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La cerise et autres fruits des Rosacées

vendredi 2 avril 2010, par Allerdata

Les fruits des Rosacées les plus étudiés sur le plan allergologique sont la pomme et la pêche. Le présent article traite des autres fruits :

Le Réseau d’Allergo-Vigilance avait colligé peu de cas de réactions sévères sur les 900 déclarations en mai 2010 : poire 2 cas, fraise 1 cas, mûres 1 cas.

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

La cerise

Surtout étudiée en Suisse et en Allemagne, l’allergie à la cerise est probablement sous estimée dans d’autres régions d’Europe où la pollinose au bouleau est courante.

On rencontre aussi des réactions avec la cerise dans le pourtour méditerranéen. L’origine de cette allergie est alors une sensibilisation aux LTP, a priori initiée par la pêche.

Le tableau ci-dessous donne un aperçu des fréquences relatives d’allergie cerise / pêche / pomme selon les régions d’Europe et le mode de recrutement des patients.

RecrutementPatientsPollinoseCerisePêchePommeRef
.
Allemagne
cerise 101 bouleau 100% 100% 57% 96%
TPODA+ 104 graminées 34%, bouleau 31% 26% 21% 68%
cerise 117 100% 64% 96%
France
céleri 20 bouleau 2 7
Rosacées 12 bouleau+ 6 7 11
10 bouleau - 0 8 4
Portugal
pêche (LTP+ 95%) 30 pollinose 57% 13% 100% 57%
Italie
pêche 21 graminées 16, bouleau 14 10 21
Rosacées, LTP - 212 bouleau 99%, .. 40% 52% 87%
Rosacées, LTP+ 25 bouleau 12%, .. 16% 88% 44%
abricot 14 bouleau+ 7 11 1
9 bouleau - 3 7 4
Rosacées, LTP+ 37 9 30 16
prune (LTP+ 80%) 23 graminées 14, bouleau 11 65% 100%
Rosacées mono-LTP 20 polliniques 10 (bouleau non) 5 20 7
mono-Bet v 1+ (non LTP+) 25 58% 100%
mono-Bet v 2+ (non LTP) 18 11% 39%
Espagne
Rosacées 11 non polliniques 4 10 10
22 graminées, olivier, .. 6 22 15
orange (LTP+ 7) 27 gram 26, olivier, .. 2 19 9
cerise 22 100% 90% 45%
allergie alim. 16 platane, .. 2 2
Grêce
raisin 11 2 10 6


L’homologue de Bet v 1 dans la cerise est Pru av 1 . La cerise contient aussi une LTP (Pru av 3) et une profiline (Pru av 4) .

Une protéine thaumatine-like (Pru av 2) , glycosylée , s’accumule avec le mûrissement du fruit .

Le recombinant de Pru av 2 synthétisé dans E. coli, et donc non glycosylé, est reconnu par moins de patients que l’allergène naturel .

La place prise par Pru av 2 dans l’allergénicité de la cerise n’est pas encore bien définie.

Une étude n’a pas trouvé de différence importante d’allergénicité entre cultivars de cerise, même si certains d’entre eux avaient des taux plus élevés de LTP (ex. cerise Napoléon) .

La cerise aigre, Prunus cerasifera, contient aussi une PR-10 IgE-réactive : Pru c 1.

En digestion gastrique simulée, Pru av 1 et la profiline Pru av 4 ne résistent pas, contrairement à Pru av 3 .

Une bande de 24 kDa, qui ne semble pas être Pru av 2, résiste aussi .

La chaleur détruit en grande partie l’IgE-réactivité de Pru av 1 dans les jus, conserves et confitures , tandis que la LTP résiste .

Pour les fruits au sirop, le stade de pelage chimique des cerises semble important dans la perte d’allergénicité du produit, y compris pour les patients sensibilisés aux LTP . La présence de sucres induit aussi des réactions de Maillard, contribuant parallèlement à faire chuter l’IgE-réactivité pour Pru av 1 .

La poire

On possède peu de données concernant l’allergie à la poire.

Celle-ci a été principalement étudiée en Allemagne.

La poire est clairement un fruit associé au syndrome bouleau-Rosacées, tous les sujets allergiques à la poire et polliniques au bouleau étant positifs pour rBet v 1 .

La poire contient quelques allergènes bien identifiés :

  • Pyr c 1, l’homologue de Bet v 1
  • Pyr c 3, une LTP
  • Pyr c 4, une profiline
  • Pyr c 5 qui appartient à la famille des isoflavone réductases.

Pyr c 5 a été trouvé fréquemment positif chez des sujets allergiques à la poire (syndrome oral) et polliniques au bouleau .

Une isoflavone réductase IgE-réactive est présente aussi dans le pollen de bouleau : Bet v 6. Mais il est difficile de déterminer la part prise par Pyr c 5 dans les réactions cliniques à la poire, tous les sujets étant aussi réactifs pour Pyr c 1.

Des tests d’inhibition ont suggéré la présence d’autres isoflavone réductases (IFR) dans différents fruits (ex. Rosacées, litchi, orange) ou pollens (ex. armoise) .

Cependant d’autres travaux seraient nécessaires pour valider ces réactivités croisées et déterminer s’il existe un "syndrome IFR".

Le tableau ci-dessous donne un aperçu des fréquences relatives d’allergie poire / pêche / pomme selon les régions d’Europe et le mode de recrutement des patients. Schématiquement :

  • en environnement PR-10 (bouleau) : poire = pêche < pomme
  • en environnement LTP : poire < pomme < pêche
RecrutementPatientsPollinosePoirePêchePommeRef
.
Allemagne
cerise 101 bouleau 100% 53% 57% 96%
poire 7 bouleau 7 5 6
TPODA+ 104 graminées 34%, bouleau 31% 12% 21% 68%
Italie
Rosacées, LTP - 212 bouleau 99%, .. 40% 52% 87%
Rosacées, LTP+ 25 bouleau 12%, .. 16% 88% 44%
Rosacées, LTP+ 37 6 30 16
Espagne
Rosacées 11 non polliniques 3 10 10
22 graminées, olivier, .. 12 22 15
TC+ Rosacées 28 polliniques 16 1 22 6
TPO+ 95 polliniques 1 14 3
orange (LTP+ 7) 27 graminées 26, olivier, .. 8 19 9

L’abricot

Ce fruit est rarement cité dans le cadre du syndrome bouleau-Rosacées. Il a été plus fréquemment associé au syndrome LTP rencontré en zones méditerranéennes.

La LTP Pru ar 3 a été identifiée par Pastorello . Cette LTP de 9 kDa a 91 % d’identité avec celle de pêche, Pru p 3. Une autre LTP, celle-ci de 7 kDa, a été trouvée dans l’abricot mais son IgE-réactivité éventuelle n’a pas été étudiée.

A noter que, contrairement à la pêche, pulpe et peau d’abricot ont des concentrations similaires en LTP .

L’abricot, comme les autres Rosacées, possède une profiline et une PR-10.

Une polyphénol-oxydase s’accumule dans l’abricot avec le mûrissement  : est-ce un facteur d’instabilité de la PR-10 (Pru ar 1) dans le fruit et, de ce fait, d’une moindre allergénicité que celle vue pour la pêche chez les polliniques au bouleau ?

Le tableau ci-dessous donne un aperçu des fréquences relatives d’allergie abricot / pêche / pomme selon les régions d’Europe et le mode de recrutement des patients. Schématiquement :

  • en environnement PR-10 (bouleau) : abricot << pêche < pomme
  • en environnement LTP : abricot < pomme < pêche
RecrutementPatientsPollinoseAbricotPêchePommeRef
.
Allemagne
cerise 101 bouleau 100% 8% 57% 96%
TPODA+ 104 graminées 34%, bouleau 31% 4% 21% 68%
Italie
pêche 21 graminées 16, bouleau 14 6 21
Rosacées, LTP - 212 bouleau 99%, .. 20% 52% 87%
Rosacées, LTP+ 25 bouleau 12%, .. 15% 88% 44%
abricot 14 bouleau+ 14 11 1
9 bouleau - 9 7 4
Rosacées, LTP+ 37 11 30 16
prune (LTP+ 80%) 23 graminées 14, bouleau 11 52% 100%
Espagne
Rosacées 11 non polliniques 2 10 10
22 graminées, olivier, .. 5 22 15
TC+ Rosacées 28 polliniques 16 5 22 6
allergie fruits (TPODA ou A) 65 polliniques 65% 5 38 10
orange (LTP+ 7) 27 graminées 26, olivier, .. 1 19 9

La prune

L’allergénicité des prunes (prune, quetsche, mirabelle) est plus faible que celle des pêches si l’on en croit les données épidémiologiques.

Dans l’étude de Pastorello qui a permis l’isolement de la LTP, Pru d 3, tous les sujets allergiques à la prune étaient allergiques à la pêche .

Cette moindre allergénicité clinique est similaire à celle de l’abricot, une autre Rosacée prunoïdée. Elle ne s’explique pourtant pas par un déficit d’homologie entre allergènes de prune et de pêche. Ni par un contact moins allergisant, les fruits étant consommés de façon équivalente.

Par contre, des concentrations en LTP plus faibles dans la prune que dans la pêche ont été relevées . Qui plus est sans écart important entre la pulpe et la peau.

On connaît aussi une PR-10 (Pru d 1) et une profiline IgE-réactives dans la prune

Le tableau ci-dessous donne un aperçu des fréquences relatives d’allergie prune / pêche / pomme selon les régions d’Europe et le mode de recrutement des patients. Schématiquement :

  • en environnement PR-10 (bouleau) : prune < pêche < pomme
  • en environnement LTP : prune < pomme < pêche
RecrutementPatientsPollinosePrunePêchePommeRef
.
Allemagne
cerise 101 bouleau 100% 9% 57% 96%
TPODA+ 104 graminées 34%, bouleau 31% 15% 21% 68%
Portugal
pêche (LTP+ 95%) 30 pollinose 57% 13% 57%
Italie
pêche 21 graminées 16, bouleau 14 8 21
Rosacées, LTP - 212 bouleau 99%, .. 11% 52% 87%
Rosacées, LTP+ 25 bouleau 12%, .. 20% 88% 44%
abricot 14 bouleau+ 2 11 1
9 bouleau - 2 7 4
Rosacées, LTP+ 37 8 30 16
Rosacées mono-LTP 20 polliniques 10 (bouleau non) 4 20 7
Espagne
Rosacées 11 non polliniques 4 10 10
22 graminées, olivier, .. 5 22 15
TC+ Rosacées 28 polliniques 16 4 22 6
TPO+ 95 polliniques 2 14 3
allergie fruits (TPODA ou A) 65 polliniques 65% 4 38 10

Asero a récemment décrit un cas d’anaphylaxie à la prune chez un patient allergique au latex et montrant une réactivité pour un nouvel allergène dans le latex, l’UDP glucose pyrophosphorylase .

En blot une réaction croisée est vue entre prune et latex, mais la responsabilité directe de l’UDP glucose phosphorylase dans cette réactivité croisée n’a pas été testée.

La fraise

La fraise a une place assez limitée dans les réactions allergiques aux fruits des Rosacées, loin derrière la pêche ou la pomme, comme le montre le tableau ci-dessous :

RecrutementPatientsPollinoseFraisePêchePommeRef
.
Allemagne
cerise 101 bouleau 100% 25% 57% 96%
Espagne
Rosacées 22 graminées, olivier, .. 3 22 15
TC+ Rosacées 28 polliniques 16 1 22 6
noisette 18 3 18 12
Grêce
raisin 37 graminées, olivier, .. 84% 2 8 3

Pourtant on connaît différentes protéines IgE-réactives dans la fraise :

  • une PR-10, Fra a 1 (18 et 20 kDa ), homologue de Bet v 1
  • une LTP, Fra a 3, qui a 63 % d’identité avec Pru p 3 (pêche)
  • une probable isoflavone réductase croisant avec Pyr c 5 (poire) , se présentant en blot comme une bande de 35 kDa et montrant un pourcentage d’identité de 53-67 % avec Pyr c 5 et l’isoflavone réductase du bouleau, Bet v 6
  • on a aussi détecté une alpha expansine de 20 kDa et un homologue d’Ole e 1 (pollen d’olivier) (39-45 % d’identité) .

Comme pour d’autres Rosacées, l’allergie peut provenir d’une réactivité croisée avec des PR-10 (bouleau) ou des LTP (pêche).

S’agissant de la LTP de fraise, Fra a 3, on a noté des différences dans les acides aminés exposés en surface de l’allergène comparativement à Pru p 3, y compris au niveau des épitopes de Pru p 3 .

Une réactivité croisée Pru p 3-Fra a 3 est possible, de même qu’une positivité pour Fra a 3 chez des sujets allergiques aux Rosacées.

Cependant certains auteurs estiment que, contrairement à la plupart des autres LTP de Rosacées, Fra a 3 n’a pas de relevance clinique .

Pour la fraise, la profiline Fra a 4 aurait une certaine importance en cas de poly-pollinose hors d’un environnement riche en bouleaux.

La framboise

Les Rosacées du genre Rubus fournissent au moins 2 fruits classés dans les fruits rouges, mais dont la consommation est moindre que celles des cerises et des fraises : la framboise (Rubus idaeus) et la mûre (Rubus fruticosus).

Si l’on se réfère à la littérature médicale, les cas d’allergie aux framboises sont excessivement rares :

  • un cas récemment décrit en Suisse (où une réactivité de type LTP était suggérée in vitro)
  • deux cas où la framboise est évoquée dans les antécédents :
    • chez une patiente Espagnole ayant présenté un œdème de Quincke avec la groseille et le cassis
    • et chez un patient Espagnol exploré après une anaphylaxie avec la mûre (du genre Morus, famille des Moracées)
  • enfin, dans l’étude menée par Marzban en vue de caractériser les allergènes dans la framboise, aucun patient ne rapportait de réactions à l’ingestion de ces fruits, même si 4 des 6 testés étaient positifs en prick natif pour la framboise

Les allergènes dénommés dans la framboise sont Rub i 1 (PR-10) et Rub i 3 (LTP).

Sont aussi IgE-réactives  :

  • une chitinase de classe 3 (30 kDa, glycosylée), proche de l’hévamine du latex (74% d’identité). Mais il ne semble pas que la framboise soit à classer parmi les « aliments latex » : en effet, parmi 7 patients présentant cette bande 30 kDa en blot framboise, seuls 2 étaient positifs pour le latex in vitro.
  • une cyclophiline de 17 kDa

Une protéine thaumatine-like a été évoquée dans la framboise du fait d’une réaction croisée notée avec celle de cerise, Pru av 2 . Mais il y a tout lieu de penser que cela recouvrait la participation de CCD. Cette thaumatine-like est donc à confirmer dans la framboise.

La mûre, le fruit des ronces

Il ne faut pas confondre ce fruit des Rosacées du genre Rubus avec celui de la famille des Moracées (mûre noire ou blanche, Morus spp.).

Une PR-10 et une LTP sont présentes dans la mûre sauvage (Rubus fruticosus), le fruit des ronces .

Un cas d’anaphylaxie avec ce fruit a été décrit chez un patient ayant eu précédemment des épisodes de syndrome oral également avec des fraises et des framboises. En blot il n’y avait pas de bande au niveau des LTP. Et l’allergène en cause n’a pas été caractérisé .

