et ayant conservé une peau (et un duvet) plus allergisante ?

Nectarine et brugnon

Peu de données sont disponibles au sujet de ces fruits.

Ils contiennent des LTP, bien sûr . Une profiline et une isoflavone réductase sont suggérées IgE-réactives également du fait de résultats montrant une réactivité croisée avec des protéines équivalentes.

Rares sont les observations cliniques où la nectarine est citée . Ces patients réagissent également à la pêche et/ou à diverses Rosacées.

Asero cite la nectarine à l’appui de son hypothèse sur l’origine des sensibilisations aux LTP en milieu méditerranéen : il remarque notablement plus d’urticaires de contact avec la pêche qu’avec la nectarine chez des ouvriers travaillant dans la récolte de ces fruits. L’absence de poils sur la peau de la nectarine est supposée rendre ce fruit moins allergisant par contact. On sait en effet que ce duvet sur la pêche est fort riche en LTP.

Ceci étant, cette absence de poils sur l’épicarpe n’implique pas que moins de LTP soit présente dans l’épicarpe lui-même. Et donc une allergénicité qui serait moindre pour le consommateur. D’autant que la nectarine se consomme plus volontiers avec la peau que la pêche.

Une étude ayant analysé le contenu en LTP de la nectarine et de la pêche n’a pas révélé des taux différents entre ces deux fruits au point que la nectarine soit considérée comme moins allergisante que la pêche pour les personnes réagissant aux LTP . De plus, cette similitude était retrouvée dans la pulpe et aussi pour les PR-10.

[1] - Garcia BE, Lombardero M, Echechipia S, Olaguibel JM, Diaz-Perales A, Sanchez-Monge R, et al. Respiratory allergy to peach leaves and lipid-transfer proteins. Clin Exp Allergy 2004;34:291-295

BACKGROUND: Several lipid-transfer proteins (LTPs) have been identified as important food allergens, especially in fruits of the Rosaceae family. The major peach (Prunus persica) allergen has been identified, sequenced and designated Pru p 3 . OBJECTIVE: To present Pru p 3 as an aeroallergen able to induce occupational asthma . METHODS: A thorough investigation was performed in a fruit grower with occupational asthma. Skin prick-prick tests with peach leaves and prick tests with perennial respiratory allergens and pollens, fruits and peach leaf extracts were done. Serum-specific IgE was tested for peach leaf, peach fruit, peach skin and respiratory allergens that were positive in skin prick tests. Specific bronchial provocation tests (BPTs) with extracts of peach leaf were also done. Before and 24 h after the BPT, BPTs with methacholine and sputum induction were done. The IgE reactivity pattern to peach leaf and fruit extracts and to Pru p 3 was identified by using SDS-PAGE and immunoblotting. Blotting inhibition of peach leaf extract by Pru p 3 was also performed. The putative allergen was quantified in leaf and fruit skin extracts with ELISA based on an anti-Pru p 3 antibody . RESULTS: Skin tests were positive for peach leaf and fruit. The BPT was positive, with immediate and delayed response. This test induced a decrease in PD20 (dose of agonist that induces a 20% fall in FEV1) methacholine and an increase in eosinophils and eosinophil cationic protein in sputum. Peach leaf extract contained concentrations of Pru p 3 similar to those found in peach skin. Specific IgE immunodetection showed that patient's sera reacted with Pru p 3, and with a single major band from the peach leaf extract fully inhibited by Pru p 3 . CONCLUSION: Pru p 3 from peach leaves can act as a respiratory allergen and cause occupational rhinoconjunctivitis and asthma.

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[2] - Garcia-Sellés J, Merida C, Navarro M, Garcia-Alonso L, Lombardero M. Occupational asthma with sensitization to Pru p 3. Allergy 2007;62(suppl. 83):280

Background: The objective of this case is to present Pru p 3 allergen responsible of occupational asthma. Method: Twenty ˜ five year old woman with history of ocular, nasal and bronquial symptoms related to laboral exposure. She worked in a laboratory manipulating leaves of peach tree. Results: Skin prick test to perennial respiratory allergens and pollens: negative. Cytology series of nasal exudation: negative. Prick test of peach extract, peach leaf and Pru p 3: Positive. Spirometry during symptoms: CVF 103%, FEV1 71%, FEV1/CVF 99% and without symptoms: CVF 103%, FEV1 85%, FEV1/CVF 83%. Methacoline test positive with PD20 of 17UA. Control exposure with peach tree leaves(specific bronchial provocation tests): immediate response (FEV1 decrease 25%) and delayed response. Methacoline test repeated after with PD20 of 9.5 UA. Specific Ig E to Pru p 3 positive 0.75 KU/L and negative to Bet v 1, Art v 1 and profilin (Centaur ADVIA System). Conclusions: The majority allergen of peach, Pru p 3 is a relevant panallergen related with fruit allergy (peach skin and leaf) and occupational rhinoconjunctivitis and asthma by direct and continuous exposure, reported in this case.

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[3] - Ahrazem O, Jimeno L, López-Torrejón G, Herrero M, Espada JL, Sánchez-Monge R, et al. Assessing allergen levels in peach and nectarine cultivars. Ann Allergy Asthma Immunol 2007;99:42-47

BACKGROUND: The lipid transfer protein Pru p 3 has been identified as a major peach fruit allergen. However, the putative peach member of the Bet v 1 family, Pru p 1, has been neither identified nor characterized. OBJECTIVES: To determine the distribution and solubility properties of the main peach allergens and to quantify Pru p 3 and Pru p 1 levels in peach and nectarine cultivars. METHODS: Peach peel and pulp were extracted using different buffers, and extracts were analyzed by means of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunodetection using polyclonal antibodies against lipid transfer proteins, profilins, and Bet v 1 homologues. Pru p 3 was quantified in peach and nectarine cultivars using a sandwich enzyme-linked immunosorbent assay method. A similar method was developed to quantify Pru p 1. RESULTS: A differential distribution between peel and pulp and different solubility properties were found for Pru p 3, Pru p 1, and peach profilin. Mean Pru p 3 levels were 132.86, 0.61, and 16.92 microg/g of fresh weight of peels, pulps, and whole fruits, respectively. The corresponding mean Pru p 1 levels were 0.62, 0.26, and 0.09 microg/g of fresh weight. Most US cultivars showed higher levels of both allergens than Spanish cultivars. CONCLUSIONS: The different distribution and solubility properties of the main peach allergens can determine the quality of fruit extracts used as diagnostic tools. These differences, together with the natural variation of Pru p 3 and Pru p 1 levels among peach and nectarine cultivars, can be exploited to reduce peach allergenicity by means of industrial processing and plant breeding.

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[4] - Marknell DeWitt A, Andersson K, Vieths S, Ballmer-Weber B, Lidholm J. Cloning and characterization of the major peach allergen Pru p 1. Allergy Clin Immunol Int 2005;17(Suppl. 1):341

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[5] - Botton A, Andreotti C, Costa G, Ramina A. Peach (Prunus persica L. Batsch) Allergen-Encoding Genes Are Developmentally Regulated and Affected by Fruit Load and Light Radiation. J Agric Food Chem 2009;57:724-734

Abstract The fruits of Rosaceae species may frequently induce allergic reactions in both adults and children, especially in the Mediterranean area. In peach, true allergens and cross-reactive proteins may cause hypersensitive reactions involving a wide diversity of symptoms. Three known classes of allergenic proteins, namely, Pru p 1, Pru p 3, and Pru p 4, have been reported to be mostly involved, but an exhaustive survey of the proteins determining the overall allergenic potential, their biological functions, and the factors affecting the expression of the related genes is still missing. In the present study, the expression profiles of some selected genes encoding peach allergen isoforms were studied during fruit growth and development and upon different fruit load and light radiation regimens. The results indicate that the majority of allergen-encoding genes are expressed at their maximum during the ripening stage, therefore representing a potential risk for peach consumers. Nevertheless, enhancing the light radiati on and decreasing the fruit load achieved a reduction of the transcription rate of most genes and a possible decrease of the overall allergenic potential at harvest. According to these data, new growing practices could be set up to obtain hypoallergenic peach fruits and eventually combined with the cultivation of hypoallergenic genotypes to obtain a significant reduction of the allergenic potential

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[6] - Botton A, Andreotti C, Costa G, Ramina A. Peach (Prunus persica L. Batsch) Allergen-Encoding Genes Are Developmentally Regulated and Affected by Fruit Load and Light Radiation. J Agric Food Chem 2009;57:724-734

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[7] - Palacín A, Tordesillas L, Gamboa P, Sanchez-Monge R, Cuesta-Herranz J, Sanz ML et al. Characterization of peach thaumatin-like proteins and their identification as major peach allergens. Clin Exp Allergy 2010;40:1422-1430

Background Peach is the most important fruit related to food allergy in the Mediterranean area. Pru p 3, its lipid transfer protein, has been described as the principal allergen responsible for cross-reactivities with other foods and pollen and the severity of clinical symptoms. However, the involvement of other allergenic families cannot be ruled out. Thaumatin-like proteins (TLPs) have been described as food allergen in several fruits, such as apple, cherry, kiwi and banana, and pollen. Objective To identify members of the TLP family in peach fruit and to characterize putative allergens. Methods Through two-dimensional (2D) electrophoresis of peach extract and immunodetections with a pool of peach-allergic patients, IgE-binding spots were identified and the corresponding proteins purified and characterized as allergens by in vitro and in vivo assays. Three isoforms, belonging to the TLP family, were purified by different chromatographic systems and characterized by N-terminal amino acid sequences, molecular weight determination (MALDI) and enzymatic activity analysis (beta-1,3-gluconase test and inhibition growth of fungi). In the same way, their IgE-binding capacity and allergenic activity were tested by ELISA assays, basophil activation tests and skin prick tests (SPT). Results Two peach-TLPs, Pru p 2.0101 and Pru p 2.0201, were identified as IgE-binding spots by 2D electrophoresis. Another peach-TLP, Pru p 2.0301, was cloned and produced as recombinant protein in a yeast system. The three isoforms were purified and characterized as TLPs by immunoblotting with anti-chestnut TLP antibodies and anti-plant N-asparagine complex glycan (anti-cross-reactive carbohydrate determinant). All of them showed beta-1,3-glucanase activity and inhibition of fungal growth. The three TLPs were recognized by around 50% of the sera from 31 patients analysed in ELISA experiments. All three gave a positive response to an SPT and/or in basophil activation experiments. Conclusion Three isoforms, belonging to the TLP family, were identified in peach as principal allergens. Their prevalence, observed in in vitro, ex vivo and in vivo analyses, suggests that they are important allergens and should therefore be included in the routine diagnosis of peach allergy, at least in the Mediterranean area.

