Les fruits des Rosacées : généralités

Recrutement	PR 10	LTP	Profilines	Thaum.-like	Ref
..........	........	..................	............	...............	.
Allemagne
Cerise	Bet v 1 17/19 Mal d 1 16/19 Pru av 1 17/19		Bet v 2 5/19
Cerise	Pru av 1 96%	Pru av 3 3%	Pru av 4 16%
Poire (et bouleau)	Bet v 1 100%		Bet v 2 50%
Poire (et bouleau)	Bet v 1 7/7 Pyr c 1 7/7		Bet v 2 2/7
Pays-Bas
Pomme	Mal d 1 70%	Mal d 3 9% Pru p 3 16%	Mal d 4 19%	Mal d 2 23%
Pomme	Bet v 1 90%	Mal d 3 1%	Mal d 4 12%	Mal d 2 5%
Autriche
Pomme	Bet v 1 92%	Mal d 3 2%	Mal d 4 10%	Mal d 2 0%
Pêche (bouleau+)	Pru p 1 41/41 (15 mono)	Pru p 3 16/41	Bet v 2 18/41
Pêche . (bouleau -)	Pru p 1 6/13	Pru p 3 10/13 (3 mono)	Bet v 2 8/13
Suisse
Cerise	Pru av 1 92%	Pru av 3 4%	Pru av 4 17%
France
Rosacées	Bet v 1 29/29		Bet v 2 4/29
Portugal
Pêche	Pru p 1 1/30	Pru p 3 29/30	Pru p 4 4/30
Italie
Pomme	Bet v 1 99%	Mal d 3 10%	Mal d 4 34%	Mal d 2 7%
Espagne
S.O. fruits + pollinose	Bet v 1 0/10		Bet v 2 9/10
Cerise	Pru av 1 0%	Pru av 3 89%	Pru av 4 22%
Cerise	Pru av 1 1/8	Pru av 3 6/8	Pru av 4 0/8
Pomme	Mal d 1 14%	Mal d 3 59%	Mal d 4 41%	Mal d 2 25%
Pomme	Bet v 1 5%	Mal d 3 45%	Mal d 4 40%	Mal d 2 15%
Pomme TPODA+	PR-10 6%	Mal d 3 70%	Profiline 30%
Pomme Anaph.		Mal d 3 100%
Pêche TPODA+	PR-10 6%	Pru p 3 62%	Profiline 30%
Pêche Anaph.		Pru p 3 100%
Pêche	Bet v 1 7%		Bet v 2 32%
Pêche		Mal d 3 19/24 Pru p 3 21/24	Mal d 4 6/24 Bet v 2 6/24
Pêche	Bet v 1 7%	Pru p 3 69%	Bet v 2 44%
Pêche		Pru p 3 positifs	Pru p 4 0/14
Pêche		Pru p 3 négatifs	Pru p 4 15/15
Pêche		Pru p 3 62%	Bet v 2 34%
Fraise	Mal d 1 10/20	Mal d 3 10/20 Fra a 3 4/16	Mal d 4 15/20	Mal d 2 8/20

Allergie à	Nb sujets	Pays	Réactions locales uniquement (ex. syndrome oral)	Réactions systémiques (avec ou sans réactions locales)
Pêche	70	Espagne	74	26
150	Espagne	69	31
57	Espagne	28	72
Prune	12	50	50
Abricot	14	35	65
Pomme	20	26	74
Pomme	94	Autriche	96	4
97	Italie	90	10
99	Pays-Bas	99	1
99	Espagne	65	35
Cerise	12	Espagne	67	33
10	Allemagne	83	17
24	Suisse	66	34
9	Espagne	34	66

Recrutement	Patients	Pollinose	Pêche	Pomme	Ref
					.
Suède
bouleau	100			63%
poll. non bouleau	74			2%
Allemagne
cerise	101	bouleau 100%	57%	96%
isolat soja	20	bouleau 16	5	12
soja	22		14	20
TPODA+	104	graminées 34%, bouleau 31%	21%	68%
poire	7	bouleau	5	6
cerise	117		64%	96%
pousses soja	10	bouleau	6	9
Pays-bas
Bet v 1+ Prof -	14		5	10
Prof+ Bet v 1 -	7		0	1
noisette	31	bouleau 29		25
bouleau	79		29%	44%
Autriche
bouleau	426		33%	56%
France
céleri	20	bouleau	0	7
polliniques	25		8	12
Rosacées	12	bouleau+	7	11
10	bouleau -	8	4
Portugal
pêche (LTP+ 95%)	30	pollinose 57%		57%
Italie
pêche	21	graminées 16, bouleau 14	21
Rosacées, LTP -	212	bouleau 99%, ..	52%	87%
Rosacées, LTP+	25	bouleau 12%, ..	88%	44%
abricot	14	bouleau+	11	1
9	bouleau -	7	4
Rosacées, LTP+	37		30	16
prune (LTP+ 80%)	23	graminées 14, bouleau 11	100%
Rosacées mono-LTP	20	polliniques 10 (bouleau non)	20	7
mono-Bet v 1+ (non LTP+)	25			25
mono-Bet v 2+ (non LTP)	18			7
polliniques	14	platane, pariétaire, ..	5	4
Espagne
Rosacées	16	graminées 16	11	8
Rosacées	11	non polliniques	10	10
22	graminées, olivier, ..	22	15
TC+ Rosacées	28	polliniques 16	22	6
polliniques	95	graminées, olivier, ..	3	12
TPODA+ ou A	65	polliniques 65%	38	10
noisette	19		18	12
orange (LTP+ 7)	27	graminées 26, olivier, ..	19	9
cerise	22		20	10
allergie alim.	16	platane, ..	2
Grêce
raisin	11		10	6
raisin	37	graminées, olivier, .. 31	8	3

[1] - Hannuksela M, Lahti A. Immediate reactions to fruits and vegetables. Contact Dermatitis 1977;3:79-84

Common fruits and vegetables were tested as such with the "scratch-chamber" method in 388 patients with various atopic disorders. Of the patients with hypersensitivity to birch pollen, 36% showed immediate positive responses to these natural, fresh materials. Apple, carrot, parsnip and potato elicited reactions more often than, e.g. swede, tomato, onion and parsley. On the basis of clinical data, the relevancy of the results of the skin tests was 80-90%. Both "false positive" and "false negative" responses were seen. Itching and tingling with or without oedema of the lips, mouth and tongue were the most common complaints after eating raw fruits and vegetables. In addition, laryngeal and abdominal disturbances, rhinitis and hand dermatitis were recorded. Among patients without allergy to birch pollen, reactions to fruits and vegetables were rare. The nature of the allergens could not be determined.

↩

[2] - Lahti A, Hannuksela M. Hypersensitivity to apple and carrot can be reliably detected with fresh material. Allergy 1978;33:143-146

Hypersensitivity to birch pollen was linked with allergy to apple and carrot, apple seeds and carrot seeds and also with their fractions. Skin reactions among other atopics were rare, and non-atopic patients showed no reactions to fresh fruits and vegetables. Apple and carrot as well as apple seeds and carrot seeds were fractionated using column chromatography. Two major fractions were obtained, one with sugar and proteins, and one with proteins, the latter giving skin reactions more often than the former. The correlation between clinical symptoms and skin test results with the fractionated material was somewhat lower than that obtained with the non-fractionated fresh material. The allergens of fruits and vegetables might be proteins (enzymes).

↩

[3] - Björksten F, Halmepuro L, Hannuksela M, Lahti A. Extraction and properties of apple allergens. Allergy 1980;35:671-677

It is shown that apple allergens are probably proteins and that they can be extracted in an active form only if reactions with phenolic compounds present in apple are inhibited. This is accomplished by incorporating chelators and solid polyvinylpolypyrrolidone in the extraction medium. Phenol, commonly used as a preservative, should not be added. With the RAST, serum-IgE antibodies capable of reacting with apple allergens were detected in 90% of patients with clinical apple allergy, in 44% of patients with clinical birch-pollen allergy and in 5-10% of patients with other atopic allergies. RAST inhibition indicated that apple and birch-pollen allergens cross-react.

↩

[4] - 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.

↩

[5] - Pastorello EA, Pravettoni V, Farioli L, Ispano M, Fortunato D, Monza M, et al. Clinical role of a lipid transfer protein that acts as a new apple-specific allergen. J Allergy Clin Immunol 1999;104:1099-1106

Allergy to apple is commonly associated with birch pollinosis because the two share homologous allergens. However, some patients have apple allergy but no birch pollinosis, suggesting that there are allergens that do not cross-react with birch. OBJECTIVE: The aim of the study was to evaluate the IgE reactivity pattern to an apple extract in subjects with allergic reactions to apple, with and without birch hay fever. METHODS: Forty-three patients with oral allergy syndrome for apple and positive open food challenge, skin prick test, and serum specific IgE antibodies to apple were admitted to the study. Thirty-two had birch pollinosis (documented by specific IgE for birch) and 11 were not allergic to birch. The IgE reactivity pattern to apple extract was identified by SDS-PAGE and immunoblotting. The consistent allergen, a 9-kd protein, was then purified by HPLC and characterized by periodic acid-Schiff staining, isoelectric point, and N-terminal amino acid sequencing. RESULTS: The sera from 28% of patients allergic to apple with birch pollinosis, but from all patients allergic only to apple, recognized the 9-kd protein. This protein has an isoelectric point of 7.5 and is not glycosylated. Determination of its partial amino acid sequence showed that it belongs to the family of lipid transfer proteins, which act as major allergens in Prunoideae fruits. CONCLUSIONS: These results indicate that a lipid transfer protein is an important allergen in patients allergic to apple but not to birch pollen. The prevalent IgE reactivity to this allergen in subjects with no birch pollinosis and the physicochemical characteristics of this protein suggest that sensitization may occur through the oral route.

↩

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

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

↩

[7] - 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.

↩

[8] - 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.

↩

[9] - 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.

↩

[10] - Fernández-Rivas M, Bolhaar S, González-Mancebo E, Asero R, van Leeuwen A, Bohle B, et al. Apple allergy across Europe: How allergen sensitization profiles determine the clinical expression of allergies to plant foods. J Allergy Clin Immunol 2006;118:481-488

BACKGROUND: Allergy to a plant food can either result from direct sensitization to that food or from primary sensitization to pollen, latex, or another food . OBJECTIVE: We sought to investigate the primary sensitizers in apple allergy across Europe, the individual allergens involved, and whether these differences determine the clinical presentation . METHODS: Patients (n = 389) with positive case histories and skin prick test responses to fresh apple were selected in the Netherlands, Austria, Italy, and Spain. Skin prick tests and RASTs to a panel of pollens and plant foods were performed, as well as RASTs to Bet v 1 and the apple allergens Mal d 1, 2, 3, and 4 . RESULTS: In the Netherlands, Austria, and Italy apple allergy is mild (>90% isolated oral symptoms) and related to birch pollinosis and sensitization to Bet v 1 and its apple homologue, Mal d 1, which has an odds ratio of local reactions of 2.85 (95% CI, 1.47-5.55). In Spain apple allergy is severe (>35% systemic reactions) and related to peach allergy and sensitization to Mal d 3 (nonspecific lipid transfer protein), which has an odds ratio of systemic reactions of 7.76 (95% CI, 3.87-15.56) . CONCLUSION: The analysis of individual apple allergens in a clinical context has provided insight into the sensitization pathway and into the intrinsic risk an allergen bears to induce mild or severe food allergy. CLINICAL IMPLICATIONS: Information on the sensitization pathway is essential to develop preventive strategies in food allergy. The application of individual food allergens with a known intrinsic risk will improve the prognostic value of diagnostic tests.

↩

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

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

↩

[12] - 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) .

↩

[13] - Pastorello EA, Pravettoni V, Farioli L, Ispano M, Fortunato D, Monza M, et al. Clinical role of a lipid transfer protein that acts as a new apple-specific allergen. J Allergy Clin Immunol 1999;104:1099-1106

Allergy to apple is commonly associated with birch pollinosis because the two share homologous allergens. However, some patients have apple allergy but no birch pollinosis, suggesting that there are allergens that do not cross-react with birch. OBJECTIVE: The aim of the study was to evaluate the IgE reactivity pattern to an apple extract in subjects with allergic reactions to apple, with and without birch hay fever. METHODS: Forty-three patients with oral allergy syndrome for apple and positive open food challenge, skin prick test, and serum specific IgE antibodies to apple were admitted to the study. Thirty-two had birch pollinosis (documented by specific IgE for birch) and 11 were not allergic to birch. The IgE reactivity pattern to apple extract was identified by SDS-PAGE and immunoblotting. The consistent allergen, a 9-kd protein, was then purified by HPLC and characterized by periodic acid-Schiff staining, isoelectric point, and N-terminal amino acid sequencing. RESULTS: The sera from 28% of patients allergic to apple with birch pollinosis, but from all patients allergic only to apple, recognized the 9-kd protein. This protein has an isoelectric point of 7.5 and is not glycosylated. Determination of its partial amino acid sequence showed that it belongs to the family of lipid transfer proteins, which act as major allergens in Prunoideae fruits. CONCLUSIONS: These results indicate that a lipid transfer protein is an important allergen in patients allergic to apple but not to birch pollen. The prevalent IgE reactivity to this allergen in subjects with no birch pollinosis and the physicochemical characteristics of this protein suggest that sensitization may occur through the oral route.

↩

[14] - Fernández-Rivas M. Alergia a frutas y hortalizas. in "Alergia a alimentos", Sociedad Española de Alergología e Inmunología Clínica (SEAIC) Ed., Euromedice, Badalona 2004, pp.143-67

↩

[15] - Fernández-Rivas M. [Cross-reactivity between fruit and vegetables]. Allergol Immunopathol (Madr) 2003;31:141-146

Vegetable foods are the most frequent cause of food allergy after the age of 5 years. The most commonly implicated foods are fruit and dried fruits, followed in Spain by legumes and fresh garden produce. In patients allergic to fruit and garden produce, multiple sensitizations to other vegetable products, whether from the same family or taxonomically unrelated, are frequent, although they do not always share the same clinical expression. Furthermore, more than 75 % of these patients are allergic to pollen, the type of pollen varying in relation to the aerobiology of the area. The basis of these associations among vegetable foods and with pollens lies in the existence of IgE antibodies against "panallergens", which determines cross-reactivity. Panallergens are proteins that are spread throughout the vegetable kingdom and are implicated in important biological functions (generally defense) and consequently their sequences and structures are highly conserved. The three best-known groups are allergens homologous to Bet v 1, profilins, and lipid transfer proteins (LTP).Allergens homologous to Bet v 1 (major birch pollen allergen) constitute a group of defense proteins (PR-10), with a molecular weight of 17 kDa, which behave as major allergens in patients from northern and central Europe with allergy to vegetables associated with birch pollen allergy. In these patients, the primary sensitization seems to be produced through the inhalation route on exposure to birch pollen. The symptomatology characteristically associated with sensitization to this family of allergens is oral allergy syndrome (OAS).Profilins are highly conserved proteins in all eukaryotic organisms and are present in pollen and a wide variety of vegetable foods. They have a molecular weight of 14 kDa and present a high degree of structural homology as well as marked cross-reactivity among one another. The presence of anti-profilin IgE broadens the spectrum of sensitizations to vegetable foods detected through skin tests and/or in vitro tests but whether it correlates with the clinical expression of food allergy is unclear.LTPs are the most commonly implicated allergens in allergy to Rosaceae fruits in patients from the Mediterranean area without birch pollen sensitization. LTPs are a family of 9kDA polypeptides, widely found in the vegetable kingdom and implicated in cuticle formation and defense against pathogens (PR-14). They are thermostable and resistant to pepsin digestion, which makes them potent food allergens and explains the frequent development of systemic symptoms (urticaria, anaphylaxis) in patients allergic to Rosaceae fruits in Spain. LTPs have also been identified in other vegetable foods and in pollens and a marked degree of cross-reactivity among them has been demonstrated, which may explain (together with profilin) the frequency of individuals sensitized to vegetable foods in the Mediterranean area.

↩

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

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

↩

[17] - Rodriguez-Perez R, Fernández-Rivas M, González-Mancebo E, Sánchez-Monge R, Diaz-Perales A, Salcedo G. Peach profilin: cloning, heterologous expression and cross-reactivity with Bet v 2. Allergy 2003;58:635-640

BACKGROUND: Peach is among the main foods causing allergic reactions in the Mediterranean adult population. Only a single peach allergen, named Pru p 3, has been characterized. However, a potential role of profilin has also been suggested in grass pollen-associated allergy to peach . METHODS: Complementary DNA clones for two different peach profilin isoforms were obtained by reverse transcriptase polymerase chain reaction using non-degenerated primers. Expression of recombinant peach profilin was performed in Escherichia coli, and confirmed using rabbit polyclonal antibodies to sunflower pollen profilin. Twenty-nine individual sera from patients with peach allergy proved by double-blind, placebo-controlled food challenges (DBPCFC), either with (n = 15) or without (n = 14) specific IgE to Bet v 2, were used in immunodetection assays to test recombinant peach profilin reactivity . RESULTS: Each peach profilin cDNA included an open reading frame coding for a 131 amino acid protein. The peach profilin isoforms, designated Pru p 4.01 and Pru p 4.02, showed 80% of amino acid sequence identity, and were very similar (>70% identity) to allergenic profilins from plant foods and pollens. Recombinant Pru p 4.01 was expressed in E. coli as a nonfusion protein, displaying the expected molecular size and reacting with anti-profilin antibodies. rPru p 4.01 was recognized by all sera (15 of 15) with specific IgE to Bet v 2, whereas no sera (zero of 14) without IgE to this birch allergen reacted with rPru p 4.01 . CONCLUSIONS: Peach profilin Pru p 4 is very closed to other allergenic profilins from plant foods and pollens. A complete correlation between reactivity to rPru p 4 and rBet v 2 has been found in sera from peach allergic patients.

↩

[18] - 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.

↩

[19] - Cuesta J, Blanco C, Fernández-Rivas M, Villalba M. Vegetalia. J Investig Allergol Clin Immunol 2006;16(Suppl. 2):40-44

↩

[20] - Scheurer S, Metzner K, Haustein D, Vieths S. Molecular cloning, expression and characterization of Pru a 1, the major cherry allergen. Mol Immunol 1997;34:619-629

A high percentage of birch pollen allergic patients experiences food hypersensitivity reactions after ingestion of several fruits and vegetables. Previous work demonstrated common epitopes on an allergen of Mr 18,000 from sweet cherry (Prunus avium) and Bet v 1, the major allergen from birch pollen. N-terminal amino acid sequencing showed a sequence identity of 67% with Bet v 1. Here we report the cloning and cDNA sequencing of this cherry allergen. The entire deduced amino acid sequence described a protein of Mr 17,700 with 59.1% identity to Bet v 1. High degrees of identity in the range of 40 to 60% were also found with related allergens from other kinds of tree pollen and plant foods as well as with stress-induced proteins from food plants such as parsley, potato and soya. The coding DNA of the cherry protein was cloned into vector pET-16b and expressed in E. coli strain BL21(DE3) as a His-tag fusion protein. As shown by SDS-PAGE, the apparent molecular masses of the nonfusion protein and the natural allergen were identical. The fusion protein showed high IgE binding potency when sera from patients allergic to cherry were tested by immunoblotting and enzyme allergosorbent tests. Moreover, it cross-reacted strongly with IgE specific for the natural counterpart and for Bet v 1. The high biological activity of the recombinant fusion protein was further confirmed by the induction of a strong histamine release in basophils from cherry-allergic patients. Since sera from 17/19 of such patients contained IgE against this allergen it was classified as a major allergen and named Pru a 1. Recombinant Pru a 1 mimics most of the allergenic potency of cherry extract and hence could be a useful tool for studying the molecular and immunological properties of pollen related food allergens.

↩

[21] - 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.

↩

[22] - 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.

↩

[23] - 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.

↩

[24] - 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.