[1] - Scheurer S, Pastorello EA, Wangorsch A, Kästner M, Haustein D, Vieths S. Recombinant allergens Pru av 1 and Pru av 4 and a newly identified lipid transfer protein in the in vitro diagnosis of cherry allergy. J Allergy Clin Immunol 2001;107:724-731
BACKGROUND: In central and northern Europe food allergy to fruits of the Rosaceae family is strongly associated with birch pollinosis because of the existence of IgE cross-reactive homologous allergens in birch pollen and food. By contrast, in the Mediterranean population allergic reactions to these fruits frequently are not related to birch pollen allergy and are predominantly elicited by lipid transfer proteins (LTPs). OBJECTIVE: We sought to determine the prevalence of IgE sensitization to the recombinant cherry allergens Pru av 1 and Pru av 4 in comparison with cherry extract within a representative group of patients who were allergic to cherries recruited in Germany and to compare the relevance of IgE to cherry LTPs in Italian patients. METHODS: Recombinant Pru av 1 and rPru av 4 were available from earlier studies. The cDNA of the cherry LTPs was obtained by using a PCR-cloning strategy. The protein was expressed in Escherichia coli and purified by means of metal chelate affinity chromatography. Sera from 101 German patients with birch pollinosis and oral allergy syndrome to cherry and sera from 7 Italian patients with cherry allergy were investigated by using enzyme allergosorbent tests for IgE reactivity with cherry extract, rPru av 1, rPru av 4, and the recombinant cherry LTP. Inhibition experiments were performed to compare the IgE reactivity of natural and recombinant cherry LTPs and to investigate potential cross-reactivity with birch pollen allergens. RESULTS: The LTP from cherry comprises 91 amino acids and a 26 amino acid signal peptide. The mature cherry LTP shows high amino acid sequence identity with allergenic LTPs from peach (Pru p 3, 88%), apricot (Pru ar 3, 86%), and maize (Zea m 14, 59%) and displays no IgE cross-reactivity with birch pollen. The IgE prevalences in the German patients were as follows: LTP, 3 of 101 (3%); rPru av 1, 97 of 101 (96.0%); rPru av 4, 16 of 101 (16.2%); and cherry extract, 98 of 101 (97%). All 7 Italian patients had IgE against the cherry LTP. CONCLUSIONS: Recombinant allergens are useful tools for a more accurate in vitro IgE-based diagnosis of cherry allergy. Taken together, they mimic the allergenic activity of cherry extract, having slightly higher biologic activity. Sensitization to the cherry LTP is relevant for a minority of patients recruited in Germany, but our data indicate that it may be a major allergen in Italy.
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[2] - Zuberbier T, Edenharter G, Worm M, Ehlers I, Reimann S, Hantke T, et al. Prevalence of adverse reactions to food in Germany – a population study. Allergy 2004;59:338-345
OBJECTIVE: A population study was performed to identify the prevalence of all kinds of adverse reactions to food . METHODS: In a representative cross-sectional survey performed in 1999 and 2000 in Berlin, 13 300 inhabitants of all ages were addressed by questionnaire. This questionnaire was answered by 4093 persons. All respondents mentioning any sign of food intolerance or the existence of allergic diseases (n = 2298) were followed up by telephone and, in case food intolerance could not be ruled out by patient history, were invited to attend to the clinic for personal investigation including double-blind, placebo-controlled food challenge tests (DBPCFC) . RESULTS: The self-reported lifetime prevalence of any adverse reaction to food in the Berlin population (mean age 41 years) was 34.9%. Eight hundred and fourteen individuals were personally investigated according to the guidelines. The point prevalence of adverse reactions to food confirmed by DBPCFC tests in the Berlin population as a mean of all age groups was 3.6% (95% confidence interval [3.0-4.2%]) and 3.7% in the adult population (18-79 years, 95% confidence interval [3.1-4.4.%]). Two and a half percent were IgE-mediated and 1.1% non-IgE-mediated, females were more frequently affected (60.6%). Based on a statistical comparison with available data of adults from the nationwide German Health Survey from 1998, adverse reactions to food in the adult population of Germany (age 18-79) were calculated with 2.6% [2.1-3.2%]) . CONCLUSIONS: The study gives for the first time information about the point prevalence of both immunological and nonimmunological adverse reactions to food and underlines the relevance of this issue in public health. The data also show that an individualized stepwise approach including provocation tests is mandatory to confirm the diagnosis.
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[3] - 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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[4] - Pauli G, Bessot JC, Braun PA, Dietemann-Molard A, Kopferschmitt-Kubler MC, Thierry R. Celery allergy: clinical and biological study of 20 cases. Ann Allergy 1988;60:243-246
In 20 patients, the ingestion of celery was responsible for mucocutaneous symptoms (generalized urticaria and angioedema) (18/20) and respiratory disorders (7/20). Four cases of systemic anaphylaxis were observed. The main associated allergic disorder was pollinosis (16/20). Food allergy to other vegetable products, mainly other Umbelliferae and apples, coexisted with celery allergy in 12 cases. It was found that celery allergy is mediated by IgE antibodies: it is easily diagnosed by skin tests (fresh extracts of celery may be used) and by adequate RAST (17 positive results). Cosensitization with mugwort pollen (14 cases) and birch pollen (9 cases) was found. Celery allergens responsible for clinical sensitization originate chiefly in the tuber and are at least partly thermally labile. The frequent association with pollen sensitization suggests the existence of common antigenic epitopes in celery extracts and mugwort and birch pollens. The immunologic investigations carried out so far (RAST inhibition and immunoprint) seem to support this hypothesis.
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[5] - Giovannini L, Bourrier T, Noormahomed MT, Albertini M, Boutté P. L'allergie aux rosacées chez l'enfant: à propos de vingt-deux cas. Rev Fr Allergol Immunol Clin 2004;44:625-633
Objectif. - L'allergie aux Rosacées est peu fréquente chez l'enfant en France. A contrario, elle est au 4e rang des allergies alimentaires de l'enfant en Espagne. Notre étude se propose d'analyser les caractéristiques de l'allergie aux Rosacées dans une population d'enfants français en milieu méditerranéen et de mettre en évidence ses spécificités. Patients et méthodes. - Cette étude rétrospective a colligé 22 cas d'allergie alimentaire pédiatrique aux fruits de la famille des Rosacées. Le diagnostic évoqué sur l'anamnèse clinique a été confirmé par la réalisation de prick-tests natifs, de prick-tests commerciaux et de CAP-RAST Pharmacia®. La sensibilisation au bouleau a également été recherchée par prick-test. Résultats. - Deux groupes ont été individualisés : 12 enfants sensibilisés au bouleau et huit enfants non sensibilisés. Pour deux enfants cette sensibilisation n'a pu être déterminée. La faible valeur diagnostique du prick-test commercial est commune aux deux groupes. Le groupe bouleau négatif a débuté son allergie par la pêche et est sensibilisé systématiquement à l'extrait natif cuit des fruits auxquels il est allergique. Le nombre de réactions sévères a été plus fréquent. Les prick-tests commerciaux ne sont positifs que dans ce groupe. Le groupe bouleau positif est plus fréquemment allergique à la pomme et le syndrome oral prédomine. Il présente une médiane de sensibilisations aux fruits crus plus élevée. Conclusion. - La région méditerranéenne française possède un profil spécifique de sensibilisations aux Rosacées avec 60 % de sensibilisations type Europe du Nord (pomme-bouleau) et 40 % de sensibilisation type Espagne-Italie (lipid-transfer-protein) .
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[6] - Rodrigues-Alves R, Pereira-Santos MC, Lopes A, Lopes-Silva S, Spinola-Santos A, Costa C, et al. Clinical, anamnestic and serological features of peach allergy in Portugal. Int Arch Allergy Immunol 2008;149:65-73
BACKGROUND: Peach is a common food allergen source throughout Europe. The aim of this study was to characterize peach allergy in a Portuguese patient population . METHODS: Thirty peach-allergic patients confirmed by double-blind placebo-controlled food challenges and 29 controls were included. All subjects completed a standardized questionnaire regarding symptoms and epidemiologic characteristics, skin prick tests with inhalant allergens and foods as well as specific IgE antibodies to peach, recombinant peach allergens rPru p 1, rPru p 3, rPru p 4 and cross-reactive carbohydrate determinants . RESULTS: Thirty-seven percent of patients reported only oral allergy syndrome, while 37% reported generalized urticaria and/or angioedema, 17% localized contact urticaria and 10% anaphylaxis with peach. Sensitization to other Rosaceae fruits and tree nuts was present in 90 and 77% of the patients, respectively. Respiratory allergy history was associated with less severe symptoms (oral allergy syndrome or contact urticaria; p < 0.01) and positive skin prick test to peach peel or plum with more severe symptoms (urticaria and/or angioedema or anaphylaxis; p < 0.05). Ninety-seven percent were sensitized to Pru p 3, 13% to Pru p 4, 3% to Pru p 1 and 10% to cross-reactive carbohydrate determinants. Pru p 3 specific IgE was associated with Artemisia vulgaris sensitization and tree nut allergy (p < 0.05) but not with clinical severity . CONCLUSIONS: Half the patients reported systemic reactions to peach. Peach allergy appeared predominantly mediated by Pru p 3 but some patients were sensitized to Pru p 4. Applying a 0.10 kU(A)/l cutoff level, the diagnostic value of combining the 3 recombinant allergens was noteworthy, with 100% sensitivity and 90% specificity.
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[7] - Pastorello EA, Ortolani C, Farioli L, Pravettoni V, Ispano M, Borga A, et al. Allergenic cross-reactivity among peach, apricot, plum, and cherry in patients with oral allergy syndrome: an in vivo and in vitro study. J Allergy Clin Immunol 1994;94:699-707
Oral allergy syndrome in response to fruits and vegetables frequently occurs as clusters of hypersensitivity to members of the same botanical family, for which the immunologic basis lies in a number of common allergens, most of them still unidentified. OBJECTIVE: This study was designed to assess the in vivo and in vitro cross-reactivity between fruits of the Prunoideae subfamily (i.e., peach, cherry, apricot, and plum) and to identify their major allergens and the cross-reactivity of the peach extract with grass and birch pollen. METHODS: The in vivo study was conducted by skin prick tests and open food challenges with fresh fruits in 23 patients with oral allergy syndrome for peach and positive skin prick test and RAST results for the other Prunoideae. In vitro sodium dodecylsulfate-polyacrylamide gel electrophoresis was followed by immunoblotting and immunoblotting-inhibition. RESULTS: A 13 kd component was identified as the only major allergen common to all the Prunoideae, the other major allergens were found at 14 kd in peach and at 30 kd in cherry. Immunoblotting inhibition showed wide cross-reactivity within the Prunoideae, whereas grass and birch pollen partially inhibited the peach blotting. CONCLUSIONS: Clinical cross-reactivity to Prunoideae is essentially due to a common 13 kd IgE-binding component, which seems to be the most important major allergen of this subfamily, not shared with grass and birch pollen.
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[8] - Asero R. Detection and clinical characterization of patients with oral allergy syndrome caused by stable allergens in Rosaceae and nuts. Ann Allergy Asthma Immunol 1999;83:377-383
A minority of patients with oral allergy syndrome (OAS) induced by Rosaceae or nuts are positive on skin prick tests with commercial food extracts. This suggests reactivity against distinct stable allergens. OBJECTIVES: (1) To define the prevalence of subjects positive on skin prick tests with commercial extracts among patients with OAS caused by Rosaceae and/or nuts and (2) To investigate whether commercial extracts-positive subjects show some peculiar clinical feature and may represent a specific subset with food allergy. METHODS: Skin prick tests were carried out with a large panel of commercial extracts of airborne allergens (Allergopharma) and of vegetable foods (Dome/Hollister-Stier) in 298 adults with OAS caused by Rosaceae (n = 237) and or nuts (n = 161), positive on skin prick tests with fresh offending foods. RESULTS: 25/237 (11%) patients were positive on prick tests with commercial plum extract. This subgroup showed a higher incidence of systemic symptoms (64% versus 6%; P < .001) and a lower incidence of birch pollen allergy (12% versus 99%; P < .001) than commercial extract-negative patients; moreover, 36% versus 0%, respectively, did not have respiratory allergy (P < .001). Apple and peach were the main offending foods among commercial extract-negative and commercial extract-positive patients, respectively (87% versus 44% for apple, P < .001; and 52% versus 88% for peach, P < .005). Eight of one hundred sixty-one (5%) nuts-sensitive patients were positive on prick test with commercial walnut extract. This subgroup showed a higher proportion of patients who experienced systemic symptoms (63% versus 6%, P < .001), a lower prevalence of birch pollen allergy (13% versus 97%, P < .001), and a higher prevalence of grass pollen allergy (88% versus 41%, P < .05) than commercial extract-negative subjects. Further, reactivity against commercial walnut extract was associated with skin reactivity against commercial extracts of peanut (88% versus 37%, P < .005), tomato (75% versus 5%, P < .001), and plum (63% versus 8%, P < .001), and inversely related with skin reactivity against fresh apple (P < .001). In most cases, high levels of IgE specific for peach, apple, and hazelnut were associated with peanut reactivity rather than with clinical sensitivity to specific foods. In a preliminary investigation, most commercial extract-positive patients reacted against a 10-kDa protein characterized as a lipid transfer protein (LTP). CONCLUSIONS: Skin prick tests with commercial extracts of plum and walnut may be usefully employed to detect patients with OAS reacting against stable allergens. The high prevalence of systemic symptoms in these patients suggests that allergens' stability is associated with a higher resistance to the gastrointestinal environment and strongly influences the clinical expression of vegetable food allergy. At least some stable allergens, namely lipid transfer protein might be shared by botanically unrelated fruits such as nuts, peanuts, legumes, tomato, and Prunoideae.
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[9] - Pastorello EA, D'Ambrosio FP, Pravettoni V, Farioli L, Giuffrida G, Monza M, et al. Evidence for a lipid transfer protein as the major allergen of apricot. J Allergy Clin Immunol 2000;105:371-377
Apricots are widely grown in Europe, and allergic reactions are becoming more common, especially oral allergy syndrome. Apricot belongs to the botanical subfamily of Prunoideae, which includes peach, the major allergen of which was identified as a 9-kd protein, a lipid transfer protein (LTP). OBJECTIVE: The aim of the study was to evaluate the IgE reactivity pattern to an apricot extract in subjects with allergic reactions to apricot, as demonstrated by a positive oral challenge response. METHODS: Thirty patients were investigated. All the patients displayed oral allergy syndrome (2 with systemic reactions) to apricot, with positive open food challenge responses, skin prick test responses, and serum-specific IgE antibodies to apricot. The IgE reactivity pattern to apricot extract was identified by using SDS-PAGE and immunoblotting. The major allergen, a 9-kd protein, was then purified by HPLC and characterized by periodic acid-Schiff stain, isoelectric point determination, and N-terminal amino acid sequencing. RESULTS: The sera from all patients allergic to apricot recognized the 9-kd protein, whereas none of the other allergens, with molecular weights from 15 to 80 kd, acted as a major allergen. The 9-kd allergen has an isoelectric point of 8.7 and is not glycosylated. Determination of the N-terminal 34 amino acid sequence showed that it belongs to the LTP family, with a 94% homology with the LTP from peach. IgE blotting of the apricot extract was completely inhibited by the 9-kd purified LTP from peach. CONCLUSIONS: The major allergen of apricot is an LTP, which is highly cross-reactive with the LTP from peach.
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[10] - Asero R, Mistrello G, Roncarolo D, de Vries SC, Gautier MF, Ciurana CL, et al. Lipid transfer protein: a pan-allergen in plant-derived foods that is highly resistant to pepsin digestion. Int Arch Allergy Immunol 2000;122:20-32
Lipid transfer proteins (LTPs) are small molecules of approximately 10 kD that demonstrate high stability. They have recently been identified as allergens in the Rosaceae subfamilies of the Prunoideae (peach, apricot, plum) and of the Pomoideae (apple). They belong to a family of structurally highly conserved proteins that are also present in non-Rosaceae vegetable foods. OBJECTIVE: The aim of this study was to investigate the cross-reactivity to non-Rosaceae LTPs, and to study the role of protein stability in allergenicity. METHODS: Thirty-eight patients with a positive SPT to Rosaceae fruit extracts enriched for LTP were characterized by interview and SPT. To investigate IgE cross-reactivity between Rosaceae and non-Rosaceae LTPs, RAST and RAST inhibition as well as ELISA and ELISA inhibition were performed, using whole food extracts and purified LTPs. Both purified natural LTPs (peach, carrot and broccoli) and Pichia pastoris recombinant LTPs (carrot and wheat) were included. Pepsin digestion was used to address the role of stability in the allergenicity of LTPs. RESULTS: IgE antibodies to Rosaceae LTPs reacted to a broad range of vegetable foods, including Gramineae (cereals), Leguminosae (peanut), Juglandaceae (walnut), Anacardiaceae (pistachio), Brassicaceae (broccoli), Umbelliferae (carrot, celery), Solanaceae (tomato), Cucurbitaceae (melon), and Actinidiaceae (kiwi). Binding and inhibition studies with purified natural and recombinant LTPs confirmed their role in this cross-reactivity. Many of these cross-reactivities were accompanied by clinical food allergy, frequently including systemic reactions. Antibody binding to LTP was shown to be resistant to pepsin treatment of whole extract or purified LTP. CONCLUSION: LTP is a pan-allergen with a degree of cross-reactivity comparable to profilin. Due to its extreme resistance to pepsin digestion, LTP is a potentially severe food allergen.
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[11] - Pastorello EA, Farioli L, Pravettoni V, Giufridda MG, Ortolani C, Fortunato D, et al. Characterization of the major allergen of plum as a lipid transfer protein. J Chromatogr B Biomed Appl 2001;756:95-103
BACKGROUND: Allergy to Prunoideae fruit (plum, peach, cherry and apricot) is one of the most frequent food allergies in southern Europe. All these fruits cross-react in vivo and in vitro, as they share their major allergen, a 9 kD lipid transfer protein (LTP). OBJECTIVE: The aim of the study was the identification and molecular characterization of the major allergen of plum. METHODS: The IgE pattern of reactivity to plums was investigated by SDS-PAGE and immunoblotting with the sera of 23 patients. The identified major allergen was purified by HPLC, using a cationic-exchange column followed by gel-filtration. Further characterization was achieved by periodic-Schiff stain, isoelectrofocusing and N-terminal amino acid sequencing. RESULTS AND CONCLUSIONS: The major allergen of plum is a 9 kD lipid transfer protein, not glycosylated and with a basic character (pI>9), highly homologous to the major allergen of peach.
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[12] - Asero R, Mistrello G, Roncarolo D, Amato S, Caldironi G, Barocci F, et al. Immunological cross-reactivity between lipid transfer proteins from botanically unrelated plant-derived foods: a clinical study. Allergy 2002;57:900-906
BACKGROUND: Lipid transfer proteins (LTP) are highly conserved and widely distributed throughout the plant kingdom. Recent studies demonstrated immunological cross-reactivity between LTP from many botanically unrelated fruits and vegetables and concluded that LTP are pan-allergens. This study aimed to evaluate the clinical relevance of such cross-reactivity in a group of subjects monosensitized to LTP . METHODS: Twenty LTP-hypersensitive patients were selected from a population of about 600 subjects with history of Rosaceae allergy by means of: 1) negative skin prick test (SPT) with a commercial birch pollen extract; 2) positive SPT with a commercial plum extract, rich in LTP but virtually lacking both Bet v 1-like proteins and profilin; 3) in-vitro IgE reactivity to the 9-10 kDa fraction of peach peel or immunoblot with peach peel showing a single band at 10 kDa; and 4) total inhibition of reactivity to whole peach extract (containing Bet v 1-related allergen, profilin, and LTP) by purified peach LTP on enzyme-linked immunoassay (ELISA). Allergy to foods other than Rosaceae was ascertained by careful interview and analysis of medical recordings. SPT with a large series of plant-derived foods were carried out as well. The cross reactivity between LTPs from botanically unrelated plant-derived foods was assessed by ELISA inhibition tests using walnut and peanut extracts as substrate, and peach LTP as inhibitor . RESULTS: All patients reported allergic reactions after the ingestion of at least one from a large number of vegetable foods other than Rosaceae, and in several cases clinical reactions were very severe (anaphylaxis, asthma, urticaria/angioedema). Nuts and peanuts were the most frequently reported causes of allergic reactions (80% and 40% of patients, respectively). All patients showed positive SPT to several non-Rosaceae food extracts. SPT with nuts, peanut, legumes, celery, rice, and corn were positive in the majority of patients. In ELISA inhibition studies, absorption of sera with peach LTP caused complete inhibition of IgE reactivity to walnut and peanut in all cases . CONCLUSION: LTP is a clinically relevant pan-allergen. Most Rosaceae-allergic, LTP-hypersensitive patients experience adverse reactions after ingestion of botanically unrelated plant-derived foods as well. In view of the high prevalence and severity of the allergic reactions induced, hazelnut, walnut, and peanut should be regarded as potentially hazardous for these patients.
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[13] - Asero R, Mistrello G, Roncarolo D, Amato S, Zanoni D, Barocci F, et al. Detection of clinical markers of sensitization to profilin in patients allergic to plant-derived foods. J Allergy Clin Immunol 2003;112:427-432