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[8] - Pastorello EA, Farioli L, Pravettoni V, Ortolani C, Ispano M, Monza M, et al. The major allergen of peach (Prunus persica) is a lipid transfer protein. J Allergy Clin Immunol 1999;103:520-526

BACKGROUND: Allergy to fresh fruits and vegetables is mostly observed in subjects with pollinosis, especially from birch, because of cross-reacting allergens in vegetable foods and pollens. However, allergic reactions to fruits, specifically Rosaceae fruits, have been reported in subjects without pollinosis. OBJECTIVE: This study evaluated the pattern of IgE reactivity, identifying the allergen responsible in 2 groups of patients with oral allergy syndrome to peach with or without birch pollinosis. METHODS: The allergenic components of peach were detected by SDS-PAGE and immunoblotting. The major peach allergen was purified by HPLC with a cation-exchange column followed by gel filtration chromatography. Its IgE-binding capacity and its homology with the protein of the crude extract were demonstrated by immunoblotting inhibition techniques. To better characterize this allergen, periodic acid-Schiff stain and isoelectrofocusing were used. The amino acid sequencing was done with a gas-phase sequencer. RESULTS: SDS-PAGE and immunoblotting of the 15 patients allergic to peach, 8 without and 7 with birch pollinosis, showed that they all recognized a protein with a molecular weight of 9 kd. This was the only allergen recognized by patients not sensitized to pollen, whereas the birch pollen-sensitive patients had IgE binding to other allergenic proteins at higher molecular weights. The purified 9-kd protein retained its IgE-binding capacity, was negative to periodic acid-Schiff stain, and had an isoelectric point value of greater than 9. A search in the Swiss Prot Bank showed this was a lipid transfer protein, belonging to a group of molecules involved in the defensive system of plants. CONCLUSIONS: The major allergen of peach is a 9-kd protein belonging to the group of lipid transfer proteins. This is the only allergen recognized by patients allergic to peach but not sensitized to birch pollen.

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[9] - Botton A, Begheldo M, Rasori A, Bonghi C, Tonutti P. Differential expression of two lipid transfer protein genes in reproductive organs of peach (Prunus persica L. Batsch). Plant Sci 2002;163:993-1000

Two cDNA clones (named Pp-LTP1 and Pp-LTP2) corresponding to different lipid transfer protein (LTP) genes have been isolated from peach (Prunus persica cv. Springcrest) epicarp and ovary, respectively. Sequence analysis revealed that the two fragments share 54% identity at nucleotide level and show common features of plant LTP genes, such as conserved cysteine residues and lipid-binding motifs. Phylogenetic analysis grouped Pp-LTP1 and Pp-LTP2 in two distinct clusters, the former with most of LTP genes sequenced in the Rosaceae family, the latter only with one almond LTP. Genomic Southern data indicated that a small LTP gene family is present in peach. Pp-LTP1 and Pp-LTP2 have been used as gene-specific probes to describe expression in flowers and fruits throughout development. In petals, sepals and stamen only Pp-LTP1 was expressed whereas transcripts of Pp-LTP2 strongly accumulated in non-pollinated and pollinated ovary with a decreasing trend in the period of four weeks after pollination. In fruits, a dramatic accumulation of Pp-LTP1 mRNA was detected in epicarp at all stages of fruit development and, with the exception of the early growth stage, no Pp-LTP1 transcripts have been detected in mesocarp. When Pp-LTP2 was used as a probe in the same fruit tissues, a faint hybridization signal was observed only in epicarp of fruitlets collected at an early growth stage. Infection with Monilia induced only a slight increase of Pp-LTP1 transcript in epicarp of pre-climacteric and climacteric fruits. These results support the hypothesis of multiple roles played by LTPs and, considering that LTPs have been recognized as the major allergen of peach, indicate that Pp-LTP1 could be related to the allergenicity of peach.

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[10] - Botton A, Begheldo M, Rasori A, Bonghi C, Tonutti P, Scott-Johnson R, et al. Factors affecting gene expression of lipid transfer protein (LTP), the major allergen of peach fruit. Acta Hortic 2002;592:237-243

AB: Considering the increase of fruit allergy diseases, a specific research on some molecular aspects of lipid transfer protein (LTP), the major allergen of peach, has been carried out. Fruits of four peach cultivars (Sentry, Royal Gemm, Summered and Tardiva Zuliani) were harvested at commercial ripeness and stored at 20 or 4°C. A 269-bp cDNA fragment (named Pp-LTP1) was PCR-synthesized from poly A+ mRNA extracted from epicarp of Sentry using degenerated primers. Sequencing analysis revealed that Pp-LTP1 encodes a protein belonging to the LTP family. Southern analysis indicated that Pp-LTP1 belongs to a small multigenic family of at least three members. Expression analysis showed that Pp-LTP1 mRNA is present in the epicarp but not in the mesocarp of ripe fruit and that specific transcript accumulation remains unchanged throughout postharvest ripening at 20°C. Storage at low temperatures induced a decrease of Pp-LTP1 transcript accumulation in Sentry and Tardiva Zuliani but not in Royal Gemm and Summered.

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[11] - Carnés J, Fernandez-Caldas E, Gallego MT, Ferrer A, Cuesta-Herrans J. Pru p 3 (LTP) content in peach extracts. Allergy 2002;57:1071-1075

BACKGROUND: Lipid transfer proteins are molecules widely distributed in fruits. Sensitization to LTP is frequent in fruit sensitive patients. The aims of this study were to purify LTP and to assess the content of LTP in ripe peach peel and pulp extracts by ELISA inhibition using polyclonal antibodies . METHODS: LTP was purified from ripe yellow peach peel by two different column chromatography methods. A polyclonal antibody was produced by injecting purified LTP into two New Zealand white rabbits. ELISA inhibition and rabbit monospecific polyclonal antibody were used to calculate the LTP content in Springcrest and Miraflores varieties of peach peel and pulp extracts. Purified LTP (2.5 mg/ml) was used to skin test 24 peach-sensitive patients . RESULTS: The purified LTP showed a single band at approximately 9 kDa. The polyclonal antibody raised anti LTP recognized only the LTP molecule in the peach extracts. LTP content, expressed in micro g/mg of freeze-dried extract in four extracts were: yellow peach peel, 15.48; yellow peach pulp 2.25; red peach peel 14.67 and red peach pulp 1.84. Twenty patients (83.3%) had a positive skin test with purified LTP . CONCLUSIONS: We have developed a system to determine the concentration of LTP in peach extracts. LTP in peel extracts is approximately seven times greater than in pulp.

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[12] - Cavatorta V, Sforza S, Mastrobuoni G, Pieraccini G, Francese S, Moneti G et al. Unambiguous characterization and tissue localization of Pru P 3 peach allergen by electrospray mass spectrometry and MALDI imaging. J Mass Spectrom 2009;44:891-897

The lipid transfer protein (LTP), Pru p 3, has been identified as the major allergen present in peach, and its sequence obtained by direct amino acid sequencing has been previously reported. However, several sequences, obtained from c-DNA and available in databases, show differences among them and from the originally proposed structure. In this paper, we report the fast and unambiguous determination of the structure of Pru p 3 protein, extracted from three different varieties of peach, by electrospray ionization mass spectrometry (ESI-MS), both coupled to single stage (quadrupole) or advanced (FT-HRMS) analyzers. The structure was identical to one of the cDNA-derived sequences and different in two positions from the previously reported structure obtained by amino acid sequencing. Moreover, the exclusive localization of the protein in the outer part of the fruits was assessed by Matrix-Assisted Laser Desorption Ionization Mass Spectrometry Imaging (MALDI MSI). The results reported here demonstrate the full potential of mass spectrometry for rapidly obtaining high quality structural data of relevant food proteins.

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[13] - Barre A, Brulé C, Borges JP, Culerrier R, Jauneau A, Didier A et al. Concentration des LTP dans la peau et la pulpe des fruits. Rev Fr Allergol 2009;49:166-169

Les principaux allergènes des fruits de Rosacées (Prunoidées) correspondent à des protéines de défense (protéines pathogenesis-related [PR]) qui sont essentiellement exprimées en réponse à l‚attaque de la plante par un champignon phytopathogène. Il s‚agit des protéines de transfert des lipides (LTP), des 1,3ß-glucanases et des protéines thaumatin-like (TLP), qui appartiennent respectivement aux familles des protéines PR-14, PR-2 et PR-5. La LTP de pomme (Mal d 3) est principalement localisée dans la peau du fruit et la peau de certaines variétés de pomme (Granny Smith, Golden delicious, Fuji) est particulièrement riche en LTP. La chair du fruit en renferme beaucoup moins. La LTP de pêche (Pru p 3) s‚accumule essentiellement dans les poils formant le revêtement duveteux du fruit. En pratique, l‚épluchage de ces fruits réduit considérablement l‚apport de LTP aux personnes sensibilisées. En revanche, la cuisson des fruits n‚offre aucun intérêt en raison de l‚extrême résistance des LTP à la dénaturation thermique. Dans d‚autres fruits, prune (Pru d 3) et abricot (Pru ar 3), les LTP sont aussi abondantes dans la peau que dans la chair et l‚épluchage de ces fruits ne réduit en rien leur allergénicité. Des résultats préliminaires semblent indiquer que les fruits issus de l‚agriculture biologique possèdent des teneurs en LTP supérieures à celles observées dans les fruits issus de l‚agriculture traditionnelle. Dans les pays du pourtour Méditerranéen, l‚Espagne et l‚Italie en particulier, l‚allergie à la pêche (Pru p 3) s‚accompagne souvent de réactions anaphylactiques sévères.

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[14] - Ahrazem O, Jimeno L, López-Torrejón G, Herrero M, Espada JL, Sánchez-Monge R, et al. Assessing allergen levels in peach and nectarine cultivars. Ann Allergy Asthma Immunol 2007;99:42-47

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[15] - Borges JP, Jauneau A, Brule C, Culerrier R, Barre A, Didier A, et al. The lipid transfer proteins (LTP) essentially concentrate in the skin of Rosaceae fruits as cell surface exposed allergens. Plant Physiol Biochem 2006;44:535-542

The localization and distribution of non-specific lipid transfer proteins (nsLTP) allergens in the skin and pulp of Rosaceae fruits (apple, peach, apricot, plum) has been investigated. nsLTP essentially concentrate in the pericarp of the fruits whereas the pulp contains lower amounts of allergens. Immunolocalization showed they are primarily located in the cytosol but are subsequently excreted and finally accumulate at the plasmalemma-cell wall interface and in the cell wall. However, high discrepancies were observed in the content of allergens among, e.g. different cultivars of apple. As a consequence, the consumption of peeled-off fruits is recommended to reduce the risk of severe allergic reactions (anaphylactic shock) in individuals sensitized to Rosaceae fruits.

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[16] - Tordesillas L, Cuesta-Herranz J, Gonzalez-Muñoz M, Pacios LF, Compés E, Garcia-Carrasco B, et al. T-cell epitopes of the major peach allergen, Pru p 3: Identification and differential T-cell response of peach-allergic and non-allergic subjects. Mol Immunol 2009;46:722-728

Lipid transfer proteins (LTPs), particularly peach Pru p 3, are the most relevant plant food allergens in the South of Europe, and, therefore, their allergic properties have been extensively studied. However, neither T-cell epitopes nor their effect on the patients' T-cell response has been investigated in any member of the LTP panallergen family. The objective of the present study was to map the major T-cell epitopes of Pru p 3, as well as to evaluate their induced T-cell response in peach-allergic versus control subjects. Thus, peripheral blood mononuclear cells (PBMCs) from 18 peach-allergic patients and Pru p 3-specific T-cell lines (TCLs) from 9 of them were cultured with Pru p 3 and with a panel of 17 derived peptides (10-mer overlapping in 5 amino acids representing the full sequence of Pru p 3). Proliferation in 5-day assays was carried out via tritiated-thymidine incorporation, while IL4 and IFNgamma production was assessed via sandwich enzyme-linked immunosorbent tests (ELISA) of TCL culture supernatants. The results were compared to those obtained from 10 non-peach allergic control volunteers. Two consecutive peptides showed the highest activation capacity. About 74% of PBMCs and TCLs recognized them, forming a single T-epitope: Pru p 3(65-80). Additionally, other specific T-cell epitopes were observed. Pru p 3(25-35) was detected by more than 60% of TCLs from peach-allergic patients, and Pru p 3(45-55) only activated PBMCs from control subjects. Interestingly, TCLs from patients were associated with a Th2-type, whereas control TCLs presented a Th1-type cytokine response. The major immunogenic T-cell epitope identified in Pru p 3, Pru p 3(65-80), is a good candidate to develop new vaccines for hypersensitivity reactions associated with LTP allergens from Rosaceae fruits.