↩

[25] - Fernández-Rivas M, Bolhaar S, González-Mancebo E, Asero R, van Leeuwen A, Bohle B, et al. Apple allergy across Europe: How allergen sensitization profiles determine the clinical expression of allergies to plant foods. J Allergy Clin Immunol 2006;118:481-488

BACKGROUND: Allergy to a plant food can either result from direct sensitization to that food or from primary sensitization to pollen, latex, or another food . OBJECTIVE: We sought to investigate the primary sensitizers in apple allergy across Europe, the individual allergens involved, and whether these differences determine the clinical presentation . METHODS: Patients (n = 389) with positive case histories and skin prick test responses to fresh apple were selected in the Netherlands, Austria, Italy, and Spain. Skin prick tests and RASTs to a panel of pollens and plant foods were performed, as well as RASTs to Bet v 1 and the apple allergens Mal d 1, 2, 3, and 4 . RESULTS: In the Netherlands, Austria, and Italy apple allergy is mild (>90% isolated oral symptoms) and related to birch pollinosis and sensitization to Bet v 1 and its apple homologue, Mal d 1, which has an odds ratio of local reactions of 2.85 (95% CI, 1.47-5.55). In Spain apple allergy is severe (>35% systemic reactions) and related to peach allergy and sensitization to Mal d 3 (nonspecific lipid transfer protein), which has an odds ratio of systemic reactions of 7.76 (95% CI, 3.87-15.56) . CONCLUSION: The analysis of individual apple allergens in a clinical context has provided insight into the sensitization pathway and into the intrinsic risk an allergen bears to induce mild or severe food allergy. CLINICAL IMPLICATIONS: Information on the sensitization pathway is essential to develop preventive strategies in food allergy. The application of individual food allergens with a known intrinsic risk will improve the prognostic value of diagnostic tests.

↩

[26] - Fernández-Rivas M, Bolhaar S, González-Mancebo E, Asero R, van Leeuwen A, Bohle B, et al. Apple allergy across Europe: How allergen sensitization profiles determine the clinical expression of allergies to plant foods. J Allergy Clin Immunol 2006;118:481-488

BACKGROUND: Allergy to a plant food can either result from direct sensitization to that food or from primary sensitization to pollen, latex, or another food . OBJECTIVE: We sought to investigate the primary sensitizers in apple allergy across Europe, the individual allergens involved, and whether these differences determine the clinical presentation . METHODS: Patients (n = 389) with positive case histories and skin prick test responses to fresh apple were selected in the Netherlands, Austria, Italy, and Spain. Skin prick tests and RASTs to a panel of pollens and plant foods were performed, as well as RASTs to Bet v 1 and the apple allergens Mal d 1, 2, 3, and 4 . RESULTS: In the Netherlands, Austria, and Italy apple allergy is mild (>90% isolated oral symptoms) and related to birch pollinosis and sensitization to Bet v 1 and its apple homologue, Mal d 1, which has an odds ratio of local reactions of 2.85 (95% CI, 1.47-5.55). In Spain apple allergy is severe (>35% systemic reactions) and related to peach allergy and sensitization to Mal d 3 (nonspecific lipid transfer protein), which has an odds ratio of systemic reactions of 7.76 (95% CI, 3.87-15.56) . CONCLUSION: The analysis of individual apple allergens in a clinical context has provided insight into the sensitization pathway and into the intrinsic risk an allergen bears to induce mild or severe food allergy. CLINICAL IMPLICATIONS: Information on the sensitization pathway is essential to develop preventive strategies in food allergy. The application of individual food allergens with a known intrinsic risk will improve the prognostic value of diagnostic tests.

↩

[27] - Gaier S, Oberhuber C, Hemmer W, Radauer C, Rigby NM, Marsh JY et al. Pru p 3 as a marker for symptom severity for patients with peach allergy in a birch pollen environment. J Allergy Clin Immunol 2009;124:166-167

↩

[28] - Ballmer-Weber BK, Scheurer S, Fritsche P, Enrique E, Cistero-Bahima A, Haase T, et al. Component-resolved diagnosis with recombinant allergens in patients with cherry allergy. J Allergy Clin Immunol 2002;110:167-173

BACKGROUND: In pollen-related food allergy, extracts for skin prick tests (SPTs) are often not standardized, and the test reliability is affected by false-negative reactions . OBJECTIVE: We sought to evaluate a panel of recombinant allergens (RAs) derived from one allergenic food for use in component-resolved in vivo diagnosis, taking cherry as a model food . METHODS: Seventy-nine subjects were included in the study: 24 Swiss patients (group 1) with a positive double-blind placebo-controlled food challenge result to cherries, 23 patients with birch pollen allergy but without cherry allergy (group 2), 23 nonatopic subjects (group 3), and 9 Spanish patients with a history of a cherry allergy (group 4). SPTs were performed in duplicate by using recombinant cherry allergens (Bet v 1-related allergen: recombinant (r) Pru av 1; profilin: rPru av 4; and lipid transfer protein: rPru av 3) in concentrations of 10, 50, and 100 microg/mL. Furthermore, IgE reactivity to rPru av 1, rPru av 4, and rPru av 3 was assessed by means of immunoblot analysis . RESULTS: SPT responses with rPru av 1, rPru av 4, and rPru av 3 were positive in 92%, 17%, and 4% of the patients in group 1; in 74%, 30%, and 0% of the patients in group 2; in 0%, 22%, and 89% of the patients in group 4; and negative for all nonatopic subjects (group 3). Thus the sensitivity of a positive SPT response to at least one of the 3 RAs was 96%. The specificities, negative predictive values, and positive predictive values with the 3 RAs were 100%, 96%, and 100% if calculated in relation to the nonatopic control group but 17%, 79%, and 60% when calculated in relation to the control group with birch pollen allergy. The correlation between SPT and immunoblotting results was excellent. Sensitization to rPru av 3 was associated with more severe symptoms than sensitization to rPru av 1 . CONCLUSIONS: SPTs with RAs proved to be highly sensitive for diagnosis of cherry allergy. Component-resolved in vivo diagnosis with standardized amounts of stable RAs allows us to determine sensitization patterns directly, to correlate them with severity of clinical symptoms, and to analyze geographic differences.

↩

[29] - Pauli G, Oster JP, Deviller P, Heiss S, Bessot JC, Susani M, et al. Skin testing with recombinant allergens rBet v 1 and birch profilin, rBet v 2: diagnostic value for birch pollen and associated allergies. J Allergy Clin Immunol 1996;97:1100-1109

This study assesses the value of two recombinant birch allergens for diagnosis of patients sensitized to birch pollen with or without associated food allergy. METHODS: Fifty-one patients with positive skin test responses to Betulaceae and seven nonallergic control subjects were investigated; specific IgE antibodies were evaluated by specific immunoassay and blot immunodetection. RESULTS: Among 51 patients, 47 reacted to rBet v 1 and 10 to rBet v 2. Seven patients reacted to both recombinant allergens. In skin prick tests we found a correlation between the wheal produced by the commercial birch extract and the wheal produced by rBet v 1. Among 47 patients with positive test responses to rBet v 1, 83% had IgE binding to the Bet v 1 protein as determined by immunoblotting. Among 10 patients sensitized to rBet v 2, six had IgE binding to Bet v 2. Eleven patients with negative results, as determined by immunoblotting, had low levels of birch IgE in the sera (less than 10 kU/L) and low concentrations of IgE to rBet v 1 or rBet v 2 in ELISA. The nonallergic control subjects (n = 7) did not react to rBet v 1 or rBet v 2 in skin prick tests, nor did they have detectable amounts of specific IgE to rBet v 1 or rBet v 2. Histamine release tests confirmed sensitization to Bet v 1 in two patients with discordant results; for Bet v 2, one patient had positive results only at a high concentration, and one had results that remained negative. Thirty-four patients had birch pollinosis, and all reacted to rBet v 1. Patients who were monosensitized to birch never reacted to rBet v 2. Sensitization to rBet v 2 was only found in patients who reacted to other pollens (mainly grass). Twenty-nine patients demonstrated allergy to apples, cherries, or hazelnuts; and all reacted to rBet v 1. Among 11 patients with allergy to Umbelliferae, only three reacted to rBet v 2. CONCLUSIONS: Use of the two recombinant allergens (rBet v 1 and rBet v 2) always permits the diagnosis of birch sensitization. Sensitization to rBet v 1 is specific for birch and Rosaceae allergies, whereas sensitization to birch profilin, Bet v 2, is encountered in multisensitized subjects and is not always related to Umbelliferae allergy.

↩

[30] - 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.

↩

[31] - Fernández-Rivas M, Bolhaar S, González-Mancebo E, Asero R, van Leeuwen A, Bohle B, et al. Apple allergy across Europe: How allergen sensitization profiles determine the clinical expression of allergies to plant foods. J Allergy Clin Immunol 2006;118:481-488

BACKGROUND: Allergy to a plant food can either result from direct sensitization to that food or from primary sensitization to pollen, latex, or another food . OBJECTIVE: We sought to investigate the primary sensitizers in apple allergy across Europe, the individual allergens involved, and whether these differences determine the clinical presentation . METHODS: Patients (n = 389) with positive case histories and skin prick test responses to fresh apple were selected in the Netherlands, Austria, Italy, and Spain. Skin prick tests and RASTs to a panel of pollens and plant foods were performed, as well as RASTs to Bet v 1 and the apple allergens Mal d 1, 2, 3, and 4 . RESULTS: In the Netherlands, Austria, and Italy apple allergy is mild (>90% isolated oral symptoms) and related to birch pollinosis and sensitization to Bet v 1 and its apple homologue, Mal d 1, which has an odds ratio of local reactions of 2.85 (95% CI, 1.47-5.55). In Spain apple allergy is severe (>35% systemic reactions) and related to peach allergy and sensitization to Mal d 3 (nonspecific lipid transfer protein), which has an odds ratio of systemic reactions of 7.76 (95% CI, 3.87-15.56) . CONCLUSION: The analysis of individual apple allergens in a clinical context has provided insight into the sensitization pathway and into the intrinsic risk an allergen bears to induce mild or severe food allergy. CLINICAL IMPLICATIONS: Information on the sensitization pathway is essential to develop preventive strategies in food allergy. The application of individual food allergens with a known intrinsic risk will improve the prognostic value of diagnostic tests.

↩

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

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

↩

[33] - Ballmer-Weber BK, Scheurer S, Fritsche P, Enrique E, Cistero-Bahima A, Haase T, et al. Component-resolved diagnosis with recombinant allergens in patients with cherry allergy. J Allergy Clin Immunol 2002;110:167-173

BACKGROUND: In pollen-related food allergy, extracts for skin prick tests (SPTs) are often not standardized, and the test reliability is affected by false-negative reactions . OBJECTIVE: We sought to evaluate a panel of recombinant allergens (RAs) derived from one allergenic food for use in component-resolved in vivo diagnosis, taking cherry as a model food . METHODS: Seventy-nine subjects were included in the study: 24 Swiss patients (group 1) with a positive double-blind placebo-controlled food challenge result to cherries, 23 patients with birch pollen allergy but without cherry allergy (group 2), 23 nonatopic subjects (group 3), and 9 Spanish patients with a history of a cherry allergy (group 4). SPTs were performed in duplicate by using recombinant cherry allergens (Bet v 1-related allergen: recombinant (r) Pru av 1; profilin: rPru av 4; and lipid transfer protein: rPru av 3) in concentrations of 10, 50, and 100 microg/mL. Furthermore, IgE reactivity to rPru av 1, rPru av 4, and rPru av 3 was assessed by means of immunoblot analysis . RESULTS: SPT responses with rPru av 1, rPru av 4, and rPru av 3 were positive in 92%, 17%, and 4% of the patients in group 1; in 74%, 30%, and 0% of the patients in group 2; in 0%, 22%, and 89% of the patients in group 4; and negative for all nonatopic subjects (group 3). Thus the sensitivity of a positive SPT response to at least one of the 3 RAs was 96%. The specificities, negative predictive values, and positive predictive values with the 3 RAs were 100%, 96%, and 100% if calculated in relation to the nonatopic control group but 17%, 79%, and 60% when calculated in relation to the control group with birch pollen allergy. The correlation between SPT and immunoblotting results was excellent. Sensitization to rPru av 3 was associated with more severe symptoms than sensitization to rPru av 1 . CONCLUSIONS: SPTs with RAs proved to be highly sensitive for diagnosis of cherry allergy. Component-resolved in vivo diagnosis with standardized amounts of stable RAs allows us to determine sensitization patterns directly, to correlate them with severity of clinical symptoms, and to analyze geographic differences.

↩

[34] - Scheurer S, Haase T, Schocker F, Cisteró-Bahíma A, Enrique E, Becker WM, et al. Prevalence of IgE-Sensitization and Cross-Reactivity of Pru av 3, the Lipid Transfer Protein From Cherry. AAAAI 58th Annual Meeting, New York, 1-6 March, 2002, Poster n°940

In Central and Northern Europe, food allergy to Rosaceae fruits is highly associated with birch pollinosis, owing to the existence of IgE cross-reactive homologous allergens in birch pollen and food. By contrast, in the Mediterranean area, severe allergic reactions to these fruits frequently are not related to birch pollen allergy, predominantly elicted by lipid transfer proteins (LTP). Recently, Pru av 3, the LTP from cherry has been identified as major allergen in Italian cherry-allergic subjects. The aim of this study was to evaluate the sensitization to Pru av 3 in a group of cherry allergic patients recruitated in Spain and to investigate the IgE-cross-reactivity of Pru av 3 with hazelnut LTP (Cor a 8) and with pollen extracts. All cherry-allergic patients (n=8) were allergic to plane tree pollen (Platanus acerifolia), and 4 out of all subjects investigated were allergic to grass pollen. The prevalence of IgE sensitization was investigated by EAST: Six out of 8 cherry-allergic patients were sensitized to rPru av 3, none was positive to the cherry profilin Pru av 4. Only one patient was sensitized to birch-related Pru av 1, identified as major allergen in the middle- and north European population. Two out of 8 cherry-allergic patients reported systemic reactions (angioedema) after ingestion of the corresponding food. IgE-Immunoblot and/or IgE-EAST (Enzyme Allergosorbent Test) inhibition experiments indicated a partial cross-reactivity between rPru av 3 and extracts from plane tree, ragweed, mugwort, and Parietaria pollen. No Inhibition of IgE-binding was found with the recombinant (r) hazelnut LTP Cor a 8, birch pollen and olive pollen extract as inhibitors. By contrast, IgE binding to recombinant hazelnut LTP was almost completely inhibited by rPru av 3. Preincubation of patient sera with cherry extract showed that natural and recombinant Pru av 3 shared identical IgE-epitopes. LTPs are potentially servere food allergens. The IgE cross-reactivity between food LTPs and pollen may indicate the presence of homologus pollen LTPs and their potential relevance in triggering food allergies by a primary sensitization by the inhalative route. Moreover, epitope similarities and differences of LTP-epitopes have to be taken into consideration in studies of cross-reactions in patients with multiple food allergies

↩

[35] - Fernández-Rivas M, Gonzalez-Mancebo E, Van Leeuwen A, Zuidmeer L, Rigby N, Ma Y, et al. Apple lipid transfer protein (Mal d 3) and profilin (Mal d 4) are the clinically relevant allergens in Spanish patients allergic to apple. Allergy Clin Immunol Int 2005;17(Suppl. 1):339

Research, Physical Biochemistry unit, Norwich, United Kingdom; 4University of Vienna, Pathophysiology department, Vienna, Austria Background. Four apple allergens have been identified so far: Mal d 1 ( Bet v 1 homologue), Mal d 2 (thaumatin-like protein), Mal d 3 (non-specific lipid transfer protein), and Mal d 4 (profilin). Mal d 1 is the major allergen in patients with apple allergy linked to birch pollinosis, and Mal d 3 is the major allergen in patients from Spain and Italy without birch pollinosis. Aim. To investigate the clinical relevance of the 4 apple allergens in allergic patients from Spain. Methods. Medical history, skin prick tests (SPTs) with fresh apple, and RAST to apple, nMal d 1, rMal d 2, nMal d 3, and rMal d 4, were carried out in patients reporting adverse reactions to apple. Clinical reactivity was confirmed by a double-blind placebo-controlled apple challenge. Results. Apple allergy was confirmed in 77 patients, 49 females, 28 males, mean age 23.3 years. Pollen allergy was present in 86%. Isolated oropharyngeal symptoms (OAS) were reported by 61% of the patients, whereas 39% had systemic symptoms (urticaria, anaphylaxis). All subjects presented positive SPTs to apple. Mean RAST values to apple, nMal d 1, rMal d 2, nMal d 3, and rMal d 4 were 5.17, 0.63, 0.78, 3.46, and 2.56 IU/ml, respectively. Positive (“0.8 IU/ ml) RAST results to nMal d 1, rMal d 2, nMal d 3, and rMal d 4 were found in 14.5%, 25%, 59.2% and 40.8% of patients, respectively. With the combination of Mal d 3 and 4, 86.8% of allergic patients were detected; with the further addition of Mal d 2, 89.5% of allergic subjects were identified. The inclusion of Mal d 1 did not increase this figure. A significant association was found between systemic symptoms induced by apple and sensitisation to Mal d 3, with an OR of 2.95 (1.13-7.67), and between OAS and sensitisation to rMal d 4, with an OR of 5.34 (1.94-14.62). Furthermore, 26 patients were monosensitised to nMal d 3, and 12 subjects to rMald 4. In this latter group isolated mild OAS was observed in 92% of patients, whereas it was only present in 31% of the subjects monosensitised to nMal d 3 (p<0.001). No associations were found between the sensitisation to Mal d 1 and 2 and the clinical presentation of apple allergy. Conclusions. Mal d 3 and Mal d 4 are the relevant allergens in Spanish patients allergic to apple. Mal d 3 is related to severe systemic reactions, whereas Mal d 4 is involved in mild local OAS. Mal d 1 and 2 do not seem to be clinically relevant apple allergens in Spanish patients.

↩

[36] - Fernández-Rivas M, Bolhaar S, González-Mancebo E, Asero R, van Leeuwen A, Bohle B, et al. Apple allergy across Europe: How allergen sensitization profiles determine the clinical expression of allergies to plant foods. J Allergy Clin Immunol 2006;118:481-488

BACKGROUND: Allergy to a plant food can either result from direct sensitization to that food or from primary sensitization to pollen, latex, or another food . OBJECTIVE: We sought to investigate the primary sensitizers in apple allergy across Europe, the individual allergens involved, and whether these differences determine the clinical presentation . METHODS: Patients (n = 389) with positive case histories and skin prick test responses to fresh apple were selected in the Netherlands, Austria, Italy, and Spain. Skin prick tests and RASTs to a panel of pollens and plant foods were performed, as well as RASTs to Bet v 1 and the apple allergens Mal d 1, 2, 3, and 4 . RESULTS: In the Netherlands, Austria, and Italy apple allergy is mild (>90% isolated oral symptoms) and related to birch pollinosis and sensitization to Bet v 1 and its apple homologue, Mal d 1, which has an odds ratio of local reactions of 2.85 (95% CI, 1.47-5.55). In Spain apple allergy is severe (>35% systemic reactions) and related to peach allergy and sensitization to Mal d 3 (nonspecific lipid transfer protein), which has an odds ratio of systemic reactions of 7.76 (95% CI, 3.87-15.56) . CONCLUSION: The analysis of individual apple allergens in a clinical context has provided insight into the sensitization pathway and into the intrinsic risk an allergen bears to induce mild or severe food allergy. CLINICAL IMPLICATIONS: Information on the sensitization pathway is essential to develop preventive strategies in food allergy. The application of individual food allergens with a known intrinsic risk will improve the prognostic value of diagnostic tests.

↩

[37] - Gonzalez-Mancebo E, Fernandez-Rivas M. Outcome and safety of double-blind, placebocontrolled food challenges in 111 patients sensitized to lipid transfer proteins. J Allergy Clin Immunol 2008;121:1507-1508

↩

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

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

↩

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

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

↩

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

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

↩

[41] - Rodriguez-Perez R, Fernández-Rivas M, González-Mancebo E, Sánchez-Monge R, Diaz-Perales A, Salcedo G. Peach profilin: cloning, heterologous expression and cross-reactivity with Bet v 2. Allergy 2003;58:635-640

BACKGROUND: Peach is among the main foods causing allergic reactions in the Mediterranean adult population. Only a single peach allergen, named Pru p 3, has been characterized. However, a potential role of profilin has also been suggested in grass pollen-associated allergy to peach . METHODS: Complementary DNA clones for two different peach profilin isoforms were obtained by reverse transcriptase polymerase chain reaction using non-degenerated primers. Expression of recombinant peach profilin was performed in Escherichia coli, and confirmed using rabbit polyclonal antibodies to sunflower pollen profilin. Twenty-nine individual sera from patients with peach allergy proved by double-blind, placebo-controlled food challenges (DBPCFC), either with (n = 15) or without (n = 14) specific IgE to Bet v 2, were used in immunodetection assays to test recombinant peach profilin reactivity . RESULTS: Each peach profilin cDNA included an open reading frame coding for a 131 amino acid protein. The peach profilin isoforms, designated Pru p 4.01 and Pru p 4.02, showed 80% of amino acid sequence identity, and were very similar (>70% identity) to allergenic profilins from plant foods and pollens. Recombinant Pru p 4.01 was expressed in E. coli as a nonfusion protein, displaying the expected molecular size and reacting with anti-profilin antibodies. rPru p 4.01 was recognized by all sera (15 of 15) with specific IgE to Bet v 2, whereas no sera (zero of 14) without IgE to this birch allergen reacted with rPru p 4.01 . CONCLUSIONS: Peach profilin Pru p 4 is very closed to other allergenic profilins from plant foods and pollens. A complete correlation between reactivity to rPru p 4 and rBet v 2 has been found in sera from peach allergic patients.