BACKGROUND: A proper classification of patients allergic to plant-derived foods is of pivotal importance because the clinical features of allergic reactions to fruits and vegetables depend on the nature and characteristics of proteins responsible for sensitization. However, in normal clinical settings this is presently impossible . OBJECTIVE: We sought to detect clinical markers of sensitization to profilin . METHODS: Seventy-one patients allergic to fruits and vegetables but not sensitized to lipid transfer protein or natural rubber latex were studied. Food allergy was ascertained on the basis of clinical history and positive skin prick test responses with fresh foods, commercial extracts, or both. Allergies to foods that had caused less than 2 adverse reactions were confirmed by means of open oral challenge. IgE reactivity to rBet v 1/rBet v 2 and to natural Phleum species profilin were detected. Moreover, IgE to the 30- to 40-kd and 60- to 90-kd birch pollen-enriched fractions, which also can be involved in cross-reactivity phenomena, were measured in sera from 52 patients by means of ELISA . RESULTS: On the basis of in vitro tests, 24, 18, and 25 patients turned out to be sensitized to Bet v 1, Bet v 2, or both, respectively. Four patients had negative test results for both allergens. Hypersensitivity to Bet v 2 was strongly associated with clinical allergy to citrus fruits (39% in patients monosensitized to Bet v 2 vs 4% in patients monosensitized to Bet v 1, P <.025), melon or watermelon (67% vs 0%, P <.001), banana (66% vs 8%, P <.001), and tomato (33% vs 0%, P <.05), whereas Bet v 1 sensitivity was associated with clinical allergy to apple (100% vs 39%, P <.001) and hazelnut (56% vs 0%, P <.001). The sensitivity of a history of allergy to gourd fruits, citrus fruits, tomato, banana, or a combination thereof as a means to detect profilin-hypersensitive patients was 85% (41/48). The specificity of an allergy to any of these fruits exceeded 85%, with positive predictive values ranging between 68% and 91% . CONCLUSION: In clinical settings in which laboratory investigations are not easily accessible, allergy to melon, watermelon, citrus fruits, tomato, and banana can be used as a marker of profilin hypersensitivity once a sensitization to natural rubber latex and lipid transfer protein is ruled out.
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[14] - Fernández-Rivas M, Van Ree R, Cuevas M. Allergy to Rosaceae fruits without related pollinosis. J Allergy Clin Immunol 1997;100:728-733
BACKGROUND: Rosaceae fruit allergy is frequently associated with birch pollinosis in Central and Northern Europe and with grass pollen allergy in Central Spain. The main cross-reactive structures involved for birch pollinosis are Bet v 1 and profilin, and for grass pollinosis they are profilin and carbohydrate determinants. Rosaceae fruit allergy can occasionally be observed in patients without pollinosis . OBJECTIVE: We investigated the clinical presentation and the allergens involved in allergy to Rosaceae fruit without pollinosis . METHODS: Eleven patients from Central Spain allergic to apples, peaches, and/or pears but not to pollens were compared with 22 control subjects with combined grass pollen and fruit allergy. Skin prick tests and RASTs to apple, peach, and pear were performed. Cross-allergenicity was studied by RAST inhibition. Bet v 1 was tested with an indirect RAST, and profilin was tested in skin prick tests, histamine release, and RAST . RESULTS: Rosaceae fruit allergy without pollinosis is severe with 82% of patients reporting systemic symptoms, mainly anaphylaxis (73%), whereas oral symptoms are less frequent (64%). Anaphylactic shock was observed in 36% of patients. The fruit allergens involved showed cross-reactivity among Rosaceae species but were not related to profilin or Bet v 1. Ninety-one percent of patients with combined grass pollinosis and fruit allergy reported oral allergy, 45% reported systemic symptoms, 18% reported anaphylaxis, and 9% reported anaphylactic shock . CONCLUSION: Allergy to Rosaceae fruits in patients without a related pollen allergy is a severe clinical entity. Profilin- and Bet v 1-related structures are not involved in Rosaceae fruit allergy without pollinosis.
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[15] - Ahrazem O, Ibáñez D, López-Torrejón G, Sánchez-Monge R, Sastre J, Lombardero M, et al. Lipid Transfer Proteins and Allergy to Oranges. Int Arch Allergy Immunol 2005;137:201-210
BACKGROUND: Lipid transfer proteins (LTPs) are relevant fruit allergens, recently proposed as model plant food allergens. No citrus fruit allergen has been characterized to date. We sought to identify and isolate citrus fruit LTPs and to explore their relevance in orange allergy . METHODS: Twenty-seven patients, showing mainly oral allergy syndrome after orange ingestion, as well as positive prick responses and serum-specific IgE levels to orange, were selected. Natural orange and lemon LTPs, as well as a recombinant orange LTP isoform expressed in Pichia pastoris, were isolated by chromatographic methods and characterized by N-terminal amino acid sequencing and matrix-assisted laser desorption/ionizaion mass spectrometry, and DNA sequencing of the corresponding cDNA in the case of the recombinant allergen. Specific IgE determination, immunodetection, ELISA-inhibition assays and in vivo skin prick tests (SPTs) were performed with all three purified allergens and with the major peach LTP allergen, Pru p 3 . RESULTS: The natural allergens purified from orange (nCit s 3) and lemon (nCit l 3) showed very similar N-terminal amino acid sequences (18 out of 20 identical residues), typical of LTPs, and molecular masses of 9,610 and 9,618 Da, respectively. The recombinant orange isoform (rCit s 3) expressed in P. pastoris (16 out of 20 residues identical to its natural counterpart in the N-terminal region) presented 92 amino acid residues and 9,463 Da, and 67% sequence identity with rPru p 3. Of the 27 sera analyzed, specific IgE to the purified allergens was found in 54% for nCit l 3, 48% for nCit s 3, 46% for rCit s 3 and 37% for rPru p 3. Positive SPT responses were obtained in 7 out of 26 patients tested for nCit s 3, 3 out of 8 for nCit l 3 and 10 out of 26 for nPru p 3. ELISA-inhibition assays showed an equivalent IgE-binding pattern for the natural and recombinant orange LTPs, and IgE cross-reactivity among the purified orange, lemon and peach LTP allergens . CONCLUSIONS: Members of the LTP allergen family are involved in allergy to oranges, displaying positive in vitro and in vivo reactions in 30-50% of the patients studied. Both orange and lemon allergens show cross-reactivity with the major peach allergen Pru p 3.
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[16] - 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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[17] - Alonso R, Enrique E, Pineda F, Basagaña M, San Miguel-Moncín MM, Bartra J, et al. An Observational Study on Outgrowing Food Allergy during Non-Birch Pollen-Specific, Subcutaneous Immunotherapy. Int Arch Allergy Immunol 2007;143:185-189
BACKGROUND: Birch pollen-specific immunotherapy (SIT) decreases allergy to foods containing birch pollen-homologous allergens. Cross-reactivity was also observed between plane tree pollen and some vegetable foods . OBJECTIVE: The aim of this study was to evaluate the outgrowing of food allergy by patients suffering from vegetable food allergy associated with plane tree pollinosis (rhinoconjunctivitis and/or asthma) during plane tree pollen SIT . METHODS: An observational and prospective study was conducted in 16 adult patients suffering from vegetable food allergy (hazelnut, walnut, lettuce, peach and cherry) and from plane tree pollinosis receiving plane tree pollen SIT for 1 year. Open oral challenges with the implicated food were performed before and after SIT. Blood samples were drawn for measurement of pollen- and food-specific IgE and IgG4 before and after treatment . RESULTS: Plane tree SIT resulted in a significant decrease in food allergy, since the mean food quantity provoking objective symptoms increased from 2.19 to 13.74 g (p < 0.05), and 6 of the 11 patients tolerated the highest level (25 g) of the challenged food after plane tree SIT. Laboratory data also showed a decrease in IgE levels and an increase in IgG4 levels after immunotherapy . CONCLUSION: SIT with plane tree pollen has a positive impact on food allergy in plane tree pollen-allergic subjects.
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[18] - Kalogeromitros DC, Makris MP, Gregoriou SG, Mousatou VG, Lyris NG, Tarassi KE, et al. Grape anaphylaxis: a study of 11 adult onset cases. Allergy Asthma Proc 2005;26:53-58
Reports of immunoglobulin E (IgE)-mediated allergic reactions to grapes and wine are limited in the literature. Nevertheless, grapes are widely grown and consumed in Mediterranean countries. The object of this prospective study was to present clinical features, in vivo and in vitro allergy testing, and human leukocyte antigen (HLA) serotyping in patients with recurring reactions to grapes and grape products. Eleven unrelated Greek patients, six men and five women (aged 16-44 years; mean, 26.9 years) were enrolled based on a documented history of IgE-mediated reactions to grapes, wine, or other grape products. Their evaluation included full history, reaction severity, clinical examination, skin-prick tests with food allergens and molds, serum IgE, specific IgEs to the same allergen battery, and HLA typing. Patients reported 35 grape-induced anaphylaxis episodes ranging from moderate (more than one system involved but not prominent respiratory or cardiovascular symptoms; 45.5%) to severe (serious respiratory obstruction and/or hypotension and loss of consciousness; 54.5%). A causative agent was identified: wine, 10/35 (28.6%); red grapes, 9/35 (25.7%); stuffed vine leaves, 8/35 (22.9%); raisins, 3/35 (8.6%); white grapes, 2/35 (5.7%); wine vinegar, 2/35 (5. 7%); and grape juice, 1/35 (2.9%). Other foods that induced anaphylaxis were apples (54.5%), cherries (18.6%), peaches (18.6%), and bananas (9.3%). Specific IgE values were in accordance with skin-prick tests reactivity. Concerning HLA typing, 9/11 possessed HLA-DR11(5) and -DQ7(3) and the remaining two possessed HLA-DR17(3) and -DQ2 antigens. Grapes, wine and other grape products might cause serious allergic reactions in sensitized individuals. The cosensitization and reaction incidence to other fruit allergens could be a basis for further investigation of panallergens of fruits. HLA class II antigens may contribute in genetic predisposition to these allergic reactions.
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[19] - Schöning B, Vieths S, Petersen A, Baltes W. Identification and characterization of allergens related to Bet v1 in apple, cherry, celery, and carrot by two-dimensional immunoblotting, and N-terminal micro-sequencing. J Sci Food Agric 1995;65:431-440
Possible relationships between Bet v I, the major birch pollen allergen, and the corresponding allergens in fruits and vegetables were studied.] Low temp. extracts taken from Granny Smith and Braeburn apples, cherries, celery and carrots were separated by 2-dimensional electrophoresis. Bet v I-related allergens in apple, cherry, celery and carrot extracts were detected by means of 2-dimensional immunoblotting with patients' sera containing IgE antibodies specific to Bet v I, a rabbit polyclonal antiserum raised against Bet v I, and two Bet v I specific mouse monoclonal antibodies. The major cross-reacting allergen spots were observed with mol. wt./isoelectric points of 18.0 kDa/pI 5.5 for apple (Granny Smith and Braeburn) and 18.0 kDa/pI 5.8 for cherry, 15.5 kDa/pI 4.4-4.6 for celery and 16.0 kDa/pI 4.4 for carrot extract. Additional antibody reactivities with certain isoprotein spots were observed, which may indicate the presence of Bet v I-related epitopes on these proteins. Based on the first 15 N-terminal amino acid residues, the major allergen spots revealed 53% sequence identity between Bet v I and the Granny Smith apple allergen, 50% between Braeburn apple allergen and Bet v I, 67% between Bet v I and the cherry allergen, and 40 and 28% for celery and carrot, respectively. The N-terminal sequences showed identities ranging from 40% (apple/cherry) to 66% (celery) with PcPR l-l, a pathogenesis-related protein in parsley.
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[20] - Scheurer S, Pastorello EA, Wangorsch A, Kästner M, Haustein D, Vieths S. Recombinant allergens Pru av 1 and Pru av 4 and a newly identified lipid transfer protein in the in vitro diagnosis of cherry allergy. J Allergy Clin Immunol 2001;107:724-731
BACKGROUND: In central and northern Europe food allergy to fruits of the Rosaceae family is strongly associated with birch pollinosis because of the existence of IgE cross-reactive homologous allergens in birch pollen and food. By contrast, in the Mediterranean population allergic reactions to these fruits frequently are not related to birch pollen allergy and are predominantly elicited by lipid transfer proteins (LTPs). OBJECTIVE: We sought to determine the prevalence of IgE sensitization to the recombinant cherry allergens Pru av 1 and Pru av 4 in comparison with cherry extract within a representative group of patients who were allergic to cherries recruited in Germany and to compare the relevance of IgE to cherry LTPs in Italian patients. METHODS: Recombinant Pru av 1 and rPru av 4 were available from earlier studies. The cDNA of the cherry LTPs was obtained by using a PCR-cloning strategy. The protein was expressed in Escherichia coli and purified by means of metal chelate affinity chromatography. Sera from 101 German patients with birch pollinosis and oral allergy syndrome to cherry and sera from 7 Italian patients with cherry allergy were investigated by using enzyme allergosorbent tests for IgE reactivity with cherry extract, rPru av 1, rPru av 4, and the recombinant cherry LTP. Inhibition experiments were performed to compare the IgE reactivity of natural and recombinant cherry LTPs and to investigate potential cross-reactivity with birch pollen allergens. RESULTS: The LTP from cherry comprises 91 amino acids and a 26 amino acid signal peptide. The mature cherry LTP shows high amino acid sequence identity with allergenic LTPs from peach (Pru p 3, 88%), apricot (Pru ar 3, 86%), and maize (Zea m 14, 59%) and displays no IgE cross-reactivity with birch pollen. The IgE prevalences in the German patients were as follows: LTP, 3 of 101 (3%); rPru av 1, 97 of 101 (96.0%); rPru av 4, 16 of 101 (16.2%); and cherry extract, 98 of 101 (97%). All 7 Italian patients had IgE against the cherry LTP. CONCLUSIONS: Recombinant allergens are useful tools for a more accurate in vitro IgE-based diagnosis of cherry allergy. Taken together, they mimic the allergenic activity of cherry extract, having slightly higher biologic activity. Sensitization to the cherry LTP is relevant for a minority of patients recruited in Germany, but our data indicate that it may be a major allergen in Italy.
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[21] - Inschlag C, Hoffmann-Sommergruber K, O'Riordain G, Ahorn H, Ebner C, Scheiner O, et al. Biochemical characterization of Pru a 2, a 23-kD thaumatin-like protein representing a potential major allergen in cherry (Prunus avium). Int Arch Allergy Immunol 1998;116:22-28
BACKGROUND: The prevalence of allergy to fruits and vegetables increased with pollinosis over the last 10 years. So far, clusters of hypersensitivity have been established and corroborated by the molecular characterization of individual cross-reacting allergens. Several case studies demonstrated the existence of allergic reactions to fruits of the subfamily Prunoideae (apricots, cherries, plums and peaches). Here, we present the characterization of a major allergen in cherry. METHODS: Characterization was performed using IgE immunoblotting and immunoblot inhibition, N-terminal sequencing, mass spectroscopy analysis and PCR-based cDNA cloning. RESULTS: A 23-kD protein was identified as IgE-binding component. As all cherry-extract-reactive sera displayed IgE-binding to this band, it was designated a major allergen from Prunus avium (Pru a 2). Sequencing the corresponding cDNA identified Pru a 2 as a thaumatin-like protein belonging to the group 5 of pathogenesis-related proteins. CONCLUSIONS: A thaumatin-like protein in cherry has been identified as a major allergen (Pru a 2). Homologous proteins from the thaumatin family share sequence similarities and should therefore be checked for the capability to elicit an IgE-mediated allergic reaction.
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[22] - Fuchs HC, Radauer C, Mari A, Altmann F, Bublin M, Scheiner O, et al. Pru av 2, the Major Allergen From Sweet Cherry, Possesses Cross-reactive Carbohydrate IgE-Epitopes. AAAAI 60th Annual Meeting, San Francisco, 19-23 March 2004, Poster n°511 (113(2 suppl):S153)
Rationale The 23 kDa thaumatin-like protein (TLP) from cherry was identified as a major allergen and designated Pru av 2. Its amino acid sequence contains a single potential N-glycosylation site. The glycosylation of Pru av 2 and its role in IgE-binding was investigated in this study. Method s : Natural (n)Pru av 2 was purified from fresh cherries by ion exchange chromatography. An anti-horse radish peroxidase (HRP) antibody was used to detect glycosylation. 20 sera from cherry allergic patients from Italy were selected by high IgE-binding to TLP and were used to study IgE-binding to nPru av 2 and to carbohydrate moieties of HRP and Api g 5, a glycoprotein from celery. ELISA inhibition experiments were performed with 10 µg/ml HRP. Result s : 180 mg nPru av 2 were purified from 1.5 kg ripe cherries. The anti-HRP-antibody as well as IgE from all sera bound to nPru av 2. Three of 20 sera were strongly reactive with the glycoproteins HRP and Api g 5, two sera were weakly reactive with both proteins, and three sera were only positive to Api g 5. IgE-binding to nPru av 2 was inhibited by preincubation with HRP when using HRP positive sera but also with some of the sera laking reactivity in the HRP-ELISA. Conclusions : The cherry TLP very likely contains simple N-glycans similar to those of HRP or Api g 5. The correct conformation and the glycosylation of Pru av 2 have an influence on its IgE-binding capacity.
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[23] - Fils-Lycaon BR, Wiersma PA, Eastwell KC, Sautière P. A cherry protein and its gene, abundantly expressed in ripening fruit, have been identified as thaumatin-like. Plant Physiol 1996;111:269-273
A 29-kD polypeptide is the most abundant soluble protein in ripe cherry fruit (Prunus avium L); accumulation begins at the onset of ripening as the fruit turns from yellow to red. This protein was extracted from ripe cherries and purified by size-exclusion and ion-exchange chromatography. Antibodies to the purified protein were used to screen a cDNA library from ripe cherries. Numerous recombinant plaques reacted positively with the antibodies; the DNA sequence of representative clones encoded a polypeptide of 245 amino acid residues. A signal peptide was indicated, and the predicted mature protein corresponded to the purified protein in size (23.3 kD, by mass spectrometry) and isoelectric point (4.2). A search of known protein sequences revealed a strong similarity between this polypeptide and the thaumatin family of pathogenesis-related proteins. The cherry thaumatin-like protein does not have a sweet taste, and no antifungal activity was seen in preliminary assays. Expression of the protein appears to be regulated at the gene level, with mRNA levels at their highest in the ripe fruit.
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[24] - Fuchs HC, Bohle B, Dall'Antonia Y, Radauer C, Hoffmann-Sommergruber K, Mari A, et al. Natural and recombinant molecules of the cherry allergen Pru av 2 show diverse structural and B cell characteristics but similar T cell reactivity. Clin Exp Allergy 2006;36:359-368
BACKGROUND: Cherry allergy is often reported in the context of allergy to other fruits of the Rosaceae family and pollinosis to trees because of cross-reactive allergens. Allergic reactions to cherry are reported by 19-29% of birch pollen-allergic patients. Pru av 2, identified as a thaumatin-like protein (TLP) from sweet cherry, was recognized by the majority of cherry-allergic patients in immunoblotting . OBJECTIVES: In order to investigate the structural characteristics and the immunoglobulin (Ig)E- and T cell reactivity of cherry-derived TLP, recombinant Pru av 2 was expressed in Escherichia coli and natural Pru av 2 was purified . METHODS: Parallel-His and FLAG expression vectors were used for recombinant production of Pru av 2 in the cytoplasm and the periplasm of E. coli. Natural Pru av 2 was purified from fresh cherries and verified by N-terminal sequencing. Structural characterization was performed using circular dichroism (CD) measurements, and the biologic activity was measured in a glucanase assay. Using cherry-specific sera, the IgE-binding ability of recombinant and natural Pru av 2 was investigated in IgE-ELISA and the T cell reactivity was studied in proliferation assays. Results Natural Pru av 2 revealed thaumatin-like structural features and bound IgE of 50% of cherry-allergic patients. It was demonstrated to be enzymatically active. Recombinant Pru av 2 expressed in the cytoplasm of E. coli exhibited a slightly different folding compared with the natural protein. It was not recognized by IgE from cherry-allergic subjects, but retained the ability to stimulate T lymphocytes. Periplasmic recombinant Pru av 2 was able to bind an anti-grape TLP antibody and cherry-specific IgE . CONCLUSIONS: We prepared two recombinant model TLPs from cherry, and compared their molecular characteristics as well as their IgE-binding activity and T cell interactions in relation to the natural counterpart. The cytoplasmic recombinant Pru av 2 can be used as a hypoallergenic variant in allergen-specific immunotherapy, whereas the periplasmic protein can be included in a component-resolved diagnosis.
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[25] - Primavesi L, Brenna OV, Pompei C, Pravettoni V, Farioli L, Pastorello EA. Influence of Cultivar and Processing on Cherry (Prunus avium) Allergenicity. J Agric Food Chem 2006;54:9930-9935
Oral allergy syndrome is an immediate food allergic event that affects lips, mouth, and pharynx, is often triggered by fruits and vegetables, and may be associated with pollinosis. Here, we report on the allergenic pattern of different varieties of cherry (Prunus avium) and results obtained by applying several technological processes to the selected varieties. Whole cherries were submitted to chemical peeling, thermal treatment, and syruping processes, and the relative protein extracts were analyzed by in vitro (sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting analysis) and in vivo tests (skin prick test). Electrophoretic analyses demonstrated that there was no marked difference among cherry cultivars. Chemical peeling successfully removed Pru av 3, a lipid transfer protein (LTP) responsible for oral allergy syndrome in patients without pollinosis, leading to the industrial production of cherry hypoallergenic derivatives. Furthermore, the syruping process removed almost all allergenic proteins to whom patients with pollinosis are responsive. In vivo tests confirmed electrophoretic results. Keywords: Prunus avium; allergy; fruit processing; lipid transfer protein; syruping.
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[26] - Scheurer S, Lauer I, Foetisch K, Moncin MS, Retzek M, Hartz C, et al. Strong allergenicity of Pru av 3, the lipid transfer protein from cherry, is related to high stability against thermal processing and digestion. J Allergy Clin Immunol 2004;114:900-907
Background Nonspecific lipid transfer proteins (nsLTPs) have been identified as major fruit allergens in patients from the Mediterranean area. Sensitization to nsLTPs is accompanied by severe reactions, possibly because of specific biophysical and biochemical properties of this allergen family. Objective To assess the protein stability and allergenic potency of nsLTP from fruits in comparison with birch pollen-related allergens from the same allergenic source. Methods Stability of natural and recombinant cherry allergens Pru av 3 (nsLTP), Pru av 1 (Bet v 1 homologue), and Pru av 4 (profilin) to pepsin digestion and to thermal processing and stability of allergens in skin prick test reagents was investigated by immunoblotting and/or circular dichroism spectroscopy. Moreover, allergenicity of processed and fresh fruits in regard to Pru av 1 and Pru av 3 was analyzed by histamine release assays. Results Lipid transfer proteins showed the highest resistance to digestion by pepsin (rPru av 3 > rPru av 1 > rPru av 4). Immunologically active Pru av 3 was detectable after 2 hours of digestion by pepsin, whereas IgE reactivity of Pru av 1 and Pru av 4 was abolished within less than 60 minutes. In contrast with Pru av 1, IgE reactivity to nsLTPs was not diminished in thermally processed fruits, and secondary structures of purified Pru av 3 were more resistant to heating. Moreover, nsLTPs were stable components in skin prick test reagents. Histamine release assays confirmed the strong allergenicity of nsLTPs, which was not affected by protease treatment or thermal processing of fruits. Conclusion In contrast with birch pollen-related allergens, nsLTPs are highly stable to pepsin treatment and thermal processing and show higher allergenic potency. Therefore, nsLTPs have the potential to act as true food allergens, probably eliciting severe systemic reactions by reaching the intestinal mucosa in an intact and fully active form.