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[17] - Pacios LF, Tordesilla L, Cuesta-Herranz J, Compes E, Sánchez-Monge R, Palacín A, et al. Mimotope mapping as a complementary strategy to define allergen IgE-epitopes: Peach Pru p 3 allergen as a model. Mol Immunol 2008;45:2269-2276

Lipid transfer proteins (LTPs) are the major allergens of Rosaceae fruits in the Mediterranean area. Pru p 3, the LTP and major allergen of peach, is a suitable model for studying food allergy and amino acid sequences related with its IgE-binding capacity. In this work, we sought to map IgE mimotopes on the structure of Pru p 3, using the combination of a random peptide phage display library and a three-dimensional modelling approach. Pru p 3-specific IgE was purified from 2 different pools of sera from peach allergic patients grouped by symptoms (OAS-pool or SYS-pool), and used for screening of a random dodecapeptide phage display library. Positive clones were further confirmed by ELISA assays testing individual sera from each pool. Three-dimensional modelling allowed location of mimotopes based on analysis of electrostatic properties and solvent exposure of the Pru p 3 surface. Twenty-one phage clones were selected using Pru p 3-specific IgE, 9 of which were chosen using OAS-specific IgE while the other 12 were selected with systemic-specific IgE. Peptide alignments revealed consensus sequences for each pool: L37 R39 T40 P42 D43 R44 A46 P70 S76 P78 Y79 for OAS-IgE, and N35 N36 L37 R39 T40 D43 A46 S76 I77 P78 for systemic-IgE. These 2 consensus sequences were mapped on the same surface of Pru p 3, corresponding to the helix 2-loop-helix 3 region and part of the non-structured C-terminal coil. Thus, 2 relevant conformational IgE-binding regions of Pru p 3 were identified using a random peptide phage display library. Mimotopes can be used to study the interaction between allergens and IgE, and to accelerate the process to design new vaccines and new immunotherapy strategies.

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[18] - Brenna O, Pompei C, Pravettoni V, Farioli L, Pastorello E.A. Variability of allergen patterns in peach cultivar. 8th International Symposium on Problems of Food Allergy, Venice 2001, March 11-13

Introduction. Recently it has been reported that different cultivar of bell peppers contain different allergenic proteins. Since peach is a fruit of growing relevance as a source of allergens causing OAS, we checked the possibility that different peach cultivar might contain different amount of the Pru p3 allergen, the main allergen to whom OAS to peaches may be ascribed, and to confirm that this allergen might be differently distributed between the peel and the flesh in the different cultivar. Furthermore, our aim was to prepare peach-derived products with reduced Pru p3 content. Materials and Methods. We collected peaches of ten cultivar; some fruits were chemically peeled, sealed under light vacuum in polyethylene bags and kept at 20 °C. The chemical peeling was carried out with a first treatment with 10 % sodium hydroxide, followed by a further one with 2 % sodium hydroxide; in both cases the fruits were washed with distilled water and then rinsed with diluted hydrochloric acid. Extract from each couple of differently treated fruits (the untreated and the chemical peeled fruits) of ten different cultivars of peaches were prepared and samples were analysed by SDS-PAGE, followed by immunoblotting. Results. Coomassie Blue stain revealed only a band corresponding to the mobility of Pru p3 in all the tested cultivar. This band was evident only in the lanes corresponding to the extract of the whole, not peeled fruits, whilst all the samples derived from chemical peeled fruits stained negatively. These results seem to clearly define that the chemical peeling can free the fruits from the Pru p3 allergenic protein. The relative immunoblottig showed a more complex situation, since a rather high number of allergenic band is revealed, not shown by the protein staining. Furthermore, a white flesh cultivar didn't show the Pru p3 band, whereas a very intense band with a MW of about 45 kDa, not shown by protein staining, appeared. Chemical peeling practically removed all these bands. Among the different cultivar tested, Percoca (a cultivar mainly used to prepare syrupy peaches) showed the lightest spots, among which the 45 kDa band was the major one. This band was the major band that keeps showing in all the cultivar tested, even if the chemical peeling led to its weakening. At the tested concentration, no protein band was associated to this immunoblotting revealed band. Conclusions. The different allergenic content of some different cultivar of peaches was demonstrated. Two cultivar among those analysed revealed the presence of a protein band corresponding to the mobility of the Pru p3 allergen, not responding to allergic sera. The possibility of obtaining hypoallergenic peach products, like nectar, jam and jelly starting from chemically pealed fruits is demonstrated.

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[19] - Karamloo F, Wangorsch A, Kasahara H, Davin LB, Haustein D, Lewis NG, et al. Phenylcoumaran benzylic ether and isoflavonoid reductases are a new class of cross-reactive allergens in birch pollen, fruits and vegetables. Eur J Biochem 2001;268:5310-5320

We investigated the biochemical function of the birch pollen allergen Bet v 6 and its role in the IgE-cross-reactivity between birch pollen and plant foods, and characterized Pyr c 5, a Bet v 6-related food allergen, from pear; the proteins were expressed as His-Tag fusion proteins in Eschershia coli and purified by Ni-chelate affinity chromatography under native conditions. Nonfusion proteins were obtained by factor Xa protease treatment. The highest degree of amino-acid sequence identity of Pyr c 5 and Bet v 6 was found with a plant protein related to a defense mechanism, which we have named phenylcoumaran benzylic ether reductase (PCBER) based on its ability to catalyze the NADPH-dependent reduction of 8-5' linked lignans such as dehydrodiconiferyl alcohol to give isodihydrodehydrodiconiferyl alcohol. Enzymatic assays with recombinant Pyr c 5 and Bet v 6 showed PCBER catalytic activity for both recombinant allergens. Both Pyr c 5 and Bet v 6 allergens had similar IgE binding characteristics in immunoblotting and enzyme allergosorbent tests (EAST), and bound IgE from 10 sera of birch-pollen-allergic patients including six pear-allergic subjects. EAST inhibition experiments with Pyr c 5 as the solid phase antigen suggested that homologous allergens may be present in many vegetable foods such as apple, peach, orange, lychee fruit, strawberry, persimmon, zucchini (courgette), and carrot. In extracts of pear, apple, orange, and persimmon, the presence of proteins of approximately 30-35 kDa containing Bet v 6 cross-reactive epitopes was demonstrated with two Bet v 6-specific monoclonal antibodies. Recombinant Pyr c 5 triggered a strong, dose-dependent mediator release from basophils of a pear-allergic subject, suggesting that Pyr c 5 has the potential to elicit type I allergic reactions.

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[20] - San Miguel-Moncin M, Krail M, Scheurer S, Enrique E, Alonso R, Conti A, et al. Lettuce anaphylaxis: identification of a lipid transfer protein as the major allergen. Allergy 2003;58:511-517

BACKGROUND: Allergy to plant-derived foods is associated with birch pollinosis in central and northern Europe. Symptoms elicited are usually limited to the oropharyngeal system. By contrast, in the Mediterranean area, allergy to the same foods manifests more frequently with systemic reactions caused by nonspecific lipid transfer proteins (nsLTP), independently of an associated pollinosis . OBJECTIVE: We sought to investigate the pattern of immunoglobulin E (IgE) binding protein bands implicated in lettuce allergy, in particular the presence of an nsLTP . METHODS: Consecutive lettuce allergic patients were selected. Determination of serum-specific IgE, immunoblot, and inhibition experiments were performed in order to study the pattern of IgE binding proteins and the potential cross-reactivity to pollens. Inhibition studies with recombinant allergens were conducted to identify the lettuce allergens. The major allergen was subjected to N-terminal amino acid sequencing . RESULTS: Fourteen patients were diagnosed as being allergic to lettuce. All were sensitized to Platanus pollen. Ten of them showed specific IgE to a lettuce protein of 9-kDa. The IgE binding to this protein was completely inhibited by the cherry-LTP and peach extract. The N-terminal sequence of the 9-kDa protein showed a high degree of amino acid sequence identity to other nsLTPs. A clear partial cross-reactivity was observed between lettuce-LTP and Platanus-pollen extract . CONCLUSIONS: An LTP has been demonstrated to be a major allergen in patients suffering from lettuce allergy.

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[21] - Vassilopoulou E, Zuidmeer L, Akkerdaas J, Tassios I, Rigby NR, Mills ENC, et al. Severe Immediate Allergic Reactions to Grapes: Part of a Lipid Transfer Protein-Associated Clinical Syndrome. Int Arch Allergy Immunol 2007;143:92-102

BACKGROUND: Grape allergy is considered rare; grape lipid transfer protein (LTP; Vit v 1), an endochitinase and a thaumatin-like protein (TLP) have been reported as grape allergens. A considerable number of patients have referred to our department for severe reactions to grapes, and several IgE binding proteins were detected . OBJECTIVES: The aim of this study was to identify and characterise the allergens involved in severe allergic reactions to grapes and describe the population in which they occur . METHODS: Patients with reported severe allergic reactions to grapes (n = 37) are described. Grape allergens were purified/fractionated by a combination of chromatographic techniques, identified by proteomic analysis and biochemically characterised. Immunoreactivity was assessed by blot (inhibitions) and RAST (inhibitions), and skin prick tests were performed with the isolated allergens . RESULTS: All subjects were polyallergic, sensitised and reactive to several additional foods and pollen. All patients were sensitised to grape LTP. A 28-kDa expansin, a 37.5-kDa polygalacturonase-inhibiting protein, a 39-kDa beta-1,3-glucanase and a 60-kDa protein were identified as minor grape allergens. Endochitinase and TLP did not play a role. Inhibition experiments revealed the possible cross-reactive role of LTP for clinical sensitivities to other LTP-containing plant foods, but also the involvement of cross-reactive carbohydrate determinants of minor allergens in IgE cross-reactivity . CONCLUSIONS: LTP is the major grape allergen, while additional minor allergens may contribute to clinical reactivity. Severe grape allergy presents in atopic patients who frequently react to other LTP-containing, plant-derived foods. The 'LTP syndrome' is the appropriate term to describe this condition.

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[22] - Reuter A, Lidholm J, Andersson K, Ostling J, Lundberg M, Scheurer S, et al. A critical assessment of allergen component-based in vitro diagnosis in cherry allergy across Europe. Clin Exp Allergy 2006;36:815-823

BACKGROUND: Food allergy to cherry occurs throughout Europe, typically with restricted oral reactions in the central and northern parts but with frequent systemic reactions in the Mediterranean region. Previous studies have demonstrated insufficient sensitivity of commercially available cherry extract reagents in the diagnosis of cherry allergy . OBJECTIVE: To assess the diagnostic performance of specific IgE tests based on recombinant cherry allergens in comparison with an extract-based assay and to skin prick test (SPT). A secondary objective was to analyse the frequency of systemic reactions in cherry-allergic subjects across Europe, including the largest population of LTP-sensitized subjects from central Europe studied to date . METHODS: A total of 186 subjects from central Europe and Spain were studied. Serum IgE was analysed with ImmunoCAP tests carrying rPru av 1, 3 and 4, combined and separately, and cherry extract . RESULTS: Among the central European cherry allergics, the mix of rPru av 1, 3 and 4 had a sensitivity of 95%, compared with 65% for cherry extract, and the IgE binding capacity of the recombinant mix was considerably higher. The sensitivity of the two tests was more comparable in the Spanish population, 95% and 86%, respectively. The recombinant allergen ImmunoCAP equalled SPT in terms of sensitivity and specificity. Consistent with previous reports, major geographic differences in sensitization pattern and prevalence of systemic reactions were found. A significantly higher rate of systemic reactions was found in Spanish patients sensitized to Pru av 3 whereas German patients sensitized to LTP only had oral allergy syndrome . CONCLUSIONS: The recombinant cherry allergen ImmunoCAP is a highly sensitive diagnostic tool, clearly superior to any diagnostic method based on cherry extract. Three cherry allergens are sufficient for detecting sensitization in 95% of cherry-allergic subjects. Systemic reactions are common in LTP-sensitized individuals but seem to require at least one additional causative factor.