↩

[42] - Fernández-Rivas M, González-Mancebo E, Rodríguez-Pérez R, Benito C, Sánchez-Monge R, Salcedo G, et al. Clinically relevant peach allergy is related to peach lipid transfer protein, Pru p 3, in the Spanish population. J Allergy Clin Immunol 2003;112:789-795

BACKGROUND: Sensitization to peach and related Rosaceae fruits without clinical expression is commonly observed as the result of the extensive cross-reactivity of IgE antibodies directed toward lipid transfer proteins (LTPs), Bet v 1 homologues, profilins, and carbohydrate determinants . OBJECTIVE: We aimed to study whether there are any clinical or immunologic differences between patients allergic to peach and those who have a current clinically irrelevant sensitization to this fruit . METHODS: One hundred subjects with adverse reactions to peach were evaluated by medical history, skin prick tests with fresh peach and purified peach LTP (Pru p 3), and specific IgE determinations to peach, rBet v 1, and rBet v 2 (birch profilin). Clinical reactivity to peach was established by double-blind, placebo-controlled food challenges. The clinical characteristics and the in vivo and in vitro tests were compared between allergic and nonallergic patients . RESULTS: Peach allergy was confirmed in 76 patients and ruled out in 16; 2 patients dropped out, and the study was not conclusive in 6 individuals (placebo reactors). Pollen allergy was found in 76% of the allergic patients and in 100% of the nonallergic patients. Positive responses to Pru p 3, rBet v 1, and rBet v 2 were observed in 62%, 7%, and 34% of patients allergic to peach, respectively. The sensitization rate to Pru p 3 was significantly higher among subjects allergic than nonallergic to peach (62% vs 31%, P =.02). IgE responses to rBet v 2 were more frequent among subjects allergic to pollen, but no difference was observed in the presence or absence of peach allergy . CONCLUSIONS: Pru p 3 is the major allergen of peach in our population, and the IgE response to this allergen is related to the clinical expression of peach allergy. Sensitization to profilin is observed in those patients with an associated pollen allergy but does not appear to be related to the clinical reactivity to peach.

↩

[43] - 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.

↩

[44] - Fernández Rivas M. Food Allergy in Alergológica-2005. J Investig Allergol Clin Immunol 2009;19(Suppl. 2):37-44

BACKGROUND: Alergológica-2005 is a cross-sectional study carried out in Spain in 4991 new patients prospectively recruited in allergy clinics . OBJECTIVE: To describe the characteristics of food allergic patients included in Alergológica-2005 . RESULTS: Food allergy was diagnosed in 369 subjects (7.4%, 95% CI, 6.7%-8.1%). The most common offending foods were fruits (33.3% of cases), nuts (26%), shellfish (22%), egg (16%), milk (13.9%) and fish (9.8%). Rosaceae fruits and crustaceans elicited 23.6% and 18.7% of the reactions, respectively. Milk and egg were the most common foods in patients < 5 years of age, whereas fruits and nuts were the most prevalent foods in patients over 5 years. The most frequent manifestations included skin reactions (65.3%), oral allergy syndrome (33.6%), digestive symptoms (24.7%) and anaphylaxis (17.9%). The clinical presentation differed among foods. Prick-tests were carried out more frequently than serum immunoglobulin E (IgE) determinations (95.9% vs. 65%). Oral challenges were performed in 13% of patients, and 72.3% of these were open. Food allergy was diagnosed on the basis of medical history and positive IgE in 60.2% of the patients, ranging from 13.7% for milk to 75% for cereals. The self perception of patients' quality of life was lower than that of the 75% of Spanish reference population . CONCLUSION: Food allergy is diagnosed in 7.4% of the patients seen in allergy clinics across Spain, and has an important impact on the quality of life of patients. The foods involved in reactions change with age. The clinical presentation changes with the food, although the skin is the most frequently affected organ.

↩

[45] - Fernández-Rivas M. [Cross-reactivity between fruit and vegetables]. Allergol Immunopathol (Madr) 2003;31:141-146

Vegetable foods are the most frequent cause of food allergy after the age of 5 years. The most commonly implicated foods are fruit and dried fruits, followed in Spain by legumes and fresh garden produce. In patients allergic to fruit and garden produce, multiple sensitizations to other vegetable products, whether from the same family or taxonomically unrelated, are frequent, although they do not always share the same clinical expression. Furthermore, more than 75 % of these patients are allergic to pollen, the type of pollen varying in relation to the aerobiology of the area. The basis of these associations among vegetable foods and with pollens lies in the existence of IgE antibodies against "panallergens", which determines cross-reactivity. Panallergens are proteins that are spread throughout the vegetable kingdom and are implicated in important biological functions (generally defense) and consequently their sequences and structures are highly conserved. The three best-known groups are allergens homologous to Bet v 1, profilins, and lipid transfer proteins (LTP).Allergens homologous to Bet v 1 (major birch pollen allergen) constitute a group of defense proteins (PR-10), with a molecular weight of 17 kDa, which behave as major allergens in patients from northern and central Europe with allergy to vegetables associated with birch pollen allergy. In these patients, the primary sensitization seems to be produced through the inhalation route on exposure to birch pollen. The symptomatology characteristically associated with sensitization to this family of allergens is oral allergy syndrome (OAS).Profilins are highly conserved proteins in all eukaryotic organisms and are present in pollen and a wide variety of vegetable foods. They have a molecular weight of 14 kDa and present a high degree of structural homology as well as marked cross-reactivity among one another. The presence of anti-profilin IgE broadens the spectrum of sensitizations to vegetable foods detected through skin tests and/or in vitro tests but whether it correlates with the clinical expression of food allergy is unclear.LTPs are the most commonly implicated allergens in allergy to Rosaceae fruits in patients from the Mediterranean area without birch pollen sensitization. LTPs are a family of 9kDA polypeptides, widely found in the vegetable kingdom and implicated in cuticle formation and defense against pathogens (PR-14). They are thermostable and resistant to pepsin digestion, which makes them potent food allergens and explains the frequent development of systemic symptoms (urticaria, anaphylaxis) in patients allergic to Rosaceae fruits in Spain. LTPs have also been identified in other vegetable foods and in pollens and a marked degree of cross-reactivity among them has been demonstrated, which may explain (together with profilin) the frequency of individuals sensitized to vegetable foods in the Mediterranean area.

↩

[46] - Asero R, Antonicelli L, Arena A, Bommarito L, Caruso B, Colombo G et al. Causes of Food-Induced Anaphylaxis in Italian Adults: A Multi-Centre Study. Int Arch Allergy Immunol 2009;150:271-277

↩

[47] - Fernández-Rivas M, Bolhaar S, González-Mancebo E, Asero R, van Leeuwen A, Bohle B, et al. Apple allergy across Europe: How allergen sensitization profiles determine the clinical expression of allergies to plant foods. J Allergy Clin Immunol 2006;118:481-488

BACKGROUND: Allergy to a plant food can either result from direct sensitization to that food or from primary sensitization to pollen, latex, or another food . OBJECTIVE: We sought to investigate the primary sensitizers in apple allergy across Europe, the individual allergens involved, and whether these differences determine the clinical presentation . METHODS: Patients (n = 389) with positive case histories and skin prick test responses to fresh apple were selected in the Netherlands, Austria, Italy, and Spain. Skin prick tests and RASTs to a panel of pollens and plant foods were performed, as well as RASTs to Bet v 1 and the apple allergens Mal d 1, 2, 3, and 4 . RESULTS: In the Netherlands, Austria, and Italy apple allergy is mild (>90% isolated oral symptoms) and related to birch pollinosis and sensitization to Bet v 1 and its apple homologue, Mal d 1, which has an odds ratio of local reactions of 2.85 (95% CI, 1.47-5.55). In Spain apple allergy is severe (>35% systemic reactions) and related to peach allergy and sensitization to Mal d 3 (nonspecific lipid transfer protein), which has an odds ratio of systemic reactions of 7.76 (95% CI, 3.87-15.56) . CONCLUSION: The analysis of individual apple allergens in a clinical context has provided insight into the sensitization pathway and into the intrinsic risk an allergen bears to induce mild or severe food allergy. CLINICAL IMPLICATIONS: Information on the sensitization pathway is essential to develop preventive strategies in food allergy. The application of individual food allergens with a known intrinsic risk will improve the prognostic value of diagnostic tests.

↩

[48] - Fernández-Rivas M, Bolhaar S, González-Mancebo E, Asero R, van Leeuwen A, Bohle B, et al. Apple allergy across Europe: How allergen sensitization profiles determine the clinical expression of allergies to plant foods. J Allergy Clin Immunol 2006;118:481-488

BACKGROUND: Allergy to a plant food can either result from direct sensitization to that food or from primary sensitization to pollen, latex, or another food . OBJECTIVE: We sought to investigate the primary sensitizers in apple allergy across Europe, the individual allergens involved, and whether these differences determine the clinical presentation . METHODS: Patients (n = 389) with positive case histories and skin prick test responses to fresh apple were selected in the Netherlands, Austria, Italy, and Spain. Skin prick tests and RASTs to a panel of pollens and plant foods were performed, as well as RASTs to Bet v 1 and the apple allergens Mal d 1, 2, 3, and 4 . RESULTS: In the Netherlands, Austria, and Italy apple allergy is mild (>90% isolated oral symptoms) and related to birch pollinosis and sensitization to Bet v 1 and its apple homologue, Mal d 1, which has an odds ratio of local reactions of 2.85 (95% CI, 1.47-5.55). In Spain apple allergy is severe (>35% systemic reactions) and related to peach allergy and sensitization to Mal d 3 (nonspecific lipid transfer protein), which has an odds ratio of systemic reactions of 7.76 (95% CI, 3.87-15.56) . CONCLUSION: The analysis of individual apple allergens in a clinical context has provided insight into the sensitization pathway and into the intrinsic risk an allergen bears to induce mild or severe food allergy. CLINICAL IMPLICATIONS: Information on the sensitization pathway is essential to develop preventive strategies in food allergy. The application of individual food allergens with a known intrinsic risk will improve the prognostic value of diagnostic tests.

↩

[49] - Cheynel A, Malandain H, Leduc V. Un cas insolite d’allergie croisée. Rev Fr Allergol Immunol Clin 2007;47:284-285

↩

[50] - 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) .

↩

[51] - Fernández-Rivas M, Bolhaar S, González-Mancebo E, Asero R, van Leeuwen A, Bohle B, et al. Apple allergy across Europe: How allergen sensitization profiles determine the clinical expression of allergies to plant foods. J Allergy Clin Immunol 2006;118:481-488

BACKGROUND: Allergy to a plant food can either result from direct sensitization to that food or from primary sensitization to pollen, latex, or another food . OBJECTIVE: We sought to investigate the primary sensitizers in apple allergy across Europe, the individual allergens involved, and whether these differences determine the clinical presentation . METHODS: Patients (n = 389) with positive case histories and skin prick test responses to fresh apple were selected in the Netherlands, Austria, Italy, and Spain. Skin prick tests and RASTs to a panel of pollens and plant foods were performed, as well as RASTs to Bet v 1 and the apple allergens Mal d 1, 2, 3, and 4 . RESULTS: In the Netherlands, Austria, and Italy apple allergy is mild (>90% isolated oral symptoms) and related to birch pollinosis and sensitization to Bet v 1 and its apple homologue, Mal d 1, which has an odds ratio of local reactions of 2.85 (95% CI, 1.47-5.55). In Spain apple allergy is severe (>35% systemic reactions) and related to peach allergy and sensitization to Mal d 3 (nonspecific lipid transfer protein), which has an odds ratio of systemic reactions of 7.76 (95% CI, 3.87-15.56) . CONCLUSION: The analysis of individual apple allergens in a clinical context has provided insight into the sensitization pathway and into the intrinsic risk an allergen bears to induce mild or severe food allergy. CLINICAL IMPLICATIONS: Information on the sensitization pathway is essential to develop preventive strategies in food allergy. The application of individual food allergens with a known intrinsic risk will improve the prognostic value of diagnostic tests.

↩

[52] - 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.

↩

[53] - Delmas A, Batard T, Pradalier A. Allergie à la prune. Rev Fr Allergol Immunol Clin 2006;46:374-375

↩

[54] - Cheynel A, Malandain H, Leduc V. Un cas insolite d’allergie croisée. Rev Fr Allergol Immunol Clin 2007;47:284-285

↩

[55] - 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) .

↩

[56] - 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.

↩

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

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

↩

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

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

↩

[59] - Fernández-Rivas M. Alergia a frutas y hortalizas. in "Alergia a alimentos", Sociedad Española de Alergología e Inmunología Clínica (SEAIC) Ed., Euromedice, Badalona 2004, pp.143-67

↩

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

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

↩

[61] - Gaier S, Oberhuber C, Hemmer W, Radauer C, Rigby NM, Marsh JY et al. Pru p 3 as a marker for symptom severity for patients with peach allergy in a birch pollen environment. J Allergy Clin Immunol 2009;124:166-167

↩

[62] - Rodriguez-Perez R, Fernández-Rivas M, González-Mancebo E, Sánchez-Monge R, Diaz-Perales A, Salcedo G. Peach profilin: cloning, heterologous expression and cross-reactivity with Bet v 2. Allergy 2003;58:635-640

BACKGROUND: Peach is among the main foods causing allergic reactions in the Mediterranean adult population. Only a single peach allergen, named Pru p 3, has been characterized. However, a potential role of profilin has also been suggested in grass pollen-associated allergy to peach . METHODS: Complementary DNA clones for two different peach profilin isoforms were obtained by reverse transcriptase polymerase chain reaction using non-degenerated primers. Expression of recombinant peach profilin was performed in Escherichia coli, and confirmed using rabbit polyclonal antibodies to sunflower pollen profilin. Twenty-nine individual sera from patients with peach allergy proved by double-blind, placebo-controlled food challenges (DBPCFC), either with (n = 15) or without (n = 14) specific IgE to Bet v 2, were used in immunodetection assays to test recombinant peach profilin reactivity . RESULTS: Each peach profilin cDNA included an open reading frame coding for a 131 amino acid protein. The peach profilin isoforms, designated Pru p 4.01 and Pru p 4.02, showed 80% of amino acid sequence identity, and were very similar (>70% identity) to allergenic profilins from plant foods and pollens. Recombinant Pru p 4.01 was expressed in E. coli as a nonfusion protein, displaying the expected molecular size and reacting with anti-profilin antibodies. rPru p 4.01 was recognized by all sera (15 of 15) with specific IgE to Bet v 2, whereas no sera (zero of 14) without IgE to this birch allergen reacted with rPru p 4.01 . CONCLUSIONS: Peach profilin Pru p 4 is very closed to other allergenic profilins from plant foods and pollens. A complete correlation between reactivity to rPru p 4 and rBet v 2 has been found in sera from peach allergic patients.

↩

[63] - Fernández-Rivas M, Barreales L, Gonzalez-Mancebo E, Zuidmeer L, Mills C, Hofmann-Sommergruber K, et al. Specific IgE Responses To Lipid Transfer Protein (LTP) And Profilin Have Interactive Effects On The Outcome Of Doubleblind Placebo-controlled Food Challenges (DBPCFC) With Apple In Allergic Patients From Spain. J Allergy Clin Immunol 2008;121:S240

RATIONALE: We aimed to study the relationship of the IgE responses to the apple allergens Mal d 1-2-3-4, and the outcome of DBPCFC with apple. METHODS: In 92 Spanish patients who reported immediate reactions to apple and had positive prick-prick tests with fresh apple, the IgE responses to Mal d 1-2-3-4 were analysed by RAST, and reactivity to apple was assessed by DBPCFC. The relationship between RAST to apple allergens and outcome of DBPCFC was analysed by logistic regression. RESULTS: IgE responses to Mal d 3 (LTP) and Mal d 4 (profilin) were significantly associated with DBPCFC outcome, although an interaction was found. The reference category for risk estimation was the presence of negative (<0.30 IU/ml) RASTs to Mal d 3 and 4 that carried over a probability of a positive DBPCFC of 25%. With a negative Mal d 3 and a positive Mal d 4 the probability of a positive DBPCFC was 85.7% (risk ratio [RR] 3.43, 95% confidence interval [CI]: 1.01-11.60; p = 0.008). With a positive Mal d 3 and a negative Mal d 4 the probability of a positive DBPCFC was 90.3% (RR 3.61, 95% CI: 1.08-12.06; p = 0.001). With positive Mal d 3 and 4 the probability of a positive DBPCFC was 91.7% (RR 3.25, 95% CI: 1.13-9.39; p = 0.0003). CONCLUSIONS: Apple LTP and profilin are clinically relevant allergens in Spanish patients, and they have an interactive effect on the clinical reactivity to apple since the presence of specific IgE to them separately or in combination carries over a similar probability of positive DBPCFC.

↩

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

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

↩

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

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

↩

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

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

↩

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

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

↩

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

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

↩

[69] - Fernández-Rivas M, Bolhaar S, González-Mancebo E, Asero R, van Leeuwen A, Bohle B, et al. Apple allergy across Europe: How allergen sensitization profiles determine the clinical expression of allergies to plant foods. J Allergy Clin Immunol 2006;118:481-488

BACKGROUND: Allergy to a plant food can either result from direct sensitization to that food or from primary sensitization to pollen, latex, or another food . OBJECTIVE: We sought to investigate the primary sensitizers in apple allergy across Europe, the individual allergens involved, and whether these differences determine the clinical presentation . METHODS: Patients (n = 389) with positive case histories and skin prick test responses to fresh apple were selected in the Netherlands, Austria, Italy, and Spain. Skin prick tests and RASTs to a panel of pollens and plant foods were performed, as well as RASTs to Bet v 1 and the apple allergens Mal d 1, 2, 3, and 4 . RESULTS: In the Netherlands, Austria, and Italy apple allergy is mild (>90% isolated oral symptoms) and related to birch pollinosis and sensitization to Bet v 1 and its apple homologue, Mal d 1, which has an odds ratio of local reactions of 2.85 (95% CI, 1.47-5.55). In Spain apple allergy is severe (>35% systemic reactions) and related to peach allergy and sensitization to Mal d 3 (nonspecific lipid transfer protein), which has an odds ratio of systemic reactions of 7.76 (95% CI, 3.87-15.56) . CONCLUSION: The analysis of individual apple allergens in a clinical context has provided insight into the sensitization pathway and into the intrinsic risk an allergen bears to induce mild or severe food allergy. CLINICAL IMPLICATIONS: Information on the sensitization pathway is essential to develop preventive strategies in food allergy. The application of individual food allergens with a known intrinsic risk will improve the prognostic value of diagnostic tests.

↩

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

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

↩

[71] - 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.

↩

[72] - Ballmer-Weber BK, Scheurer S, Fritsche P, Enrique E, Cistero-Bahima A, Haase T, et al. Component-resolved diagnosis with recombinant allergens in patients with cherry allergy. J Allergy Clin Immunol 2002;110:167-173

BACKGROUND: In pollen-related food allergy, extracts for skin prick tests (SPTs) are often not standardized, and the test reliability is affected by false-negative reactions . OBJECTIVE: We sought to evaluate a panel of recombinant allergens (RAs) derived from one allergenic food for use in component-resolved in vivo diagnosis, taking cherry as a model food . METHODS: Seventy-nine subjects were included in the study: 24 Swiss patients (group 1) with a positive double-blind placebo-controlled food challenge result to cherries, 23 patients with birch pollen allergy but without cherry allergy (group 2), 23 nonatopic subjects (group 3), and 9 Spanish patients with a history of a cherry allergy (group 4). SPTs were performed in duplicate by using recombinant cherry allergens (Bet v 1-related allergen: recombinant (r) Pru av 1; profilin: rPru av 4; and lipid transfer protein: rPru av 3) in concentrations of 10, 50, and 100 microg/mL. Furthermore, IgE reactivity to rPru av 1, rPru av 4, and rPru av 3 was assessed by means of immunoblot analysis . RESULTS: SPT responses with rPru av 1, rPru av 4, and rPru av 3 were positive in 92%, 17%, and 4% of the patients in group 1; in 74%, 30%, and 0% of the patients in group 2; in 0%, 22%, and 89% of the patients in group 4; and negative for all nonatopic subjects (group 3). Thus the sensitivity of a positive SPT response to at least one of the 3 RAs was 96%. The specificities, negative predictive values, and positive predictive values with the 3 RAs were 100%, 96%, and 100% if calculated in relation to the nonatopic control group but 17%, 79%, and 60% when calculated in relation to the control group with birch pollen allergy. The correlation between SPT and immunoblotting results was excellent. Sensitization to rPru av 3 was associated with more severe symptoms than sensitization to rPru av 1 . CONCLUSIONS: SPTs with RAs proved to be highly sensitive for diagnosis of cherry allergy. Component-resolved in vivo diagnosis with standardized amounts of stable RAs allows us to determine sensitization patterns directly, to correlate them with severity of clinical symptoms, and to analyze geographic differences.