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[27] - Scheurer S, Lauer I, Foetisch K, Moncin MS, Retzek M, Hartz C, et al. Strong allergenicity of Pru av 3, the lipid transfer protein from cherry, is related to high stability against thermal processing and digestion. J Allergy Clin Immunol 2004;114:900-907
Background Nonspecific lipid transfer proteins (nsLTPs) have been identified as major fruit allergens in patients from the Mediterranean area. Sensitization to nsLTPs is accompanied by severe reactions, possibly because of specific biophysical and biochemical properties of this allergen family. Objective To assess the protein stability and allergenic potency of nsLTP from fruits in comparison with birch pollen-related allergens from the same allergenic source. Methods Stability of natural and recombinant cherry allergens Pru av 3 (nsLTP), Pru av 1 (Bet v 1 homologue), and Pru av 4 (profilin) to pepsin digestion and to thermal processing and stability of allergens in skin prick test reagents was investigated by immunoblotting and/or circular dichroism spectroscopy. Moreover, allergenicity of processed and fresh fruits in regard to Pru av 1 and Pru av 3 was analyzed by histamine release assays. Results Lipid transfer proteins showed the highest resistance to digestion by pepsin (rPru av 3 > rPru av 1 > rPru av 4). Immunologically active Pru av 3 was detectable after 2 hours of digestion by pepsin, whereas IgE reactivity of Pru av 1 and Pru av 4 was abolished within less than 60 minutes. In contrast with Pru av 1, IgE reactivity to nsLTPs was not diminished in thermally processed fruits, and secondary structures of purified Pru av 3 were more resistant to heating. Moreover, nsLTPs were stable components in skin prick test reagents. Histamine release assays confirmed the strong allergenicity of nsLTPs, which was not affected by protease treatment or thermal processing of fruits. Conclusion In contrast with birch pollen-related allergens, nsLTPs are highly stable to pepsin treatment and thermal processing and show higher allergenic potency. Therefore, nsLTPs have the potential to act as true food allergens, probably eliciting severe systemic reactions by reaching the intestinal mucosa in an intact and fully active form.
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[28] - Polovic N, Blanusa M, Gavrovic-Jankulovic M, Atanaskovic-Markovic M, Burazer L, Jankov R, et al. A matrix effect in pectin-rich fruits hampers digestion of allergen by pepsin in vivo and in vitro. Clin Exp Allergy 2007;37:764-771
Abstract Background It is a general belief that a food allergen should be stable to gastric digestion. Various acidic plant polysaccharides, including pectin, are ubiquitous in fruit matrixes and can form hydrogels under low-pH conditions. Objective The purpose of this study was to investigate the effect of hydrogel forming polysaccharide-rich fruit matrixes on in vivo gastric and in vitro pepsic digestion of fruit allergens. Methods Fruit extract proteins (kiwi, banana, apple and cherry) and a purified major kiwi allergen Act c 2 were digested with simulated gastric fluid in accordance with the US Pharmacopeia. In vivo experiments on kiwi fruit digestion were performed on four healthy non-atopic volunteers by examining the gastric content 1 h after ingestion of kiwi fruit. The Act c 2 and kiwi proteins were detected in immunoblots using monoclonal anti-Act c 2 antibodies and rabbit polyclonal antisera. Results Crude fruit extracts were resistant to digestion by pepsin when compared with commonly prepared extracts. In the gastric content of all volunteers, following kiwi fruit ingestion and immunoblotting, intact Act c 2 was detected with anti-Act c 2 monoclonal antibodies, while kiwi proteins of higher molecular weights were detected using rabbit polyclonal antisera. Addition of apple fruit pectin (1.5% and 3%) to the purified kiwi allergen was able to protect it from pepsin digestion in vitro. Conclusion The matrix effect in pectin-rich fruits can influence the digestibility of food proteins and thereby the process of allergic sensitization in atopic individuals.
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[29] - Scheurer S, Lauer I, Foetisch K, Moncin MS, Retzek M, Hartz C, et al. Strong allergenicity of Pru av 3, the lipid transfer protein from cherry, is related to high stability against thermal processing and digestion. J Allergy Clin Immunol 2004;114:900-907
Background Nonspecific lipid transfer proteins (nsLTPs) have been identified as major fruit allergens in patients from the Mediterranean area. Sensitization to nsLTPs is accompanied by severe reactions, possibly because of specific biophysical and biochemical properties of this allergen family. Objective To assess the protein stability and allergenic potency of nsLTP from fruits in comparison with birch pollen-related allergens from the same allergenic source. Methods Stability of natural and recombinant cherry allergens Pru av 3 (nsLTP), Pru av 1 (Bet v 1 homologue), and Pru av 4 (profilin) to pepsin digestion and to thermal processing and stability of allergens in skin prick test reagents was investigated by immunoblotting and/or circular dichroism spectroscopy. Moreover, allergenicity of processed and fresh fruits in regard to Pru av 1 and Pru av 3 was analyzed by histamine release assays. Results Lipid transfer proteins showed the highest resistance to digestion by pepsin (rPru av 3 > rPru av 1 > rPru av 4). Immunologically active Pru av 3 was detectable after 2 hours of digestion by pepsin, whereas IgE reactivity of Pru av 1 and Pru av 4 was abolished within less than 60 minutes. In contrast with Pru av 1, IgE reactivity to nsLTPs was not diminished in thermally processed fruits, and secondary structures of purified Pru av 3 were more resistant to heating. Moreover, nsLTPs were stable components in skin prick test reagents. Histamine release assays confirmed the strong allergenicity of nsLTPs, which was not affected by protease treatment or thermal processing of fruits. Conclusion In contrast with birch pollen-related allergens, nsLTPs are highly stable to pepsin treatment and thermal processing and show higher allergenic potency. Therefore, nsLTPs have the potential to act as true food allergens, probably eliciting severe systemic reactions by reaching the intestinal mucosa in an intact and fully active form.
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[30] - Primavesi L, Brenna OV, Pompei C, Pravettoni V, Farioli L, Pastorello EA. Influence of Cultivar and Processing on Cherry (Prunus avium) Allergenicity. J Agric Food Chem 2006;54:9930-9935
Oral allergy syndrome is an immediate food allergic event that affects lips, mouth, and pharynx, is often triggered by fruits and vegetables, and may be associated with pollinosis. Here, we report on the allergenic pattern of different varieties of cherry (Prunus avium) and results obtained by applying several technological processes to the selected varieties. Whole cherries were submitted to chemical peeling, thermal treatment, and syruping processes, and the relative protein extracts were analyzed by in vitro (sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting analysis) and in vivo tests (skin prick test). Electrophoretic analyses demonstrated that there was no marked difference among cherry cultivars. Chemical peeling successfully removed Pru av 3, a lipid transfer protein (LTP) responsible for oral allergy syndrome in patients without pollinosis, leading to the industrial production of cherry hypoallergenic derivatives. Furthermore, the syruping process removed almost all allergenic proteins to whom patients with pollinosis are responsive. In vivo tests confirmed electrophoretic results. Keywords: Prunus avium; allergy; fruit processing; lipid transfer protein; syruping.
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[31] - Primavesi L, Brenna OV, Pompei C, Pravettoni V, Farioli L, Pastorello EA. Influence of Cultivar and Processing on Cherry (Prunus avium) Allergenicity. J Agric Food Chem 2006;54:9930-9935
Oral allergy syndrome is an immediate food allergic event that affects lips, mouth, and pharynx, is often triggered by fruits and vegetables, and may be associated with pollinosis. Here, we report on the allergenic pattern of different varieties of cherry (Prunus avium) and results obtained by applying several technological processes to the selected varieties. Whole cherries were submitted to chemical peeling, thermal treatment, and syruping processes, and the relative protein extracts were analyzed by in vitro (sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting analysis) and in vivo tests (skin prick test). Electrophoretic analyses demonstrated that there was no marked difference among cherry cultivars. Chemical peeling successfully removed Pru av 3, a lipid transfer protein (LTP) responsible for oral allergy syndrome in patients without pollinosis, leading to the industrial production of cherry hypoallergenic derivatives. Furthermore, the syruping process removed almost all allergenic proteins to whom patients with pollinosis are responsive. In vivo tests confirmed electrophoretic results. Keywords: Prunus avium; allergy; fruit processing; lipid transfer protein; syruping.
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[32] - Gruber P, Vieths S, Wangorsch A, Nerkamp J, Hofmann T. Maillard reaction and enzymatic browning affect the allergenicity of Pru av 1, the major allergen from cherry (Prunus avium). J Agric Food Chem 2004;52:4002-4007
The influence of thermal processing and nonenymatic as well as polyphenoloxidase-catalyzed browning reaction on the allergenicity of the major cherry allergen Pru av 1 was investigated. After thermal treatment of the recombinant protein rPru av 1 in the absence or presence of carbohydrates, SDS-PAGE, enzyme allergosorbent tests, and inhibition assays revealed that thermal treatment of rPru av 1 alone did not show any influence on the IgE-binding activity of the protein at least for 30 min, thus correlating well with the refolding of the allergen in buffer solution as demonstrated by CD spectroscopic experiments. Incubation of the protein with starch and maltose also showed no effect on IgE-binding activity, whereas reaction with glucose and ribose and, even more pronounced, with the carbohydrate breakdown products glyceraldehyde and glyoxal induced a strong decrease of the IgE-binding capacity of rPru av 1. In the second part of the study, the effect of polyphenoloxidase-catalyzed oxidation of polyphenols on food allergen activity was investigated. Incubation of rPru av 1 with epicatechin in the presence of tyrosinase led to a drastic decrease in IgE-binding activity of the protein. Variations of the phenolic compound revealed caffeic acid and epicatechin as the most active inhibitors of the IgE-binding activity of rPru av 1, followed by catechin and gallic acid, and, finally, by quercetin and rutin, showing significantly lower activity. On the basis of these data, reactive intermediates formed during thermal carbohydrate degradation as well as during enzymatic polyphenol oxidation are suggested as the active chemical species responsible for modifying nucleophilic amino acid side chains of proteins, thus inducing an irreversible change in the tertiary structure of the protein and resulting in a loss of conformational epitopes of the allergen.
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[33] - Karamloo F, Scheurer S, Wangorsch A, May S, Haustein D, Vieths S. Pyr c 1, the major allergen from pear (Pyrus communis), is a new member of the Bet v 1 allergen family. J Chromatogr B Biomed Appl 2001;756:281-293
Pear is known as an allergenic food involved in the 'oral allergy syndrome' which affects a high percentage of patients allergic to birch pollen. The aim of this study was to clone the major allergen of this fruit, to express it as bacterial recombinant protein and to study its allergenic properties in relation to homologous proteins and natural allergen extracts. The coding region of the cDNA was obtained by a PCR strategy, cloned, and the allergen was expressed as His-Tag fusion protein. The fusion peptide was removed by treatment with cyanogen bromide. Purified non-fusion protein was subjected to allergenicity testing by the enzyme allergosorbent test (EAST), Western blotting, competitive inhibition assays, and basophil histamine release. The deduced protein sequence shared a high degree of identity with other major allergens from fruits, nuts, vegetables, and pollen, and with a family of PR-10 pathogenesis related proteins. The recombinant (r) protein was recognised by specific IgE from sera of all pear-allergic patients (n = 16) investigated in this study. Hence, the allergen was classified as a major allergen and named Pyr c 1. The IgE binding characteristics of rPyr c 1 appeared to be similar to the natural pear protein, as was demonstrated by EAST-inhibition and Western blot-inhibition experiments. Moreover, the biological activity of rPyr c 1 was equal to that of pear extract, as indicated by basophil histamine release in two patients allergic to pears. The related major allergens Bet v 1 from birch pollen and Mal d 1 from apple inhibited to a high degree the binding of IgE to Pyr c 1, whereas Api g 1 from celery, also belonging to this family, had little inhibitory effects, indicating epitope differences between Bet v 1-related food allergens. Unlimited amounts of pure rPyr c 1 are now available for studies on the structure and epitopes of pollen-related food allergens. Moreover, the allergen may serve as stable and standardised diagnostic material.
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[34] - 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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[35] - 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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[36] - 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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[37] - Scheurer S, Pastorello EA, Wangorsch A, Kästner M, Haustein D, Vieths S. Recombinant allergens Pru av 1 and Pru av 4 and a newly identified lipid transfer protein in the in vitro diagnosis of cherry allergy. J Allergy Clin Immunol 2001;107:724-731
BACKGROUND: In central and northern Europe food allergy to fruits of the Rosaceae family is strongly associated with birch pollinosis because of the existence of IgE cross-reactive homologous allergens in birch pollen and food. By contrast, in the Mediterranean population allergic reactions to these fruits frequently are not related to birch pollen allergy and are predominantly elicited by lipid transfer proteins (LTPs). OBJECTIVE: We sought to determine the prevalence of IgE sensitization to the recombinant cherry allergens Pru av 1 and Pru av 4 in comparison with cherry extract within a representative group of patients who were allergic to cherries recruited in Germany and to compare the relevance of IgE to cherry LTPs in Italian patients. METHODS: Recombinant Pru av 1 and rPru av 4 were available from earlier studies. The cDNA of the cherry LTPs was obtained by using a PCR-cloning strategy. The protein was expressed in Escherichia coli and purified by means of metal chelate affinity chromatography. Sera from 101 German patients with birch pollinosis and oral allergy syndrome to cherry and sera from 7 Italian patients with cherry allergy were investigated by using enzyme allergosorbent tests for IgE reactivity with cherry extract, rPru av 1, rPru av 4, and the recombinant cherry LTP. Inhibition experiments were performed to compare the IgE reactivity of natural and recombinant cherry LTPs and to investigate potential cross-reactivity with birch pollen allergens. RESULTS: The LTP from cherry comprises 91 amino acids and a 26 amino acid signal peptide. The mature cherry LTP shows high amino acid sequence identity with allergenic LTPs from peach (Pru p 3, 88%), apricot (Pru ar 3, 86%), and maize (Zea m 14, 59%) and displays no IgE cross-reactivity with birch pollen. The IgE prevalences in the German patients were as follows: LTP, 3 of 101 (3%); rPru av 1, 97 of 101 (96.0%); rPru av 4, 16 of 101 (16.2%); and cherry extract, 98 of 101 (97%). All 7 Italian patients had IgE against the cherry LTP. CONCLUSIONS: Recombinant allergens are useful tools for a more accurate in vitro IgE-based diagnosis of cherry allergy. Taken together, they mimic the allergenic activity of cherry extract, having slightly higher biologic activity. Sensitization to the cherry LTP is relevant for a minority of patients recruited in Germany, but our data indicate that it may be a major allergen in Italy.
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[38] - 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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[39] - Zuberbier T, Edenharter G, Worm M, Ehlers I, Reimann S, Hantke T, et al. Prevalence of adverse reactions to food in Germany – a population study. Allergy 2004;59:338-345
OBJECTIVE: A population study was performed to identify the prevalence of all kinds of adverse reactions to food . METHODS: In a representative cross-sectional survey performed in 1999 and 2000 in Berlin, 13 300 inhabitants of all ages were addressed by questionnaire. This questionnaire was answered by 4093 persons. All respondents mentioning any sign of food intolerance or the existence of allergic diseases (n = 2298) were followed up by telephone and, in case food intolerance could not be ruled out by patient history, were invited to attend to the clinic for personal investigation including double-blind, placebo-controlled food challenge tests (DBPCFC) . RESULTS: The self-reported lifetime prevalence of any adverse reaction to food in the Berlin population (mean age 41 years) was 34.9%. Eight hundred and fourteen individuals were personally investigated according to the guidelines. The point prevalence of adverse reactions to food confirmed by DBPCFC tests in the Berlin population as a mean of all age groups was 3.6% (95% confidence interval [3.0-4.2%]) and 3.7% in the adult population (18-79 years, 95% confidence interval [3.1-4.4.%]). Two and a half percent were IgE-mediated and 1.1% non-IgE-mediated, females were more frequently affected (60.6%). Based on a statistical comparison with available data of adults from the nationwide German Health Survey from 1998, adverse reactions to food in the adult population of Germany (age 18-79) were calculated with 2.6% [2.1-3.2%]) . CONCLUSIONS: The study gives for the first time information about the point prevalence of both immunological and nonimmunological adverse reactions to food and underlines the relevance of this issue in public health. The data also show that an individualized stepwise approach including provocation tests is mandatory to confirm the diagnosis.
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[40] - Asero R. Detection and clinical characterization of patients with oral allergy syndrome caused by stable allergens in Rosaceae and nuts. Ann Allergy Asthma Immunol 1999;83:377-383
A minority of patients with oral allergy syndrome (OAS) induced by Rosaceae or nuts are positive on skin prick tests with commercial food extracts. This suggests reactivity against distinct stable allergens. OBJECTIVES: (1) To define the prevalence of subjects positive on skin prick tests with commercial extracts among patients with OAS caused by Rosaceae and/or nuts and (2) To investigate whether commercial extracts-positive subjects show some peculiar clinical feature and may represent a specific subset with food allergy. METHODS: Skin prick tests were carried out with a large panel of commercial extracts of airborne allergens (Allergopharma) and of vegetable foods (Dome/Hollister-Stier) in 298 adults with OAS caused by Rosaceae (n = 237) and or nuts (n = 161), positive on skin prick tests with fresh offending foods. RESULTS: 25/237 (11%) patients were positive on prick tests with commercial plum extract. This subgroup showed a higher incidence of systemic symptoms (64% versus 6%; P < .001) and a lower incidence of birch pollen allergy (12% versus 99%; P < .001) than commercial extract-negative patients; moreover, 36% versus 0%, respectively, did not have respiratory allergy (P < .001). Apple and peach were the main offending foods among commercial extract-negative and commercial extract-positive patients, respectively (87% versus 44% for apple, P < .001; and 52% versus 88% for peach, P < .005). Eight of one hundred sixty-one (5%) nuts-sensitive patients were positive on prick test with commercial walnut extract. This subgroup showed a higher proportion of patients who experienced systemic symptoms (63% versus 6%, P < .001), a lower prevalence of birch pollen allergy (13% versus 97%, P < .001), and a higher prevalence of grass pollen allergy (88% versus 41%, P < .05) than commercial extract-negative subjects. Further, reactivity against commercial walnut extract was associated with skin reactivity against commercial extracts of peanut (88% versus 37%, P < .005), tomato (75% versus 5%, P < .001), and plum (63% versus 8%, P < .001), and inversely related with skin reactivity against fresh apple (P < .001). In most cases, high levels of IgE specific for peach, apple, and hazelnut were associated with peanut reactivity rather than with clinical sensitivity to specific foods. In a preliminary investigation, most commercial extract-positive patients reacted against a 10-kDa protein characterized as a lipid transfer protein (LTP). CONCLUSIONS: Skin prick tests with commercial extracts of plum and walnut may be usefully employed to detect patients with OAS reacting against stable allergens. The high prevalence of systemic symptoms in these patients suggests that allergens' stability is associated with a higher resistance to the gastrointestinal environment and strongly influences the clinical expression of vegetable food allergy. At least some stable allergens, namely lipid transfer protein might be shared by botanically unrelated fruits such as nuts, peanuts, legumes, tomato, and Prunoideae.
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[41] - Asero R, Mistrello G, Roncarolo D, de Vries SC, Gautier MF, Ciurana CL, et al. Lipid transfer protein: a pan-allergen in plant-derived foods that is highly resistant to pepsin digestion. Int Arch Allergy Immunol 2000;122:20-32