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[23] - Vassilopoulou E, Zuidmeer L, Akkerdaas J, Tassios I, Rigby NR, Mills ENC, et al. Severe Immediate Allergic Reactions to Grapes: Part of a Lipid Transfer Protein-Associated Clinical Syndrome. Int Arch Allergy Immunol 2007;143:92-102

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[24] - Lombardero M, Garcia-Sellés FJ, Polo F, Jimeno L, Chamorro MJ, Garcia-Casado G, et al. Prevalence of sensitization to Artemisia allergens Art v 1, Art v 3 and Art v 60 kDa. Cross-reactivity among Art v 3 and other relevant lipid-transfer protein allergens. Clin Exp Allergy 2004;34:1415-1421

BACKGROUND: Artemisia vulgaris is a widespread weed in the Mediterranean area and several allergens have been detected in its pollen. One of them, Art v 3, belongs to the lipid-transfer protein (LTP) family and its prevalence in Artemisia-sensitized patients or its relationship with other LTP allergens is not clear . OBJECTIVE: To assess the pattern of sensitization to an array of mugwort allergens in a Mediterranean population, and to study the cross-reactivity of Art v 3 with Pru p 3 and Par j 1, relevant LTP allergens in the area . METHODS: Skin prick test was performed with whole extracts (A. vulgaris, Parietaria judaica and peach) and pure natural allergens Art v 1, Art v 3, Art v 60 kDa and Par j 1 in 24 mugwort-allergic patients from a Mediterranean area. In vitro assays included measurement of specific IgE and ELISA inhibition among LTP allergens . RESULTS: The three Artemisia allergens elicited a positive skin response in 70-80% of the patients. Seven patients were clearly sensitized to Par j 1 and 11 to Pru p 3. There was no correlation between Par j 1 and Pru p 3 sensitization, but a highly significant correlation was found between peach extract and Art v 3 as regards the skin response. No IgE cross-reactivity was observed between Art v 3/Par j 1 or Pru p 3/Par j 1. In contrast, Art v 3 significantly inhibited the binding to Pru p 3 of IgE from three patients' sera out of six studied, but Pru p 3 was not able to inhibit the IgE binding to Art v 3 . CONCLUSION: Art v 3 is a major mugwort allergen and in some patients with IgE to both Art v 3 and Pru p 3, Art v 3 behaves as the primary sensitizing agent.

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[25] - Pastorello EA, Pravettoni V, Farioli L, Rivolta F, Conti A, Ispano M, et al. Hypersensitivity to mugwort (Artemisia vulgaris) in patients with peach allergy is due to a common lipid transfer protein allergen and is often without clinical expression. J Allergy Clin Immunol 2002;110:310-317

Background: The observation of mugwort-specific IgE antibodies in patients with peach allergy suggests that mugwort sensitization might play a role in sensitization to peach. Objective: We sought to study the clinical manifestations of mugwort hypersensitivity in patients with peach allergy, identify the common allergens, and evaluate their IgE crossreactivity. Methods: Patients with oral allergy syndrome for peach and specific IgE antibodies to mugwort were investigated for respiratory symptoms during the mugwort season. Peach and mugwort allergens were identified by means of SDS-PAGE and IgE immunoblotting. Immunoblotting inhibition experiments were done to study cross-reactivity between peach and mugwort and other pollens. Results: Seventeen patients were studied, 10 with no seasonal respiratory symptoms and 7 with clear late summer respiratory symptoms. In IgE immunoblotting the 10 asymptomatic patients reacted only to a 9-kd allergen of both mugwort and peach, whereas the 7 patients with pollinosis reacted to other allergens. Ten patients with mugwort allergy, no history of allergy to peach, and negative results for peach-specific IgE antibodies were also studied. The mugwort 9-kd protein was identified as a lipid transfer protein (LTP) homologous to peach LTP. Immunoblotting inhibition showed that IgE binding to the peach 9-kd band was totally inhibited by 4 µg of peach LTP but only by 400 µg of mugwort LTP, whereas 4 µg of both mugwort and peach LTP totally inhibited the mugwort immunoblotting. The results were similar with other pollens. Conclusions: Patients sensitized only to the 9-kd LTP of mugwort do not present hay fever symptoms, and this sensitization is a consequence of the peach sensitization.

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[26] - Pastorello EA, Pravettoni V, Farioli L, Rivolta F, Conti A, Ispano M, et al. Hypersensitivity to mugwort (Artemisia vulgaris) in patients with peach allergy is due to a common lipid transfer protein allergen and is often without clinical expression. J Allergy Clin Immunol 2002;110:310-317

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[27] - Lombardero M, Garcia-Sellés FJ, Polo F, Jimeno L, Chamorro MJ, Garcia-Casado G, et al. Prevalence of sensitization to Artemisia allergens Art v 1, Art v 3 and Art v 60 kDa. Cross-reactivity among Art v 3 and other relevant lipid-transfer protein allergens. Clin Exp Allergy 2004;34:1415-1421

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[28] - Fernández-Rivas M. The place of lipid transfer proteins (LTP) in the cross-reactivity of plant foods. Rev Fr Allergol 2009;49:433-436

Plant non-specific lipid transfer proteins (nsLTP) are a family of small polypeptides involved in plant defence mechanisms. They possess a compact structure stabilized by four disulphide bridges that confer them a high stability to both thermal treatment and proteolytic digestion, properties that make them true food allergens. Several members of this family have been identified as clinically relevant plant food allergens. Rosaceae fruits are the foods most frequently involved in allergic reactions in patients sensitized to nsLTP, and Pru p 3, the peach nsLTP, seems to be the primary sensitizer in most cases, and can be used as a marker allergen. Since this family of allergens is widely distributed in the plant kingdom and a high degree of IgE cross-reactivity exists among their members, it is common that patients allergic to nsLTP react to a wide variety of plant foods including non-Rosaceae fruits, tree nuts, and vegetables. Due to their high stability, nsLTP are able to induce systemic reactions, especially frequent in those patients who do not have an associated pollen allergy. There is a specific and still unexplained geographical distribution pattern of allergies to nsLTP, that are almost exclusively found in the non-birch pollen areas of the Mediterranean, where nsLTP are the major plant food panallergens.

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[29] - Florido Lopez JF, Quiralte Enriquez J, Arias de Saavedra Alías JM, Saenz de San Pedro B, Martin Casañez E. An allergen from Olea europaea pollen (Ole e 7) is associated with plant-derived food anaphylaxis. Allergy 2002;57(suppl. 71):53-59

Background: Several cross-reacting proteins have been identified as responsible of the co-occurrence of pollinosis and plant-derived food allergy. This association has been mainly described in the birch-apple syndrome but other pollens such as Olea europaea and other fruits may also contain homologous proteins. Objective: To evaluate the associations between sensitization to allergens of Olea europaea pollen and confirmed plant-derived food allergy, in addition to investigate if any pattern of clinical hypersensitivity of food allergy reaction (oral allergy syndrome (OAS) or anaphylaxis) and/or any fresh fruit or nut allergy, are associated to one or several Olea pollen allergen(s). Methods: 134 consecutive patients diagnosed with pollinosis by Olea were studied. Of these patients only 40, reported adverse reaction to plant-derived food. Twenty-one (group A) were classified as OAS and 19 (group B) as anaphylaxis. Skin-tests with six Olea pollen allergens and several groups of fruits, were performed. Double-blind placebo-controlled food challenge (DBPCFC), confirmed the diagnostics of food allergy with the exception of patients who suffered previous anaphylactic reaction. Results: All patients, showed a positive skin prick test (SPT), against one or more of Olea europaea allergens. Sensitization to Ole e 7, was more frequent (P = 0.02) in patients from group B. A total of 84 DBPCFC were performed with 44% positive results. Challenge confirmed at least the 50% of positive SPT in any case (peach: 68.42%; pear: 50%; melon: 71.42% and kiwi: 53.84%). In patients from group B, significant association with O. europaea pollen allergens were found between positive SPT to Rosaceae fruits and Ole e 3 (P = 0.045) and Ole e 7 (P = 0.03); Cucurbitaceae and Ole e 7 (P = 0.03) and Actinidiaceae with Ole e 3 (P = 0.04). Conclusions: The results of this study, establish a new spectrum of associations between pollens and plant-derived foods: sensitization to olive profilin (Ole e 2) is not more frequent in OAS patients. Patients with anaphylactic reaction after eating fruit are also sensitized to Ole e 7, a LTP present in Olea pollen, and suffer pollinic symptoms. Finally a polcalcin (Ole e 3) could be also associated to Olea pollen respiratory and food allergy.

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[30] - Hugues B, Didierlaurent A, Charpin D. Cross-reactivity between cypress pollen and peach: a report of seven cases. Allergy 2006;61:1241-1243

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[31] - Delimi B, Dhivert-Donnadieu H, Demoly P. Allergies cyprès-pêche : allergie croisée ou simple coïncidence ? Rev Fr Allergol Immunol Clin 2007;47:350-354

Plusieurs cas d‚allergies croisées cyprès-pêche ont été rapportés récemment. Dans ce travail, nous avons cherché à confirmer cette possibilité dans une population de 33 patients du pourtour méditerranéen allergiques aux pollens de Cupressacées. L‚analyse des données de ces patients nous a permis de distinguer deux groupes : I) un groupe de neuf patients qui avaient une sensibilisation à la pêche, soit 27,3 % (quatre patients présentaient une allergie vraie à la pêche et cinq patients avaient une simple sensibilisation à ce fruit sans symptômes lors de son ingestion) ; II) un deuxième groupe de 24 patients allergiques aux pollens de cyprès, non sensibilisés à la pêche. Ces résultats permettent de démontrer que le syndrome « cyprès-pêche » existe au même titre que le syndrome « pomme-bouleau », confirmant ainsi des études récentes. Des analyses par immunoblot des sérums de ces patients permettront la caractérisation des allergènes croisants.

