Les Apiacées : généralités

	LTP	PR10	Profiline	Autres
Anis vert		Pim a 1	Pim a 2
Carotte	LTP (contesté)	Dau c 1	Dau c 4	Cyclophiline (20kDa)
Céleri	Api g 2	Api g 1	Api g 4	Api g 3 = chlorophyll Ab-binding protein Api g 5 (58kD) = FAD-containing oxidase
Coriandre		Cor s 1	Cor s 2
Cumin		Cum c 1	Cum c 2
Fenouil	LTP ? ?	Foe v 1	Foe v 2
Persil	LTP	Pet c 1	Pet c 2	RubisCO

[1] - Pauli G, Bessot JC, Dietemann-Molard A, Braun PA, Thierry R. Celery sensitivity: clinical and immunological correlations with pollen allergy. Clin Allergy 1985;15:273-279

The authors studied twenty patients with celery allergy and concomitant hypersensitivity to certain pollens (mugwort, birch). The specific symptoms induced by eating celery were attacks of urticaria and angio oedema (seventeen out of twenty) respiratory complaints (eight out of twenty), systemic anaphylaxis with vascular collapse (three out of twenty). A strong association between clinical reactions to celery and mugwort sensitization, and to a lesser degree between celery allergy and birch pollen sensitization was established. Celery allergy is mediated by IgE antibodies and can be easily diagnosed by cutaneous tests using fresh material and/or by adequate RAST test. RAST inhibitions performed on individual sera suggest the existence of common antigens in celery and mugwort, and in celery and birch pollen. However, the exact nature of these common antigens has not yet been determined.

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[2] - Wüthrich B, Hofer T. [Food allergy: the celery-mugwort-spice syndrome. Association with mango allergy ?]. Dtsch Med Wochenschr 1984;109:981-986

Celery allergies are commonly observed in mugwort pollen-allergic patients. This situation is frequently associated with an allergy to spices of the umbelliferous family. Thus a "celery-mugwort-spice syndrome" has been established. In 31 patients (27 females, 4 males) allergy to celery was diagnosed between 1978 and 1982. They were followed up in 1983 and complementary tests (skin and RAST) with various raw vegetables, spices and mugwort pollen were done. For specific diagnosis of allergy to celery skin testing (scratch or prick) with fresh celery bulb was reliable. In contrast, RAST was not sufficiently sensitive, 87% of patients allergic to celery had pollinosis in the form of mugwort pollen sensitisation established by skin testing and RAST. Allergy to carrots was clinically and by testing currently associated in 52%. In contrast, a clinically relevant hypersensitivity to caraway (26%), parsley (16%), fennel (13%), green pepper (10%) and aniseed (3%) was found less frequently although these allergens showed a much larger positivity in skin testing. This signifies group sensitisation within the umbelliferous plants. The association of celery-mugwort allergy with allergy to mango fruit described here for the first time cannot be explained as group sensitisation within a botanic relation, as mango, and pistachio-nut, belong to the family of anacardiaceae.

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[3] - Vallier P, Dechamp C, Vial O, Deviller P. A study of allergens in celery with cross-sensitivity to mugwort and birch pollens. Clin Allergy 1988;18:491-500

Sixty-one sera with positive RAST to mugwort pollen (Artemisiae vulgaris) were submitted to RASTs for birch pollen (Betula verrucosa) and celery (Apium graveolens). In 36 cases RAST results were positive for celery. In addition, 23 sera presented specific IgE to birch pollen. The binding of specific IgE to individual allergens in celery, mugwort pollen and birch pollen was studied by the immunoblotting technique. This involved electrophoretic separation of allergenic extracts, electrotransfer of proteins onto nitrocellulose sheets and sensitive immunoenzymatic detection. Eighteen sera had specific IgE binding to two celery components of molecular weight around 15 kD. All these sera also detected a 15 kD allergen in mugwort and two allergens in birch of 14 kD and 16 kD molecular weight. The sera that did not detect the 15 kD bands in celery failed to react with both the 15 kD mugwort component and the 14 and 16 kD birch components. Specific cross-inhibitions of the detection of these allergens on immunoblots were obtained by pre-incubation of the sera with crude extract of the three species. These results strongly suggest that such allergens display some structural identity and that they could be at the origin of some cases of crossed hypersensitivity to celery, mugwort pollen and birch pollen

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[4] - Helbling A, Lopez M, Schwartz HJ, Lehrer SB. Reactivity of carrot-specific IgE antibodies with celery, apiaceous spices, and birch pollen. Ann Allergy 1993;70:495-499

Although carrot allergy is not well recognized in North America, the celery-carrot-mugwort-spice syndrome is well known in Europe. In the current study, cross reactivity of carrot, stalk celery, and spices, all members of the Apiaceae and birch pollen was assessed with serum from a patient reporting raw carrot-induced, raw stalk celery-induced, or spice-induced laryngeal edema and bronchospasm. By RAST inhibition, some cross-reactivity was demonstrated between raw carrot and stalk celery and other members of the Apiaceae. Immunoprint inhibition revealed common allergic epitopes on a 17-kD band shared by carrot, celery, and birch pollen. The results suggest that subjects sensitized to carrot may also have allergic reaction to other vegetables of spices of the Apiaceous family. Furthermore, carrot hypersensitivity can be associated with birch pollen allergy.