Lipid transfer proteins (LTPs) are small molecules of approximately 10 kD that demonstrate high stability. They have recently been identified as allergens in the Rosaceae subfamilies of the Prunoideae (peach, apricot, plum) and of the Pomoideae (apple). They belong to a family of structurally highly conserved proteins that are also present in non-Rosaceae vegetable foods. OBJECTIVE: The aim of this study was to investigate the cross-reactivity to non-Rosaceae LTPs, and to study the role of protein stability in allergenicity. METHODS: Thirty-eight patients with a positive SPT to Rosaceae fruit extracts enriched for LTP were characterized by interview and SPT. To investigate IgE cross-reactivity between Rosaceae and non-Rosaceae LTPs, RAST and RAST inhibition as well as ELISA and ELISA inhibition were performed, using whole food extracts and purified LTPs. Both purified natural LTPs (peach, carrot and broccoli) and Pichia pastoris recombinant LTPs (carrot and wheat) were included. Pepsin digestion was used to address the role of stability in the allergenicity of LTPs. RESULTS: IgE antibodies to Rosaceae LTPs reacted to a broad range of vegetable foods, including Gramineae (cereals), Leguminosae (peanut), Juglandaceae (walnut), Anacardiaceae (pistachio), Brassicaceae (broccoli), Umbelliferae (carrot, celery), Solanaceae (tomato), Cucurbitaceae (melon), and Actinidiaceae (kiwi). Binding and inhibition studies with purified natural and recombinant LTPs confirmed their role in this cross-reactivity. Many of these cross-reactivities were accompanied by clinical food allergy, frequently including systemic reactions. Antibody binding to LTP was shown to be resistant to pepsin treatment of whole extract or purified LTP. CONCLUSION: LTP is a pan-allergen with a degree of cross-reactivity comparable to profilin. Due to its extreme resistance to pepsin digestion, LTP is a potentially severe food allergen.
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[42] - Fernández-Rivas M, Cuevas M. Peels of Rosaceae fruits have a higher allergenicity than pulps. Clin Exp Allergy 1999;29:1239-1247
It is not uncommon that patients allergic to fruits such as apple, pear, and peach, refer adverse reactions after the ingestion of the whole fruit, but subsequently tolerate the pulp. OBJECTIVE: This study aimed to compare the allergenicity of peels and pulps of apple, peach, and pear in 33 patients allergic to these fruits. METHODS: Clinical reactivity to the ingestion of whole fruit (peel + pulp) and pulp was established by medical history. Peels and pulps were tested separately in skin prick tests (SPTs), histamine release tests (HRTs) and RASTs. Cross-allergenicity between peel and pulp of apple and peach was studied by RAST inhibition. RESULTS: Adverse reactions appeared more frequently and were more severe when the whole fruit was eaten. More than 40% of patients allergic to apple and pear tolerated the ingestion of the pulp of these fruits, and reactions were only elicited by the intake of the whole fruit. Peels induced higher SPTs, HRTs and RASTs than pulps. An important cross-allergenicity was found between the peel and pulp of apple and peach, although the amount of the shared allergenic epitopes seemed to be higher in peels. CONCLUSION: Our results suggest that peels of Rosaceae fruits such as apple, peach, and pear, have a higher allergenicity than pulps, which is clinically relevant. This aspect should be considered in the evaluation of patients allergic to Rosaceae fruits, and in the production of diagnostic materials.
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[43] - Rodriguez J, Crespo JF, López-Rubio A, De La Cruz-Bertolo J, Ferrando-Vivas P, Vives R, et al. Clinical cross-reactivity among foods of the Rosaceae family. J Allergy Clin Immunol 2000;106:183-189
Foods from the Rosaceae botanical family have been increasingly reported as causes of allergic reaction. Patients frequently have positive skin tests or radioallergosorbent test results for multiple members of this botanical family. OBJECTIVE: Our purpose was to investigate the clinical cross-reactivity assessed by double-blind, placebo-controlled food challenge (DBPCFC) of Rosaceae foods (apricot, almond, plum, strawberry, apple, peach, and pear). METHODS: Thirty-four consecutive adult patients complaining of adverse reactions to Rosaceae were included in the study. Skin prick tests and CAP System (FEIA) were performed with Rosaceae foods in all patients. Clinical reactivity to Rosaceae was systematically evaluated by open food challenges (OFCs), unless there was a convincing history of a recent severe anaphylaxis. Positive reactions on OFCs were subsequently evaluated by DBPCFCs. RESULTS: Twenty-six and 24 patients had positive skin prick tests and CAP FEIA with Rosaceae, respectively; from these 88% and 100% had positive tests with >/=2. No evidence of clinical reactivity was found in 66% percent of positive skin prick tests and 63% of positive specific IgE determinations to fruits. A total of 226 food challenges (including OFC and DBPCFC) were performed in the 28 patients with positive skin prick tests or CAP System FEIA. Of 182 initial OFCs carried out, 26 (14%) reactions were confirmed by DBPCFCs. Overall, 40 reactions were considered positive in 22 patients with positive skin tests or CAP FEIA. Thirty-eight reactions had been previously reported, the remaining two were detected by systematic challenges. Most reactions were caused by peach (22 patients), apple (6), and apricot (5). Ten patients (46%) were clinically allergic to peach and other Rosaceae. CONCLUSION: Positive skin test and CAP System FEIA should not be taken as the only guide for multi-species dietary restrictions. Nevertheless, the potential clinical allergy to other Rosaceae should not be neglected. If the reported reaction is confirmed, current tolerance to other Rosaceae should be precisely established unless there has been ingestion without symptoms after the reaction.
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[44] - Cuesta-Herranz J, Lazaro M, Figueredo E, Igea JM, Umpierrez A, De Las Heras M. Allergy to plant-derived fresh foods in a birch- and ragweed-free area. Clin Exp Allergy 2000;30:1411-1416
BACKGROUND: Allergy to plant-derived fresh foods has often been reported in geographical areas where birch or ragweed pollens are frequent and has been attributed to cross-reactivity to pollens. OBJECTIVE: The aim of this study has been to evaluate allergy to plant-derived fresh foods among pollen-allergic patients from a birch and ragweed-free area. METHODS: Ninety-five pollen-allergic patients took part in the study. The study consisted of a questionnaire, skin prick tests and challenge tests. Pollen skin tests to five grasses, eight trees and seven weeds were performed in duplicate. Prick tests (prick by prick) and challenge tests were carried out with the fresh foods. RESULTS: Most patients allergic to pollens were sensitized to grass (Lolium and Phleum; 97.9%), followed by tree (Olea; 82.1%) and weed pollens (Plantago; 64.2%). 35 of the 95 pollen-allergic patients had positive skin test responses to some plant-derived fresh foods, the highest percentage corresponding to several fruits in the Rosaceae family (peach and pear, 26.3%), followed by Cucurbitacea fruits (melon, 13.7%). The 21. 05% of the pollen-allergic patients were allergic to some type of plant-derived fresh food. Peach was the plant-derived fresh food which most frequently elicited allergy symptoms (12.6%), followed by melon (7.36%). The cluster of positive responses to Rosaceae fruits was higher for skin testing than for challenge testing. CONCLUSION: Peach was the most important allergy provoking fruit in a birch and ragweed free-area where apples were consumed at a rate of two times more than peaches and the patients allergic to pollen were principally sensitized to grass pollens.
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[45] - Ahrazem O, Ibáñez D, López-Torrejón G, Sánchez-Monge R, Sastre J, Lombardero M, et al. Lipid Transfer Proteins and Allergy to Oranges. Int Arch Allergy Immunol 2005;137:201-210
BACKGROUND: Lipid transfer proteins (LTPs) are relevant fruit allergens, recently proposed as model plant food allergens. No citrus fruit allergen has been characterized to date. We sought to identify and isolate citrus fruit LTPs and to explore their relevance in orange allergy . METHODS: Twenty-seven patients, showing mainly oral allergy syndrome after orange ingestion, as well as positive prick responses and serum-specific IgE levels to orange, were selected. Natural orange and lemon LTPs, as well as a recombinant orange LTP isoform expressed in Pichia pastoris, were isolated by chromatographic methods and characterized by N-terminal amino acid sequencing and matrix-assisted laser desorption/ionizaion mass spectrometry, and DNA sequencing of the corresponding cDNA in the case of the recombinant allergen. Specific IgE determination, immunodetection, ELISA-inhibition assays and in vivo skin prick tests (SPTs) were performed with all three purified allergens and with the major peach LTP allergen, Pru p 3 . RESULTS: The natural allergens purified from orange (nCit s 3) and lemon (nCit l 3) showed very similar N-terminal amino acid sequences (18 out of 20 identical residues), typical of LTPs, and molecular masses of 9,610 and 9,618 Da, respectively. The recombinant orange isoform (rCit s 3) expressed in P. pastoris (16 out of 20 residues identical to its natural counterpart in the N-terminal region) presented 92 amino acid residues and 9,463 Da, and 67% sequence identity with rPru p 3. Of the 27 sera analyzed, specific IgE to the purified allergens was found in 54% for nCit l 3, 48% for nCit s 3, 46% for rCit s 3 and 37% for rPru p 3. Positive SPT responses were obtained in 7 out of 26 patients tested for nCit s 3, 3 out of 8 for nCit l 3 and 10 out of 26 for nPru p 3. ELISA-inhibition assays showed an equivalent IgE-binding pattern for the natural and recombinant orange LTPs, and IgE cross-reactivity among the purified orange, lemon and peach LTP allergens . CONCLUSIONS: Members of the LTP allergen family are involved in allergy to oranges, displaying positive in vitro and in vivo reactions in 30-50% of the patients studied. Both orange and lemon allergens show cross-reactivity with the major peach allergen Pru p 3.
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[46] - Pastorello EA, D'Ambrosio FP, Pravettoni V, Farioli L, Giuffrida G, Monza M, et al. Evidence for a lipid transfer protein as the major allergen of apricot. J Allergy Clin Immunol 2000;105:371-377
Apricots are widely grown in Europe, and allergic reactions are becoming more common, especially oral allergy syndrome. Apricot belongs to the botanical subfamily of Prunoideae, which includes peach, the major allergen of which was identified as a 9-kd protein, a lipid transfer protein (LTP). OBJECTIVE: The aim of the study was to evaluate the IgE reactivity pattern to an apricot extract in subjects with allergic reactions to apricot, as demonstrated by a positive oral challenge response. METHODS: Thirty patients were investigated. All the patients displayed oral allergy syndrome (2 with systemic reactions) to apricot, with positive open food challenge responses, skin prick test responses, and serum-specific IgE antibodies to apricot. The IgE reactivity pattern to apricot extract was identified by using SDS-PAGE and immunoblotting. The major allergen, a 9-kd protein, was then purified by HPLC and characterized by periodic acid-Schiff stain, isoelectric point determination, and N-terminal amino acid sequencing. RESULTS: The sera from all patients allergic to apricot recognized the 9-kd protein, whereas none of the other allergens, with molecular weights from 15 to 80 kd, acted as a major allergen. The 9-kd allergen has an isoelectric point of 8.7 and is not glycosylated. Determination of the N-terminal 34 amino acid sequence showed that it belongs to the LTP family, with a 94% homology with the LTP from peach. IgE blotting of the apricot extract was completely inhibited by the 9-kd purified LTP from peach. CONCLUSIONS: The major allergen of apricot is an LTP, which is highly cross-reactive with the LTP from peach.
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[47] - Conti A, Fortunato D, Ortolani C, Giufridda MG, Pravettoni V, Napolitano L, et al. Determination of the primary structure of two lipid transfer proteins from apricot (Prunus armeniaca). J Chromatogr B Biomed Appl 2001;756:123-129
It has been recently demonstrated that the major allergen of apricot is a protein of molecular mass (Mr) 9000 belonging to the family of Lipid Transfer Protein. The aim of this study was the determination of the primary structure of apricot LTP by micro-sequencing and mass spectrometric analyses. Apricot LTP is a 91 amino acids protein like peach and almond LTPs with a sequence identity of 91% and 94%, respectively. Like for the peach LTP, out of the 25 amino acids forming the inner surface of the tunnel-like hydrophobic cavity in maize ns-LTP, 16 are identical and 7 similar in the apricot LTP, supporting the hypothesis of a similar function.
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[48] - 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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[49] - Chevalier T, de Rigal D, Mbéguié-A-Mbéguié D, Gauillard F, Richard-Forget F, Fils-Lycaon BR. Molecular cloning and characterization of apricot fruit polyphenol oxidase. Plant Physiol 1999;119:1261-1270
A reverse transcriptase-polymerase chain reaction experiment was done to synthesize a homologous polyphenol oxidase (PPO) probe from apricot (Prunus armeniaca var Bergeron) fruit. This probe was further used to isolate a full-length PPO cDNA, PA-PPO (accession no. AF020786), from an immature-green fruit cDNA library. PA-PPO is 2070 bp long and contains a single open reading frame encoding a PPO precursor peptide of 597 amino acids with a calculated molecular mass of 67.1 kD and an isoelectric point of 6.84. The mature protein has a predicted molecular mass of 56.2 kD and an isoelectric point of 5.84. PA-PPO belongs to a multigene family. The gene is highly expressed in young, immature-green fruit and is turned off early in the ripening process. The ratio of PPO protein to total proteins per fruit apparently remains stable regardless of the stage of development, whereas PPO specific activity peaks at the breaker stage. These results suggest that, in addition to a transcriptional control of PPO expression, other regulation factors such as translational and posttranslational controls also occur.
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[50] - Scheurer S, Pastorello EA, Wangorsch A, Kästner M, Haustein D, Vieths S. Recombinant allergens Pru av 1 and Pru av 4 and a newly identified lipid transfer protein in the in vitro diagnosis of cherry allergy. J Allergy Clin Immunol 2001;107:724-731
BACKGROUND: In central and northern Europe food allergy to fruits of the Rosaceae family is strongly associated with birch pollinosis because of the existence of IgE cross-reactive homologous allergens in birch pollen and food. By contrast, in the Mediterranean population allergic reactions to these fruits frequently are not related to birch pollen allergy and are predominantly elicited by lipid transfer proteins (LTPs). OBJECTIVE: We sought to determine the prevalence of IgE sensitization to the recombinant cherry allergens Pru av 1 and Pru av 4 in comparison with cherry extract within a representative group of patients who were allergic to cherries recruited in Germany and to compare the relevance of IgE to cherry LTPs in Italian patients. METHODS: Recombinant Pru av 1 and rPru av 4 were available from earlier studies. The cDNA of the cherry LTPs was obtained by using a PCR-cloning strategy. The protein was expressed in Escherichia coli and purified by means of metal chelate affinity chromatography. Sera from 101 German patients with birch pollinosis and oral allergy syndrome to cherry and sera from 7 Italian patients with cherry allergy were investigated by using enzyme allergosorbent tests for IgE reactivity with cherry extract, rPru av 1, rPru av 4, and the recombinant cherry LTP. Inhibition experiments were performed to compare the IgE reactivity of natural and recombinant cherry LTPs and to investigate potential cross-reactivity with birch pollen allergens. RESULTS: The LTP from cherry comprises 91 amino acids and a 26 amino acid signal peptide. The mature cherry LTP shows high amino acid sequence identity with allergenic LTPs from peach (Pru p 3, 88%), apricot (Pru ar 3, 86%), and maize (Zea m 14, 59%) and displays no IgE cross-reactivity with birch pollen. The IgE prevalences in the German patients were as follows: LTP, 3 of 101 (3%); rPru av 1, 97 of 101 (96.0%); rPru av 4, 16 of 101 (16.2%); and cherry extract, 98 of 101 (97%). All 7 Italian patients had IgE against the cherry LTP. CONCLUSIONS: Recombinant allergens are useful tools for a more accurate in vitro IgE-based diagnosis of cherry allergy. Taken together, they mimic the allergenic activity of cherry extract, having slightly higher biologic activity. Sensitization to the cherry LTP is relevant for a minority of patients recruited in Germany, but our data indicate that it may be a major allergen in Italy.
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[51] - Zuberbier T, Edenharter G, Worm M, Ehlers I, Reimann S, Hantke T, et al. Prevalence of adverse reactions to food in Germany – a population study. Allergy 2004;59:338-345
OBJECTIVE: A population study was performed to identify the prevalence of all kinds of adverse reactions to food . METHODS: In a representative cross-sectional survey performed in 1999 and 2000 in Berlin, 13 300 inhabitants of all ages were addressed by questionnaire. This questionnaire was answered by 4093 persons. All respondents mentioning any sign of food intolerance or the existence of allergic diseases (n = 2298) were followed up by telephone and, in case food intolerance could not be ruled out by patient history, were invited to attend to the clinic for personal investigation including double-blind, placebo-controlled food challenge tests (DBPCFC) . RESULTS: The self-reported lifetime prevalence of any adverse reaction to food in the Berlin population (mean age 41 years) was 34.9%. Eight hundred and fourteen individuals were personally investigated according to the guidelines. The point prevalence of adverse reactions to food confirmed by DBPCFC tests in the Berlin population as a mean of all age groups was 3.6% (95% confidence interval [3.0-4.2%]) and 3.7% in the adult population (18-79 years, 95% confidence interval [3.1-4.4.%]). Two and a half percent were IgE-mediated and 1.1% non-IgE-mediated, females were more frequently affected (60.6%). Based on a statistical comparison with available data of adults from the nationwide German Health Survey from 1998, adverse reactions to food in the adult population of Germany (age 18-79) were calculated with 2.6% [2.1-3.2%]) . CONCLUSIONS: The study gives for the first time information about the point prevalence of both immunological and nonimmunological adverse reactions to food and underlines the relevance of this issue in public health. The data also show that an individualized stepwise approach including provocation tests is mandatory to confirm the diagnosis.
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[52] - Pastorello EA, Ortolani C, Farioli L, Pravettoni V, Ispano M, Borga A, et al. Allergenic cross-reactivity among peach, apricot, plum, and cherry in patients with oral allergy syndrome: an in vivo and in vitro study. J Allergy Clin Immunol 1994;94:699-707
Oral allergy syndrome in response to fruits and vegetables frequently occurs as clusters of hypersensitivity to members of the same botanical family, for which the immunologic basis lies in a number of common allergens, most of them still unidentified. OBJECTIVE: This study was designed to assess the in vivo and in vitro cross-reactivity between fruits of the Prunoideae subfamily (i.e., peach, cherry, apricot, and plum) and to identify their major allergens and the cross-reactivity of the peach extract with grass and birch pollen. METHODS: The in vivo study was conducted by skin prick tests and open food challenges with fresh fruits in 23 patients with oral allergy syndrome for peach and positive skin prick test and RAST results for the other Prunoideae. In vitro sodium dodecylsulfate-polyacrylamide gel electrophoresis was followed by immunoblotting and immunoblotting-inhibition. RESULTS: A 13 kd component was identified as the only major allergen common to all the Prunoideae, the other major allergens were found at 14 kd in peach and at 30 kd in cherry. Immunoblotting inhibition showed wide cross-reactivity within the Prunoideae, whereas grass and birch pollen partially inhibited the peach blotting. CONCLUSIONS: Clinical cross-reactivity to Prunoideae is essentially due to a common 13 kd IgE-binding component, which seems to be the most important major allergen of this subfamily, not shared with grass and birch pollen.
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[53] - Asero R. Detection and clinical characterization of patients with oral allergy syndrome caused by stable allergens in Rosaceae and nuts. Ann Allergy Asthma Immunol 1999;83:377-383
A minority of patients with oral allergy syndrome (OAS) induced by Rosaceae or nuts are positive on skin prick tests with commercial food extracts. This suggests reactivity against distinct stable allergens. OBJECTIVES: (1) To define the prevalence of subjects positive on skin prick tests with commercial extracts among patients with OAS caused by Rosaceae and/or nuts and (2) To investigate whether commercial extracts-positive subjects show some peculiar clinical feature and may represent a specific subset with food allergy. METHODS: Skin prick tests were carried out with a large panel of commercial extracts of airborne allergens (Allergopharma) and of vegetable foods (Dome/Hollister-Stier) in 298 adults with OAS caused by Rosaceae (n = 237) and or nuts (n = 161), positive on skin prick tests with fresh offending foods. RESULTS: 25/237 (11%) patients were positive on prick tests with commercial plum extract. This subgroup showed a higher incidence of systemic symptoms (64% versus 6%; P < .001) and a lower incidence of birch pollen allergy (12% versus 99%; P < .001) than commercial extract-negative patients; moreover, 36% versus 0%, respectively, did not have respiratory allergy (P < .001). Apple and peach were the main offending foods among commercial extract-negative and commercial extract-positive patients, respectively (87% versus 44% for apple, P < .001; and 52% versus 88% for peach, P < .005). Eight of one hundred sixty-one (5%) nuts-sensitive patients were positive on prick test with commercial walnut extract. This subgroup showed a higher proportion of patients who experienced systemic symptoms (63% versus 6%, P < .001), a lower prevalence of birch pollen allergy (13% versus 97%, P < .001), and a higher prevalence of grass pollen allergy (88% versus 41%, P < .05) than commercial extract-negative subjects. Further, reactivity against commercial walnut extract was associated with skin reactivity against commercial extracts of peanut (88% versus 37%, P < .005), tomato (75% versus 5%, P < .001), and plum (63% versus 8%, P < .001), and inversely related with skin reactivity against fresh apple (P < .001). In most cases, high levels of IgE specific for peach, apple, and hazelnut were associated with peanut reactivity rather than with clinical sensitivity to specific foods. In a preliminary investigation, most commercial extract-positive patients reacted against a 10-kDa protein characterized as a lipid transfer protein (LTP). CONCLUSIONS: Skin prick tests with commercial extracts of plum and walnut may be usefully employed to detect patients with OAS reacting against stable allergens. The high prevalence of systemic symptoms in these patients suggests that allergens' stability is associated with a higher resistance to the gastrointestinal environment and strongly influences the clinical expression of vegetable food allergy. At least some stable allergens, namely lipid transfer protein might be shared by botanically unrelated fruits such as nuts, peanuts, legumes, tomato, and Prunoideae.
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[54] - Pastorello EA, D'Ambrosio FP, Pravettoni V, Farioli L, Giuffrida G, Monza M, et al. Evidence for a lipid transfer protein as the major allergen of apricot. J Allergy Clin Immunol 2000;105:371-377