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[32] - de Groot H, de Jong NW, Vuijk MH, Gerth van Wijk R. Birch pollinosis and atopy caused by apple, peach, and hazelnut: comparison of three extraction procedures with two apple strains. Allergy 1996;51:712-718

This study aimed, first, to study the prevalence in The Netherlands of atopy caused by apple, peach, and hazelnut in patients with tree pollinosis, and, second, to compare three extraction procedures for skin prick testing with two different apple strains. Skin prick tests and RAST were performed on 79 consecutive patients with tree pollinosis, visiting the department of allergology during spring 1995. In skin prick tests, we used three different extracts (juice, freeze-dried extract, and low-temperature acetone powder extract) of two apple strains, Golden Delicious and Granny Smith. Case histories for apple, peach, and hazelnut were positive in 35 (44.3%), 23 (29%), and 35 (44.3%) patients, respectively. More than two-thirds of the patients had symptoms characteristic of oral allergy syndrome. Skin prick tests for apple, peach, and hazelnut were positive in 51 (64.6%), 61 (77.2%), and 71 (89.9%) patients, respectively. Granny Smith showed more positive skin reactions and a better agreement with clinical history than Golden Delicious, and juice was superior to the two other extraction procedures for both apple strains. RAST for apple, peach, and hazelnut was positive in 53 (68.8%), 13 (16.9%), and 31 (40.3%) patients, respectively. Concordance between skin prick test and case history was found in 77%, 52%, and 54%, for apple, peach, and hazelnut, respectively. We found a high percentage of concurrence of clinical allergy to birch pollen and apple, peach, and hazelnut, confirmed by both skin prick testing and RAST. Approximately half of these patients had symptoms (especially oral allergy syndrome) after eating these products. We also found an easy extraction procedure (juice extract) suitable for apple skin prick testing, superior even to freeze-dried extraction or the low-temperature acetone powder technique.

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[33] - 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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[34] - Mittag D, Vieths S, Vogel L, Becker WM, Rihs HP, Helbling A, et al. Soybean allergy in patients allergic to birch pollen: Clinical investigation and molecular characterization of allergens. J Allergy Clin Immunol 2004;113:148-154

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[35] - Cuesta-Herranz J, Lazaro M, Martinez A, Figueredo E, Palacios R, Delhaseras M, et al. Pollen allergy in peach-allergic patients: sensitization and cross-reactivity to taxonomically unrelated pollens. J Allergy Clin Immunol 1999;104:688-694

BACKGROUND: Fruit allergy has been attributed to cross-reactive IgE to pollens and has been associated with a particular pollen sensitization . OBJECTIVE: The aim of the study was to evaluate sensitization to several taxonomically unrelated pollens in peach- and pollen-allergic patients and to study cross-reactivity between them . METHODS: One hundred sixty-five patients were evaluated: 70 peach- allergic patients together with 95 pollen-allergic patients (control group). Pollen skin tests in duplicate were performed to 5 grasses, 8 trees, and 7 weeds. Cross-reactivity between peach and taxonomically diverse pollens was determined by radioallergosorbent inhibition and Western blot inhibition tests. Experiments were also carried out after preadsorption of the sera with purified natural profilin . RESULTS: The skin test results revealed that peach-allergic patients frequently reacted to most pollens-grasses, weeds, and trees-even when some of these are not found in our geographic area. There was a statistically significant increase in sensitization frequency to most trees and weeds, with a statistically higher occurrence of asthma (odds ratio 2.98, 95% confidence interval 1.46-6.09). Inhibition test results provided evidence that taxonomically unrelated grasses, weeds, and trees produced various and substantial degrees of inhibition in specific IgE to peach and that the peach extract elicited strong inhibitions to those pollens. Profilin was found to be a relevant cross-reactive antigen in these patients . CONCLUSION: The results of this study provide evidence that peach allergy is linked to sensitization to several taxonomically unrelated pollens. This is attributable to the ubiquitous nature of the IgE binding determinants-such as profilins-between peach and taxonomically unrelated pollens.

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[36] - van Ree R, Fernández-Rivas M, Cuevas M, van Wijngaarden M, Aalberse RC. Pollen-related allergy to peach and apple: an important role for profilin. J Allergy Clin Immunol 1995;95:726-734

Birch pollinosis is often accompanied by allergy to fruits such as peach and apple. Bet v I is of major importance as cross-reactive allergen for this combined allergy. We studied a group of patients with combined grass pollinosis and fruit allergy from an area virtually without birch trees. OBJECTIVE: The aim of this study was to investigate the possible involvement of profilin and carbohydrate groups as cross-reactive structures in pollen and fruits. METHODS: RAST inhibition was performed to measure cross-reactive IgE to pollen and fruits. The presence of IgE against profilin was determined in a RAST with purified grass profilin, and IgE against carbohydrate structures was determined in a RAST with proteinase K-digested grass pollen extract. The biologic activity of IgE in response to profilin was tested by in vitro histamine release and skin prick tests. RESULTS: IgE against fruits was shown to be largely cross-reactive with grass pollen. The majority of the patients had IgE against profilin (12 of 16) and carbohydrate structures (9 of 10). Profilin was shown to have biologic activity, in both histamine release and skin prick tests. CONCLUSION: Profilin is an important allergen for patients with combined grass pollen/fruit allergy in areas without birch trees.

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[37] - Daschner A, Crespo JF, Pascual CY. Specific IgE to recombinant vegetal panallergen (rBet v 2) and fruit allergy in pollinic patients. Allergy 1999;53:614-618

The presence of IgE antibodies to the allergens rBet v 1 and rBet v 2 was investigated in patients with known sensitization to rye grass (Lolium perenne) and/or olive (Olea europaea) pollen, by comparing a group of 10 patients who had allergic symptoms after ingestion of fruits [including bananas, figs, kiwifruit, muskmelons, oranges and peaches] with a group of 17 patients who had only seasonal respiratory symptoms. There was no significant difference between the 2 groups for total IgE. All patients showed specific IgE to both L. perenne and O. europaea. No specific IgE binding to rBet v 1 was detected in any patient. The incidence of the presence of IgE antibodies to rBet v 2 was 90% in the group with fruit allergy and 35% in the group [without fruit allergy]. There was a significant association between presence of IgE antibodies to rBet v 2 and fruit allergy (P = 0.007). Specific IgE values to O. europaea pollen were higher in the fruit-allergy group than the group without fruit allergy (P = 0.032). It is concluded that pollen-allergic patients with specific IgE to birch pollen profilin show a significantly elevated frequency of fruit allergy.

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[38] - Gonzalez-Mancebo E, Rodriguez-Pérez R, Alonso MD, Rosado A, Tejedor MA, Vila C, et al. Peach allergy. Clinical characteristics and allergen sensitization in 84 patients evaluated by DBPCFC. EAACI 21th Congress, Naples, 1-5 June, 2002, Poster n°58

Background. Allergy to Rosaceae fruits is a common food allergy in the adult population of the Mediterranean area and among them peach is the most prevalent. We have studied a group of 84 patients referred to our Allergy Unit for adverse reactions to peach in order to establish the clinical characteristics and the allergens involved. Method. The diagnosis of peach allergy was established by double-blind placebo-controlled food challenges (DBPCFC) except in those patiens referring anaphylaxis after the ingestion of peach in the 2 previous years. Skin prick tests (SPTs) were performed with fresh peach peel and peach lipid transfer protein (Pru p 3) at 20 µg/mL. Specific IgE to peach, rBet v1 and rBet v 2 were measured by Pharmacia CAP. The study was performed with the approval of the ethics committee, and written informed consent was given by all the patients. Results. Allergy to peach was confirmed in 62 patients (74%) , whereas 14 subjects did not react in the DBPCFC. Two patients dropped-out. The 62 reactors included 24 males and 38 females, with a mean age of 23 years. An associated pollen allergy was present in 49 patients (79%). Clinical presentation: oral allergy syndrome (OAS) in 57% of patients, isolated systemic involvement (anaphylaxis or urticaria) in 36%, isolated oropharyngeal symptoms in 42 % and contact urticaria in 66%. Positive SPTs with peach peel and Pru p 3 were observed in 94 % and 65 % of the patients, respectively. Positive CAP results for peach, r Bet v1 and r Bet v 2 were obtained in 61%, 7% and 32% of subjects, respectively. No significant differences were observed between the 62 allergics and the 14 tolerants in sex distribution, age, pollen allergy, clinical presentation, and SPT to peach and CAP results. However SPT responses to Pru p 3 were significatively greater in the group of allergics. Conclusions. Peach allergy is a severe food allergy in our population. Although OAS was the most frequent manifestation, systemic involvement was observed in more than 1/3 of patients. The major allergen involved is the lipid transfer protein of peach (Pru p 3). Profilin is a minor allergen and sensitization to rBet v1 is exceptionally detected. The only significant difference observed between allergics and tolerants is a higher skin response to Pru p3 in the former group, suggesting that the IgE response to this major allergen could be related to the clinical reactivity to peach

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[39] - Fernández-Rivas M, González -Mancebo E, Alonso-Díaz de Durana MD, Rodríguez-Pérez R, Benito C, Sánchez-Monge R, et al. The natural history of peach allergy. EAACI 22th Congress, Paris, 7-11 June, 2003, Poster n°131

Background: The natural history of peach allergy is unknown. This study aimed to approach the natural history of peach allergy by analysing the clinical and allergen profiles of peach allergy at different age groups. METHODS: The study comprised 100 patients, 40 males and 60 females, age range 6 to 69 years, who reported adverse reactions to peach. The allergological evaluation included: medical history, skin prick tests (SPTs) with fresh peach (prick-prick method) and Pru p 3 (at 20 mg/ml), and specific IgE to peach and rBet v 2 (Pharmacia CAP System). The actual reactivity to peach was established by a double-blind placebo-controlled food challenge (DBPCFC). For the analysis the patients were divided into 4 categories according to their age at onset of peach allergy: <5, 6-10, 11-15, and >15 years (y). RESULTS: < 5 y 6-10 y 11-15 y > 15 y PN 21 21 20 38 Isolated OAS/Systemic involvement (%) 21/79 64/36 75/25 56/44 0.02 SPT peach (mean, mm”) 78 55 44 33 <0.001 CAP peach (mean, kU/L) 7.18 2.84 4.37 2.14 NS Positive SPTs to Pru p 3 (%) 91 76 35 32 <0.001 Positive CAP to rBet v 2 (%) 25 19 50 51 0.03 Pollen allergy (%) 24 43 67 84 <0.001 Peach avoidance (years) 13.19 11.71 6.35 5.74 <0.001 Positive DBPCFCs (%) 81 86 65 74 NS Conclusions: Peach allergy started before 16 years of age in 62% of patients. The appearance of systemic symtomps after peach intake tends to decrease with age, as well as the skin response to peach, the serum specific IgE, and the frequency of sensitisation to Pru p 3. The appearance of isolated oral symptoms after peach intake increases with age, as well as the frequencies of pollen allergy, and sensitisation to rBet v 2 (birch profilin). Peach allergy appears to be a persistent food allergy, and the time of avoidance does not seem to favour the development of oral tolerance.

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[40] - Gonzalez-Mancebo E, Benito C, Casas ML, Sanchez-Monge R, Salcedo G, Fernandez-Rivas M. Component resolved diagnosis in peach allergy in Spain. Allergy Clin Immunol Int 2005;17(Suppl. 1):342

Background The advantages of recombinant allergen-based diagnosis have been demonstrated in several studies. Using recombinant allergens in vivo or in vitro, a patient's individual IgE reactivity profile can be quantitatively established, this is the so-called component-resolved diagnosis (CRD). The purpose of this study was to apply the CRD to a Spanish population with peach allergy. Method One hundred and fifty-one patients with actual peach allergy, established by double-blind placebo-controlled peach challenges were studied. Sixtynine percent of these patients presented only local symptoms, 7% only systemic symptoms and 24% local and systemic symptoms. Pollen allergy was demonstrated in 79% of the subjects. The group included 102 (67%) females and 49 (32%) males, with a mean age of 23.45 years. IgE determinations to nPru p 3, rBet v 2 and rBet v 1 were carried out. Results IgE determinations to nPru p 3, rBet v 2 and rBet v1 were positive in 69%, 44% and 6% of the patients, respectively. So, using only nPru p 3 specific IgE, 69% of the patients were identified. If nPru p 3 and rBet v 2 positive results were combined, more than 92% of the patients were identified. Adding rBet v 1 specific IgE positive results, only one more patient was identified. Ten patients (6.62%) allergic to peach were not identified by any of these allergens. Systemic symptoms were referred by 90% of the patients sensitized to nPru p 3 and 36% of the patients sensitized to rBet v2. Conclusion Using CRD with nPru p 3 and rBet v 2, we have been able to diagnose 92,7% of our peach allergic population, so we can conclude that purified allergens are useful for diagnosis of peach allergy. CRD allows us to determine sensitization patterns directly and to correlate them with severity of clinical symptoms.