↩

[73] - Borges JP, Barre A, Culerrier R, Granier C, Didier A, Rougé P. Lipid transfer proteins from Rosaceae fruits share consensus epitopes responsible for their IgE-binding cross-reactivity. Biochem Biophys Res Commun 2008;365:685-690

Four IgE-binding epitopes have been characterized that cover a large area (40%) of the molecular surface of lipid transfer protein allergens of Rosaceae (apple, peach, apricot, and plum). They mainly correspond to electropositively charged regions protruding on the molecular surface of the modeled apple (Mal d 3), apricot (Pru ar 3), and plum (Pru d 3) allergens. Two of these epitopes consist of consensus epitopes structurally conserved among the lipid transfer protein allergens from the Rosaceae. Their occurrence in different lipid transfer protein allergens presumably accounts for the IgE-binding cross-reactivity often observed among different Rosaceae fruits. In this respect, LTP consist of phylogenetically- and structurally-related pan allergens. However, the IgE-binding cross-reactivity due to fruit lipid transfer protein has varying degrees of clinical relevance and this cross-reactivity is not necessarily accompanied by a cross-allergenicity to the corresponding fruits.

↩

[74] - 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.

↩

[75] - García-Sellés FJ, Díaz-Perales A, Sánchez-Monge R, Alcántara M, Lombardero M, Barber D, et al. Patterns of reactivity to lipid transfer proteins of plant foods and Artemisia pollen: an in vivo study. Int Arch Allergy Immunol 2002;128:115-122

Background: Lipid transfer proteins (LTPs) are major allergens of Rosaceae fruits in the Mediterranean area. IgE-cross-reactivity has been demonstrated in vitro among LTPs from peach, apple, chestnut and Artemisia pollen. The aim of this study was to evaluate the reactivity to LTPs from peach, apple, chestnut and Artemisia pollen by means of skin prick tests (SPTs). Methods: Forty-seven patients allergic to peach (peach group), 20 patients sensitized to Artemisia pollen with no food allergies (Artemisia group), and 12 control subjects were skin tested with fresh peach, as well as with whole extracts and purified LTPs of peach, apple, chestnut and Artemisia pollen. Results: The rates of positive SPTs for peach, apple, chestnut and Artemisia LTPs were, respectively, 91, 77, 23, and 36% in the peach group, and 30, 5, 15 and 40% in the Artemisia group. No response was observed in the control subjects. SPTs with peach LTP strongly correlated with SPTs conducted with fresh peach. In the peach group, the most frequent pattern of reactivity to LTPs was the combination peach-apple (45%), followed by peach-apple-Artemisia-chestnut (21%). Significant correlations were found between peach and apple LTPs, and between Artemisia and chestnut LTPs. Positive SPTs to chestnut LTP were only observed in patients with positive SPTs to Artemisia LTP. All the patients with positive case histories to chestnut reacted to chestnut LTP. Conclusions: LTPs are plant panallergens with different patterns of cross-reactivity. They are major allergens of Rosaceae fruits and seem to be involved in allergic reactions to unrelated foodstuffs such as chestnut, probably through sensitization to the cross-reactive Artemisia LTP. Rosaceae LTPs could be useful tools for in vivo diagnosis of Rosaceae fruit allergy.

↩

[76] - 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.

↩

[77] - Asero R, Bolhaar STH, Gonzalez-Mancebo E, Fernandez-Rivas M, Knulst AC, Hoffmann-Sommergruber K, et al. Are the different prevalences of sensitization to lipid transfer protein throughout Europe related to food consumption habits ? EAACI 23th Congress, Amsterdam, 12-16 June, 2004, Poster n°1003

Background Allergy to lipid transfer protein (LTP) is frequently observed in the Mediterranean countries but is very rare in Northern and Central Europe. Objective This study aimed to assess whether such discrepancy may be explained by differences in consumption habits of both fresh and processed peach and apple, the two foods most frequently implicated in allergy to LTP, in various European countries. Methods Consumption data of fresh peach and apple by a small sample in 4 European countries (Austria, Italy, Netherlands, and Spain) were collected in a fruit consumption study, and consumption data by the general population of 3 countries (Austria, Italy, and Spain) were obtained from official national agencies; moreover, the availability of infant foods containing these two fruits in supermarkets and general stores of the 4 countries was directly checked by the participants of this study. All information obtained were plotted against the prevalence of allergy to LTP recorded in about 400 apple-allergic patients seen at 4 clinical centers in Austria, Italy, Netherlands, and Spain (about 100 patients per center) within the EU-funded project SAFE. LTP hypersensitivity was ascertained by RAST using natural apple LTP as allergen. Results As expected, RAST showed that allergy to LTP was frequent in Spain and quite common in Italy, but virtually absent In Austria and in the Netherlands. Some differences in consumption per person of peach and apple and in the amount of infant foods containing peach or apple were observed between the 4 countries, but these differences could not explain the different patterns of LTP allergy. Conclusion The higher prevalence of hypersensitivity to LTP in Southern Europe does not seem related to differences in consumption of fresh and/or processed peach and apple; possibly other environmental factors play a role in sensitization to this protein.

↩

[78] - 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.

↩

[79] - 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.

↩

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

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

↩

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

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

↩

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

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

↩

[83] - 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.

↩

[84] - 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.

↩

[85] - Scheurer S, Haase T, Schocker F, Cisteró-Bahíma A, Enrique E, Becker WM, et al. Prevalence of IgE-Sensitization and Cross-Reactivity of Pru av 3, the Lipid Transfer Protein From Cherry. AAAAI 58th Annual Meeting, New York, 1-6 March, 2002, Poster n°940

In Central and Northern Europe, food allergy to Rosaceae fruits is highly associated with birch pollinosis, owing to the existence of IgE cross-reactive homologous allergens in birch pollen and food. By contrast, in the Mediterranean area, severe allergic reactions to these fruits frequently are not related to birch pollen allergy, predominantly elicted by lipid transfer proteins (LTP). Recently, Pru av 3, the LTP from cherry has been identified as major allergen in Italian cherry-allergic subjects. The aim of this study was to evaluate the sensitization to Pru av 3 in a group of cherry allergic patients recruitated in Spain and to investigate the IgE-cross-reactivity of Pru av 3 with hazelnut LTP (Cor a 8) and with pollen extracts. All cherry-allergic patients (n=8) were allergic to plane tree pollen (Platanus acerifolia), and 4 out of all subjects investigated were allergic to grass pollen. The prevalence of IgE sensitization was investigated by EAST: Six out of 8 cherry-allergic patients were sensitized to rPru av 3, none was positive to the cherry profilin Pru av 4. Only one patient was sensitized to birch-related Pru av 1, identified as major allergen in the middle- and north European population. Two out of 8 cherry-allergic patients reported systemic reactions (angioedema) after ingestion of the corresponding food. IgE-Immunoblot and/or IgE-EAST (Enzyme Allergosorbent Test) inhibition experiments indicated a partial cross-reactivity between rPru av 3 and extracts from plane tree, ragweed, mugwort, and Parietaria pollen. No Inhibition of IgE-binding was found with the recombinant (r) hazelnut LTP Cor a 8, birch pollen and olive pollen extract as inhibitors. By contrast, IgE binding to recombinant hazelnut LTP was almost completely inhibited by rPru av 3. Preincubation of patient sera with cherry extract showed that natural and recombinant Pru av 3 shared identical IgE-epitopes. LTPs are potentially servere food allergens. The IgE cross-reactivity between food LTPs and pollen may indicate the presence of homologus pollen LTPs and their potential relevance in triggering food allergies by a primary sensitization by the inhalative route. Moreover, epitope similarities and differences of LTP-epitopes have to be taken into consideration in studies of cross-reactions in patients with multiple food allergies

↩

[86] - Rodriguez-Perez R, Fernández-Rivas M, González-Mancebo E, Sánchez-Monge R, Diaz-Perales A, Salcedo G. Peach profilin: cloning, heterologous expression and cross-reactivity with Bet v 2. Allergy 2003;58:635-640

BACKGROUND: Peach is among the main foods causing allergic reactions in the Mediterranean adult population. Only a single peach allergen, named Pru p 3, has been characterized. However, a potential role of profilin has also been suggested in grass pollen-associated allergy to peach . METHODS: Complementary DNA clones for two different peach profilin isoforms were obtained by reverse transcriptase polymerase chain reaction using non-degenerated primers. Expression of recombinant peach profilin was performed in Escherichia coli, and confirmed using rabbit polyclonal antibodies to sunflower pollen profilin. Twenty-nine individual sera from patients with peach allergy proved by double-blind, placebo-controlled food challenges (DBPCFC), either with (n = 15) or without (n = 14) specific IgE to Bet v 2, were used in immunodetection assays to test recombinant peach profilin reactivity . RESULTS: Each peach profilin cDNA included an open reading frame coding for a 131 amino acid protein. The peach profilin isoforms, designated Pru p 4.01 and Pru p 4.02, showed 80% of amino acid sequence identity, and were very similar (>70% identity) to allergenic profilins from plant foods and pollens. Recombinant Pru p 4.01 was expressed in E. coli as a nonfusion protein, displaying the expected molecular size and reacting with anti-profilin antibodies. rPru p 4.01 was recognized by all sera (15 of 15) with specific IgE to Bet v 2, whereas no sera (zero of 14) without IgE to this birch allergen reacted with rPru p 4.01 . CONCLUSIONS: Peach profilin Pru p 4 is very closed to other allergenic profilins from plant foods and pollens. A complete correlation between reactivity to rPru p 4 and rBet v 2 has been found in sera from peach allergic patients.

↩

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

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

↩

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

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

↩

[89] - Fernández-Rivas M, González-Mancebo E, Rodríguez-Pérez R, Benito C, Sánchez-Monge R, Salcedo G, et al. Clinically relevant peach allergy is related to peach lipid transfer protein, Pru p 3, in the Spanish population. J Allergy Clin Immunol 2003;112:789-795

BACKGROUND: Sensitization to peach and related Rosaceae fruits without clinical expression is commonly observed as the result of the extensive cross-reactivity of IgE antibodies directed toward lipid transfer proteins (LTPs), Bet v 1 homologues, profilins, and carbohydrate determinants . OBJECTIVE: We aimed to study whether there are any clinical or immunologic differences between patients allergic to peach and those who have a current clinically irrelevant sensitization to this fruit . METHODS: One hundred subjects with adverse reactions to peach were evaluated by medical history, skin prick tests with fresh peach and purified peach LTP (Pru p 3), and specific IgE determinations to peach, rBet v 1, and rBet v 2 (birch profilin). Clinical reactivity to peach was established by double-blind, placebo-controlled food challenges. The clinical characteristics and the in vivo and in vitro tests were compared between allergic and nonallergic patients . RESULTS: Peach allergy was confirmed in 76 patients and ruled out in 16; 2 patients dropped out, and the study was not conclusive in 6 individuals (placebo reactors). Pollen allergy was found in 76% of the allergic patients and in 100% of the nonallergic patients. Positive responses to Pru p 3, rBet v 1, and rBet v 2 were observed in 62%, 7%, and 34% of patients allergic to peach, respectively. The sensitization rate to Pru p 3 was significantly higher among subjects allergic than nonallergic to peach (62% vs 31%, P =.02). IgE responses to rBet v 2 were more frequent among subjects allergic to pollen, but no difference was observed in the presence or absence of peach allergy . CONCLUSIONS: Pru p 3 is the major allergen of peach in our population, and the IgE response to this allergen is related to the clinical expression of peach allergy. Sensitization to profilin is observed in those patients with an associated pollen allergy but does not appear to be related to the clinical reactivity to peach.

↩

[90] - Fernández-Rivas M, Gonzalez-Mancebo E, Van Leeuwen A, Zuidmeer L, Rigby N, Ma Y, et al. Apple lipid transfer protein (Mal d 3) and profilin (Mal d 4) are the clinically relevant allergens in Spanish patients allergic to apple. Allergy Clin Immunol Int 2005;17(Suppl. 1):339

Research, Physical Biochemistry unit, Norwich, United Kingdom; 4University of Vienna, Pathophysiology department, Vienna, Austria Background. Four apple allergens have been identified so far: Mal d 1 ( Bet v 1 homologue), Mal d 2 (thaumatin-like protein), Mal d 3 (non-specific lipid transfer protein), and Mal d 4 (profilin). Mal d 1 is the major allergen in patients with apple allergy linked to birch pollinosis, and Mal d 3 is the major allergen in patients from Spain and Italy without birch pollinosis. Aim. To investigate the clinical relevance of the 4 apple allergens in allergic patients from Spain. Methods. Medical history, skin prick tests (SPTs) with fresh apple, and RAST to apple, nMal d 1, rMal d 2, nMal d 3, and rMal d 4, were carried out in patients reporting adverse reactions to apple. Clinical reactivity was confirmed by a double-blind placebo-controlled apple challenge. Results. Apple allergy was confirmed in 77 patients, 49 females, 28 males, mean age 23.3 years. Pollen allergy was present in 86%. Isolated oropharyngeal symptoms (OAS) were reported by 61% of the patients, whereas 39% had systemic symptoms (urticaria, anaphylaxis). All subjects presented positive SPTs to apple. Mean RAST values to apple, nMal d 1, rMal d 2, nMal d 3, and rMal d 4 were 5.17, 0.63, 0.78, 3.46, and 2.56 IU/ml, respectively. Positive (“0.8 IU/ ml) RAST results to nMal d 1, rMal d 2, nMal d 3, and rMal d 4 were found in 14.5%, 25%, 59.2% and 40.8% of patients, respectively. With the combination of Mal d 3 and 4, 86.8% of allergic patients were detected; with the further addition of Mal d 2, 89.5% of allergic subjects were identified. The inclusion of Mal d 1 did not increase this figure. A significant association was found between systemic symptoms induced by apple and sensitisation to Mal d 3, with an OR of 2.95 (1.13-7.67), and between OAS and sensitisation to rMal d 4, with an OR of 5.34 (1.94-14.62). Furthermore, 26 patients were monosensitised to nMal d 3, and 12 subjects to rMald 4. In this latter group isolated mild OAS was observed in 92% of patients, whereas it was only present in 31% of the subjects monosensitised to nMal d 3 (p<0.001). No associations were found between the sensitisation to Mal d 1 and 2 and the clinical presentation of apple allergy. Conclusions. Mal d 3 and Mal d 4 are the relevant allergens in Spanish patients allergic to apple. Mal d 3 is related to severe systemic reactions, whereas Mal d 4 is involved in mild local OAS. Mal d 1 and 2 do not seem to be clinically relevant apple allergens in Spanish patients.

↩

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

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

↩

[92] - 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.

↩

[93] - 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.

↩

[94] - 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.

↩

[95] - 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.

↩

[96] - Gaier S, Oberhuber C, Hemmer W, Radauer C, Rigby NM, Marsh JY et al. Pru p 3 as a marker for symptom severity for patients with peach allergy in a birch pollen environment. J Allergy Clin Immunol 2009;124:166-167

↩

[97] - Fernández-Rivas M, Bolhaar S, González-Mancebo E, Asero R, van Leeuwen A, Bohle B, et al. Apple allergy across Europe: How allergen sensitization profiles determine the clinical expression of allergies to plant foods. J Allergy Clin Immunol 2006;118:481-488

BACKGROUND: Allergy to a plant food can either result from direct sensitization to that food or from primary sensitization to pollen, latex, or another food . OBJECTIVE: We sought to investigate the primary sensitizers in apple allergy across Europe, the individual allergens involved, and whether these differences determine the clinical presentation . METHODS: Patients (n = 389) with positive case histories and skin prick test responses to fresh apple were selected in the Netherlands, Austria, Italy, and Spain. Skin prick tests and RASTs to a panel of pollens and plant foods were performed, as well as RASTs to Bet v 1 and the apple allergens Mal d 1, 2, 3, and 4 . RESULTS: In the Netherlands, Austria, and Italy apple allergy is mild (>90% isolated oral symptoms) and related to birch pollinosis and sensitization to Bet v 1 and its apple homologue, Mal d 1, which has an odds ratio of local reactions of 2.85 (95% CI, 1.47-5.55). In Spain apple allergy is severe (>35% systemic reactions) and related to peach allergy and sensitization to Mal d 3 (nonspecific lipid transfer protein), which has an odds ratio of systemic reactions of 7.76 (95% CI, 3.87-15.56) . CONCLUSION: The analysis of individual apple allergens in a clinical context has provided insight into the sensitization pathway and into the intrinsic risk an allergen bears to induce mild or severe food allergy. CLINICAL IMPLICATIONS: Information on the sensitization pathway is essential to develop preventive strategies in food allergy. The application of individual food allergens with a known intrinsic risk will improve the prognostic value of diagnostic tests.

↩

[98] - Fernández-Rivas M, Bolhaar S, González-Mancebo E, Asero R, van Leeuwen A, Bohle B, et al. Apple allergy across Europe: How allergen sensitization profiles determine the clinical expression of allergies to plant foods. J Allergy Clin Immunol 2006;118:481-488

BACKGROUND: Allergy to a plant food can either result from direct sensitization to that food or from primary sensitization to pollen, latex, or another food . OBJECTIVE: We sought to investigate the primary sensitizers in apple allergy across Europe, the individual allergens involved, and whether these differences determine the clinical presentation . METHODS: Patients (n = 389) with positive case histories and skin prick test responses to fresh apple were selected in the Netherlands, Austria, Italy, and Spain. Skin prick tests and RASTs to a panel of pollens and plant foods were performed, as well as RASTs to Bet v 1 and the apple allergens Mal d 1, 2, 3, and 4 . RESULTS: In the Netherlands, Austria, and Italy apple allergy is mild (>90% isolated oral symptoms) and related to birch pollinosis and sensitization to Bet v 1 and its apple homologue, Mal d 1, which has an odds ratio of local reactions of 2.85 (95% CI, 1.47-5.55). In Spain apple allergy is severe (>35% systemic reactions) and related to peach allergy and sensitization to Mal d 3 (nonspecific lipid transfer protein), which has an odds ratio of systemic reactions of 7.76 (95% CI, 3.87-15.56) . CONCLUSION: The analysis of individual apple allergens in a clinical context has provided insight into the sensitization pathway and into the intrinsic risk an allergen bears to induce mild or severe food allergy. CLINICAL IMPLICATIONS: Information on the sensitization pathway is essential to develop preventive strategies in food allergy. The application of individual food allergens with a known intrinsic risk will improve the prognostic value of diagnostic tests.

↩

[99] - Ebo DG, Ahrazem O, Lopez-Torrejon G, Bridts CH, Salcedo G, Stevens WJ. Anaphylaxis from Mandarin (Citrus reticulata) : Identification of Potential Responsible Allergens. Int Arch Allergy Immunol 2007;144:39-43

We report on a patient with anaphylaxis from mandarin. Temporal relationship between consumption of the fruit, the presence of positive specific IgE, the positive skin test and the basophil activation test for mandarin strongly supported the diagnosis of an IgE-mediated allergy from mandarin. The lipid transfer protein allergen from mandarin fruit was isolated and characterized. Specific IgE levels and IgE immunodetection data indicated the patient's sensitization to orange (Cit s 3) and mandarin (Cit r 3) lipid transfer protein allergens, as well as to germin-like (Cit s 1) allergen. These results were fully confirmed by skin prick test and basophil activation test (BAT) for lipid transfer proteins, and a BAT for Cit s 1. This case report has several particularities. First, in Central and Northern Europe, it is not widely appreciated that citrus fruits, particularly mandarin, can elicit anaphylaxis. Second, this case report re-emphasizes sensitization from lipid transfer proteins to predispose for severe allergic reactions. Finally, it provides an opportunity to summarize the applications of flow cytometry-assisted analysis and quantification of in vitro activated basophils in the diagnostic approach of anaphylaxis from food.