Apricots are widely grown in Europe, and allergic reactions are becoming more common, especially oral allergy syndrome. Apricot belongs to the botanical subfamily of Prunoideae, which includes peach, the major allergen of which was identified as a 9-kd protein, a lipid transfer protein (LTP). OBJECTIVE: The aim of the study was to evaluate the IgE reactivity pattern to an apricot extract in subjects with allergic reactions to apricot, as demonstrated by a positive oral challenge response. METHODS: Thirty patients were investigated. All the patients displayed oral allergy syndrome (2 with systemic reactions) to apricot, with positive open food challenge responses, skin prick test responses, and serum-specific IgE antibodies to apricot. The IgE reactivity pattern to apricot extract was identified by using SDS-PAGE and immunoblotting. The major allergen, a 9-kd protein, was then purified by HPLC and characterized by periodic acid-Schiff stain, isoelectric point determination, and N-terminal amino acid sequencing. RESULTS: The sera from all patients allergic to apricot recognized the 9-kd protein, whereas none of the other allergens, with molecular weights from 15 to 80 kd, acted as a major allergen. The 9-kd allergen has an isoelectric point of 8.7 and is not glycosylated. Determination of the N-terminal 34 amino acid sequence showed that it belongs to the LTP family, with a 94% homology with the LTP from peach. IgE blotting of the apricot extract was completely inhibited by the 9-kd purified LTP from peach. CONCLUSIONS: The major allergen of apricot is an LTP, which is highly cross-reactive with the LTP from peach.
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[55] - Asero R, Mistrello G, Roncarolo D, de Vries SC, Gautier MF, Ciurana CL, et al. Lipid transfer protein: a pan-allergen in plant-derived foods that is highly resistant to pepsin digestion. Int Arch Allergy Immunol 2000;122:20-32
Lipid transfer proteins (LTPs) are small molecules of approximately 10 kD that demonstrate high stability. They have recently been identified as allergens in the Rosaceae subfamilies of the Prunoideae (peach, apricot, plum) and of the Pomoideae (apple). They belong to a family of structurally highly conserved proteins that are also present in non-Rosaceae vegetable foods. OBJECTIVE: The aim of this study was to investigate the cross-reactivity to non-Rosaceae LTPs, and to study the role of protein stability in allergenicity. METHODS: Thirty-eight patients with a positive SPT to Rosaceae fruit extracts enriched for LTP were characterized by interview and SPT. To investigate IgE cross-reactivity between Rosaceae and non-Rosaceae LTPs, RAST and RAST inhibition as well as ELISA and ELISA inhibition were performed, using whole food extracts and purified LTPs. Both purified natural LTPs (peach, carrot and broccoli) and Pichia pastoris recombinant LTPs (carrot and wheat) were included. Pepsin digestion was used to address the role of stability in the allergenicity of LTPs. RESULTS: IgE antibodies to Rosaceae LTPs reacted to a broad range of vegetable foods, including Gramineae (cereals), Leguminosae (peanut), Juglandaceae (walnut), Anacardiaceae (pistachio), Brassicaceae (broccoli), Umbelliferae (carrot, celery), Solanaceae (tomato), Cucurbitaceae (melon), and Actinidiaceae (kiwi). Binding and inhibition studies with purified natural and recombinant LTPs confirmed their role in this cross-reactivity. Many of these cross-reactivities were accompanied by clinical food allergy, frequently including systemic reactions. Antibody binding to LTP was shown to be resistant to pepsin treatment of whole extract or purified LTP. CONCLUSION: LTP is a pan-allergen with a degree of cross-reactivity comparable to profilin. Due to its extreme resistance to pepsin digestion, LTP is a potentially severe food allergen.
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[56] - Pastorello EA, Farioli L, Pravettoni V, Giufridda MG, Ortolani C, Fortunato D, et al. Characterization of the major allergen of plum as a lipid transfer protein. J Chromatogr B Biomed Appl 2001;756:95-103
BACKGROUND: Allergy to Prunoideae fruit (plum, peach, cherry and apricot) is one of the most frequent food allergies in southern Europe. All these fruits cross-react in vivo and in vitro, as they share their major allergen, a 9 kD lipid transfer protein (LTP). OBJECTIVE: The aim of the study was the identification and molecular characterization of the major allergen of plum. METHODS: The IgE pattern of reactivity to plums was investigated by SDS-PAGE and immunoblotting with the sera of 23 patients. The identified major allergen was purified by HPLC, using a cationic-exchange column followed by gel-filtration. Further characterization was achieved by periodic-Schiff stain, isoelectrofocusing and N-terminal amino acid sequencing. RESULTS AND CONCLUSIONS: The major allergen of plum is a 9 kD lipid transfer protein, not glycosylated and with a basic character (pI>9), highly homologous to the major allergen of peach.
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[57] - Fernández-Rivas M, Van Ree R, Cuevas M. Allergy to Rosaceae fruits without related pollinosis. J Allergy Clin Immunol 1997;100:728-733
BACKGROUND: Rosaceae fruit allergy is frequently associated with birch pollinosis in Central and Northern Europe and with grass pollen allergy in Central Spain. The main cross-reactive structures involved for birch pollinosis are Bet v 1 and profilin, and for grass pollinosis they are profilin and carbohydrate determinants. Rosaceae fruit allergy can occasionally be observed in patients without pollinosis . OBJECTIVE: We investigated the clinical presentation and the allergens involved in allergy to Rosaceae fruit without pollinosis . METHODS: Eleven patients from Central Spain allergic to apples, peaches, and/or pears but not to pollens were compared with 22 control subjects with combined grass pollen and fruit allergy. Skin prick tests and RASTs to apple, peach, and pear were performed. Cross-allergenicity was studied by RAST inhibition. Bet v 1 was tested with an indirect RAST, and profilin was tested in skin prick tests, histamine release, and RAST . RESULTS: Rosaceae fruit allergy without pollinosis is severe with 82% of patients reporting systemic symptoms, mainly anaphylaxis (73%), whereas oral symptoms are less frequent (64%). Anaphylactic shock was observed in 36% of patients. The fruit allergens involved showed cross-reactivity among Rosaceae species but were not related to profilin or Bet v 1. Ninety-one percent of patients with combined grass pollinosis and fruit allergy reported oral allergy, 45% reported systemic symptoms, 18% reported anaphylaxis, and 9% reported anaphylactic shock . CONCLUSION: Allergy to Rosaceae fruits in patients without a related pollen allergy is a severe clinical entity. Profilin- and Bet v 1-related structures are not involved in Rosaceae fruit allergy without pollinosis.
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[58] - Rodriguez J, Crespo JF, López-Rubio A, De La Cruz-Bertolo J, Ferrando-Vivas P, Vives R, et al. Clinical cross-reactivity among foods of the Rosaceae family. J Allergy Clin Immunol 2000;106:183-189
Foods from the Rosaceae botanical family have been increasingly reported as causes of allergic reaction. Patients frequently have positive skin tests or radioallergosorbent test results for multiple members of this botanical family. OBJECTIVE: Our purpose was to investigate the clinical cross-reactivity assessed by double-blind, placebo-controlled food challenge (DBPCFC) of Rosaceae foods (apricot, almond, plum, strawberry, apple, peach, and pear). METHODS: Thirty-four consecutive adult patients complaining of adverse reactions to Rosaceae were included in the study. Skin prick tests and CAP System (FEIA) were performed with Rosaceae foods in all patients. Clinical reactivity to Rosaceae was systematically evaluated by open food challenges (OFCs), unless there was a convincing history of a recent severe anaphylaxis. Positive reactions on OFCs were subsequently evaluated by DBPCFCs. RESULTS: Twenty-six and 24 patients had positive skin prick tests and CAP FEIA with Rosaceae, respectively; from these 88% and 100% had positive tests with >/=2. No evidence of clinical reactivity was found in 66% percent of positive skin prick tests and 63% of positive specific IgE determinations to fruits. A total of 226 food challenges (including OFC and DBPCFC) were performed in the 28 patients with positive skin prick tests or CAP System FEIA. Of 182 initial OFCs carried out, 26 (14%) reactions were confirmed by DBPCFCs. Overall, 40 reactions were considered positive in 22 patients with positive skin tests or CAP FEIA. Thirty-eight reactions had been previously reported, the remaining two were detected by systematic challenges. Most reactions were caused by peach (22 patients), apple (6), and apricot (5). Ten patients (46%) were clinically allergic to peach and other Rosaceae. CONCLUSION: Positive skin test and CAP System FEIA should not be taken as the only guide for multi-species dietary restrictions. Nevertheless, the potential clinical allergy to other Rosaceae should not be neglected. If the reported reaction is confirmed, current tolerance to other Rosaceae should be precisely established unless there has been ingestion without symptoms after the reaction.
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[59] - Crespo JF, Rodriguez J, James JM, Daroca P, Reaño M, Vives R. Reactivity to potential cross-reactive foods in fruit-allergic patients: implications for prescribing food avoidance. Allergy 2002;57:946-949
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[60] - Ahrazem O, Ibáñez D, López-Torrejón G, Sánchez-Monge R, Sastre J, Lombardero M, et al. Lipid Transfer Proteins and Allergy to Oranges. Int Arch Allergy Immunol 2005;137:201-210
BACKGROUND: Lipid transfer proteins (LTPs) are relevant fruit allergens, recently proposed as model plant food allergens. No citrus fruit allergen has been characterized to date. We sought to identify and isolate citrus fruit LTPs and to explore their relevance in orange allergy . METHODS: Twenty-seven patients, showing mainly oral allergy syndrome after orange ingestion, as well as positive prick responses and serum-specific IgE levels to orange, were selected. Natural orange and lemon LTPs, as well as a recombinant orange LTP isoform expressed in Pichia pastoris, were isolated by chromatographic methods and characterized by N-terminal amino acid sequencing and matrix-assisted laser desorption/ionizaion mass spectrometry, and DNA sequencing of the corresponding cDNA in the case of the recombinant allergen. Specific IgE determination, immunodetection, ELISA-inhibition assays and in vivo skin prick tests (SPTs) were performed with all three purified allergens and with the major peach LTP allergen, Pru p 3 . RESULTS: The natural allergens purified from orange (nCit s 3) and lemon (nCit l 3) showed very similar N-terminal amino acid sequences (18 out of 20 identical residues), typical of LTPs, and molecular masses of 9,610 and 9,618 Da, respectively. The recombinant orange isoform (rCit s 3) expressed in P. pastoris (16 out of 20 residues identical to its natural counterpart in the N-terminal region) presented 92 amino acid residues and 9,463 Da, and 67% sequence identity with rPru p 3. Of the 27 sera analyzed, specific IgE to the purified allergens was found in 54% for nCit l 3, 48% for nCit s 3, 46% for rCit s 3 and 37% for rPru p 3. Positive SPT responses were obtained in 7 out of 26 patients tested for nCit s 3, 3 out of 8 for nCit l 3 and 10 out of 26 for nPru p 3. ELISA-inhibition assays showed an equivalent IgE-binding pattern for the natural and recombinant orange LTPs, and IgE cross-reactivity among the purified orange, lemon and peach LTP allergens . CONCLUSIONS: Members of the LTP allergen family are involved in allergy to oranges, displaying positive in vitro and in vivo reactions in 30-50% of the patients studied. Both orange and lemon allergens show cross-reactivity with the major peach allergen Pru p 3.
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[61] - Pastorello EA, Farioli L, Pravettoni V, Giufridda MG, Ortolani C, Fortunato D, et al. Characterization of the major allergen of plum as a lipid transfer protein. J Chromatogr B Biomed Appl 2001;756:95-103
BACKGROUND: Allergy to Prunoideae fruit (plum, peach, cherry and apricot) is one of the most frequent food allergies in southern Europe. All these fruits cross-react in vivo and in vitro, as they share their major allergen, a 9 kD lipid transfer protein (LTP). OBJECTIVE: The aim of the study was the identification and molecular characterization of the major allergen of plum. METHODS: The IgE pattern of reactivity to plums was investigated by SDS-PAGE and immunoblotting with the sera of 23 patients. The identified major allergen was purified by HPLC, using a cationic-exchange column followed by gel-filtration. Further characterization was achieved by periodic-Schiff stain, isoelectrofocusing and N-terminal amino acid sequencing. RESULTS AND CONCLUSIONS: The major allergen of plum is a 9 kD lipid transfer protein, not glycosylated and with a basic character (pI>9), highly homologous to the major allergen of peach.
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[62] - 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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[63] - Scheurer S, Pastorello EA, Wangorsch A, Kästner M, Haustein D, Vieths S. Recombinant allergens Pru av 1 and Pru av 4 and a newly identified lipid transfer protein in the in vitro diagnosis of cherry allergy. J Allergy Clin Immunol 2001;107:724-731
BACKGROUND: In central and northern Europe food allergy to fruits of the Rosaceae family is strongly associated with birch pollinosis because of the existence of IgE cross-reactive homologous allergens in birch pollen and food. By contrast, in the Mediterranean population allergic reactions to these fruits frequently are not related to birch pollen allergy and are predominantly elicited by lipid transfer proteins (LTPs). OBJECTIVE: We sought to determine the prevalence of IgE sensitization to the recombinant cherry allergens Pru av 1 and Pru av 4 in comparison with cherry extract within a representative group of patients who were allergic to cherries recruited in Germany and to compare the relevance of IgE to cherry LTPs in Italian patients. METHODS: Recombinant Pru av 1 and rPru av 4 were available from earlier studies. The cDNA of the cherry LTPs was obtained by using a PCR-cloning strategy. The protein was expressed in Escherichia coli and purified by means of metal chelate affinity chromatography. Sera from 101 German patients with birch pollinosis and oral allergy syndrome to cherry and sera from 7 Italian patients with cherry allergy were investigated by using enzyme allergosorbent tests for IgE reactivity with cherry extract, rPru av 1, rPru av 4, and the recombinant cherry LTP. Inhibition experiments were performed to compare the IgE reactivity of natural and recombinant cherry LTPs and to investigate potential cross-reactivity with birch pollen allergens. RESULTS: The LTP from cherry comprises 91 amino acids and a 26 amino acid signal peptide. The mature cherry LTP shows high amino acid sequence identity with allergenic LTPs from peach (Pru p 3, 88%), apricot (Pru ar 3, 86%), and maize (Zea m 14, 59%) and displays no IgE cross-reactivity with birch pollen. The IgE prevalences in the German patients were as follows: LTP, 3 of 101 (3%); rPru av 1, 97 of 101 (96.0%); rPru av 4, 16 of 101 (16.2%); and cherry extract, 98 of 101 (97%). All 7 Italian patients had IgE against the cherry LTP. CONCLUSIONS: Recombinant allergens are useful tools for a more accurate in vitro IgE-based diagnosis of cherry allergy. Taken together, they mimic the allergenic activity of cherry extract, having slightly higher biologic activity. Sensitization to the cherry LTP is relevant for a minority of patients recruited in Germany, but our data indicate that it may be a major allergen in Italy.
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[64] - Zuberbier T, Edenharter G, Worm M, Ehlers I, Reimann S, Hantke T, et al. Prevalence of adverse reactions to food in Germany – a population study. Allergy 2004;59:338-345
OBJECTIVE: A population study was performed to identify the prevalence of all kinds of adverse reactions to food . METHODS: In a representative cross-sectional survey performed in 1999 and 2000 in Berlin, 13 300 inhabitants of all ages were addressed by questionnaire. This questionnaire was answered by 4093 persons. All respondents mentioning any sign of food intolerance or the existence of allergic diseases (n = 2298) were followed up by telephone and, in case food intolerance could not be ruled out by patient history, were invited to attend to the clinic for personal investigation including double-blind, placebo-controlled food challenge tests (DBPCFC) . RESULTS: The self-reported lifetime prevalence of any adverse reaction to food in the Berlin population (mean age 41 years) was 34.9%. Eight hundred and fourteen individuals were personally investigated according to the guidelines. The point prevalence of adverse reactions to food confirmed by DBPCFC tests in the Berlin population as a mean of all age groups was 3.6% (95% confidence interval [3.0-4.2%]) and 3.7% in the adult population (18-79 years, 95% confidence interval [3.1-4.4.%]). Two and a half percent were IgE-mediated and 1.1% non-IgE-mediated, females were more frequently affected (60.6%). Based on a statistical comparison with available data of adults from the nationwide German Health Survey from 1998, adverse reactions to food in the adult population of Germany (age 18-79) were calculated with 2.6% [2.1-3.2%]) . CONCLUSIONS: The study gives for the first time information about the point prevalence of both immunological and nonimmunological adverse reactions to food and underlines the relevance of this issue in public health. The data also show that an individualized stepwise approach including provocation tests is mandatory to confirm the diagnosis.
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[65] - Rodrigues-Alves R, Pereira-Santos MC, Lopes A, Lopes-Silva S, Spinola-Santos A, Costa C, et al. Clinical, anamnestic and serological features of peach allergy in Portugal. Int Arch Allergy Immunol 2008;149:65-73
BACKGROUND: Peach is a common food allergen source throughout Europe. The aim of this study was to characterize peach allergy in a Portuguese patient population . METHODS: Thirty peach-allergic patients confirmed by double-blind placebo-controlled food challenges and 29 controls were included. All subjects completed a standardized questionnaire regarding symptoms and epidemiologic characteristics, skin prick tests with inhalant allergens and foods as well as specific IgE antibodies to peach, recombinant peach allergens rPru p 1, rPru p 3, rPru p 4 and cross-reactive carbohydrate determinants . RESULTS: Thirty-seven percent of patients reported only oral allergy syndrome, while 37% reported generalized urticaria and/or angioedema, 17% localized contact urticaria and 10% anaphylaxis with peach. Sensitization to other Rosaceae fruits and tree nuts was present in 90 and 77% of the patients, respectively. Respiratory allergy history was associated with less severe symptoms (oral allergy syndrome or contact urticaria; p < 0.01) and positive skin prick test to peach peel or plum with more severe symptoms (urticaria and/or angioedema or anaphylaxis; p < 0.05). Ninety-seven percent were sensitized to Pru p 3, 13% to Pru p 4, 3% to Pru p 1 and 10% to cross-reactive carbohydrate determinants. Pru p 3 specific IgE was associated with Artemisia vulgaris sensitization and tree nut allergy (p < 0.05) but not with clinical severity . CONCLUSIONS: Half the patients reported systemic reactions to peach. Peach allergy appeared predominantly mediated by Pru p 3 but some patients were sensitized to Pru p 4. Applying a 0.10 kU(A)/l cutoff level, the diagnostic value of combining the 3 recombinant allergens was noteworthy, with 100% sensitivity and 90% specificity.
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[66] - Pastorello EA, Ortolani C, Farioli L, Pravettoni V, Ispano M, Borga A, et al. Allergenic cross-reactivity among peach, apricot, plum, and cherry in patients with oral allergy syndrome: an in vivo and in vitro study. J Allergy Clin Immunol 1994;94:699-707
Oral allergy syndrome in response to fruits and vegetables frequently occurs as clusters of hypersensitivity to members of the same botanical family, for which the immunologic basis lies in a number of common allergens, most of them still unidentified. OBJECTIVE: This study was designed to assess the in vivo and in vitro cross-reactivity between fruits of the Prunoideae subfamily (i.e., peach, cherry, apricot, and plum) and to identify their major allergens and the cross-reactivity of the peach extract with grass and birch pollen. METHODS: The in vivo study was conducted by skin prick tests and open food challenges with fresh fruits in 23 patients with oral allergy syndrome for peach and positive skin prick test and RAST results for the other Prunoideae. In vitro sodium dodecylsulfate-polyacrylamide gel electrophoresis was followed by immunoblotting and immunoblotting-inhibition. RESULTS: A 13 kd component was identified as the only major allergen common to all the Prunoideae, the other major allergens were found at 14 kd in peach and at 30 kd in cherry. Immunoblotting inhibition showed wide cross-reactivity within the Prunoideae, whereas grass and birch pollen partially inhibited the peach blotting. CONCLUSIONS: Clinical cross-reactivity to Prunoideae is essentially due to a common 13 kd IgE-binding component, which seems to be the most important major allergen of this subfamily, not shared with grass and birch pollen.
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[67] - Asero R. Detection and clinical characterization of patients with oral allergy syndrome caused by stable allergens in Rosaceae and nuts. Ann Allergy Asthma Immunol 1999;83:377-383
A minority of patients with oral allergy syndrome (OAS) induced by Rosaceae or nuts are positive on skin prick tests with commercial food extracts. This suggests reactivity against distinct stable allergens. OBJECTIVES: (1) To define the prevalence of subjects positive on skin prick tests with commercial extracts among patients with OAS caused by Rosaceae and/or nuts and (2) To investigate whether commercial extracts-positive subjects show some peculiar clinical feature and may represent a specific subset with food allergy. METHODS: Skin prick tests were carried out with a large panel of commercial extracts of airborne allergens (Allergopharma) and of vegetable foods (Dome/Hollister-Stier) in 298 adults with OAS caused by Rosaceae (n = 237) and or nuts (n = 161), positive on skin prick tests with fresh offending foods. RESULTS: 25/237 (11%) patients were positive on prick tests with commercial plum extract. This subgroup showed a higher incidence of systemic symptoms (64% versus 6%; P < .001) and a lower incidence of birch pollen allergy (12% versus 99%; P < .001) than commercial extract-negative patients; moreover, 36% versus 0%, respectively, did not have respiratory allergy (P < .001). Apple and peach were the main offending foods among commercial extract-negative and commercial extract-positive patients, respectively (87% versus 44% for apple, P < .001; and 52% versus 88% for peach, P < .005). Eight of one hundred sixty-one (5%) nuts-sensitive patients were positive on prick test with commercial walnut extract. This subgroup showed a higher proportion of patients who experienced systemic symptoms (63% versus 6%, P < .001), a lower prevalence of birch pollen allergy (13% versus 97%, P < .001), and a higher prevalence of grass pollen allergy (88% versus 41%, P < .05) than commercial extract-negative subjects. Further, reactivity against commercial walnut extract was associated with skin reactivity against commercial extracts of peanut (88% versus 37%, P < .005), tomato (75% versus 5%, P < .001), and plum (63% versus 8%, P < .001), and inversely related with skin reactivity against fresh apple (P < .001). In most cases, high levels of IgE specific for peach, apple, and hazelnut were associated with peanut reactivity rather than with clinical sensitivity to specific foods. In a preliminary investigation, most commercial extract-positive patients reacted against a 10-kDa protein characterized as a lipid transfer protein (LTP). CONCLUSIONS: Skin prick tests with commercial extracts of plum and walnut may be usefully employed to detect patients with OAS reacting against stable allergens. The high prevalence of systemic symptoms in these patients suggests that allergens' stability is associated with a higher resistance to the gastrointestinal environment and strongly influences the clinical expression of vegetable food allergy. At least some stable allergens, namely lipid transfer protein might be shared by botanically unrelated fruits such as nuts, peanuts, legumes, tomato, and Prunoideae.