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[41] - Gamboa PM, Caceres O, Antepara I, Sanchez-Monge R, Ahrazem O, Salcedo G, Barber D, Lombardero M, Sanz ML. Two different profiles of peach allergy in the north of Spain. Allergy 2007;62:408-414

BACKGROUND: Peach allergy has two different patterns: central Europe with oral allergy syndrome (OAS) related to a primary sensitization to birch pollen Bet v 1 and profilins and southern Europe with mostly systemic symptoms, in many cases due to sensitization to lipid-transfer proteins . METHODS: Thirty peach-allergic patients with positive skin and food challenge tests and 29 control subjects were included. Skin prick tests (SPT) with inhalant allergens, commercial peach and apple extracts and native Pru p 3 were performed. In vitro specific immunoglobulin (Ig) E to grass pollen, birch pollen, peach, apple, rBet v 1, rBet v 2 and rPhl p 12 was determined by CAP, and rBet v 1, rMal d 1, rMal d 4, rMal d 3 and rPru p 3 using the ADVIA-Centaur platform. Basophil activation test (BAT) with commercial peach extract, commercial apple extract, nPru p 3, rMal d 3, rMal d 1 and rMal d 4 was also performed . RESULTS: Pru p 3 was the major allergen in the patient group from northern Spain. Sensitization to this allergen was found in 100% of the patients with systemic symptoms or contact urticaria. Only 60% of OAS patients were sensitized to Pru p 3, being all of them sensitized to profilins and 60% of them to allergens of the Bet v 1 family. Specific IgE determination and BAT using recombinant allergens (rPru p 3) show specificity and sensitivity values close to 100% . CONCLUSIONS: Most peach-allergic patients coming from the north of Spain present systemic symptoms after ingestion of peach, Pru p 3 being the main allergen. Patients with OAS present profilin-Bet v 1-related sensitization. Thus, in the north of Spain our patients show a mixed central-south Europe pattern with LTP-profilin-Bet v 1 sensitization depending on the symptoms presented. The use of natural and recombinant plant allergens, allows establishing the sensitization patterns to the different allergens studied.

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[42] - Gallo R, Roncarolo D, Mistrello G. Cross-reactivity between latex and sweet pepper due to prohevein. Allergy 1998;53:1007-1008

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[43] - Fontaine JF, Didierlaurent A, Lavaud F, Deslée G, Lebargy F. Latex allergy and cross-reactivity with rosaceae fruits: study of one case by immunoblot. EAACI 22th Congress, Paris, 7-11 June, 2003, Poster n°703

If few cases of cosensitisation were described with natural latex and rosaceae fruits, there is no evidence for cross-reactivity between these allergens. We report the case of a 58-year-old woman, allergic to latex, who presented several anaphylactic shocks after ingestion of avocados, chesnuts, but also cherry plums, peaches or apricots. Skin prick-tests (SPT) were negative for usual aeroallergens, including betula mix pollens, and most of the food allergens. However, SPT were clearly positive for latex, fresh peach, apricot, cherry plum, plum and apple. The specific IgE level measured by CAP Pharmacia° technique were 6,22 kU/l for latex. No specific IgE were found for birch pollen, rBet v1, rBet v2, apple, peach and plum. An immunoblot performed with latex extract and our patient's serum showed a 25 kD protein, which was not found with sera from pool of patients allergic to latex. Immunoblot inhibition with fruits juices resulted in a disappearance of this 25 kD band, and showed a new 28 kD band with peach juice, plum juice and apricot juice; no difference was found after inhibition with apple juice. These results showed, in our case, a cross-reactivity between latex and some fruits belonging to Rosaceae family. A 25 kD protein, which could be a chitinase or a superoxide-dismutase, can be considered as the cross-allergen in latex extract.

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[44] - Blanco C, Carillo T, Castillo R, Quiralte J, Cuevas M. Avocado hypersensivity. Allergy 1994;49:454-459

The avocado (Av) is a fruit that belongs to the Lauraceae family. We report 17 patients with immediate hypersensitivity to avocado. Clinical manifestations in relation to avocado ingestion were as follows: systemic anaphylaxis in seven patients, angioedema/urticaria in six, vomiting in two, bronchial asthma in one, and rhinoconjunctivitis in one. Skin prick test (SPT) with fresh avocado was positive in all patients with the Strong avocado variety (SAv) and in 14 patients with the Hass avocado variety (HAv). Our patient-associated sensitizations were as follows: 10 to latex, eight to chestnut, eight to banana, four to kiwi, and four to walnut. Avocado-sensitized patients with latex allergy were typically middle-aged women, professionally exposed to latex, who also exhibited frequent associated sensitizations to chestnut, banana, and other fruits. Specific IgE against avocado was demonstrated in 11 of our patients, by both commercial CAP and RAST with avocado extract coupled to nitrocellulose disks. Despite its lower protein content, SAv seems to be more allergenic than HAv, both in vivo and in vitro. On incubating a pool of sera from our patients with avocado, latex, chestnut, and banana extracts, a progressive RAST inhibition was obtained, with SAv- and chestnut-marked disks. This suggests the existence of common antigenic determinants among these allergens.

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[45] - Levy DA, Mounedji N, Noirot C, Leynadier F. Allergic sensitization and clinical reactions to latex, food and pollen in adult patients. Clin Exp Allergy 2000;30:270-275

Many latex-allergic patients are sensitized to one or more foods. Patients allergic to tree and/or grass pollens are also often sensitized to plant-derived foods. Atopy, defined in most studies as sensitivity to an aeroallergen, is a risk factor for latex allergy. The relative importance of pollen sensitivity, a sign of atopy, as a risk factor for food allergy in latex-allergic patients has not, however, been examined. OBJECTIVE: To investigate the relationship between pollen sensitivity and sensitivity to food in latex-allergic patients. METHODS: Forty-four latex-allergic patients (Groups 1 and 2), 24 of whom were also allergic to tree and/or grass pollen (Group 1) and 25 pollinosis patients who were not allergic to latex (Group 3) were studied. We obtained a history of reactions to food and skin tested them with 12 fresh-frozen fruits. RESULTS: All 12 foods induced a skin test reaction in at least one patient in each of the three Groups. There were, however, twice as many positive skin test reactions to food in patients with pollinosis, whether or not they were allergic to latex, as there were in patients allergic to latex but not to pollen. Latex-allergic patients were most likely to have a positive skin test and a history of a reaction to avocado or banana whereas patients with pollinosis only were most likely to have a positive skin test and a history of a reaction to apple, peach or celery. CONCLUSIONS: These results suggest that concomitant allergy to pollen is an important risk factor in determining which plant-derived foods sensitize latex-allergic patients.

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[46] - Somoza ML, Rico P, Feliu A, Jiménez A, Rodriguez J, Crespo JF. Banana allergy confirmed by double-blind placebo-controlled food challenge (DBPCFC). EAACI 22th Congress, Paris, 7-11 June, 2003, Poster n°132

Background: Banana, a member of the Musaceae family, is widely consumed all over the world. It is available almost all year round in temperate climates due to heavy exporting from banana-growing countries. Banana allergy has been reported mainly associated to latex allergy. The aim of the study was to investigate clinical features of acute allergic reactions to banana confirmed by double-blind, placebo-controlled, food challenge (DBPCFC). Methods: Case series of 23 adult patients diagnosed with clinical banana allergy in the Food Allergy Unit from 'Hospital Universitario 12 de Octubre' (Madrid). Diagnostic procedure including a clinical questionnaire, skin testing by prick-prick with fresh fruit and detection of specific IgE (CAP FEIA) were performed in all patients reporting adverse reactions to foods. Patients first underwent an open food challenge (OFC), unless they had a convincing history of severe anaphylaxis. Positive OFC reactions were subsequently evaluated by DBPCFCs. All negative results of DBPCFCs were followed by an open feeding. Results: The age of patients ranged from 10 to 66 yr. (median= 24 yr.) with a female/male ratio of 2.8. Most patients (62%) experienced multiple allergic reactions after banana ingestion before being diagnosed with banana allergy. The most common clinical manifestation was the oral allergy syndrome (52%), followed by gastrointestinal anaphylaxis (8%) and acute respiratory symptoms (8%). The results of allergy testing were positive in 94% of the patients; however, an IgE-mediated mechanism could not be demostrated in two patients. In addition, 54 adverse reactions to other 20 different foods of vegetable origin were confirmed by DBPCFC‚s in the banana allergic patients; including melon, 12 patients; chestnut, 6 pt.; and avocado and kiwi, 5 pt. each. Latex allergy was diagnosed in 8 out of 23 (35%). Seventy-eight percent of the patients had pollen allergy. Conclusions: Banana can induce severe anaphylaxis, although most adverse reactions consist of oral allergy syndrome. Isolated banana allergy is uncommon, being associated frequently to melon allergy and latex allergy.

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[47] - Gaspar A, Raulf-Heimsoth M, Pires G, Rihs HP, Yeang HY, Matos V, et al. Latex allergen sensitization patterns in different risk groups and latex-fruit syndrome patients from Portugal. EAACI 23th Congress, Amsterdam, 12-16 June, 2004, Poster n°1188

Background: Latex allergy has been recognized as a clinically important health problem, mainly in some risk groups: spina bifida (SB), other congenital malformations submitted to multiple surgeries (MS) and health care workers (HCW). Purpose: To study IgE-binding reactivity to latex allergens, by using recombinant and natural allergens, in latex-allergic patients from different risk groups including patients with latex-fruit syndrome (LFS). Material and Methods: We selected sera of 51 latex-allergic patients within different risk groups: 20 with SB, 10 MS and 21 HCW; all these patients had positive skin prick tests with commercial latex extract (ALK-Abelló) and serum latex-specific IgE determined by UniCAP®(Pharmacia Diagnostics). Sixteen out of the 51 patients had LFS. A panel of single recombinant latex allergens was coupled to ImmunoCAPs (CAP system®) and the isolated natural allergen nHev b 2 was coupled on paper disks (EAST testing). This panel comprised rHev b 1, nHev b 2, rHev b 3, rHev b 5, rHev b 6.01, rHev b 7, rHev b 8, rHev b 9, rHev b 10 and rHev b 11. The recombinant allergens were produced as fusion protein with maltose-binding protein (MBP) in E. coli. MBP coupled on ImmunoCAPs served as control. Specific IgE values of >0.35kU/l were considered positive. Major allergen is defined if produces a positive IgE response in more than 50% of the tested group. Results: Recombinant Hev b 1 specific IgE antibodies were detected in 70% sera from SB, 30% from MS, 5% from HCW, and in 13% sera from LFS patients. For nHev b 2: SB-69%, MS-71%, HCW-71% and LFS-75%. For rHev b 3: SB-50%, MS-20%, HCW-10% and LFS-13%. For rHev b 5: SB-55%, MS-40%, HCW-62% and LFS-75%. For rHev b 6.01: SB-45%, MS-30%, HCW-76% and LFS-81%. For rHev b 7: SB-33%, MS-0%, HCW-16% and LFS-27%. For rHev b 8: SB-10%, MS-0%, HCW-5% and LFS-6%. For rHev b 9 and rHev b 10: SB-0%, MS-0%, HCW-5% and LFS-7%. For rHev b 11: SB-7%, MS-0%, HCW-5% and LFS-7%. Conclusions: The different routes of exposure influence the IgE antibody pattern. The major latex allergens identified in SB were Hev b 1, Hev b 2, Hev b 3 and Hev b 5, being Hev b 1 the most important one. The major allergen identified in MS was Hev b 2. For occupational exposure route, the major latex allergens identified in HCW were Hev b 2, Hev b 5 and Hev b 6.01, being prohevein the most important one. Regarding cross-reactivity with foods, the major allergens identified in LFS were Hev b 2, Hev b 5 and Hev b 6.01.