↩

[100] - Egger C, Oberhuber C, Reider N, Marsh J, Focke M. Anaphylaxis to blueberry (Vaccinium myrtillus) - identification of a new lipid transfer protein. EAACI 25th Congress, Vienna, 10-14 June, 2006, Poster n°1671

A 29-year old female presented with a first-time anaphylactic reaction with pruritus, generalized urticaria, angioedema of the face and tongue, dyspnoe and dizziness. After appropriate anti-anaphylactic treatment and observation for another two days she was discharged from the hospital. Emergency medication including an epinephrine autoinjector was prescribed. History revealed the intake of "Schwarzbeernocken", a tyrolean speciality and sort of pancake containing blueberries, one hour prior to the onset of symptoms. Previous allergies were not known. Positive prick-to-prick-test and specific IgE to blueberry as determined by UniCAP confirmed allergy to blueberries. In addition, sensitisation to mugwort was noted. Tryptase, which was elevated at admission, later returned to normal levels. For allergen-specific evaluation, blueberries were extracted according to the low-temperature acetone-powder method described by Vieths et al. Extracts were separated by reducing SDS-PAGE and blotted to nitrocellulose for IgE immunoblot experiments. For IgE inhibition experiments, serum was pre-adsorbed with purified natural lipid transfer protein (LTP) from peach, Pru p 3. A single positive band of approximately 9 kD, which could be inhibited with Pru p 3, was detected. LTPs are widely distributed in the plant kingdom. Because of their high resistance to heat and digestion LTPs are important food allergens. Sensitisation to LPTs not only in Southern Europe, where it was first recognized to cause severe food allergy, is increasingly reported and should be considered when severe reactions to plant foods occur. To our knowledge, this is the first report of food allergy to blueberries and the first description of a lipid transfer protein in blueberry.

↩

[101] - Süß A, Vieths S, Treudler R, Simon JC. IgE-mediated anaphylaxis to blueberry and grape. Allergy 2007;62(suppl. 83):337

Background: Reports on the allergenicity of small fruits such as berries are rare. We investigated rare case of an IgE-mediated anapyhalactic reaction to blueberry and grape. METHODS: We investigated a 21-year old patient, who had suffered three episodes of oral pruritus followed by massive angioedema of the face, dyspnea and flushing 10 minutes after ingestion of blueberry pancake, preserved blueberry and fresh blue grapes, respectively. Each episode required emergency treatment. There was no history of rhinoconjunctivits or other atopic diseases. We performed prick-to-prick tests and in-vitro studies to confirm that the reported reactions were IgE-mediated. RESULTS: Skin prick tests with the culprit foods showed histamine equivalent reactions to fresh and cooked blueberry as well as to blue and green grapes, but not to perennial and seasonal aeroallergens. Specific IgE to blueberry and grape was 1,94 kU/l and 1,44 kU/l, respectively (CAP 2, Phadia CAP) and total IgE 122 kU/l (normal <100). Serum tryptase was normal as was skin inspection for cutaneous mastocytosis. CONCLUSION: True food allergy in adults as a result of primary sensitisation to fruit proteins is rarely found in Germany, where most allergic reactions to food in adults are caused by pollen associated cross-reactivity. Anaphylactic reactions to grapes were found to be caused by sensitization to a lipid transfer protein (LTP), Vit v 1. Blueberries also contain a LTP homologous protein. LTPs are a class of conserved plant proteins widely distributed in nature. They are thermostabile and extremely resistant to peptic digestion. Thus, LTPs can induce sensitisation and systemic symptoms through the gastrointestinal mucosa and have been claimed as the elicitors of true food allergy. Investigations on sensitisation to LTPs in our patient as well as western blot studies with blueberry extract are ongoing.

↩

[102] - Cheynel A, Malandain H, Leduc V. Un cas insolite d’allergie croisée. Rev Fr Allergol Immunol Clin 2007;47:284-285

↩

[103] - 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.

↩

[104] - Lauer I, San Miguel-Moncín M, Hartz C, Cistero-Bahima A, Conti A, Vieths S, et al. The lipid transfer protein (LTP) from Platanus acerifolia pollen, Pla a 3, is a major allergen and cross-reacts with food LTPs. EAACI 23th Congress, Amsterdam, 12-16 June, 2004, Poster n°1001

Background: Plane trees such as Platanus acerifolia are an important source of airborne allergens in many cities of Europe. Until now, only two major allergens of plane pollen, Pla a 1 and Pla a 2, have been characterised. In the Mediterranean area food-allergic patients are frequently sensitised to non-specific LTPs (nsLTPs). Sensitisation to food LTPs is not related to birch pollen allergy, and patients suffer from more severe reactions, in addition to OAS. An association between plane tree pollen allergy and plant food allergy was described, but the cross-reacting allergens have not been identified. The aim of this study was identification and characterisation of the nsLTP from plane pollen, and to investigate its immunological relationship to food LTPs from lettuce, cherry and hazelnut. Methods: Plane pollen extract was fractionated by ion exchange chromatography. Plane pollen LTP was identified by N-terminal amino acid sequence analysis. The prevalence of IgE reactivity to plane LTP was determined by immunoblotting using 45 Spanish plane pollen allergic patients with various plant food allergies and 14 plane pollen sensitised patients from Germany. In addition, sensitisation to LTPs from nuts, fruits and vegetables and their cross-reactivity to plane LTP was evaluated by immunoblotting and IgE inhibition experiments. Results: A 10 kDa IgE-reactive protein from plane pollen was partially purified and identified by N-terminal amino acid sequencing as nsLTP (Pla a 3). The N-terminal sequence of Pla a 3 showed the highest aa-ID of 58% with that of Lac s 1 (lettuce LTP). Pla a 3 was cross-reactive with food LTPs. Pla a 3 showed a prevalence of IgE reactivity of 66.6% (30/45) in Spanish plane pollen allergic patients. In contrast, Pla a 3 sensitisation was not relevant in plane pollen sensitised patients from Germany. We observed only partial correlation between sensitisation to plane pollen LTP and LTPs from lettuce, cherry and hazelnut. On the one hand, there were Spanish patients sensitised to food LTPs without plane pollen allergy, on the other hand, food LTP allergic patients without sensitisation to Pla a 3 were found. Conclusions: We have characterised the plane LTP as major allergen in plane pollen allergic patients with concomitant allergy to plant-derived foods recruited in the Mediterranean area. In Spanish patients sensitisation to food LTPs is not associated exclusively to the plane pollen LTP.

↩

[105] - Gaier S, Oberhuber C, Hemmer W, Radauer C, Rigby NM, Marsh JY et al. Pru p 3 as a marker for symptom severity for patients with peach allergy in a birch pollen environment. J Allergy Clin Immunol 2009;124:166-167

↩

[106] - Flinterman AE, Akkerdaas JH, den Hartog Jager CF, Hoekstra MO, Knulst AC, van Ree R, et al. Sensitization to Cor a 8, a Lipid Transfer Protein, in Children with Objective Reactions to Hazelnut from a Birch Endemic Country. J Allergy Clin Immunol 2007;119(1 suppl):S191

RATIONALE: Although hazelnut allergy is usually related to birch pollen allergy and limited to mild oral allergy, in particular children also report more serious reactions. In areas without birches, sensitization to lipid transfer protein (LTP) in hazelnut, Cor a 8, has been observed in subjects with severe reactions. Children from a birch-endemic area might develop sensitization to LTP at a young age independent from birch pollen. METHODS: Specific IgE to purified Cor a 1, Cor a 8, and Bet v 1 was determined in sera of 217 Dutch children aged 0-17 years. Twenty-six of these children also underwent a DBPCFC. RESULTS: In 217 hazelnut-sensitized children, sensitization to Cor a 1 was observed in 62.7%, to Cor a 8 in 31.3%, and to Bet v 1 in 55.0%. Sensitization to Cor a 8 was predominantly present at early ages, whereas sensitization to Cor a 1 and Bet v 1 was correlated with increasing age (r=0.630 and r=0.503, respectively). Children with objective reactions during DBPCFC (n=8) were all sensitized to Cor a 8 (0.51 - 23.3 kU/l), compared to only one child (1/18) without objective reactions (0.90 kU/l). Levels of IgE to Cor a 8 and Cor a 1, but not to Bet v 1, were significantly higher in the group with objective reactions. CONCLUSIONS: This is the first report of a substantial presence of LTP sensitization in children from a birch-endemic area, associated with serious reactions to hazelnut. The route of sensitization to LTP remains to be elucidated, but seems independent from birch pollen.

↩

[107] - Akkerdaas J, Flinterman A, Rigby N, Lidholm J, van Ree R, Knulst A. Cor a 8 sensitisation in the Dutch adult hazelnut allergic population. Allergy 2009;64(Suppl. 90):81

Background: Most studies have demonstrated that lipid transfer protein (the major hazelnut allergen, Cor a 8 e.g.) sensitization in Europe is largely restricted to the Mediterranean area. It was our aim to investigate whether Cor a 8 sensitization also exists among adult hazelnut allergic adults in the Netherlands, who are predominantly sensitized to Cor a 1 from hazelnut due to primary sensitization to Bet v 1. Methods: Serum from hazelnut allergic patients (n=96) was analyzed by RAST/CAP for specific IgE against birch, nCor a 1 and n/rCor a 8. Sera with a (+)ve rCor a 8 CAP (0.35 IU/ml) were selected for n/rCor a 8 immunoblot and tested in Pru p 3 (major peach allergen) CAP. Results: A (+)ve RAST for Birch/Cor a 1 was observed for 92/86.5% of the patients, respectively. To our surprise, 22.9% (n=22) were nCor a 8 RAST(+)ve which corresponded with nCor a 8 blot and rCor a 8 CAP results (with the exception of 5 sera that were exclusively nCor a 8 (+) ve). 6/10 sera (rCor a 8 score ranging from 0.9-9.5 IU/ml) showed a positive rCor a 8 immunoblot result. 9/10 sera (same group) were Pru p 3 positive with specific IgE titers ranging from 2.3-55.7 IU/ml. Conclusion: Cor a 8 sensitization can also occur in birch endemic areas like the Netherlands. The clinical implication has to be further unravelled. Blotresults, obtained with natural Cor a 8 confirmed the Cor a 8 sensitization (frequency). The structural homology between Cor a 8 and Pru p 3 must be responsible for the mutual recognition. For patients from the studied population with clinical symptoms after peach/hazelnut consumption further (inhibition) experiments must indicate which LTP was the sensitizing agent. The European Unionfunded Integrated Project EuroPrevall (contract no.514000) is acknowledged for donation of purified nCor a 8.

↩

[108] - Gaier S, Oberhuber C, Hemmer W, Radauer C, Rigby NM, Marsh JY et al. Pru p 3 as a marker for symptom severity for patients with peach allergy in a birch pollen environment. J Allergy Clin Immunol 2009;124:166-167

↩

[109] - Flinterman AE, Akkerdaas JH, den Hartog Jager CF, Rigby NM, Fernandez-Rivas M, Hoekstra MO, et al. Lipid transfer protein–linked hazelnut allergy in children from a non-Mediterranean birch-endemic area. J Allergy Clin Immunol 2008;121:423-428

BACKGROUND: Hazelnut allergy in birch pollen-exposed areas is usually due to cross-reactivity (Cor a 1 and 2) and is usually mild in nature (oral allergy). In areas without birches, severe reactions are more prevalent and linked to sensitization to the lipid transfer protein (LTP) Cor a 8 . OBJECTIVE: We sought to investigate whether sensitization to LTP plays a role in more severe (objective) hazelnut-induced symptoms in children from a birch-endemic area . METHODS: Sensitization to Cor a 8, Cor a 2, Cor a 1, and Bet v 1 was determined by means of RASTs and immunoblotting in hazelnut-sensitized children with (n = 8) and without (n = 18) objective reactions during double-blind, placebo-controlled food challenges. Additionally, samples from 191 hazelnut-sensitized nonchallenged children were analyzed . RESULTS: Children with objective reactions during double-blind, placebo-controlled food challenge had higher IgE titers to hazelnut (P < .001) and recognized more allergens on immunoblotting (P = .001) than those without such reactions. All children with objective symptoms were sensitized to Cor a 8 (0.51-23.3 IU/mL) compared with only 1 child without objective reactions (0.90 IU/mL). In a multivariate analysis only IgE against Cor a 8 remained as an independent risk factor (undefined odds ratio; P < .0001). In the group of nonchallenged children (n = 191), the prevalence of LTP sensitization was greater than 30%. Unexpectedly, sensitization to Cor a 1 was observed in children not sensitized to Bet v 1 . CONCLUSION: Sensitization to hazelnut LTP is a risk factor for objective symptoms in children from a birch-endemic area.

↩

[110] - Akkerdaas J, Flinterman A, Rigby N, Lidholm J, van Ree R, Knulst A. Cor a 8 sensitisation in the Dutch adult hazelnut allergic population. Allergy 2009;64(Suppl. 90):81

Background: Most studies have demonstrated that lipid transfer protein (the major hazelnut allergen, Cor a 8 e.g.) sensitization in Europe is largely restricted to the Mediterranean area. It was our aim to investigate whether Cor a 8 sensitization also exists among adult hazelnut allergic adults in the Netherlands, who are predominantly sensitized to Cor a 1 from hazelnut due to primary sensitization to Bet v 1. Methods: Serum from hazelnut allergic patients (n=96) was analyzed by RAST/CAP for specific IgE against birch, nCor a 1 and n/rCor a 8. Sera with a (+)ve rCor a 8 CAP (0.35 IU/ml) were selected for n/rCor a 8 immunoblot and tested in Pru p 3 (major peach allergen) CAP. Results: A (+)ve RAST for Birch/Cor a 1 was observed for 92/86.5% of the patients, respectively. To our surprise, 22.9% (n=22) were nCor a 8 RAST(+)ve which corresponded with nCor a 8 blot and rCor a 8 CAP results (with the exception of 5 sera that were exclusively nCor a 8 (+) ve). 6/10 sera (rCor a 8 score ranging from 0.9-9.5 IU/ml) showed a positive rCor a 8 immunoblot result. 9/10 sera (same group) were Pru p 3 positive with specific IgE titers ranging from 2.3-55.7 IU/ml. Conclusion: Cor a 8 sensitization can also occur in birch endemic areas like the Netherlands. The clinical implication has to be further unravelled. Blotresults, obtained with natural Cor a 8 confirmed the Cor a 8 sensitization (frequency). The structural homology between Cor a 8 and Pru p 3 must be responsible for the mutual recognition. For patients from the studied population with clinical symptoms after peach/hazelnut consumption further (inhibition) experiments must indicate which LTP was the sensitizing agent. The European Unionfunded Integrated Project EuroPrevall (contract no.514000) is acknowledged for donation of purified nCor a 8.

↩

[111] - 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.

↩

[112] - 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.

↩

[113] - 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.

↩

[114] - 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.

↩

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

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

↩

[116] - Fernández-Rivas M, González-Mancebo E, Rodríguez-Pérez R, Benito C, Sánchez-Monge R, Salcedo G, et al. Clinically relevant peach allergy is related to peach lipid transfer protein, Pru p 3, in the Spanish population. J Allergy Clin Immunol 2003;112:789-795

BACKGROUND: Sensitization to peach and related Rosaceae fruits without clinical expression is commonly observed as the result of the extensive cross-reactivity of IgE antibodies directed toward lipid transfer proteins (LTPs), Bet v 1 homologues, profilins, and carbohydrate determinants . OBJECTIVE: We aimed to study whether there are any clinical or immunologic differences between patients allergic to peach and those who have a current clinically irrelevant sensitization to this fruit . METHODS: One hundred subjects with adverse reactions to peach were evaluated by medical history, skin prick tests with fresh peach and purified peach LTP (Pru p 3), and specific IgE determinations to peach, rBet v 1, and rBet v 2 (birch profilin). Clinical reactivity to peach was established by double-blind, placebo-controlled food challenges. The clinical characteristics and the in vivo and in vitro tests were compared between allergic and nonallergic patients . RESULTS: Peach allergy was confirmed in 76 patients and ruled out in 16; 2 patients dropped out, and the study was not conclusive in 6 individuals (placebo reactors). Pollen allergy was found in 76% of the allergic patients and in 100% of the nonallergic patients. Positive responses to Pru p 3, rBet v 1, and rBet v 2 were observed in 62%, 7%, and 34% of patients allergic to peach, respectively. The sensitization rate to Pru p 3 was significantly higher among subjects allergic than nonallergic to peach (62% vs 31%, P =.02). IgE responses to rBet v 2 were more frequent among subjects allergic to pollen, but no difference was observed in the presence or absence of peach allergy . CONCLUSIONS: Pru p 3 is the major allergen of peach in our population, and the IgE response to this allergen is related to the clinical expression of peach allergy. Sensitization to profilin is observed in those patients with an associated pollen allergy but does not appear to be related to the clinical reactivity to peach.

↩

[117] - Fernández Rivas M. Food Allergy in Alergológica-2005. J Investig Allergol Clin Immunol 2009;19(Suppl. 2):37-44

BACKGROUND: Alergológica-2005 is a cross-sectional study carried out in Spain in 4991 new patients prospectively recruited in allergy clinics . OBJECTIVE: To describe the characteristics of food allergic patients included in Alergológica-2005 . RESULTS: Food allergy was diagnosed in 369 subjects (7.4%, 95% CI, 6.7%-8.1%). The most common offending foods were fruits (33.3% of cases), nuts (26%), shellfish (22%), egg (16%), milk (13.9%) and fish (9.8%). Rosaceae fruits and crustaceans elicited 23.6% and 18.7% of the reactions, respectively. Milk and egg were the most common foods in patients < 5 years of age, whereas fruits and nuts were the most prevalent foods in patients over 5 years. The most frequent manifestations included skin reactions (65.3%), oral allergy syndrome (33.6%), digestive symptoms (24.7%) and anaphylaxis (17.9%). The clinical presentation differed among foods. Prick-tests were carried out more frequently than serum immunoglobulin E (IgE) determinations (95.9% vs. 65%). Oral challenges were performed in 13% of patients, and 72.3% of these were open. Food allergy was diagnosed on the basis of medical history and positive IgE in 60.2% of the patients, ranging from 13.7% for milk to 75% for cereals. The self perception of patients' quality of life was lower than that of the 75% of Spanish reference population . CONCLUSION: Food allergy is diagnosed in 7.4% of the patients seen in allergy clinics across Spain, and has an important impact on the quality of life of patients. The foods involved in reactions change with age. The clinical presentation changes with the food, although the skin is the most frequently affected organ.

↩

[118] - Ferrer A, Carnes J, Gallego MT, Andreu C, Fernandez-Caldas E. Characterization and improvement of apple extracts for the diagnosis of apple IgE-mediated allergy. Ann Allergy Asthma Immunol 2005;95:462-467

BACKGROUND: The production process of reliable fruit extracts is not well established. OBJECTIVES: To improve the overall quality of apple extracts by reducing protein loss during the manufacturing process and to evaluate the improved extracts using in vivo and in vitro experiments. METHODS: Two types of extracts were prepared from peels of Golden Delicious apples (Malus domesticus). Extract A was extracted, 1:2 wt/vol, for 30 minutes at 40 degrees C in 0.01 M phosphate-buffered saline, and extract B was extracted, 1:2 wt/vol, in phosphate-buffered saline with 20% polyvinylpolypyrrolidone and 2-mmol/L EDTA. Both extracts were filtered, dialyzed in 3.5-kDa dialysis membranes, and lyophilized. The antigenic and allergenic profiles were analyzed using immunoblot and enzyme-linked immunosorbent assay. Nine patients clinically sensitive to apples and 12 controls underwent skin testing with both extracts. RESULTS: Extracts A and B had dry weight yields of 0.71% and 1.86% and protein contents of 104.6 and 257 microg/mg of freeze-dried material, respectively. A steady and progressive loss of protein, greater in extract A than in extract B, was detected at different intervals during the manufacturing process of both extracts. Extract B produced larger wheal sizes than extract A (P = .008). Enzyme-linked immunosorbent assay inhibition results confirmed that extract B had a greater inhibition capacity than extract A. CONCLUSIONS: A progressive loss of protein content occurs during the manufacturing of apple extracts. Wheal sizes induced by extract B were significantly larger than those induced by extract A and prick-by-prick solutions. Extract B was also more potent in vitro than extract A.