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[68] - Pastorello EA, D'Ambrosio FP, Pravettoni V, Farioli L, Giuffrida G, Monza M, et al. Evidence for a lipid transfer protein as the major allergen of apricot. J Allergy Clin Immunol 2000;105:371-377
Apricots are widely grown in Europe, and allergic reactions are becoming more common, especially oral allergy syndrome. Apricot belongs to the botanical subfamily of Prunoideae, which includes peach, the major allergen of which was identified as a 9-kd protein, a lipid transfer protein (LTP). OBJECTIVE: The aim of the study was to evaluate the IgE reactivity pattern to an apricot extract in subjects with allergic reactions to apricot, as demonstrated by a positive oral challenge response. METHODS: Thirty patients were investigated. All the patients displayed oral allergy syndrome (2 with systemic reactions) to apricot, with positive open food challenge responses, skin prick test responses, and serum-specific IgE antibodies to apricot. The IgE reactivity pattern to apricot extract was identified by using SDS-PAGE and immunoblotting. The major allergen, a 9-kd protein, was then purified by HPLC and characterized by periodic acid-Schiff stain, isoelectric point determination, and N-terminal amino acid sequencing. RESULTS: The sera from all patients allergic to apricot recognized the 9-kd protein, whereas none of the other allergens, with molecular weights from 15 to 80 kd, acted as a major allergen. The 9-kd allergen has an isoelectric point of 8.7 and is not glycosylated. Determination of the N-terminal 34 amino acid sequence showed that it belongs to the LTP family, with a 94% homology with the LTP from peach. IgE blotting of the apricot extract was completely inhibited by the 9-kd purified LTP from peach. CONCLUSIONS: The major allergen of apricot is an LTP, which is highly cross-reactive with the LTP from peach.
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[69] - Asero R, Mistrello G, Roncarolo D, de Vries SC, Gautier MF, Ciurana CL, et al. Lipid transfer protein: a pan-allergen in plant-derived foods that is highly resistant to pepsin digestion. Int Arch Allergy Immunol 2000;122:20-32
Lipid transfer proteins (LTPs) are small molecules of approximately 10 kD that demonstrate high stability. They have recently been identified as allergens in the Rosaceae subfamilies of the Prunoideae (peach, apricot, plum) and of the Pomoideae (apple). They belong to a family of structurally highly conserved proteins that are also present in non-Rosaceae vegetable foods. OBJECTIVE: The aim of this study was to investigate the cross-reactivity to non-Rosaceae LTPs, and to study the role of protein stability in allergenicity. METHODS: Thirty-eight patients with a positive SPT to Rosaceae fruit extracts enriched for LTP were characterized by interview and SPT. To investigate IgE cross-reactivity between Rosaceae and non-Rosaceae LTPs, RAST and RAST inhibition as well as ELISA and ELISA inhibition were performed, using whole food extracts and purified LTPs. Both purified natural LTPs (peach, carrot and broccoli) and Pichia pastoris recombinant LTPs (carrot and wheat) were included. Pepsin digestion was used to address the role of stability in the allergenicity of LTPs. RESULTS: IgE antibodies to Rosaceae LTPs reacted to a broad range of vegetable foods, including Gramineae (cereals), Leguminosae (peanut), Juglandaceae (walnut), Anacardiaceae (pistachio), Brassicaceae (broccoli), Umbelliferae (carrot, celery), Solanaceae (tomato), Cucurbitaceae (melon), and Actinidiaceae (kiwi). Binding and inhibition studies with purified natural and recombinant LTPs confirmed their role in this cross-reactivity. Many of these cross-reactivities were accompanied by clinical food allergy, frequently including systemic reactions. Antibody binding to LTP was shown to be resistant to pepsin treatment of whole extract or purified LTP. CONCLUSION: LTP is a pan-allergen with a degree of cross-reactivity comparable to profilin. Due to its extreme resistance to pepsin digestion, LTP is a potentially severe food allergen.
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[70] - Asero R, Mistrello G, Roncarolo D, Amato S, Caldironi G, Barocci F, et al. Immunological cross-reactivity between lipid transfer proteins from botanically unrelated plant-derived foods: a clinical study. Allergy 2002;57:900-906
BACKGROUND: Lipid transfer proteins (LTP) are highly conserved and widely distributed throughout the plant kingdom. Recent studies demonstrated immunological cross-reactivity between LTP from many botanically unrelated fruits and vegetables and concluded that LTP are pan-allergens. This study aimed to evaluate the clinical relevance of such cross-reactivity in a group of subjects monosensitized to LTP . METHODS: Twenty LTP-hypersensitive patients were selected from a population of about 600 subjects with history of Rosaceae allergy by means of: 1) negative skin prick test (SPT) with a commercial birch pollen extract; 2) positive SPT with a commercial plum extract, rich in LTP but virtually lacking both Bet v 1-like proteins and profilin; 3) in-vitro IgE reactivity to the 9-10 kDa fraction of peach peel or immunoblot with peach peel showing a single band at 10 kDa; and 4) total inhibition of reactivity to whole peach extract (containing Bet v 1-related allergen, profilin, and LTP) by purified peach LTP on enzyme-linked immunoassay (ELISA). Allergy to foods other than Rosaceae was ascertained by careful interview and analysis of medical recordings. SPT with a large series of plant-derived foods were carried out as well. The cross reactivity between LTPs from botanically unrelated plant-derived foods was assessed by ELISA inhibition tests using walnut and peanut extracts as substrate, and peach LTP as inhibitor . RESULTS: All patients reported allergic reactions after the ingestion of at least one from a large number of vegetable foods other than Rosaceae, and in several cases clinical reactions were very severe (anaphylaxis, asthma, urticaria/angioedema). Nuts and peanuts were the most frequently reported causes of allergic reactions (80% and 40% of patients, respectively). All patients showed positive SPT to several non-Rosaceae food extracts. SPT with nuts, peanut, legumes, celery, rice, and corn were positive in the majority of patients. In ELISA inhibition studies, absorption of sera with peach LTP caused complete inhibition of IgE reactivity to walnut and peanut in all cases . CONCLUSION: LTP is a clinically relevant pan-allergen. Most Rosaceae-allergic, LTP-hypersensitive patients experience adverse reactions after ingestion of botanically unrelated plant-derived foods as well. In view of the high prevalence and severity of the allergic reactions induced, hazelnut, walnut, and peanut should be regarded as potentially hazardous for these patients.
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[71] - Fernández-Rivas M, Van Ree R, Cuevas M. Allergy to Rosaceae fruits without related pollinosis. J Allergy Clin Immunol 1997;100:728-733
BACKGROUND: Rosaceae fruit allergy is frequently associated with birch pollinosis in Central and Northern Europe and with grass pollen allergy in Central Spain. The main cross-reactive structures involved for birch pollinosis are Bet v 1 and profilin, and for grass pollinosis they are profilin and carbohydrate determinants. Rosaceae fruit allergy can occasionally be observed in patients without pollinosis . OBJECTIVE: We investigated the clinical presentation and the allergens involved in allergy to Rosaceae fruit without pollinosis . METHODS: Eleven patients from Central Spain allergic to apples, peaches, and/or pears but not to pollens were compared with 22 control subjects with combined grass pollen and fruit allergy. Skin prick tests and RASTs to apple, peach, and pear were performed. Cross-allergenicity was studied by RAST inhibition. Bet v 1 was tested with an indirect RAST, and profilin was tested in skin prick tests, histamine release, and RAST . RESULTS: Rosaceae fruit allergy without pollinosis is severe with 82% of patients reporting systemic symptoms, mainly anaphylaxis (73%), whereas oral symptoms are less frequent (64%). Anaphylactic shock was observed in 36% of patients. The fruit allergens involved showed cross-reactivity among Rosaceae species but were not related to profilin or Bet v 1. Ninety-one percent of patients with combined grass pollinosis and fruit allergy reported oral allergy, 45% reported systemic symptoms, 18% reported anaphylaxis, and 9% reported anaphylactic shock . CONCLUSION: Allergy to Rosaceae fruits in patients without a related pollen allergy is a severe clinical entity. Profilin- and Bet v 1-related structures are not involved in Rosaceae fruit allergy without pollinosis.
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[72] - Rodriguez J, Crespo JF, López-Rubio A, De La Cruz-Bertolo J, Ferrando-Vivas P, Vives R, et al. Clinical cross-reactivity among foods of the Rosaceae family. J Allergy Clin Immunol 2000;106:183-189
Foods from the Rosaceae botanical family have been increasingly reported as causes of allergic reaction. Patients frequently have positive skin tests or radioallergosorbent test results for multiple members of this botanical family. OBJECTIVE: Our purpose was to investigate the clinical cross-reactivity assessed by double-blind, placebo-controlled food challenge (DBPCFC) of Rosaceae foods (apricot, almond, plum, strawberry, apple, peach, and pear). METHODS: Thirty-four consecutive adult patients complaining of adverse reactions to Rosaceae were included in the study. Skin prick tests and CAP System (FEIA) were performed with Rosaceae foods in all patients. Clinical reactivity to Rosaceae was systematically evaluated by open food challenges (OFCs), unless there was a convincing history of a recent severe anaphylaxis. Positive reactions on OFCs were subsequently evaluated by DBPCFCs. RESULTS: Twenty-six and 24 patients had positive skin prick tests and CAP FEIA with Rosaceae, respectively; from these 88% and 100% had positive tests with >/=2. No evidence of clinical reactivity was found in 66% percent of positive skin prick tests and 63% of positive specific IgE determinations to fruits. A total of 226 food challenges (including OFC and DBPCFC) were performed in the 28 patients with positive skin prick tests or CAP System FEIA. Of 182 initial OFCs carried out, 26 (14%) reactions were confirmed by DBPCFCs. Overall, 40 reactions were considered positive in 22 patients with positive skin tests or CAP FEIA. Thirty-eight reactions had been previously reported, the remaining two were detected by systematic challenges. Most reactions were caused by peach (22 patients), apple (6), and apricot (5). Ten patients (46%) were clinically allergic to peach and other Rosaceae. CONCLUSION: Positive skin test and CAP System FEIA should not be taken as the only guide for multi-species dietary restrictions. Nevertheless, the potential clinical allergy to other Rosaceae should not be neglected. If the reported reaction is confirmed, current tolerance to other Rosaceae should be precisely established unless there has been ingestion without symptoms after the reaction.
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[73] - Cuesta-Herranz J, Lazaro M, Figueredo E, Igea JM, Umpierrez A, De Las Heras M. Allergy to plant-derived fresh foods in a birch- and ragweed-free area. Clin Exp Allergy 2000;30:1411-1416
BACKGROUND: Allergy to plant-derived fresh foods has often been reported in geographical areas where birch or ragweed pollens are frequent and has been attributed to cross-reactivity to pollens. OBJECTIVE: The aim of this study has been to evaluate allergy to plant-derived fresh foods among pollen-allergic patients from a birch and ragweed-free area. METHODS: Ninety-five pollen-allergic patients took part in the study. The study consisted of a questionnaire, skin prick tests and challenge tests. Pollen skin tests to five grasses, eight trees and seven weeds were performed in duplicate. Prick tests (prick by prick) and challenge tests were carried out with the fresh foods. RESULTS: Most patients allergic to pollens were sensitized to grass (Lolium and Phleum; 97.9%), followed by tree (Olea; 82.1%) and weed pollens (Plantago; 64.2%). 35 of the 95 pollen-allergic patients had positive skin test responses to some plant-derived fresh foods, the highest percentage corresponding to several fruits in the Rosaceae family (peach and pear, 26.3%), followed by Cucurbitacea fruits (melon, 13.7%). The 21. 05% of the pollen-allergic patients were allergic to some type of plant-derived fresh food. Peach was the plant-derived fresh food which most frequently elicited allergy symptoms (12.6%), followed by melon (7.36%). The cluster of positive responses to Rosaceae fruits was higher for skin testing than for challenge testing. CONCLUSION: Peach was the most important allergy provoking fruit in a birch and ragweed free-area where apples were consumed at a rate of two times more than peaches and the patients allergic to pollen were principally sensitized to grass pollens.
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[74] - Crespo JF, Rodriguez J, James JM, Daroca P, Reaño M, Vives R. Reactivity to potential cross-reactive foods in fruit-allergic patients: implications for prescribing food avoidance. Allergy 2002;57:946-949
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[75] - Asero R, Mistrello MG, Roncarolo D, Amato S, Falagiani P. Detection of novel latex allergens associated with clinically relevant allergy to plant-derived foods. J Allergy Clin Immunol 2005;115:1312-1314
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[76] - Conti A, Giuffrida MG, Hoffmann-Sommergruber K, Wagner S, Amato S, Mistrello G, et al. Identification of latex UDP glucose pyrophosphorylase (Hev b UDPGP) as a novel cause of latex fruit allergy syndrome. Eur Ann Allergy Clin Immunol 2007;39:116-118
Based on a recently published unusual case of food allergy in a latex-allergic patients, the present study identifies Hev b UDPGP as a novel allergen in natural rubber latex able to cause latex-fruit allergy syndrome and as a novel, potential pan-allergen in vegetable foods.
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[77] - Scheurer S, Pastorello EA, Wangorsch A, Kästner M, Haustein D, Vieths S. Recombinant allergens Pru av 1 and Pru av 4 and a newly identified lipid transfer protein in the in vitro diagnosis of cherry allergy. J Allergy Clin Immunol 2001;107:724-731
BACKGROUND: In central and northern Europe food allergy to fruits of the Rosaceae family is strongly associated with birch pollinosis because of the existence of IgE cross-reactive homologous allergens in birch pollen and food. By contrast, in the Mediterranean population allergic reactions to these fruits frequently are not related to birch pollen allergy and are predominantly elicited by lipid transfer proteins (LTPs). OBJECTIVE: We sought to determine the prevalence of IgE sensitization to the recombinant cherry allergens Pru av 1 and Pru av 4 in comparison with cherry extract within a representative group of patients who were allergic to cherries recruited in Germany and to compare the relevance of IgE to cherry LTPs in Italian patients. METHODS: Recombinant Pru av 1 and rPru av 4 were available from earlier studies. The cDNA of the cherry LTPs was obtained by using a PCR-cloning strategy. The protein was expressed in Escherichia coli and purified by means of metal chelate affinity chromatography. Sera from 101 German patients with birch pollinosis and oral allergy syndrome to cherry and sera from 7 Italian patients with cherry allergy were investigated by using enzyme allergosorbent tests for IgE reactivity with cherry extract, rPru av 1, rPru av 4, and the recombinant cherry LTP. Inhibition experiments were performed to compare the IgE reactivity of natural and recombinant cherry LTPs and to investigate potential cross-reactivity with birch pollen allergens. RESULTS: The LTP from cherry comprises 91 amino acids and a 26 amino acid signal peptide. The mature cherry LTP shows high amino acid sequence identity with allergenic LTPs from peach (Pru p 3, 88%), apricot (Pru ar 3, 86%), and maize (Zea m 14, 59%) and displays no IgE cross-reactivity with birch pollen. The IgE prevalences in the German patients were as follows: LTP, 3 of 101 (3%); rPru av 1, 97 of 101 (96.0%); rPru av 4, 16 of 101 (16.2%); and cherry extract, 98 of 101 (97%). All 7 Italian patients had IgE against the cherry LTP. CONCLUSIONS: Recombinant allergens are useful tools for a more accurate in vitro IgE-based diagnosis of cherry allergy. Taken together, they mimic the allergenic activity of cherry extract, having slightly higher biologic activity. Sensitization to the cherry LTP is relevant for a minority of patients recruited in Germany, but our data indicate that it may be a major allergen in Italy.
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[78] - Fernández-Rivas M, Van Ree R, Cuevas M. Allergy to Rosaceae fruits without related pollinosis. J Allergy Clin Immunol 1997;100:728-733
BACKGROUND: Rosaceae fruit allergy is frequently associated with birch pollinosis in Central and Northern Europe and with grass pollen allergy in Central Spain. The main cross-reactive structures involved for birch pollinosis are Bet v 1 and profilin, and for grass pollinosis they are profilin and carbohydrate determinants. Rosaceae fruit allergy can occasionally be observed in patients without pollinosis . OBJECTIVE: We investigated the clinical presentation and the allergens involved in allergy to Rosaceae fruit without pollinosis . METHODS: Eleven patients from Central Spain allergic to apples, peaches, and/or pears but not to pollens were compared with 22 control subjects with combined grass pollen and fruit allergy. Skin prick tests and RASTs to apple, peach, and pear were performed. Cross-allergenicity was studied by RAST inhibition. Bet v 1 was tested with an indirect RAST, and profilin was tested in skin prick tests, histamine release, and RAST . RESULTS: Rosaceae fruit allergy without pollinosis is severe with 82% of patients reporting systemic symptoms, mainly anaphylaxis (73%), whereas oral symptoms are less frequent (64%). Anaphylactic shock was observed in 36% of patients. The fruit allergens involved showed cross-reactivity among Rosaceae species but were not related to profilin or Bet v 1. Ninety-one percent of patients with combined grass pollinosis and fruit allergy reported oral allergy, 45% reported systemic symptoms, 18% reported anaphylaxis, and 9% reported anaphylactic shock . CONCLUSION: Allergy to Rosaceae fruits in patients without a related pollen allergy is a severe clinical entity. Profilin- and Bet v 1-related structures are not involved in Rosaceae fruit allergy without pollinosis.
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[79] - Rodriguez J, Crespo JF, López-Rubio A, De La Cruz-Bertolo J, Ferrando-Vivas P, Vives R, et al. Clinical cross-reactivity among foods of the Rosaceae family. J Allergy Clin Immunol 2000;106:183-189
Foods from the Rosaceae botanical family have been increasingly reported as causes of allergic reaction. Patients frequently have positive skin tests or radioallergosorbent test results for multiple members of this botanical family. OBJECTIVE: Our purpose was to investigate the clinical cross-reactivity assessed by double-blind, placebo-controlled food challenge (DBPCFC) of Rosaceae foods (apricot, almond, plum, strawberry, apple, peach, and pear). METHODS: Thirty-four consecutive adult patients complaining of adverse reactions to Rosaceae were included in the study. Skin prick tests and CAP System (FEIA) were performed with Rosaceae foods in all patients. Clinical reactivity to Rosaceae was systematically evaluated by open food challenges (OFCs), unless there was a convincing history of a recent severe anaphylaxis. Positive reactions on OFCs were subsequently evaluated by DBPCFCs. RESULTS: Twenty-six and 24 patients had positive skin prick tests and CAP FEIA with Rosaceae, respectively; from these 88% and 100% had positive tests with >/=2. No evidence of clinical reactivity was found in 66% percent of positive skin prick tests and 63% of positive specific IgE determinations to fruits. A total of 226 food challenges (including OFC and DBPCFC) were performed in the 28 patients with positive skin prick tests or CAP System FEIA. Of 182 initial OFCs carried out, 26 (14%) reactions were confirmed by DBPCFCs. Overall, 40 reactions were considered positive in 22 patients with positive skin tests or CAP FEIA. Thirty-eight reactions had been previously reported, the remaining two were detected by systematic challenges. Most reactions were caused by peach (22 patients), apple (6), and apricot (5). Ten patients (46%) were clinically allergic to peach and other Rosaceae. CONCLUSION: Positive skin test and CAP System FEIA should not be taken as the only guide for multi-species dietary restrictions. Nevertheless, the potential clinical allergy to other Rosaceae should not be neglected. If the reported reaction is confirmed, current tolerance to other Rosaceae should be precisely established unless there has been ingestion without symptoms after the reaction.
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[80] - Akkerdaas J, Fernandez-Rivas M, Zuidmeer L, Hefle S, Aalberse R, van Ree R. IgE binding profiles to several food LTPs of Spanish patients with apple ingestion related symptoms. Allergy Clin Immunol Int 2005;17(Suppl. 1):338-339
Background Fruit allergic individuals from the Mediterranean area are predominantly sensitized by the non-specific lipid transfer protein (nsLTP). It seems that peach LTP is the most likely sensitizing allergen, however other factors (pollen related, genetic) can not be excluded yet. Objective It was our aim to identify IgE binding profiles to different natural (n) and recombinant (r) purified LTPs and to study the pair wise correlation between amino acid sequence identity and LTP specific IgE titers. Methods Serum and clinical data (after ingestion of 6 different foods) was obtained from Spanish apple allergic patients (n=19), as was proven by SPT/ DBPCFC. Seven different LTPs (rApple, rPeach, nGrape, rStrawberry, nSunflower, nHazelnut and nMaize, also analyzed for sequence homology) were applied in RAST to determine specific-IgE titers. Results Clinical symptoms after apple ingestion ranged from no- to severe systemic reactions like anaphylaxis. LTP Sequence homologies ranged from 81% (apple/peach) to 44% (hazelnut/maize). Highest correlation observed for IgE binding:0.989 (apple:peach). Lowest correlation: 0.228 (maize/grape). RAST highest/lowest mean score: apple 6.4/maize 0.4 IU/ml respectively. Conclusion Since most patients were selected on having a true apple allergy and the amino acid sequence homology between apple and peach LTP is 81%, an IgE binding-correlation of 0.989 was to be expected. Elucidation of IgE crossreactivity patterns between the LTPs applied might predict clinical symptoms. The intriguing question remains: How do we identify the sensitizing LTP and how do we prevent the development of (severe) food allergy as result of the induction of IgE against LTP?
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[81] - 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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[82] - Karlsson AL, Alm R, Ekstrand B, Fjelkner-Modig S, Schiött A, Bengtsson U, et al. Bet v 1 homologues in strawberry identified as IgE-binding proteins and presumptive allergens. Allergy 2004;59:1277-1284