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[48] - Rodriguez J, Mielgo R, Gonzalez A, Crespo JF. Allergic Reactions to Fresh Fruits: Beyond Oral Symptoms. AAAAI 62nd Annual Meeting, Miami, 3-7 March 2006, Poster n°1163

RATIONALE: Fruit allergy is frequently considered to be associated with mild oral symptoms in the context of the pollen-food allergy syndrome, while few studies have focused on fruits as a cause of severe allergic reactions METHODS: Three hundred forty-six allergic reactions to fruits were diagnosed in 194 patients (135 female), aged from 14 to 72 yrs. (median = 27 yrs). The diagnostic procedure followed the guidelines for the evaluation of the adverse reactions to foods from the EAACI, including a clinical questionnaire, skin testing, fruit-specific serum IgE (CAP-FEIA) and open challenges, followed if positive by DBPCFC, unless a convincing history of severe anaphylaxis RESULTS: One hundred-eighty (52%) reactions consisted exclusively of isolated oral symptoms. The most frequent offenders were: melon, 42(23%); kiwi, 28(15.5%) and avocado, 24 (13%). One hundred twentyeight (37%) systemic reactions were accompanied with oral symptoms The most frequent foods were peach, 29 (23%); kiwi, 21 (16%), and melon, 18 (14%). Thirty-eight (11%) systemic reactions did not show any oral symptoms. Peach, 10 reactions (26%); banana, 8 (21%); and kiwi, 4(10.5%), were the most frequent causal foods. Grades of severity were: mild, 15 reactions; moderate, 5 reactions; severe, 13 reactions; life-threatening, 5 reactions. Three patients reported having isolated oral symptoms months before their systemic reactions took place (2 peach, 1 banana) CONCLUSIONS: Half of the fresh-fruit allergic reactions consist of isolated oral symptoms, with melon being the most frequent culprit However, systemic reactions are relevant, being peach the most frequent causative agent in our area Funding: Mº Sanidad (Grant G03-094)

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[49] - Quirce S, Bombin C, Aleman A, Sastre J. Allergy to latex, fruit, and pollen. Allergy 2000;55:896-898

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[50] - Rodriguez J, Crespo JF, Burks W, Rivas-Plata C, Fernández-Anaya S, Vives R, et al. Randomized, double-blind, crossover challenge study in 53 subjects reporting adverse reactions to melon (Cucumis melo). J Allergy Clin Immunol 2000;106:968-972

BACKGROUND: Few studies have evaluated IgE-mediated hypersensitivity to melon with details of clinical reactions confirmed by double-blind, placebo-controlled, food challenges (DBPCFCs). OBJECTIVE: We sought to investigate clinical features (type and severity of reactions, age at onset, results of skin prick and in vitro tests, and incidence of other allergic diseases and associated food allergies) of acute allergic reactions to melon confirmed by DBPCFCs. METHODS: Fifty-three consecutive adult patients complaining of adverse reactions to melon were included in the study. Skin prick tests and detection of specific IgE were performed in all patients with melon, avocado, kiwi, banana, chestnut, latex, pollen, and other offending foods. Patients first underwent an open food challenge, unless they had a convincing history of severe anaphylaxis. Positive open food challenge reactions were subsequently evaluated by DBPCFCs. RESULTS: Actual clinical reactivity was confirmed in 19 (36%) of 53 patients. The most frequent symptom was oral allergy syndrome (n = 14), but two patients experienced life-threatening reactions, including respiratory symptoms and hypotension. The positive predictive value for a skin prick test was 42%, and that for specific IgE measurement was 44%. Forty-five reactions to 15 other foods were confirmed in 18 patients. The most common foods associated with melon allergy were avocado (n = 7), banana (n = 7), kiwi (n = 6), watermelon (n = 6), and peach (n = 5). Onset of melon-induced allergic symptoms occurred from 6 to 45 years (median, 20 years), preceded by seasonal rhinitis, asthma, or both in 88% (15/17). CONCLUSION: About one third of reported reactions to melon are confirmed by means of DBPCFC, which has been proven to be the most reliable procedure in the diagnosis of clinical fruit allergy. Isolated melon allergy is rare, with most patients either having allergic rhinitis, asthma, or both and associated food allergies.

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[51] - Gaier S, Oberhuber C, Rigby N, Marsh J, Hemmer W, Mills C, et al. Allergenicity of peach allergens Pru p 1 and Pru p 3 related with protein stability. Allergy 2008;63(suppl. 88):572

Background: Non-specific lipid transfer proteins (nsLTP) are known to be resistant to proteolysis and thermal treatment due to their 4 disulfide-bridges. Therefore, they were described as the elicitors of true food allergies combined with severe allergic reactions. In contrast, Bet v 1-homologues (responsible for the birch-fruit syndrome) cause frequently OAS due to their sensitivity to proteolytic and heat processing. We investigated the effects of thermal and digestive treatment on the major peach allergens, Pru p 1 and Pru p 3. Furthermore, the IgE binding frequency of Austrian Rosaceae allergic patients‚ sera to Pru p 1 and Pru p 3 linked with symptom scores was analyzed. Methods: Recombinant Pru p 1 and natural Pru p 3 were purified according to established methods. CD spectra were performed to analyze heat and pH dependent unfolding and refolding. In vitro gastric and subsequent duodenal digestion were performed under physiological conditions. IgE ELISA was performed with 54 sera from peach allergic patients. Results: Pru p 1 was degraded within seconds after incubation with pepsin and unfolded at pH 3. Whereas, heat treatment at pH 7.5 resulted in unfolding at 651C and complete refolding after cooling from 951C to 251C. Pru p 3 underwent unfolding during thermal treatment at pH 3 and 7.5 at 851C. Refolding was only observed under acidic pH. Pru p 3 remained intact after 1 h of in vitro gastric digestion. Subsequent duodenal digestion resulted in a B6 kD protein fragment displaying the intact Nterminus and positive IgE binding.Overall IgE reactivity to Pru p 1 was found in 87%, to Pru p 3 in 48%, and to profilin in 48%, respectively. Monosensitisation to Pru p 1 (28%), Pru p 3 (6%), and profilin (2%) was also observed. OAS was the most commonly reported symptom (85%). Sensitisation to Pru p 1 was associated with decreased frequency of urticaria and systemic reactions. Whereas sensitisation to Pru p 3 was associated with increased frequency of respiratory symptoms, urticaria, and systemic reactions. Conclusion: IgE reactivity to Pru p 1 and Pru p 3 of a representative number of Rosaceae allergic patients is linked with different severity of food allergic symptoms. Changes in pH and heat treatment exerts great influence on stability of Pru p 1 and Pru p 3. Thus special attention should be drawn to food processing treatment affecting structural related allergenicity.

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[52] - Gaier S, Oberhuber C, Rigby N, Marsh J, Hemmer W, Mills C, et al. Allergenicity of peach allergens Pru p 1 and Pru p 3 related with protein stability. Allergy 2008;63(suppl. 88):572

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[53] - Brenna OV, Pastorello EA, Farioli L, Pravettoni V, Pompei C. Presence of Allergenic Proteins in Different Peach (Prunus persica) Cultivars and Dependence of Their Content on Fruit Ripening. J Agric Food Chem 2004;52:7997-8000

It has been reported that various cultivars of fruits and vegetables may present a different pattern for the contained allergens. Here, we report on the different content in allergenic proteins for different peach (Prunus persica) cultivars, sampled during two consecutive harvest seasons. Fruits from six cultivars of peaches were harvested fully ripe, and the proteins extracted from whole or chemically peeled fruits were analyzed by SDS-PAGE and immunoblotting. All the protein extracts from whole fruit contained a 9 kDa protein. This protein proved to be absent in the extracts taken from chemically peeled fruit. In four cultivars, this protein corresponds to the allergen Pru p3, a lipid transfer protein that causes the oral allergy syndrome (OAS) in sensitized people. In the following year, fruits from four of the six cultivars of peaches studied previously were harvested at different times, at one and two weeks before the commercial ripening time and when fully ripe, to ascertain whether the presence of the 9 kDa allergen might be related to the ripening process. Two cultivars out of four produced an intense allergenic band corresponding to a 9 kDa protein already two weeks before the commercial ripening date, while the others showed a progressive increment of the 9 kDa allergen during ripening.

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[54] - Ahrazem O, Jimeno L, López-Torrejón G, Herrero M, Espada JL, Sánchez-Monge R, et al. Assessing allergen levels in peach and nectarine cultivars. Ann Allergy Asthma Immunol 2007;99:42-47

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[55] - Botton A, Andreotti C, Costa G, Ramina A. Peach (Prunus persica L. Batsch) Allergen-Encoding Genes Are Developmentally Regulated and Affected by Fruit Load and Light Radiation. J Agric Food Chem 2009;57:724-734

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[56] - Botton A, Begheldo M, Rasori A, Bonghi C, Tonutti P, Scott-Johnson R, et al. Factors affecting gene expression of lipid transfer protein (LTP), the major allergen of peach fruit. Acta Hortic 2002;592:237-243

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[57] - Brenna O, Pompei C, Ortolani C, Pravettoni V, Farioli L, Pastorello EA. Technological processes to decrease the allergenicity of peach juice and nectar. J Agric Food Chem 2000;48:493-497

Among vegetable foods peach (Prunus persica) has been recognized as a significant cause of allergy. The protein, which is considered to be the major peach allergen, has been named Pru p 1. Because peaches are consumed both as fresh fruits and after processing to obtain peach juice, nectar, jam, syrupy peach, etc., research was carried out to identify a technological process for production of hypo- or nonallergenic peach-based products. SDS-PAGE and immunoblotting analysis of extracts prepared from four commercial peach nectars showed that the Pru p 1 was not removed, and neither was its allergenic activity decreased by technological treatments carried out for nectar production. Some treatments oriented toward a removal of or, at least, a decrease in the allergenic power were assumed and verified at laboratory scale. A variable considered was heat treatment at 121 degrees C for 10 and 30 min: this treatment was not able to decrease the allergenicity of the Pru p 1 protein. Furthermore, the protein band was still present even after 60-min reaction with two different acidic proteases. The two technological treatments that were found to decrease the major allergen of peach were chemical lye peeling of fruits and ultrafiltration of juice through membranes with suitable cutoff. On the basis of the results obtained from this research, a processing flow sheet was defined to obtain hypoallergenic or probably nonallergenic limpid juices and nectars. These products may represent, besides finished foods, intermediates to obtain various products after addition of further ingredients such as pectins, sugars, and fiber.