↩

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

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

↩

[120] - Skypala I, Calderon M, Leeds A, Durham S. Can symptoms or skin prick wheal size predict the outcome of food challenge with apple ? Allergy 2007;62(suppl. 83):147

Apple is one of the commonest foods to cause pollen food syndrome (PFS), a manifestation of Oral Allergy Syndrome (OAS), common in people with springtime hayfever. The aim of this study was to evaluate whether symptom score or skin prick test wheal size for apple could be predictive of the outcome of open or blinded food challenge. Subjects aged 18 and over with symptoms of hay fever from March to May recorded details of any foods they had reported symptoms to and scored those symptoms on a linear scale from 0 to 10. Following this, all subjects had a skin prick test to a fresh Granny Smith apple using the prick by prick method (PPT) and an open and blinded challenge with apple. Apple was the commonest food reported to cause symptoms, affecting 39 of the 119 subjects who completed the study (mean symptom score 4.4). 72% of those affected by apple (28) also had a positive prick test to apple (<3mm). Following open challenge 33/39 (84%) had their symptoms confirmed by a combination of positive PPT and open challenge, with 24 (73%) also having either objective symptoms on open challenge (OJOC) or a positive double-blind challenge (+veBC). Reported symptoms, symptom score and PPT wheal size to apple were all significantly associated with either a +veBC or OJOC. Reported symptoms were also an independently significant predictor of a positive open challenge (p=0.006). There was no correlation between apple symptom score and PPT wheal size. As a predictor of a +veBC or OJOC, reported symptoms had a sensitivity of 75%, specificity of 94% , positive predictive value (PPV) of 86% and negative predictive value (NPV) of 89%. For PPT wheal size the best predictive values came from a wheal size of 4mm with a PPV of 75% and NPV of 88% or 5mm with a PPV of 72% and NPV of 90%. Reported symptoms and PPT wheal size to apple are both associated with a positive challenge result, but only reported symptoms are predictive of a positive challenge result. Reported symptoms have similar sensitivity and specificity to the optimal PPT wheal size but have better PPV and NPV. There was no correlation between the severity of reported symptoms to apple and the size of the PPT wheal for apple.

↩

[121] - Bolhaar STHP, van de Weg WE, van Ree R, Gonzalez-Mancebo E, Zuidmeer L, Bruijnzeel-Koomen CAFM, et al. In vivo assessment with prick-to-prick testing and double-blind, placebo-controlled food challenge of allergenicity of apple cultivars. J Allergy Clin Immunol 2005;116:1080-1086

BACKGROUND: Apple cultivars have been reported to differ in allergenicity on the basis of in vitro and skin prick tests with apple extracts . OBJECTIVES: We sought to evaluate the efficacy of the prick-to-prick method in assessing differences in allergenicity of apple cultivars and to confirm differences by means of double-blind, placebo-controlled food challenge (DBPCFC) . METHODS: Intra-assay and intracultivar variation of prick-to-prick test results were determined in 6 Dutch and 8 Spanish patients with apple allergy by using 5 apples of the cultivars Golden Delicious, Fuji, and Ecolette in duplicate. In addition, 21 cultivars were screened for allergenicity in 15 Dutch patients with birch pollen and apple allergy. Two selected cultivars (Golden Delicious and Santana) were tested with DBPCFCs. The influence of storage conditions on allergenicity was assessed in 5 cultivars . RESULTS: Intra-assay variation of skin prick testing was 3.9%, and intracultivar variation was 4.1%. A ranking of 21 cultivars was made on the basis of prick-to-prick tests in 9 patients. Apple cultivars were classified as of low, intermediate, and high allergenicity, with a significant difference between low and high allergenicity (P < .001). A significant difference in allergenicity determined between Golden Delicious and Santana cultivars (P < .05) was confirmed by means of DBPCFC. With 5 cultivars, controlled atmosphere (2.5% oxygen/1% carbon dioxide) was shown to reduce allergenicity (P < .001) by 15% compared with storage at 2 degrees C . CONCLUSIONS: Prick-to-prick testing with fresh apples is a reproducible method of assessing allergenicity. Apples can be classified as of low or high allergenicity for the majority of patients. This was confirmed by using DBPCFCs. Selection of cultivars and control of storage conditions are both viable strategies for reduction of symptoms in patients with apple allergy.

↩

[122] - van der Werff SA, Dubois AE, van der Heide S, van de Weg E, Arens P, Vlieg-Boerstra BJ. Where To Prick The Apple For Skin Testing ?. J Allergy Clin Immunol 2009;123:S29

RATIONALE: Prick-to-prick skin tests (SPTs) with fresh apple are used to assess sensitisation in subjects with oral allergy symptoms (OAS) to apple. We hypothesized that the location of pricking in the apple influences the outcome of SPT results. Aim: To study differences in SPT results taken from different locations of the apple surface in several apple cultivars. METHODS: In the fall of 2007, 20 adult apple allergic subjects were tested by SPTs with apple material obtained from both the stalk and the middle region of four different apple cultivars: Crisps Pink, Golden Delicious, Elize and Santana. These tests were repeated in the spring of 2008 in 31 subjects before the birch pollen season. Sixteen subjects participated in both test series. In the spring 2008, Santana was replaced by Modi due to the limited availability of Santana. For each apple cultivar, differences between SPTs from both regions were calculated by Wilcoxon Signed Rank test within each test series. Besides, the percentage of negative SPTs (wheal diameter < 3 mm) were assessed. RESULTS: SPTs from the stalk region gave significantly greater responses than SPTs from the middle region (p values varying from 0.001 to 0.04), except for Santana (p = 0.067) and Elize 2008 (p = 0.228). 5% of SPTs from the middle region were negative, while only 1% of the SPTs from the stalk region were negative. CONCLUSIONS: SPT response is correlated to the location of pricking. Therefore, standardization of the location of pricking is needed. We propose the stalk region as location of choice.

↩

[123] - Asero R, Mistrello G, Roncarolo D, Casarini M, Falagiani P. Allergy to nonspecific lipid transfer proteins in Rosaceae: a comparative study of different in vivo diagnostic methods. Ann Allergy Asthma Immunol 2001;87:68-71

ACKGROUND: Lipid transfer proteins (LTPs) are the major allergens in patients sensitive to Rosaceae (apple, peach, apricot, cherry, plum, and pear) who are not allergic to birch pollen . OBJECTIVE: The purpose of this study was to find a sensitive, specific, and relatively easy method for detection of LTP-sensitive patients . METHODS: We studied 36 persons who experienced oral allergy syndrome after the ingestion of fruits in the family Rosaceae. This study cohort was divided into two groups: 18 without allergy to birch pollen (patients) and 18 with birch pollen allergy (control subjects). All were tested by skin prick tests (SPTs) with fresh Golden Delicious apple, fresh peach, and extracts of peel and pulp from both fruits. Their specific IgE reactivities against peach peel extract were further investigated by immunoblot analysis . RESULTS: All 18 subjects in the control group showed strongly positive skin reactions with both fresh apple and fresh peach, whereas no skin reactivity was found with extracts from peach peel, peach pulp, or apple pulp. Extract of apple peel produced positive skin reactions in 17 of 18 control subjects; however, the wheals were generally smaller than those induced by fresh fruits. Immunoblot analysis showed no reactivity for peach peel extract. In contrast, the SPTs with fresh fruits showed that some of the 18 patients had strongly positive reactions, but others had weak reactions or negative responses. Further, in a high proportion of the patients, consecutive SPT with fresh apple yielded inconsistent results. In all patients, SPTs with extracts from apple pulp and peach pulp were negative, whereas SPTs with peel extracts were strongly positive in all patients. In most patients, the wheal area induced by SPT with peel extracts was larger than that induced by SPTs with fresh fruits. Immunoblot analysis showed that serum specimens from all 18 patients reacted with a 10-kD protein in peach peel. This is the molecular mass of LTPs . CONCLUSIONS: In birch pollen-allergic patients, the SPTs with fresh foods still remains the most reliable method of diagnosing vegetable food hypersensitivity. In contrast, in patients not allergic to birch pollen, the most reliable strategy for detection of patients sensitive to LTPs is skin prick testing with properly prepared fruit peel extracts. The loss of Bet v 1- and Bet v 2-like structures, which probably occurs during extraction, may facilitate immediate identification of the relevant allergen.

↩

[124] - 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.

↩

[125] - Asero R, Mistrello G, Roncarolo D, Amato S. Prick test with heat-processed apple peel extract to detect LTP hypersensitivity. Eur Ann Allergy Clin Immunol 2006;38:351-354

BACKGROUND: Allergen-resolved diagnosis of food allergy may be essential in the clinical practice, particularly in patients allergic to foods that may contain both labile and stable allergens. However, presently available diagnostic tests are not useful in this sense. OBJECTIVE: To assess the clinical usefulness of SPT with a heat-processed apple peel extract as an easily available means to detect hypersensitivity to lipid transfer protein (LTP), an extremely stable and potentially hazardous apple allergen. METHODS: Raw and heat-processed (100 degrees C x 10 min) apple peel extract (100 microg/ml) were used to carry out SPT in 23 patients with apple-allergy, 15 of which considered as probably sensitized to labile allergens (Maid 1, Mal d 4) and 8 to stable allergens (Mal d 3, LTP), respectively on the basis of the presence/absence of IgE reactivity to birch pollen. IgE reactivity to the heat-processed apple peel extract was further analyzed by immunoblot. RESULTS: Altogether SPT with raw apple extract scored positive in 20/23 (87%) patients, including 12/15 patients considered as probably sensitized to labile allergens and 8/8 patients considered as probably sensitized to LTP. In contrast, the heat-processed apple extract induced a wheal-and-flare reaction only in the 8 (100%) presumptive LTP reactors. Immunoblot analysis showed IgE reactivity to a 10 kDa protein (LTP) in heat-processed apple extract. CONCLUSION: The heat-resistance of stable apple allergens like LTP can be usefully exploited to prepare extracts for allergen-resolved diagnosis in-vivo.

↩

[126] - Asero R, Mistrello G, Roncarolo D, Amato S. Relationship between peach lipid transfer protein specific IgE levels and hypersensitivity to non-Rosaceae vegetable foods in patients allergic to lipid transfer protein. Ann Allergy Asthma Immunol 2004;92:268-272

BACKGROUND: Lipid transfer protein (LTP), the major allergen in Rosaceae in geographic areas where the prevalence of birch pollen allergy is low, is a widely cross-reacting pan-allergen, but the pattern of cross-reactivity to plant-derived foods botanically unrelated to Rosaceae shows much variability. OBJECTIVE: To examine the relationship between peach LTP specific IgE levels and cross-reactivity to several non-Rosaceae, plant-derived foods. METHODS: IgE specific for peach LTP was measured by enzyme-linked immunosorbent assay in serum samples from 40 patients with Rosaceae allergy monosensitized to LTP. Patients were considered monosensitized to this protein in the absence of sensitization to other cross-reacting, plant-derived foods as shown by negative skin prick test (SPT) results with both birch and mugwort pollen. SPTs with commercial extracts of walnut, hazelnut, peanut, celery, maize, rice, tomato, orange, and onion were performed to detect possible immunologic cross-reactivity to these foods. RESULTS: Patients with negative SPT results with non-Rosaceae foods showed significantly lower levels of IgE to peach LTP than patients showing skin reactivity to one or more non-Rosaceae foods (P < .001). A significant difference in specific IgE to peach LTP between patients with positive or negative SPT results was observed with each individual food (P < .001 in all cases). The level of IgE to peach LTP was strongly related to the number of positive SPT results with non-Rosaceae foods (r = 0.78; P < .001). Increasing levels of IgE to peach LTP were associated with skin reactivity to nuts (29/40 [72%]), peanut (27/40 [67%]), maize (16/39 [41%]), rice (14/39 [36%]), onion (13/37 [35%]), orange (9/32 [28%]), celery (11/40 [27%]), and tomato (8/39 [20%]). CONCLUSIONS: This study suggests that all allergenic determinants in LTP from vegetable foods other than peach cross-react with peach LTP determinants, whereas only some peach LTP epitopes cross-react with allergenic determinants on botanically unrelated, plant-derived foods. The high levels of IgE to peach LTP seem to reflect the presence of IgE targeting common allergenic determinants of LTP, causing cross-reactivity to botanically unrelated, vegetable foods. In LTP-allergic patients, increasing levels of IgE to peach LTP are paralleled by an increasing number of foods other than Rosaceae positive on SPT that cause clinical symptoms.

↩

[127] - Gamboa PM, Sanz ML, Lombardero M, Barber D, Sánchez-Monje R, Goikoetxea MJ et al. Component-resolved in vitro diagnosis in peach-allergic patients. J Investig Allergol Clin Immunol 2009;19:13-20

BACKGROUND:The in vitro diagnosis of pollen-related food allergy presents low specificity and reproducibility with many conventional extracts. This can be improved using natural purified allergens, recombinant purified allergens, or both. OBJECTIVE: We compared specific immunoglobulin (Ig) E determination (slgE), the basophil activation test (BAT), the histamine release test (HRT), and the cellular allergen stimulation test (CAST) using natural and recombinant allergens in the diagnosis of peach allergy. METHODS: Thirty-two peach allergic patients were studied. Skin prick tests were performed with commercial peach and extract with Mal d 1, nPru p 3, and profilin (nPho d 2). slgE, BAT, CAST, and HRT were determined using rPru p 3, rMal d 3, rBet v 1, rMal d 1, and rMal d 4. RESULTS: Agreement between the techniques was good with all the allergens, except HRT with rMal d 1 and rMal d 4. With rPru p 3, slgE, CAST, BAT, and HRT showed sensitivity values of 88%, 81%, 72%, and 69% and specificity values of 100%, 93%, 97%, and 83%, respectively. In patients with systemic symptoms or contact urticaria, the values were 100%, 85%, 81%, and 81%. In patients with oral allergy syndrome, sensitivity to profilins or homologues of Bet v 1 was detected in 100% of the cases by all the techniques, except by HRT with rMal d 1, which detected 66% of the cases. CONCLUSIONS: The use of single allergens in the in vitro diagnosis of peach allergy by specific IgE determination, BAT, and CAST offers high specificity and sensitivity, with better results than the HRT.

↩

[128] - Asero R, Jimeno L, Barber D. Component-resolved diagnosis of plant food allergy by SPT. Eur Ann Allergy Clin Immunol 2008;40:115-121

BACKGROUND: Fruits and vegetables may contain both labile and stable allergens. The former induce only OAS, whereas stable allergens may induce systemic reactions. Component-resolved diagnosis (CRD) of allergy to plant foods is therefore essential for the clinical management of allergic patients. METHODS: 80 adults allergic to plant foods underwent SPT with purified natural date palm profilin (Pho d 2), purified Mal d 1, a peach extract containing uniquely LTP, and with a kiwi extract containing uniquely stable allergens. RESULTS: 58 (72%) patients were monosensitized: 24 to Mal d 1, 24 to profilin, 7 to LTP, and 3 to kiwi. 22 patients were multi-sensitised: 14 to Mal d 1 and profilin, 2 to Mal d 1 and kiwi, 1 to LTP and profilin, 3 to LTP and Mal d 1, and 2 to LTP, Mal d 1 and profilin. Mal d 1 and LTP sensitisation were associated with apple and peach allergy, respectively, whereas profilin sensitisation was associated with allergy to melon, watermelon, banana, tomato and citrus fruits. 18/21 kiwi-allergic patients were sensitised to one of the cross-reacting allergens, but 2/18 reacted to kiwi-specific allergens as well. CONCLUSIONS: In patients with allergy to plant-derived foods CRD can be performed by SPT with purified allergen proteins. In the future, the availability of a larger number of purified natural or recombinant allergens for SPT will represent a simple means to classify food-allergic patients properly on the first visit.

↩

[129] - Asero R, Monsalve R, Barber D. Profilin sensitization detected in the office by skin prick test: a study of prevalence and clinical relevance of profilin as a plant food allergen. Clin Exp Allergy 2008;38:1033-1037

BACKGROUND: Profilin, a pan-allergen present in all eukaryotic cells, is one of the main causes of cross-sensitization between pollen and plant-derived foods, but its clinical relevance as a food allergen is still debated . OBJECTIVE: To investigate the prevalence of profilin sensitization in a pollen-allergic population and its clinical relevance as a food allergen . METHODS: Two hundred consecutive patients with pollen allergy underwent skin prick tests (SPT) with purified natural date palm profilin (Pho d 2; 50 microg/mL; Alk Abello, Madrid, Spain). Those reporting adverse reactions to foods (confirmed by SPT with either commercial food extracts or fresh foods) underwent SPT with an apple extract containing uniquely Mal d 1 (2 microg/mL; ALK-Abello), and with a commercial peach extract containing uniquely lipid transfer protein (LTP 30 microg/mL; ALK-Abello) . RESULTS: Sixty patients (30%) showed skin reactivity to date palm profilin, Pho d 2. All were sensitized to grass pollen, and most of them reacted to birch, mugwort, ragweed and plantain pollen as well. SPT with pellitory and cypress scored negative in a high proportion of profilin reactors [26/60 (43%) and 33/60 (55%), respectively]. More than one half (34/60 [57%]) of profilin reactors had food allergy; 21 of these were monosensitized to profilin, 11 were sensitized to both profilin and Bet v 1 homologous protein, one to both profilin and LTP, and one to all the three allergens. The large majority of profilin-allergic patients reported oral allergy syndrome as the only food-induced symptom and were able to tolerate the offending foods if they were cooked or otherwise processed. Twenty-eight of 34 reported reactivity to two or more plant-derived foods. Rosaceae, tree nuts, melon and watermelon, tomato, pineapple, citrus fruits and banana were the more frequently offending foods . CONCLUSION: Profilin should be considered a clinically relevant food allergen. Allergy to melon, watermelon, tomato, banana, pineapple and orange may be considered as a marker of profilin hypersensitivity. This study underlines the clinical importance of being able to diagnose hypersensitivity to single food allergenic proteins by SPT, particularly when the relevant food allergen sources contain several allergens that show different chemical/physical features and, hence, completely different risk profiles.

↩

[130] - Tavares B, Machado D, Loureiro G, Cemlyn-Jones J, Pereira C. Sensitization to profilin in the Central region of Portugal. Sci Total Environ 2008;407:273-278

BACKGROUND: Profilin is a panallergen found in pollens and fruits. Sensitization to this protein may explain some sensitization to multiple pollen species. We aimed to evaluate sensitization to profilin in patients suffering from respiratory allergy sensitized to pollens, in the Central region of Portugal. METHODS: Patients were evaluated for asthma symptoms, rhinitis, conjunctivitis and food allergy. Skin prick tests (SPT) to aeroallergens including 12 different pollens and profilin (nPho d 2) were performed. The patients were divided into two study groups according to the region of residence: A--inland region and B--coastal region. RESULTS: A total of 370 patients were evaluated (277-group A; 93-group B). 65.9% showed positive skin prick tests and 76.2% were positive to pollens (87.1%-group A; 42.85%-group B; p<0.0001). All the patients sensitized to pollens had rhinitis (p=0.001). Sensitization to profilin was associated with pollen sensitization (p=0.014). 43 patients were sensitized to profilin (40-group A; 3-group B; p=0.006). 21.0% of patients sensitized to pollens, were also sensitized to profilin. 39 patients were sensitized to at least two pollens (p<0.0001). Four profilin and pollen sensitized patients had oral allergy syndrome complaints to melon. This syndrome was related with profilin sensitization (p=0.001). CONCLUSIONS: It is advisable to perform SPT to profilin, particularly in the Inland region, for a better differential diagnosis between cross-reactivity and true sensitization to pollens. The results together with the medical history may support the choice for a specific immunotherapy option.

↩

[131] - Asero R, Jimeno L, Barber D. Prevalence and clinical relevance of hypersensitivity to pollen pan-allergens (polcalcin and Profilin): a SPT study. Allergy 2009;64(Suppl. 90):376-377

Background: Calcium binding proteins (polcalcins) and profilin are cross-reacting pan-allergens that sensitize a minority of pollenallergic patients. Their clinical relevance is still debated. Objective: To assess the clinical relevance of polcalcin and profilin hypersensitivity, detected for the first time by SPT, in a large group of pollen-allergic patients. Methods: 200 pollen-allergic adults (M/F 101/99; mean age 34 years) underwent SPT with 9 pollens present in this geographical area. Hypersensitivity to pan-allergens was detected by SPT with date palm polcalcin and profilin. Three symptomatic periods (from end February to mid May, from end April to mid July, and from mid August to the end of September) were considered as associated with birch and/ or cypress, grass and/or pellitory, and ragweed and/or mugwort allergy, respectively. Results: 16 (8%) patients reacted to date palm polcalcin; 5/5 (100%) co-recognized Phl p 7, the grass polcalcin, in-vitro. Clinically, only 4 (25%) reported symptoms in all 3 seasonal periods. 40 (20%) patients reacted to profilin; only 32 (80%) and 22 (55%) of them reacted to cypress and pellitory pollen, respectively. Clinically, only 4/40 (10%) had symptoms during all 3 seasonal periods. Six patients (3%) were co-sensitized to both polcalcin and profilin. Conclusions: The clinical relevance of hypersensitivity to pollen pan-allergens is often limited; many reactors have symptoms only during the central period, suggesting primary grass sensitization. Profilin reactors often do not co-recognize pellitory and cypress pollen. Invivo component-resolved diagnosis of seasonal respiratory allergies is a promising approach that might lead to reduction of costs and to a prompter definition of cases of multiple pollen reactivity.