BACKGROUND: No strawberry allergen has so far been identified and characterized . METHODS: Serum samples were collected from patients with a suggestive case history of adverse reactions to strawberry and other fruits. Extracts from fresh and frozen strawberries were analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blotting and mass spectrometry. Patient blood samples were analysed for inhibition of IgE binding and basophil degranulation . RESULTS: Several IgE-binding proteins could be detected. In more than half of the patient sera, a 20/18-kDa doublet band was observed in Western blotting. These two bands were excised and analysed by mass spectrometry showing the presence of proteins belonging to the Bet v 1 family of allergens. Inhibition of the IgE binding to the 20/18-kDa doublet was obtained by addition of two recombinantly expressed allergens belonging to the Bet v 1 family (Bet v 1 and Mal d 1) and strawberry protein extract. In a cell-based assay of patient blood samples, basophil degranulation could be induced by strawberry protein extract and by Bet v 1 and Mal d 1 . CONCLUSIONS: We conclude that strawberry homologues to Bet v 1 may be allergens of importance for adverse reactions to strawberry.
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[83] - 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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[84] - Karlsson AL, Alm R, Ekstrand B, Fjelkner-Modig S, Schiött A, Bengtsson U, et al. Bet v 1 homologues in strawberry identified as IgE-binding proteins and presumptive allergens. Allergy 2004;59:1277-1284
BACKGROUND: No strawberry allergen has so far been identified and characterized . METHODS: Serum samples were collected from patients with a suggestive case history of adverse reactions to strawberry and other fruits. Extracts from fresh and frozen strawberries were analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blotting and mass spectrometry. Patient blood samples were analysed for inhibition of IgE binding and basophil degranulation . RESULTS: Several IgE-binding proteins could be detected. In more than half of the patient sera, a 20/18-kDa doublet band was observed in Western blotting. These two bands were excised and analysed by mass spectrometry showing the presence of proteins belonging to the Bet v 1 family of allergens. Inhibition of the IgE binding to the 20/18-kDa doublet was obtained by addition of two recombinantly expressed allergens belonging to the Bet v 1 family (Bet v 1 and Mal d 1) and strawberry protein extract. In a cell-based assay of patient blood samples, basophil degranulation could be induced by strawberry protein extract and by Bet v 1 and Mal d 1 . CONCLUSIONS: We conclude that strawberry homologues to Bet v 1 may be allergens of importance for adverse reactions to strawberry.
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[85] - Zuidmeer L, Salentijn E, Fernandez-Rivas M, Gonzalez-Mancebo E, Asero R, Matos CI, et al. The role of profilin and lipid transfer protein in strawberry allergy in the Mediterranean area. Clin Exp Allergy 2006;36:666-675
BACKGROUND: In contrast to other Rosaceae fruit, only few cases of patients with adverse reactions to strawberry are listed in literature. OBJECTIVE To identify allergenic proteins in strawberry and to express and characterize recombinant strawberry lipid transfer protein (LTP; rFra a 3) . METHODS: Established apple-allergic patients were recruited on the basis of a reported allergic reaction to strawberry (n=28, confirmed by double-blind placebo-controlled food challenge in four patients) or on the basis of IgE reactivity to LTP (n=34). Sensitization to purified natural and recombinant allergens was assessed by RAST, immunoblot (inhibition) and basophil histamine release (BHR). A strawberry cDNA library was screened for genes homologous to known fruit allergens. Fra a 3 was cloned and expressed in the yeast Pichia pastoris and compared with peach and apple LTP by RAST, immunoblot-inhibition and BHR tests . RESULTS: Genes homologous to Bet v 1, Bet v 6, profilin and LTP were identified in a strawberry cDNA library. In BHR the rFra a 3 induced histamine release at a 100-fold higher concentration than peach LTP. RAST inhibition showed high cross-reactivity to peach and apple LTP, although IgE reactivity was lower by a factor 5. On strawberry immunoblot, patients' IgE showed reactivity to a Bet v 1 homologue, profilin, LTP and high-molecular weight bands . CONCLUSION: In addition to a Bet v 1 homologue, strawberry also contains IgE-binding profilin and LTP. The rFra a 3 has less allergenic potency than peach and apple LTP, and therefore is an interesting tool for future immunotherapy. Fra a 3 does not seem to be clinically relevant.
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[86] - Karlsson AL, Alm R, Ekstrand B, Fjelkner-Modig S, Schiött A, Bengtsson U, et al. Bet v 1 homologues in strawberry identified as IgE-binding proteins and presumptive allergens. Allergy 2004;59:1277-1284
BACKGROUND: No strawberry allergen has so far been identified and characterized . METHODS: Serum samples were collected from patients with a suggestive case history of adverse reactions to strawberry and other fruits. Extracts from fresh and frozen strawberries were analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blotting and mass spectrometry. Patient blood samples were analysed for inhibition of IgE binding and basophil degranulation . RESULTS: Several IgE-binding proteins could be detected. In more than half of the patient sera, a 20/18-kDa doublet band was observed in Western blotting. These two bands were excised and analysed by mass spectrometry showing the presence of proteins belonging to the Bet v 1 family of allergens. Inhibition of the IgE binding to the 20/18-kDa doublet was obtained by addition of two recombinantly expressed allergens belonging to the Bet v 1 family (Bet v 1 and Mal d 1) and strawberry protein extract. In a cell-based assay of patient blood samples, basophil degranulation could be induced by strawberry protein extract and by Bet v 1 and Mal d 1 . CONCLUSIONS: We conclude that strawberry homologues to Bet v 1 may be allergens of importance for adverse reactions to strawberry.
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[87] - Marzban G, Mansfeld A, Hemmer W, Stoyanova E, Katinger H, da Camara Machado ML. Fruit cross-reactive allergens: a theme of uprising interest for consumers' health. Biofactors 2005;23:235-241
Regular consumption of fruits has a positive influence on human health by disease prevention. However parallel to dietetic benefits, IgE-mediated fruit allergies have been shown to be an increasing health risk for children and adults in the Northern hemisphere. The spectrum of food allergies ranges from chronic symptoms to more acute problems and even anaphylaxis. Fruit proteins with high primary sequence similarity display also homologous tertiary structures, resulting in similar epitopes to IgEs and consequently in cross-reactivity. In this review we present the major allergens of stone and pome fruits and discuss the presence of homologous proteins in small fruits. Interestingly these proteins, which might pose an allergenic potential for pre-sensitised individuals are expressed also in strawberry, raspberry and blueberry, otherwise rich in beneficial biofactors.
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[88] - Zuidmeer L, Salentijn E, Fernandez-Rivas M, Gonzalez-Mancebo E, Asero R, Matos CI, et al. The role of profilin and lipid transfer protein in strawberry allergy in the Mediterranean area. Clin Exp Allergy 2006;36:666-675
BACKGROUND: In contrast to other Rosaceae fruit, only few cases of patients with adverse reactions to strawberry are listed in literature. OBJECTIVE To identify allergenic proteins in strawberry and to express and characterize recombinant strawberry lipid transfer protein (LTP; rFra a 3) . METHODS: Established apple-allergic patients were recruited on the basis of a reported allergic reaction to strawberry (n=28, confirmed by double-blind placebo-controlled food challenge in four patients) or on the basis of IgE reactivity to LTP (n=34). Sensitization to purified natural and recombinant allergens was assessed by RAST, immunoblot (inhibition) and basophil histamine release (BHR). A strawberry cDNA library was screened for genes homologous to known fruit allergens. Fra a 3 was cloned and expressed in the yeast Pichia pastoris and compared with peach and apple LTP by RAST, immunoblot-inhibition and BHR tests . RESULTS: Genes homologous to Bet v 1, Bet v 6, profilin and LTP were identified in a strawberry cDNA library. In BHR the rFra a 3 induced histamine release at a 100-fold higher concentration than peach LTP. RAST inhibition showed high cross-reactivity to peach and apple LTP, although IgE reactivity was lower by a factor 5. On strawberry immunoblot, patients' IgE showed reactivity to a Bet v 1 homologue, profilin, LTP and high-molecular weight bands . CONCLUSION: In addition to a Bet v 1 homologue, strawberry also contains IgE-binding profilin and LTP. The rFra a 3 has less allergenic potency than peach and apple LTP, and therefore is an interesting tool for future immunotherapy. Fra a 3 does not seem to be clinically relevant.
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[89] - Zuidmeer L, Salentijn E, Fernandez-Rivas M, Gonzalez-Mancebo E, Asero R, Matos CI, et al. The role of profilin and lipid transfer protein in strawberry allergy in the Mediterranean area. Clin Exp Allergy 2006;36:666-675
BACKGROUND: In contrast to other Rosaceae fruit, only few cases of patients with adverse reactions to strawberry are listed in literature. OBJECTIVE To identify allergenic proteins in strawberry and to express and characterize recombinant strawberry lipid transfer protein (LTP; rFra a 3) . METHODS: Established apple-allergic patients were recruited on the basis of a reported allergic reaction to strawberry (n=28, confirmed by double-blind placebo-controlled food challenge in four patients) or on the basis of IgE reactivity to LTP (n=34). Sensitization to purified natural and recombinant allergens was assessed by RAST, immunoblot (inhibition) and basophil histamine release (BHR). A strawberry cDNA library was screened for genes homologous to known fruit allergens. Fra a 3 was cloned and expressed in the yeast Pichia pastoris and compared with peach and apple LTP by RAST, immunoblot-inhibition and BHR tests . RESULTS: Genes homologous to Bet v 1, Bet v 6, profilin and LTP were identified in a strawberry cDNA library. In BHR the rFra a 3 induced histamine release at a 100-fold higher concentration than peach LTP. RAST inhibition showed high cross-reactivity to peach and apple LTP, although IgE reactivity was lower by a factor 5. On strawberry immunoblot, patients' IgE showed reactivity to a Bet v 1 homologue, profilin, LTP and high-molecular weight bands . CONCLUSION: In addition to a Bet v 1 homologue, strawberry also contains IgE-binding profilin and LTP. The rFra a 3 has less allergenic potency than peach and apple LTP, and therefore is an interesting tool for future immunotherapy. Fra a 3 does not seem to be clinically relevant.
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[90] - Déruaz C, Fellrath JM, Ballmer-Weber B. A delicious but dangerous raspberry (Rubus ideaus) jelly jam from a 68 year non atopic grand mam having presented a lipid transfer protein (Rub i 3) mediated anaphylactic reaction. Rev Fr Allergol 2009;49:302-303
Background.ˆ Up to 50% of birch pollen allergic patients react mildly (by the so-called oral allergic syndrome) when eating fruits of the rosaceae familiy (mainly apples, pears, peaches, rarely also berries), nuts and vegetables in relation with Bet v 1 homologous or profilin allergens. More severe anaphylactic reactions are reported after consumption of peach for example, related to a heat stable protein called lipid transfer protein (LTP: Pru p 3) an ubiquitous plant food lipid binding protein and potential panallergen. We report an anaphylactic reaction ˆ to our knowledge the first reported ˆ in a 68-year-old lady after ingestion of cooked raspberry jelly jam. History.ˆ Five minutes after eating raspberry jelly jam this 68 old patient ˆ treated for a metabolic syndrome ˆ presented palmoplantar pruritus, generalized urticaria and an enormous lip angioedema treated by i.v. corticosteroid and i.v. antihistamine. Ten years ago she presented a similar but weaker reaction after raw blueberry consumption. She tolerates well peaches and apples. Allergic workup.ˆ Revealed strong positive skin reactions to defrozen raspberry (wild and common) and blueberry extract, and cooked raspberry jelly jam whereas all other skin test (common aeroallergen and food allergens) resulted negative. Total IgE amounted 6,82 kU/l. Specific IgE (CAP: Phadia1) : positive in class 2 for recombinant peach LTP (rPru p3 = f420) 1,64 kU/l, for raspberry (f343) 0,84 kU/l and in class 1 for blueberry (f288) at 0,37 kU/l. Bet v1 (t215) was negative. Conclusion.ˆ This unique anaphylactic reaction underlines the extreme high degree of heat stability of the raspberry LTP allergen (Rub i 3). The strong positive skin test with heated raspberry jelly jam established the diagnosis, supported by a recombinant serological peach LTP ˆ probably highly homologous to raspberry LTP. Future work in the clinical and molecular field of Bet v 1 homologous (Rub i1) and LTP (Rub i 3) proteins in raspberry and other berries is warranted.
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[91] - 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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[92] - Armentia A, Barber D, Lombardero M, Martin Santos JM, Martin Gil FJ, Arranz Pena ML, et al. Anaphylaxis associated with antiphospholipid syndrome. Ann Allergy Asthma Immunol 2001;87:54-59
BACKGROUND: To our knowledge, no previously published reports have described food-induced anaphylaxis associated with the antiphospholipid syndrome. OBJECTIVE: We undertook a study of four patients with thrombosis associated with the antiphospholipid syndrome after each patient experienced anaphylaxis attributable to ingestion of vegetal foods. METHODS: IgE antibody levels to various foods were determined in serum specimens from the study patients, and skin prick tests with the same allergens were conducted to determine their in vivo responses. Hematologic, cardiopulmonary, vascular, and rheumatologic studies were also performed. IgG anticardiolipin antibody levels were determined by ELISA. RESULTS: All four patients fulfilled the criteria for antiphospholipid syndrome and had high levels of specific IgE antibodies for certain food allergens. By immunoblot analysis, the presence of serum IgE specific for a 45-kD protein band in an almond extract was detected in these four patients who experienced food-related anaphylaxis. No specific IgE was detected in sera from normal subjects. No IgE antibodies specific for the food panallergen lipid transfer proteins were detected. CONCLUSIONS: This is the first report of severe food-precipitated anaphylaxis associated with the antiphospholipid syndrome and the first description of a patient with allergy to blackberry. The possible involvement of food panallergens distinct from lipid transfer proteins is also discussed
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[93] - Marzban G, Herndl A, Kolarich D, Maghuly F, Mansfeld A, Hemmer W, et al. Identification of four IgE-reactive proteins in raspberry (Rubus ideaeus L.). Mol Nutr Food Res 2008;52:1497-1506
Abstract IgE-reactive proteins in raspberry (Rubus ideaus L.) were identified using PCR, RT-PCR, 2-DE and MS/MS peptide sequencing. Specific polyclonal antibodies and patient sera were used in Western blotting to identify crossreactive epitopes. Initially, two potential allergens Rub i 1 and Rub i 3 were detected using PCR, showing high sequence identity to proteins in Rosaceous species like Mal d 1 and Mal d 3 from apple, Pru av 1 and Pru av 3 from cherry and Pru p 1 and Pru p 3 from peach. Furthermore, de novo identified peptides of a protein band at about 30 kDa reacting with most of the patient sera tested (> 80%) revealed a high sequence homology with class III chitinases. Raspberry chitinase, when subjected to glycoproteomic analysis, showed typical complex plant-type N-glycans with a core alpha1,3 fucose and a beta1,2 xylose at least at one position, indicating the presence of crossreacting carbohydrate determinants (CCDs). Finally, MS/MS analysis revealed an IgE-reactive raspberry cyclophilin, homologous to Be t v 7. Results obtained suggest that the consumption of raspberries might be responsible for adverse reactions in sensitised individuals
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[94] - Marzban G, Herndl A, Kolarich D, Maghuly F, Mansfeld A, Hemmer W, et al. Identification of four IgE-reactive proteins in raspberry (Rubus ideaeus L.). Mol Nutr Food Res 2008;52:1497-1506
Abstract IgE-reactive proteins in raspberry (Rubus ideaus L.) were identified using PCR, RT-PCR, 2-DE and MS/MS peptide sequencing. Specific polyclonal antibodies and patient sera were used in Western blotting to identify crossreactive epitopes. Initially, two potential allergens Rub i 1 and Rub i 3 were detected using PCR, showing high sequence identity to proteins in Rosaceous species like Mal d 1 and Mal d 3 from apple, Pru av 1 and Pru av 3 from cherry and Pru p 1 and Pru p 3 from peach. Furthermore, de novo identified peptides of a protein band at about 30 kDa reacting with most of the patient sera tested (> 80%) revealed a high sequence homology with class III chitinases. Raspberry chitinase, when subjected to glycoproteomic analysis, showed typical complex plant-type N-glycans with a core alpha1,3 fucose and a beta1,2 xylose at least at one position, indicating the presence of crossreacting carbohydrate determinants (CCDs). Finally, MS/MS analysis revealed an IgE-reactive raspberry cyclophilin, homologous to Be t v 7. Results obtained suggest that the consumption of raspberries might be responsible for adverse reactions in sensitised individuals
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[95] - Zuidmeer L, Bolhaar S, Knulst A, van Leeuwen WA, Aalbers M, Krebitz M, et al. Severe symptoms caused by thaumatin-like proteins in foods: a result of primary sensitization to birch pollen ? EAACI 22th Congress, Paris, 7-11 June, 2003, Poster n°196
Background: It has almost become a dogma that birch pollen-related food allergies are always mild, because Bet v 1 and its homologues from foods as well as profilins are extremely heat and protease-sensitive. Protease-resistant birch pollen allergens have so far not been reported. Methods: A panel of 477 sera from The Netherlands with >2.0 IU/ml of specific IgE against apple were analysed by RAST for specific IgE against 4 apple allergens: nMal d 1, rMal d 2 (thaumatin-like protein [TLP]), nMal d 3 (LTP) and nMal d 4 (profilin). rMal d 2-positive sera (>0.7 IU/ml) were further analysed by RAST inhibition with (protease-digested) birch pollen extract. Serum from a patient with severe food-related anaphylaxis was studied in more detail by RAST(-inhibition) and immunoblot(- inhibition). Results: As expected most sera contained significant titers of specific IgE against Mal d 1 (70% with > 1.0 IU/ml). Frequency of recognition of Mal d 2, 3 and 4 was much lower (23%, 9% and 19% respectively). For 55 sera with a clear positive RAST for rMal d 2 IgE-binding to this allergen was inhibited with birch pollen extract. On average the Mald 2 RAST was inhibited by 75%, indicating that birch pollen extract contains a crossreactive structure with food TLP. A similar inhibition was observed with protease-digested birch pollen extract, suggesting that the putative TLP-related allergen from birch pollen is extremely protease resistant. This observation fits with the high degree of stability observed for rMal d 2 in simulated gastric fluid. Considering this stability, it is not surprising that among the patients with TLP-specific IgE antibodies, severe food allergic symptoms were observed. One of these patients was further analysed in depth. This patient experienced anaphylactic shock upon consumption of hazelnut liquor, raspberry and several other fruits on different occasions. IgE binding to raspberry and several other foods could be inhibited by birch pollen and by rMal d 2 in both RAST- and immunoblot-inhibition, indicating that the severe food allergy of this patient is caused by TLP-like allergens and that sensitisation is most likely birch pollen driven. Conclusion: Birch pollen contain an allergen that is crossreactive with food TLPs and is most likely responsible for sensitisation. In contrast to most Bet v 1-related allergens, food TLPs can cause severe food allergy.
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[96] - Marzban G, Maghuly F, Herndl A, Katinger H, Laimer M. Screening and identification of putative allergens in berry fruits of the Rosaceae family: technical challenges. Biofactors 2008;34:37-46
Cross-reactive proteins in small fruits of the Rosaceae family like strawberry, raspberry and blackberry revealed an unexpected complex IgE-reactivity pattern. Several copies of PR-10 and PR-14 proteins were detected by Southern blots in strawberry, raspberry and blackberry. In raspberry, the highest similarity at the DNA level for PR-10 and PR-14 (Rub i 1 and Rub i 3) was detected to strawberry sequences of Fra a 1 and Fra a 3. At the protein level, Rub i 1 and Rub i 3 showed more than 70% identity with homologous proteins of rosaceous fruits. Furthermore, raspberries contained additional putative allergens, e.g. class III acidic chitinases and cyclophilins. Blackberries were shown to share at least two well-known major fruit allergens with other rosaceous fruits, namely PR-10s and PR-14s homologous proteins. However the IgE-reactive proteins of small fruits are still not extensively investigated. The main challenges in studying small fruit allergens are the complexity of the fruit matrix, the diversity of physico-chemical properties of fruit proteins, the lack of appropriate protein extraction procedures and the missing information about the influence of processing treatments on food components.
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[97] - Armentia A, Barber D, Lombardero M, Martin Santos JM, Martin Gil FJ, Arranz Pena ML, et al. Anaphylaxis associated with antiphospholipid syndrome. Ann Allergy Asthma Immunol 2001;87:54-59
BACKGROUND: To our knowledge, no previously published reports have described food-induced anaphylaxis associated with the antiphospholipid syndrome. OBJECTIVE: We undertook a study of four patients with thrombosis associated with the antiphospholipid syndrome after each patient experienced anaphylaxis attributable to ingestion of vegetal foods. METHODS: IgE antibody levels to various foods were determined in serum specimens from the study patients, and skin prick tests with the same allergens were conducted to determine their in vivo responses. Hematologic, cardiopulmonary, vascular, and rheumatologic studies were also performed. IgG anticardiolipin antibody levels were determined by ELISA. RESULTS: All four patients fulfilled the criteria for antiphospholipid syndrome and had high levels of specific IgE antibodies for certain food allergens. By immunoblot analysis, the presence of serum IgE specific for a 45-kD protein band in an almond extract was detected in these four patients who experienced food-related anaphylaxis. No specific IgE was detected in sera from normal subjects. No IgE antibodies specific for the food panallergen lipid transfer proteins were detected. CONCLUSIONS: This is the first report of severe food-precipitated anaphylaxis associated with the antiphospholipid syndrome and the first description of a patient with allergy to blackberry. The possible involvement of food panallergens distinct from lipid transfer proteins is also discussed
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