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[58] - Brenna O, Pompei C, Ortolani C, Pravettoni V, Farioli L, Pastorello EA. Technological processes to decrease the allergenicity of peach juice and nectar. J Agric Food Chem 2000;48:493-497

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[59] - Brenna OV, Pastorello EA, Farioli L, Pravettoni V, Pompei C. Presence of Allergenic Proteins in Different Peach (Prunus persica) Cultivars and Dependence of Their Content on Fruit Ripening. J Agric Food Chem 2004;52:7997-8000

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[60] - Brenna O, Pompei C, Pravettoni V, Farioli L, Pastorello E.A. Variability of allergen patterns in peach cultivar. 8th International Symposium on Problems of Food Allergy, Venice 2001, March 11-13

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[61] - Cuesta-Herranz J, Lazaro M, de las Heras M, Lluch M, Figueredo E, Umpierrez A, et al. Peach allergy pattern: experience in 70 patients. Allergy 1998;53:78-82

We report the clinical characteristics of peach allergy encountered in a population of peach-allergic patients. We evaluated 165 patients. The 70 peach-allergic patients were diagnosed through clinical history, the skin prick-prick test, and open oral challenge and rub tests to peach. As a pollinic control group, 95 pollen-allergic patients were also evaluated. Some 49% of the patients were male and 51% female. The mean age was 20 +/- 8 years. Oral allergy syndrome (86%) was the most common symptom, followed by contact urticaria (61%) and systemic symptoms (26%). Some 67% of the patients were allergic to peach pulp and 36% reported symptoms related to canned peach. Canned peach and pulp symptoms were statistically associated (P < 0.01), and symptoms to canned peach were more frequently reported by patients with systemic symptoms (P < 0.05). On evaluation of the peach-allergic patients' characteristics, three risk factors--allergy to peach pulp, allergy to canned peach, and peach allergy in non-pollen-allergic patients--were found, indicating development of systemic symptoms on eating peach. Most of the peach-allergic patients (81%) also had pollen allergy, which was linked to a higher prevalence of asthma (73%) than in the pollen-allergic patients of the control group (48%); this difference was statistically significant (P < 0.01). Finally, two groups were clearly defined by the seriousness of the peach allergy--the non-pollen-allergic patients were more predisposed to the occurrence of systemic symptoms (> 50%), and the pollen-allergic patients to asthma (> 70%).

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[62] - Gaier S, Oberhuber C, Rigby N, Marsh J, Hemmer W, Mills C, et al. Allergenicity of peach allergens Pru p 1 and Pru p 3 related with protein stability. Allergy 2008;63(suppl. 88):572

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[63] - Zuidmeer L, van Leeuwen A, Kleine Budde I, Cornelissen J, Bulder I, Rafalska I, et al. Lipid Transfer Proteins from Fruit: Cloning, Expression and Quantification. Int Arch Allergy Immunol 2005;137:273-281

BACKGROUND: Lipid transfer proteins (LTP) are stable, potentially life-threatening allergens in fruits and many other vegetable foods. The aim of this study was to clone and express recombinant apple LTP (Mal d 3), as has previously been done for peach LTP (Pru p 3) and set up quantitative tests for measuring fruit LTPs. METHODS: cDNA for Mal d 3 and Pru p 3 was cloned, expressed in the yeast Pichia pastoris and the resulting proteins were purified via cation exchange chromatography. The immune reactivity of rMal d 3 was compared to nMal d 3 by RAST (inhibition), immunoblotting and basophil histamine release testing. To obtain monoclonal and monospecific polyclonal antibodies, mice and rabbits were immunized with purified nMal d 3. RESULTS: The deduced amino acid sequence of Mal d 3 was identical to the published sequence, Pru p 3 differed at two positions (S9A and S76H). The rMal d 3 had an IgE-binding potency and biological activity close to its natural counterpart. One sandwich ELISA selectively detecting apple LTP and another cross-reactive with cherry, nectarine and hazelnut LTP were developed. In addition, a competitive RIA was developed with polyclonal rabbit antiserum and labeled nMal d 3. CONCLUSION: rMal d 3 (as shown before for rPru p 3) may be a useful tool for application in component-resolved diagnosis of food allergy. Assays for the measurement of LTP will increase the traceability of this potentially dangerous allergen

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[64] - van Ree R, Voitenko V, van Leeuwen WA, Aalberse RC. Profilin is a cross-reactive allergen in pollen and vegetable foods. Int Arch Allergy Immunol 1992;98:97-104

Sera with IgE antibodies against grass pollen often contain IgE against vegetable foods. We investigated the role of the ubiquitous protein profilin in this cross-reactivity. Profilin was purified from Lolium perenne grass pollen by means of affinity purification with Sepharose-coupled poly(L-proline). This solid phase was also used as capturing agent for profilin from pollen and food extracts for application in a radioallergosorbent test. It was shown that profilin is an allergen in grass pollen and in a wide range of vegetable foods, like potato and celery. Within a grass-pollen-sensitive population, patients with IgE to vegetable foods have a high incidence of antibodies against profilin. IgE antibodies against grass pollen profilin were shown to be cross-reactive with respect to vegetable foods

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[65] - Karamloo F, Wangorsch A, Kasahara H, Davin LB, Haustein D, Lewis NG, et al. Phenylcoumaran benzylic ether and isoflavonoid reductases are a new class of cross-reactive allergens in birch pollen, fruits and vegetables. Eur J Biochem 2001;268:5310-5320

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[66] - Rodriguez del Rio P, Rodriguez-Jimenez B, Plaza A, Reig I, Sanchez-Lopez J, Vazquez-Cortes S, et al. Early onset of profilin sensitation. EAACI 25th Congress, Vienna, 10-14 June, 2006, Poster n°1513

Background: Profilin is a cross-reactive allergen in pollens and plant foods. Sensitisation to profilins of plant foods is found in pollen allergic patients who get sensitised through the inhalation route. This is related to the lability of profilin which also explains the restriction of symptoms to the oral cavity (oral allergy syndrome, OAS). The clinical relevance of profilin is questioned in birch pollen allergic patients. However, profilin is a relevant allergen in OAS to plant foods in patients allergic to grass pollen from Southern Europe. Case report: We report a 6 years-old girl with a mild atopic dermatitis who present since the age of 3 years rhinoconjunctival symptoms in spring. At 3.5 years she presented OAS after the intake of watermelon. Since then until the last visit she has presented OAS with other plant foods including apple, kiwi, apricot, plum, peach, nectarine, pear, strawberry, grape, orange, tangerine, banana, tomato, cucumber and hazelnut. She tolerates processed fruits (juices, jam) and latex contact. Skin prick tests (SPT) to inhalants were positive to grass, olive, plane tree, mugwort, and plantain pollens. SPTs with commercial extracts of fruits were negative, but positive results were observed to some of them tested fresh (prick-prick). An oral challenge with fresh plum elicited oropharyngeal pruritus and labial angioedema. Specific IgE (Pharmacia CAP, kU/L) was performed to the following allergens: 2 years 9 months 5 years 6 years Grass mixture 5.11 31.9 65.5 Phleum 2.98 23.2 43.3 rPhl p1 10.6 25.1 27.6 rPhl p5b <0.35 0.79 14.4 rPhl p7 <0.35 <0.35 <0.35 rPhl p12 <0.35 9.01 13.6 rBet v1 <0.35 <0.35 <0.35 rBet v2 <0.35 12.6 15.4 Watermelon <0.35 1.79 1.71 Orange <0.35 0.85 1.21 Apple <0.35 1.66 1.42 Tomato <0.35 - 3.66 Banana 0.39 1.38 1.08 SPT and serum specific IgE to Pru p 3 were negative. SPT with nPho d 2 (date palm profilin) was positive. CAP to latex was 3.03 kU/L, with negative results to the recombinant latex allergens 1, 3, 5, 6.01 and 6.02. CAP inhibition assays of pollens, plant foods and latex were performed with rBet v 2 with the following inhibitions: > 85% to rBet v 2 and rPhl p 12; 40- 100% to watermelon, orange, apple, tomato, banana; 100% to latex; > 70% to mugwort, plane tree and olive pollens; no inhibition to grass and Phleum. CONCLUSION: We present a 6 years old girl with a grass pollen allergy sensitised to group 1 and 5 allergens, and grass profilin, who presents and OAS to multiple fresh plant foods related to the profilin sensitisation.

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[67] - Vazquez de la Torre Gaspar M. Blackcurrant Allergy. AAAAI 62nd Annual Meeting, Miami, 3-7 March 2006, Poster n°177

RATIONALE: The blackcurrant is a temperate shrub that ussually grows by the edge of the pathways when not cultivated. It produces small edible berries, very dark purple/blue colour_almost black_hence the name. They belong to the Rosaceae family, Saxifragaceae order. Although the fruits of this family are a common cause of allergy in our enviroment (i.e. peaches, apple), allergy to blackcurrants has not been reported METHODS: A 50-year-old woman, previously diagnosed of rhinitis due to arizonic, gramineae and olive pollen allergy, presented oral pruritus and pharyngeal occupation with dysphagia while eating either fresh fruit, jam or juice of peach, apricot and nectarine. The same symptoms appear after fresh fruit and jam of blackcurrant, in several times. She tolerates other fruits of the Rosaceae family (i.e. raspberry, strawberry, plum, apple and pear) RESULTS: Peach and raspberry prick tests were possitive. Skin test to blackcurrant resulted negative. Specific IgE determination ( Pharmacia CAP System) to blackcurrant measured 5.7 kU/L, and 2.92 kU/L to peach After SDS-PAGE-Immunoblot IgE reactive bands of >100-43 kd were detected in blackcurrant extract and >100 kd in raspberry extract CONCLUSIONS: Cross-sensitivity among the different species of the Rosaceae family seems variable.We report a case of blackcurrant, peach, apricot and nectarine allergy due to immediate type hypersensitivity (IgE mediated) and subclinical sensitization to raspberry. In our knowledge, this might be the first case of blackcurrant allergy to be reported Funding: Hospital General Universitario Gregorio Maran~ón

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[68] - Asero R, Mistrello G, Amato S, Roncarolo D, Martinelli A, Zaccarini M. Peach fuzz contains large amounts of lipid transfer protein: is this the cause of the high prevalence of sensitization to LTP in Mediterranean countries ? Eur Ann Allergy Clin Immunol 2006;38:118-121

BACKGROUND: Allergy to lipid transfer protein (LTP) is quite common in the Mediterranean countries but virtually absent in Northern Europe. The reasons for this latitude-dependent distribution are unclear. One hypothesis is that peach, the primary sensitizer to LTP, may lose in part its allergenicity as a consequence of treatments (handling, brushing, washing, and packaging) preceding marketing in Northern European. Peach surface fuzz might represent a potential vehicle of LTP. OBJECTIVE: To detect LTP in peach fuzz, and compare IgE reactivity to peach fuzz and peel of sera from LTP-allergic patients. METHODS: IgE reactivity to peach peel and peach fuzz extract was measured by ELISA using sera from 2 LTP-allergic PATIENTS. Purified peach LTP was used in inhibition studies. RESULTS: Both sera strongly reacted both to peach peel and fuzz but reactivity to fuzz was stronger than to peel. Pre-absorption of one serum with peach LTP caused an 87% reduction of IgE reactivity to peach fuzz extract. CONCLUSION: Peach fuzz contains large amounts of LTP and might be a potential vehicle of this allergen causing sensitization in genetically predisposed subjects. Fuzz loss during pre-marketing handling of peaches might be at the basis of the geographic differences that characterize allergy to LTP.

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[69] - Ahrazem O, Jimeno L, López-Torrejón G, Herrero M, Espada JL, Sánchez-Monge R, et al. Assessing allergen levels in peach and nectarine cultivars. Ann Allergy Asthma Immunol 2007;99:42-47

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