↩

[132] - EXPO: Determinacion de la prevalencia de sensibilizacion a polenes mediante tecnicas de diagnostico molecular. ALK-Abello Ed., 2009, 86pp.

↩

[133] - Barber D, de la Torre F, Lombardero M, Antépara I, Colas C, Dávila I et al. Component-resolved diagnosis of pollen allergy based on skin testing with profilin, polcalcin and lipid transfer protein pan-allergens. Clin Exp Allergy 2009;39:1764-1771

Background Allergy diagnosis needs to be improved in patients suffering from pollen polysensitization due to the existence of possible confounding factors in this type of patients. Objective To evaluate new diagnostic strategies by comparing skin responses to pan-allergens and conventional allergenic extracts with specific IgE (sIgE) to purified allergen molecules. Methods One thousand three hundred and twenty-nine pollen-allergic patients were diagnosed by a combination of an in vitro method with a panel of 13 purified allergens, including major allergens and pan-allergens, using a high-capacity screening technology (ADVIA-Centaurs) and skin prick test (SPT) to pan-allergens and conventional extracts. Results There was a high concordance (k index) between in vitro (sIgE to major allergens) and in vivo (SPT to conventional extracts) methods in patients who were not sensitized to panallergens, but SPT with conventional extracts failed to diagnose patients with sensitization to pan-allergens. In patients who were simultaneously sensitized to polcalcins and profilins, there was a duplication both in the number of sensitizations to major allergens and in the years of disease evolution. There was a statistical association between sensitization to profilins and/or lipid transfer proteins and food allergy (Po0.0001). Conclusion The novel diagnostic strategy has proven to be a valuable tool in daily clinical practice. Introduction of routine SPT to pan-allergens is a simple and feasible way of improving diagnostic efficacy. Patients sensitized to pan-allergens should be tested by an adequate panel of allergenic molecules in order to identify the allergens that are responsible for the allergic disease.

↩

[134] - Fernández-Rivas M, González-Mancebo E, Rodríguez-Pérez R, Benito C, Sánchez-Monge R, Salcedo G, et al. Clinically relevant peach allergy is related to peach lipid transfer protein, Pru p 3, in the Spanish population. J Allergy Clin Immunol 2003;112:789-795

BACKGROUND: Sensitization to peach and related Rosaceae fruits without clinical expression is commonly observed as the result of the extensive cross-reactivity of IgE antibodies directed toward lipid transfer proteins (LTPs), Bet v 1 homologues, profilins, and carbohydrate determinants . OBJECTIVE: We aimed to study whether there are any clinical or immunologic differences between patients allergic to peach and those who have a current clinically irrelevant sensitization to this fruit . METHODS: One hundred subjects with adverse reactions to peach were evaluated by medical history, skin prick tests with fresh peach and purified peach LTP (Pru p 3), and specific IgE determinations to peach, rBet v 1, and rBet v 2 (birch profilin). Clinical reactivity to peach was established by double-blind, placebo-controlled food challenges. The clinical characteristics and the in vivo and in vitro tests were compared between allergic and nonallergic patients . RESULTS: Peach allergy was confirmed in 76 patients and ruled out in 16; 2 patients dropped out, and the study was not conclusive in 6 individuals (placebo reactors). Pollen allergy was found in 76% of the allergic patients and in 100% of the nonallergic patients. Positive responses to Pru p 3, rBet v 1, and rBet v 2 were observed in 62%, 7%, and 34% of patients allergic to peach, respectively. The sensitization rate to Pru p 3 was significantly higher among subjects allergic than nonallergic to peach (62% vs 31%, P =.02). IgE responses to rBet v 2 were more frequent among subjects allergic to pollen, but no difference was observed in the presence or absence of peach allergy . CONCLUSIONS: Pru p 3 is the major allergen of peach in our population, and the IgE response to this allergen is related to the clinical expression of peach allergy. Sensitization to profilin is observed in those patients with an associated pollen allergy but does not appear to be related to the clinical reactivity to peach.

↩

[135] - Lilja G, Nordlund M, Andersson K, Ostling J, Lundgren T, Marknell DeWitt A, et al. Analysis of specific IgE to vegetable foods in a cohort of birch-pollen allergic children and adolescents using recombinant allergens. Allergy Clin Immunol Int 2005;17(Suppl. 1):340

Background: Allergic reactions to certain vegetable foods are common among birch pollen sensitized individuals. The gold standard for diagnosis of such pollen-related food allergies consists of laborious challenge procedures. More widely available and cost-effective diagnostic methods, with favourable performance characteristics, are highly desired. This study was performed with the aim to assess the use of natural food extracts and recombinant food allergens for detection of specific IgE in a cohort of birch pollen allergic patients in relation to self-reported food allergy symptoms. Methods: Case history regarding symptoms of food allergy to hazelnut, apple, carrot, pear, cherry, peach, kiwi, celery and potato was obtained from 103 Swedish consecutive birch pollen allergic patients, 4-18 years of age. A serum sample from each subject was analysed for specific IgE to birch pollen and food extracts and recombinant birch and food allergens, using regular and experimental ImmunoCAP tests. Results: The proportion of subjects who had been knowingly exposed and could provide a definite answer regarding tolerance or reaction ranged from 62% to 98% between the different foods. Among those patients, reported allergic symptoms were most common to hazelnut (72%), followed by apple (69%), pear (50%), kiwi (48%), carrot (45%), peach (35%), cherry (33%), potato (12%) and celery (9%). Eighty-four percent of all subjects reported symptoms to at least one of the nine foods while 11% reported tolerance to all foods. Using natural food extracts, positive IgE test results were obtained from 4-91% of the subjects reporting symptoms. Specific IgE reactivity to at least one recombinant allergen derived from each of hazelnut, apple, carrot, pear, cherry and celery was detected in 91-100% of the subjects reporting symptoms to those foods. Among the recombinant allergens tested, IgE reactivity was most common to members of the PR-10 family. The measured levels of IgE to allergens of this family often exceeded those to natural food extract. The higher sensitivity of recombinant food allergens as assay reagents was accompanied by a more frequent detection of specific IgE also among subjects not reporting food allergy symptoms. Conclusion: Recombinant food allergens enabled improved detection and quantitation of specific IgE antibodies but did not enhance the discrimination between birch pollen allergic subjects with and without allergy to the foods studied.

↩

[136] - Malandain H, Giroux F, Cano Y. The influence of carbohydrate structures present in common allergen sources on specific IgE results. Eur Ann Allergy Clin Immunol 2007;39:216-220

BACKGROUND: Cross-reactive carbohydrate determinants (CCD) are well known interferants in specific IgE assays (sIgE). Glyco-epitopes are not restricted to CCD and extracts used to prepare in vitro tests contain many other glycoproteins able to bind glycan-specific IgE. The overall amounts of IgE-bindable glycan structures in allergen sources are unknown . OBJECTIVE: We aimed at quantifying the influence of N-glycan structures on IgE reactivity to commonly tested allergen sources . METHODS: IgE reactivity to 51 allergen extracts, one purified natural allergen and 10 recombinant allergens was measured on Phadia UniCAP system using 2 sera demonstrating significant levels of glycan-related IgE reactivity. Immobilized bromelain and horseradish peroxidase (HRP) were used to capture N-glycan-specific IgE from these sera. Residual IgE reactivity was measured for 42 allergen sources and 4 recombinant/purified allergens . RESULTS: An obviously excessive number of positive CAP-results were obtained with both sera, especially for plant-based allergen sources. Capture of glycan-specific IgE led to a decrease of serum IgE ractivity, variable among allergen sources and between sera. Among others, peanut results were proven largely interfered by the presence of glycan-specific IgE. Unexpectedly some allergen sources showed a slight influence of glycan-related reactivity, such as cockroach, mosquito, mussel, shrimp and domestic mites . CONCLUSION: In patients sensitized to pollens or to Hymenoptera venoms sIgE results should be interpreted with caution. One cannot substract the result of a glyco-reporter test (bromelain and/or HRP) in order to compute glycan-free slgE results for common allergen sources like peanuts. As long as the demonstration of a significant role for glycan structures in clinical allergic reactions is lacking, a simple pre-treatment able to discard glycan-specific IgE from serum would be useful to improve accuracy of in vitro diagnostic tests.

↩

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

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

↩

[138] - 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.

↩

[139] - Ebo DG, Hagendorens MM, Bridts CH, de Clerck LS, Stevens WJ. Sensitization to cross-reactive carbohydrate determinants and the ubiquitous protein profilin: mimickers of allergy. Clin Exp Allergy 2004;34:137-144

↩

[140] - Ballmer-Weber BK, Scheurer S, Fritsche P, Enrique E, Cistero-Bahima A, Haase T, et al. Component-resolved diagnosis with recombinant allergens in patients with cherry allergy. J Allergy Clin Immunol 2002;110:167-173

BACKGROUND: In pollen-related food allergy, extracts for skin prick tests (SPTs) are often not standardized, and the test reliability is affected by false-negative reactions . OBJECTIVE: We sought to evaluate a panel of recombinant allergens (RAs) derived from one allergenic food for use in component-resolved in vivo diagnosis, taking cherry as a model food . METHODS: Seventy-nine subjects were included in the study: 24 Swiss patients (group 1) with a positive double-blind placebo-controlled food challenge result to cherries, 23 patients with birch pollen allergy but without cherry allergy (group 2), 23 nonatopic subjects (group 3), and 9 Spanish patients with a history of a cherry allergy (group 4). SPTs were performed in duplicate by using recombinant cherry allergens (Bet v 1-related allergen: recombinant (r) Pru av 1; profilin: rPru av 4; and lipid transfer protein: rPru av 3) in concentrations of 10, 50, and 100 microg/mL. Furthermore, IgE reactivity to rPru av 1, rPru av 4, and rPru av 3 was assessed by means of immunoblot analysis . RESULTS: SPT responses with rPru av 1, rPru av 4, and rPru av 3 were positive in 92%, 17%, and 4% of the patients in group 1; in 74%, 30%, and 0% of the patients in group 2; in 0%, 22%, and 89% of the patients in group 4; and negative for all nonatopic subjects (group 3). Thus the sensitivity of a positive SPT response to at least one of the 3 RAs was 96%. The specificities, negative predictive values, and positive predictive values with the 3 RAs were 100%, 96%, and 100% if calculated in relation to the nonatopic control group but 17%, 79%, and 60% when calculated in relation to the control group with birch pollen allergy. The correlation between SPT and immunoblotting results was excellent. Sensitization to rPru av 3 was associated with more severe symptoms than sensitization to rPru av 1 . CONCLUSIONS: SPTs with RAs proved to be highly sensitive for diagnosis of cherry allergy. Component-resolved in vivo diagnosis with standardized amounts of stable RAs allows us to determine sensitization patterns directly, to correlate them with severity of clinical symptoms, and to analyze geographic differences.

↩

[141] - Lilja G, Nordlund M, Andersson K, Ostling J, Lundgren T, Marknell DeWitt A, et al. Analysis of specific IgE to vegetable foods in a cohort of birch-pollen allergic children and adolescents using recombinant allergens. Allergy Clin Immunol Int 2005;17(Suppl. 1):340

Background: Allergic reactions to certain vegetable foods are common among birch pollen sensitized individuals. The gold standard for diagnosis of such pollen-related food allergies consists of laborious challenge procedures. More widely available and cost-effective diagnostic methods, with favourable performance characteristics, are highly desired. This study was performed with the aim to assess the use of natural food extracts and recombinant food allergens for detection of specific IgE in a cohort of birch pollen allergic patients in relation to self-reported food allergy symptoms. Methods: Case history regarding symptoms of food allergy to hazelnut, apple, carrot, pear, cherry, peach, kiwi, celery and potato was obtained from 103 Swedish consecutive birch pollen allergic patients, 4-18 years of age. A serum sample from each subject was analysed for specific IgE to birch pollen and food extracts and recombinant birch and food allergens, using regular and experimental ImmunoCAP tests. Results: The proportion of subjects who had been knowingly exposed and could provide a definite answer regarding tolerance or reaction ranged from 62% to 98% between the different foods. Among those patients, reported allergic symptoms were most common to hazelnut (72%), followed by apple (69%), pear (50%), kiwi (48%), carrot (45%), peach (35%), cherry (33%), potato (12%) and celery (9%). Eighty-four percent of all subjects reported symptoms to at least one of the nine foods while 11% reported tolerance to all foods. Using natural food extracts, positive IgE test results were obtained from 4-91% of the subjects reporting symptoms. Specific IgE reactivity to at least one recombinant allergen derived from each of hazelnut, apple, carrot, pear, cherry and celery was detected in 91-100% of the subjects reporting symptoms to those foods. Among the recombinant allergens tested, IgE reactivity was most common to members of the PR-10 family. The measured levels of IgE to allergens of this family often exceeded those to natural food extract. The higher sensitivity of recombinant food allergens as assay reagents was accompanied by a more frequent detection of specific IgE also among subjects not reporting food allergy symptoms. Conclusion: Recombinant food allergens enabled improved detection and quantitation of specific IgE antibodies but did not enhance the discrimination between birch pollen allergic subjects with and without allergy to the foods studied.

↩

[142] - 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.

↩

[143] - EXPO: Determinacion de la prevalencia de sensibilizacion a polenes mediante tecnicas de diagnostico molecular. ALK-Abello Ed., 2009, 86pp.

↩

[144] - Cuesta J, Blanco C, Fernández-Rivas M, Villalba M. Vegetalia. J Investig Allergol Clin Immunol 2006;16(Suppl. 2):40-44

↩

[145] - Gamboa PM, Sanz ML, Lombardero M, Barber D, Sánchez-Monje R, Goikoetxea MJ et al. Component-resolved in vitro diagnosis in peach-allergic patients. J Investig Allergol Clin Immunol 2009;19:13-20

BACKGROUND:The in vitro diagnosis of pollen-related food allergy presents low specificity and reproducibility with many conventional extracts. This can be improved using natural purified allergens, recombinant purified allergens, or both. OBJECTIVE: We compared specific immunoglobulin (Ig) E determination (slgE), the basophil activation test (BAT), the histamine release test (HRT), and the cellular allergen stimulation test (CAST) using natural and recombinant allergens in the diagnosis of peach allergy. METHODS: Thirty-two peach allergic patients were studied. Skin prick tests were performed with commercial peach and extract with Mal d 1, nPru p 3, and profilin (nPho d 2). slgE, BAT, CAST, and HRT were determined using rPru p 3, rMal d 3, rBet v 1, rMal d 1, and rMal d 4. RESULTS: Agreement between the techniques was good with all the allergens, except HRT with rMal d 1 and rMal d 4. With rPru p 3, slgE, CAST, BAT, and HRT showed sensitivity values of 88%, 81%, 72%, and 69% and specificity values of 100%, 93%, 97%, and 83%, respectively. In patients with systemic symptoms or contact urticaria, the values were 100%, 85%, 81%, and 81%. In patients with oral allergy syndrome, sensitivity to profilins or homologues of Bet v 1 was detected in 100% of the cases by all the techniques, except by HRT with rMal d 1, which detected 66% of the cases. CONCLUSIONS: The use of single allergens in the in vitro diagnosis of peach allergy by specific IgE determination, BAT, and CAST offers high specificity and sensitivity, with better results than the HRT.

↩

[146] - Ebo DG, Bridts CH, Verweij MM, De Knop KJ, Hagendorens MM, De Clerck LS et al. Sensitization profiles in birch pollen-allergic patients with and without oral allergy syndrome to apple: lessons from multiplexed component-resolved allergy diagnosis. Clin Exp Allergy 2010;40:339-347

↩

[147] - Valenta R, Lidholm J, Niederberger V, Hayek B, Kraft D, Grönlund H. The recombinant allergen-based concept of component-resolved diagnostics and immunotherapy (CRD and CRIT). Clin Exp Allergy 1999;29:896-904

↩

[148] - Stapleton GL, Griffiths H, Karim MY, Deacock SJ. Specific IgE reactivty to birch pollen allergens in a UK population with the oral allergy syndrome: are there any pre-disposing factors for anaphylaxis ? Clin Exp Allergy 2006;36:1210-1211

Introduction Pollen-allergic patients frequently experience symptoms on ingestion of fresh plant-derived foods termed oral allergy syndrome (OAS). IgE cross-reactivity between pollen and food allergens represents the molecular basis of OAS. Symptoms range from commonly immediate oral itching to rarely lifethreatening anaphylaxis. Studies from Europe have described use of recombinant birch pollen allergens to identify birch pollen-sensitive patients and those with OAS; however this has not been evaluated in the UK or for predicting severity of symptoms in OAS. Objectives To determine whether patients with OAS-related anaphylaxis reacted to any particular birch pollen allergen and whether specific IgE (sIgE) level to any particular allergen reflected disease severity. Methods 280 patients with birch pollen sensitivity were analysed retrospectively for associations between clinical food sensitivities (e.g. apple, carrot, hazelnut etc.) and incidence of anaphylaxis. Of 280 patients, 65 serum samples were tested for sIgE to natural birch pollen extract, recombinant (r)Bet v 1, rBet v 2 and rBet v 4 by fluoroenzyme immunoassay using the Pharmacia UniCAPs 100E automated system. Results No correlation was found between specific clinical food sensitivities and incidence of anaphylaxis. No correlation was found between reactivity to particular birch pollen allergens and incidence of anaphylaxis. Likewise, no correlation was found between level of sIgE to birch pollen allergens and incidence of anaphylaxis. Conclusion Measurement of specific IgE to birch pollen allergens could not be used to predict disease severity in OAS. Further work to identify markers for OAS severity is necessary.

↩

[149] - Asero R, Jimeno L, Barber D. Component-resolved diagnosis of plant food allergy by SPT. Eur Ann Allergy Clin Immunol 2008;40:115-121

BACKGROUND: Fruits and vegetables may contain both labile and stable allergens. The former induce only OAS, whereas stable allergens may induce systemic reactions. Component-resolved diagnosis (CRD) of allergy to plant foods is therefore essential for the clinical management of allergic patients. METHODS: 80 adults allergic to plant foods underwent SPT with purified natural date palm profilin (Pho d 2), purified Mal d 1, a peach extract containing uniquely LTP, and with a kiwi extract containing uniquely stable allergens. RESULTS: 58 (72%) patients were monosensitized: 24 to Mal d 1, 24 to profilin, 7 to LTP, and 3 to kiwi. 22 patients were multi-sensitised: 14 to Mal d 1 and profilin, 2 to Mal d 1 and kiwi, 1 to LTP and profilin, 3 to LTP and Mal d 1, and 2 to LTP, Mal d 1 and profilin. Mal d 1 and LTP sensitisation were associated with apple and peach allergy, respectively, whereas profilin sensitisation was associated with allergy to melon, watermelon, banana, tomato and citrus fruits. 18/21 kiwi-allergic patients were sensitised to one of the cross-reacting allergens, but 2/18 reacted to kiwi-specific allergens as well. CONCLUSIONS: In patients with allergy to plant-derived foods CRD can be performed by SPT with purified allergen proteins. In the future, the availability of a larger number of purified natural or recombinant allergens for SPT will represent a simple means to classify food-allergic patients properly on the first visit.

↩

[150] - Rodriguez-Perez R, Fernández-Rivas M, González-Mancebo E, Sánchez-Monge R, Diaz-Perales A, Salcedo G. Peach profilin: cloning, heterologous expression and cross-reactivity with Bet v 2. Allergy 2003;58:635-640

BACKGROUND: Peach is among the main foods causing allergic reactions in the Mediterranean adult population. Only a single peach allergen, named Pru p 3, has been characterized. However, a potential role of profilin has also been suggested in grass pollen-associated allergy to peach . METHODS: Complementary DNA clones for two different peach profilin isoforms were obtained by reverse transcriptase polymerase chain reaction using non-degenerated primers. Expression of recombinant peach profilin was performed in Escherichia coli, and confirmed using rabbit polyclonal antibodies to sunflower pollen profilin. Twenty-nine individual sera from patients with peach allergy proved by double-blind, placebo-controlled food challenges (DBPCFC), either with (n = 15) or without (n = 14) specific IgE to Bet v 2, were used in immunodetection assays to test recombinant peach profilin reactivity . RESULTS: Each peach profilin cDNA included an open reading frame coding for a 131 amino acid protein. The peach profilin isoforms, designated Pru p 4.01 and Pru p 4.02, showed 80% of amino acid sequence identity, and were very similar (>70% identity) to allergenic profilins from plant foods and pollens. Recombinant Pru p 4.01 was expressed in E. coli as a nonfusion protein, displaying the expected molecular size and reacting with anti-profilin antibodies. rPru p 4.01 was recognized by all sera (15 of 15) with specific IgE to Bet v 2, whereas no sera (zero of 14) without IgE to this birch allergen reacted with rPru p 4.01 . CONCLUSIONS: Peach profilin Pru p 4 is very closed to other allergenic profilins from plant foods and pollens. A complete correlation between reactivity to rPru p 4 and rBet v 2 has been found in sera from peach allergic patients.

↩

[151] - Gamboa PM, Sanz ML, Lombardero M, Barber D, Sánchez-Monje R, Goikoetxea MJ et al. Component-resolved in vitro diagnosis in peach-allergic patients. J Investig Allergol Clin Immunol 2009;19:13-20

BACKGROUND:The in vitro diagnosis of pollen-related food allergy presents low specificity and reproducibility with many conventional extracts. This can be improved using natural purified allergens, recombinant purified allergens, or both. OBJECTIVE: We compared specific immunoglobulin (Ig) E determination (slgE), the basophil activation test (BAT), the histamine release test (HRT), and the cellular allergen stimulation test (CAST) using natural and recombinant allergens in the diagnosis of peach allergy. METHODS: Thirty-two peach allergic patients were studied. Skin prick tests were performed with commercial peach and extract with Mal d 1, nPru p 3, and profilin (nPho d 2). slgE, BAT, CAST, and HRT were determined using rPru p 3, rMal d 3, rBet v 1, rMal d 1, and rMal d 4. RESULTS: Agreement between the techniques was good with all the allergens, except HRT with rMal d 1 and rMal d 4. With rPru p 3, slgE, CAST, BAT, and HRT showed sensitivity values of 88%, 81%, 72%, and 69% and specificity values of 100%, 93%, 97%, and 83%, respectively. In patients with systemic symptoms or contact urticaria, the values were 100%, 85%, 81%, and 81%. In patients with oral allergy syndrome, sensitivity to profilins or homologues of Bet v 1 was detected in 100% of the cases by all the techniques, except by HRT with rMal d 1, which detected 66% of the cases. CONCLUSIONS: The use of single allergens in the in vitro diagnosis of peach allergy by specific IgE determination, BAT, and CAST offers high specificity and sensitivity, with better results than the HRT.

↩

[152] - Skypala I, Calderon M, Leeds A, Durham S. Can symptoms or skin prick wheal size predict the outcome of food challenge with apple ? Allergy 2007;62(suppl. 83):147

Apple is one of the commonest foods to cause pollen food syndrome (PFS), a manifestation of Oral Allergy Syndrome (OAS), common in people with springtime hayfever. The aim of this study was to evaluate whether symptom score or skin prick test wheal size for apple could be predictive of the outcome of open or blinded food challenge. Subjects aged 18 and over with symptoms of hay fever from March to May recorded details of any foods they had reported symptoms to and scored those symptoms on a linear scale from 0 to 10. Following this, all subjects had a skin prick test to a fresh Granny Smith apple using the prick by prick method (PPT) and an open and blinded challenge with apple. Apple was the commonest food reported to cause symptoms, affecting 39 of the 119 subjects who completed the study (mean symptom score 4.4). 72% of those affected by apple (28) also had a positive prick test to apple (<3mm). Following open challenge 33/39 (84%) had their symptoms confirmed by a combination of positive PPT and open challenge, with 24 (73%) also having either objective symptoms on open challenge (OJOC) or a positive double-blind challenge (+veBC). Reported symptoms, symptom score and PPT wheal size to apple were all significantly associated with either a +veBC or OJOC. Reported symptoms were also an independently significant predictor of a positive open challenge (p=0.006). There was no correlation between apple symptom score and PPT wheal size. As a predictor of a +veBC or OJOC, reported symptoms had a sensitivity of 75%, specificity of 94% , positive predictive value (PPV) of 86% and negative predictive value (NPV) of 89%. For PPT wheal size the best predictive values came from a wheal size of 4mm with a PPV of 75% and NPV of 88% or 5mm with a PPV of 72% and NPV of 90%. Reported symptoms and PPT wheal size to apple are both associated with a positive challenge result, but only reported symptoms are predictive of a positive challenge result. Reported symptoms have similar sensitivity and specificity to the optimal PPT wheal size but have better PPV and NPV. There was no correlation between the severity of reported symptoms to apple and the size of the PPT wheal for apple.

↩

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

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

↩

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

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

↩

[155] - 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.

↩

[156] - Barber D, de la Torre F, Lombardero M, Antépara I, Colas C, Dávila I et al. Component-resolved diagnosis of pollen allergy based on skin testing with profilin, polcalcin and lipid transfer protein pan-allergens. Clin Exp Allergy 2009;39:1764-1771

Background Allergy diagnosis needs to be improved in patients suffering from pollen polysensitization due to the existence of possible confounding factors in this type of patients. Objective To evaluate new diagnostic strategies by comparing skin responses to pan-allergens and conventional allergenic extracts with specific IgE (sIgE) to purified allergen molecules. Methods One thousand three hundred and twenty-nine pollen-allergic patients were diagnosed by a combination of an in vitro method with a panel of 13 purified allergens, including major allergens and pan-allergens, using a high-capacity screening technology (ADVIA-Centaurs) and skin prick test (SPT) to pan-allergens and conventional extracts. Results There was a high concordance (k index) between in vitro (sIgE to major allergens) and in vivo (SPT to conventional extracts) methods in patients who were not sensitized to panallergens, but SPT with conventional extracts failed to diagnose patients with sensitization to pan-allergens. In patients who were simultaneously sensitized to polcalcins and profilins, there was a duplication both in the number of sensitizations to major allergens and in the years of disease evolution. There was a statistical association between sensitization to profilins and/or lipid transfer proteins and food allergy (Po0.0001). Conclusion The novel diagnostic strategy has proven to be a valuable tool in daily clinical practice. Introduction of routine SPT to pan-allergens is a simple and feasible way of improving diagnostic efficacy. Patients sensitized to pan-allergens should be tested by an adequate panel of allergenic molecules in order to identify the allergens that are responsible for the allergic disease.

↩

[157] - Dreborg S, Foucard T. Allergy to apple, carrot and potato in children with birch pollen allergy. Allergy 1983;38:167-172

Skin sensitivity to apple, carrot and potato, clinically related to birch pollinosis was investigated. Different skin test techniques using fresh fruit were compared. A simple prick test (SPT) technique with a lancet piercing the apple peel just before pricking the skin was shown to be the most practical and to give reproducible results. The allergenic activity in apple was found to be heat labile and deteriorated during storage at room temperature. Apple, carrot, potato, hazelnut and birch reactivity was transferable in Prausnitz-Kustner test like IgE antibodies. SPT reactivity to fresh material from apple, carrot and potato was investigated in 174 children of whom 128 suffered from pollen allergy. Positive SPT results were obtained almost exclusively in children who were SPT positive to a birch pollen extract. Children who noticed clinical symptoms when eating apple, raw carrot or potato were found to have a significantly larger SPT reaction than children with a negative history.

↩

[158] - 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.

↩

[159] - Kleine-Tebbe J, Wangorsch A, Vogel L, Crowell DN, Haustein UF, Vieths S. Severe oral allergy syndrome and anaphylactic reactions caused by a Bet v 1-related PR-10 protein in soybean, SAM22. J Allergy Clin Immunol 2002;110:797-804

BACKGROUND: Anaphylactic reactions to soy products have been attributed to stable class 1 food allergens . OBJECTIVE: IgE- mediated reactions to a soy-containing dietary food product in patients allergic to birch pollen were investigated . METHODS: Detailed case histories were taken from 20 patients. Their sera were analyzed for IgE (UniCAP) specific for birch, grass, mugwort, the recombinant birch allergens rBet v 1 and rBet v2, and soy protein. Extracts from birch pollen, soy isolate, rBet v 1, and the recombinant PR-10 soy protein rSAM22 were coupled to paper disks or nitrocellulose for IgE measurements (enzyme allergosorbent test) or Western blot analysis. Enzyme allergosorbent testing, Western blot inhibition, and histamine release studies were performed with the same allergens . RESULTS: Most patients (17/20) experienced facial, oropharyngeal, and/or systemic allergic symptoms within 20 minutes after ingesting the soy product for the first time. Birch pollen allergy (16/20) was common, along with oral allergy syndrome to apple (12/20) or hazelnut (11/20). IgE levels to birch and Bet v 1 but not to other inhalants were high in 18 of 20 patients. Significant IgE binding to rSAM22 occurred in 17 of 20 patients. Blot experiments with the soy isolate revealed IgE-binding bands at 17 kd (15/20), 22 kd (1/20), and 35 to 38 kd (2/20); the former was inhibited by preincubation of the sera with rBet v 1 or rSAM22. Birch extract and soy isolate, rBet v 1, and rSAM22 induced dose-dependent histamine release in the nanomolar range . CONCLUSION: Immediate-type allergic symptoms in patients with birch pollen allergy after ingestion of soy protein-containing food items can result from cross-reactivity of Bet v 1 -specific IgE to homologous pathogenesis-related proteins, particularly the PR-10 protein SAM22.

↩

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

↩

[161] - 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.

↩

[162] - 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.

↩

[163] - 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.

↩

[164] - Mittag D, Vieths S, Vogel L, Wagner-Loew D, Starke A, Hunziker P, et al. Birch pollen-related food allergy to legumes: identification and characterization of the Bet v 1 homologue in mungbean (Vigna radiata), Vig r 1. Clin Exp Allergy 2005;35:1049-1055

BACKGROUND: Recently allergic reactions to legumes mediated by Bet v 1-homologous food allergens were described for soy and peanut. In this study we assessed allergic reactions to another legume, to mungbean seedlings, and identified its Bet v 1-homologous allergen Vig r 1 . METHODS: Ten patients were selected who had a history of allergic reactions to mungbean seedlings and a respiratory allergy to birch pollen. The Bet v 1 homologue in mungbean seedlings, Vig r 1, was cloned by a PCR strategy, expressed in Escherichia coli, and purified by preparative SDS-PAGE. In all sera, specific IgE against birch pollen, Bet v 1, Bet v 2, Vig r 1, and the Bet v 1 homologues in soy (Gly m 4) and cherry (Pru av 1) was determined by CAP-FEIA. Cross-reactivity of specific IgE with Vig r 1, Bet v 1, Gly m 4, and Pru av 1 was assessed by immunoblot inhibition. Expression of Vig r 1 during development of mungbean seedlings and under wounding stress was analysed by immunoblotting. The Vig r 1 double band was analysed by matrix-assisted laser desorption/ionization time-of-flight and liquid chromatography/tandem mass spectrometry (LC/MS/MS) . RESULTS: All patients were sensitized to birch pollen and Bet v 1, 20% to Bet v 2, and 90% to Gly m 4. Seventy percent of the patients showed IgE binding to a double band at 15 kDa in mungbean extract that was inhibited after pre-incubation of sera with rBet v 1. PCR cloning revealed that the mungbean homologue of Bet v 1 had a molecular weight of 16.2 kDa, a calculated pI of 4.6% and 42.8% amino acid sequence identity with Bet v 1. MS analysis confirmed similarity of the double band with the deduced Vig r 1 sequence, but also indicated the existence of other Vig r 1 isoforms. ImmunoCAP analysis detected IgE against Vig r 1 in 80% of the sera. IgE binding to Vig r 1 was inhibited with Gly m 4 in six of six and with rPru av 1 in four of six patients. Vig r 1 expression occurred during development of seedlings and was increased by wounding stress . CONCLUSIONS: Food allergy to mungbean seedlings can be caused by primary sensitization to birch pollen and is mediated by Vig r 1 in the majority of the patients with birch pollen-related allergy to mungbean seedlings.

↩

[165] - Wensing M, Akkerdaas JH, van Leeuwen A, Stapel SO, Bruijnzeel-Koomen CAFM, Aalberse RC, et al. IgE to Bet v 1 and profilin: Crossreactivity patterns and clinical relevance. J Allergy Clin Immunol 2002;110:435-442

BACKGROUND: Individuals with pollen allergy often have IgE against plant-derived foods. This can be due to cross-reactive IgE against Bet v 1 and homologues, profilins, and/or cross-reactive carbohydrate determinants . OBJECTIVE: The aim of this study was to correlate sensitization to Bet v 1 and profilin with individual recognition patterns to plant foods and clinical relevance . METHODS: Fifty-two patients with pollen allergy and IgE against at least one plant-derived food were included in the study. Adverse reactions to plant-derived foods were documented by using standardized interviews. Skin prick tests were performed for pollen (grass, birch, and mugwort) and 14 plant-derived foods. In addition, recombinant (r) Bet v 1 and rBet v 2 (profilin) were tested intracutaneously. Specific IgE against the abovementioned allergens were determined by means of RAST. Cross-reactivity was studied by means of RAST inhibition . RESULTS: Eighty-five percent of patients were sensitized to Bet v 1, and 71% were sensitized to profilin. Profilin was associated with a higher number of positive RAST results to plant-derived foods than Bet v 1. In contrast, Bet v 1 was associated with more positive skin prick test responses and more food-related symptoms. Sensitization to Bet v 1 was associated with IgE against apple, hazelnut, and peach, whereas sensitization to profilin was associated with positive RAST results to all investigated plant-derived foods except apple, peach, and melon . CONCLUSIONS: IgE antibodies against Bet v 1 have a more limited spectrum of cross-reactivity than those against profilin, but they frequently give rise to clinically relevant cross-reactivities to food. In analogy to anticarbohydrate IgE, cross-reactive IgE against food profilins have no or very limited clinical relevance.

↩

[166] - Wensing M, Penninks AH, Hefle SL, Akkerdaas JH, van Ree R, Koppelman SJ, et al. The range of minimum provoking doses in hazelnut-allergic patients as determined by double-blind, placebo-controlled food challenges. Clin Exp Allergy 2002;32:1757-1762

Background The risk for allergic reactions depends on the sensitivity of individuals and the quantities of offending food ingested. The sensitivity varies among allergic individuals, as does the threshold dose of a food allergen capable of inducing an allergic reaction. Objective : This study aimed at determining the distribution of minimum provoking doses of hazelnut in a hazelnut-allergic population. Method s : Thirty-one patients with a history of hazelnut-related allergic symptoms, a positive skin prick test to hazelnut and/or an elevated specific IgE level, were included. Double-blind, placebo-controlled food challenges (DBPCFC) were performed with seven increasing doses of dried hazelnut (1 mg to 1 g hazelnut protein) randomly interspersed with seven placebo doses. Result s : Twenty-nine patients had a positive challenge. Itching of the oral cavity and/or lips was the first symptom in all cases. Additional gastrointestinal symptoms were reported in five patients and difficulty in swallowing in one patient. Lip swelling was observed in two patients, followed by generalized urticaria in one of these. Threshold doses for eliciting subjective reactions varied from a dose of 1 mg up to 100 mg hazelnut protein (equivalent to 6.4-640 mg hazelnut meal). Extrapolation of the dose-response curve showed that 50% of our hazelnut-allergic population will suffer from an allergic reaction after ingestion of 6 mg (95% CI, 2-11 mg) of hazelnut protein. Objective symptoms were observed in two patients after 1 and 1000 mg, respectively. Conclusion : DBPCFCs demonstrated threshold doses in half of the hazelnut-allergic patients similar to doses previously described to be hidden in consumer products. This stresses the need for careful labelling and strategies to prevent and detect contamination of food products with hazelnut residues.

↩

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

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

↩

[168] - Jung P, Sesztak-Greinecker G, Wantke F, Goetz M, Jarisch R, Hemmer W. Prevalence of cross-sensitisation to soy allergens in patients with birch pollen allergy and allergenicity of different soy products. EAACI 25th Congress, Vienna, 10-14 June, 2006, Poster n°1530

Background: Soybean Gly m 4, a member of the PR-10 protein family, has been recognized as a cross-reactive food allergen in birch pollen-related food hypersensitivity with the potential to elicit severe systemic reactions. We investigated the prevalence of cross-sensitisation to soybean and the allergenicity of various soy-based food items in patients with birch pollen allergy. Method: A commercial brand of soy milk was added to our routine skin prick test panels and tested in consecutive patients with suspect inhalant or food allergy. Consumption habits concerning soy products and eventual adverse reactions were recorded by questionnaire. Additional prick-to-prick testing with different soy products was done in selected patients. Results: Among 292 patients with a positive skin prick test to birch pollen, 72 (24.7%) reacted to soy milk but only 18 (6.2%) reacted to a commercial soy skin prick test. 34/97 (35%) of birch pollen-allergic patients reported to have knowingly consumed soy milk before with 11/34 (31%) of them having experienced side effects. Mild reactions to tofu and soybean sprouts were occasionally reported. Prick-to-prick testing with different soy products in 16 patients with a positive skin test to soy milk revealed positive reactions to raw and cooked soybean sprouts (94%/50%), raw and cooked tofu (87%/40%), soy dessert (86%), and soy joghurt (21%). No significant differences in skin test responses were seen between seven different brands of soy milk (protein content 3.0-3.7g/100ml). Skin tests remained positive even after boiling soy milk für 5, 10 and 30 minutes. Conclusions: Cross-sensitisation to soy is frequent among patients with birch pollen allergy and many soy-based foods retain considerable allergenicity. As soy products are becoming increasingly popular on the market, birch pollen-allergic patients may be at growing risk of experiencing allergic reactions to these products.

↩

[169] - 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.

↩

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

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

↩

[171] - 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) .

↩

[172] - 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.

↩

[173] - 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.

↩

[174] - 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.

↩

[175] - 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.

↩

[176] - 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.

↩

[177] - 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.

↩

[178] - 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.

↩

[179] - 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.

↩

[180] - Minciullo PL, Patafi M, Marotta G, Tigano V, Barresi L, Ferlazzo B, et al. Platanus acerifolia co-sensitivity in patients with parietaria judaica allergy and oral allergic syndrome. Allergy Clin Immunol Int 2005;17(Suppl. 1):250

Plane tree pollen allergy is a clinical disorder affecting particularly populations of the United States and Western Europe, but little is known about its relevant allergens. Pla a 1 and Pla a 2 are known as major allergens of Platanus acerifolia. Recently, a new allergen, called Pla a 3 has been characterized as major allergen in plane pollen allergic patients with concomitant allergy to vegetables and/or fruits in the Mediterranean area. It is a lipid transfer protein (LTP) that cross-reacts with food LTPs, in particular with lettuce, cherry and hazelnut LTP. Other previous studies described an association between the pollen of the Platanus tree and hazelnut, peach, apple, banana, peanut, celery, maize and chickpea. This association could be explained by the in vitro IgE crossreactivity detected. No data are present about a possible cross-reactivity between plane tree pollen and other plant or thee pollens. During the period between January 2004 and November 2004, 14 patients, referred to our Operative Unit for seasonal rhinitis and/or asthma symptoms, showed positivity to skin prick test to plane tree. All patients showed co-sensitivity to other pollens and in particular 13 out of 14 patients presented skin positivity to Parietaria judaica, the most important pollen in our area. Other important co-sensitivities were with Olea europea (10 patients) and grass pollen (7 patients). No monosensitivity to platanus was found. Therefore, platanus sensitivity in our area seems no to be so important and might be related to parietaria pollen. Moreover, 5 out of 14 patients with platanus sensitivity, referred an oral allergic syndrome (OAS) and showed skin positivity for vegetables and/or fruits. All of 5 patients were sensitive to peach, 4 to apple, 4 to hazelnut, 4 to almond, 4 to beet, 3 to peanut and 3 to walnut. Since in the Mediterranean area OAS occurs independently of an associated birch pollinosis and since platanus pollinosis in our area seems no to play an important role, it may be hypothesised that the skin positivity to platanus is related to fruit LTP in patients with OAS. The role of parietaria allergens in cross-reactivity with platanus and fruit LTP should be investigate.

↩

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

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

↩

[182] - 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.

↩

[183] - 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.

↩

[184] - 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.

↩

[185] - 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

↩

[186] - 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?

↩

[187] - 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.

↩

[188] - 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.

↩

[189] - 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.

↩

[190] - 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.

↩

[191] - 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.

↩

Une même allergie avec différentes étiologies

Une chronologie variable des sensibilisations

Une allergie dont la sévérité est variable selon les pays

Un même groupe d’aliments et une allergénicité variable

L’allergie aux Rosacées due aux LTP : une allergie sans pollinose ?

Vers la fin du dogme LTP = pays méditerranéens ?

Diagnostic étiologique d’une allergie aux fruits des Rosacées

Rubriques

Dans la même rubrique

La cerise et autres fruits des Rosacées

La pêche

La pomme

	Pays-Bas	Autriche	Italie	Espagne
Pollinose	97%	96%	96%	88%
Pollen dominant	bouleau			graminées
allergie pêche	71%	70%	66%	90%
allergie noisette	74%	73%	58%	21%
âge début pêche vs pollinose		après	après	avant