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[5] - Halmepuro L, Lowenstein H. Immunological investigation of possible structural similarities between pollen antigens and antigens in apple, carrot and celery tuber. Allergy 1985;40:264-272

Crossed line immunoelectrophoresis (CLIE) was used to show that a minor allergen present in birch pollen and another present in timothy pollen shared common epitopes with antigens in apple, carrot and celery tuber. Major pollen allergens were not involved. Structural similarities were also noticed in some mugwort pollen antigens and antigens in apple, carrot and celery, but none of these mugwort antigens acted as an allergen. In crossed immunoelectrophoresis (CIE) of samples of apple, carrot or celery extracts towards antibodies against birch pollen, grass pollen or mugwort pollen, four to 14 distinct precipitates were observed and shown to be specific. In crossed radioimmunoelectrophoresis (CRIE) none of the precipitates could be shown to act as an allergen. Using allergen detection on nitrocellulose paper, the apple, carrot and celery extracts were shown to possess IgE-binding capacity.

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[6] - Wüthrich B, Hofer T. [Food allergy: the celery-mugwort-spice syndrome. Association with mango allergy ?]. Dtsch Med Wochenschr 1984;109:981-986

Celery allergies are commonly observed in mugwort pollen-allergic patients. This situation is frequently associated with an allergy to spices of the umbelliferous family. Thus a "celery-mugwort-spice syndrome" has been established. In 31 patients (27 females, 4 males) allergy to celery was diagnosed between 1978 and 1982. They were followed up in 1983 and complementary tests (skin and RAST) with various raw vegetables, spices and mugwort pollen were done. For specific diagnosis of allergy to celery skin testing (scratch or prick) with fresh celery bulb was reliable. In contrast, RAST was not sufficiently sensitive, 87% of patients allergic to celery had pollinosis in the form of mugwort pollen sensitisation established by skin testing and RAST. Allergy to carrots was clinically and by testing currently associated in 52%. In contrast, a clinically relevant hypersensitivity to caraway (26%), parsley (16%), fennel (13%), green pepper (10%) and aniseed (3%) was found less frequently although these allergens showed a much larger positivity in skin testing. This signifies group sensitisation within the umbelliferous plants. The association of celery-mugwort allergy with allergy to mango fruit described here for the first time cannot be explained as group sensitisation within a botanic relation, as mango, and pistachio-nut, belong to the family of anacardiaceae.

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[7] - Gomez M, Curiel G, Mendez J, Rodriguez M, Moneo I. Hypersensitivity to carrot associated with specific IgE to grass and tree pollens. Allergy 1996;51:425-429

This study deals with a 34-year-old female cook with no previous history of atopy, who was studied because of allergic rhinoconjunctivitis and contact urticaria in both hands associated with severe itching when she handled raw carrot. The patient had had anaphylactic episodes after accidental ingestion of raw carrots, but she tolerated cooked carrots. Skin prick tests with carrot, celery, and olive, and birch, grass, and mugwort pollens were positive. Total IgE was 411 UI/ml. Specific IgE to olive, grass, and weed pollens were 10.92, 6.17, and 2.4 AU/ml, respectively. The histamine release test was positive for carrot, celery, celeriac, and olive pollen up to a dilution of 1/10(6). Immunoblot of raw carrot showed a single IgE-binding 18-kDa band. IgE reactivity for raw carrot immunoblot was completely inhibited by carrot and by celery, but not by olive or grass pollens. Specific IgE to olive pollen was not inhibited by carrot. The existence of monosensitization to an 18-kDa protein in carrot and specific IgE to olive pollen has not been reported in the celery-carrot-mugwort-spice syndrome.

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[8] - Jensen-Jarolim E, Leitner A, Hirschwehr R, Kraft D, Wüthrich B, Scheiner O, et al. Characterization of allergens in Apiaceae spices: anise, fennel, coriander and cumin. Clin Exp Allergy 1997;27:1299-1306

BACKGROUND: Symptoms elicited by IgE-mediated food allergy range from mild local to severe systemic reactions. Allergens in spices are particularly dangerous due to their hidden presence in many dishes. OBJECTIVES AND METHODS: According to clinical observations, mugwort and birch pollen allergy, and hypersensitivity to spices are frequently associated, but the crossreacting compounds were not defined so far. We tested sera of 15 patients who experienced adverse reactions to spiced food and characterized their IgE-binding patterns on anise, fennel, coriander and cumin extracts through immunoblot and inhibition experiments. RESULTS: The use of anti-Bet v 1 (MoAb) and anti-profilin (rabbit) antibodies revealed the presence of crossreacting allergens in the tested spice extracts. Inhibition experiments showed that IgE-binding to allergens in Apiaceae spices could be blocked by preincubation of sera with rBet v 1 or rBet v 2 (birch profilin). Moreover, we detected crossreacting allergenic molecules in the molecular weight range of 60 kDa. IgE-binding to spice allergens occurred only with sera of 10/15 (66%) patients with allergy to pollen (birch, mugwort) and/or celeriac. In five out of 15 (33%) patients with a history of adverse reaction to spices, but without pollen and celeriac allergy, no IgE-binding to any spice protein could be demonstrated. It is possible that these clinical reactions could be elicited by other types of hypersensitivity (Type II, III, IV), however, as spices contain highly reactive substances, the symptoms may most likely be classified as food-intolerant. CONCLUSIONS: Bet v 1- and profilin-related allergens may, besides higher molecular weight allergenic molecules, be responsible for Type I allergy to anise, fennel, coriander or cumin, members of the Apiaceae.

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[9] - Schwartz HJ, Jones RT, Rojas AR, Squillace DL, Yunginger JW. Occupational allergic rhinoconjunctivitis and asthma due to fennel seed. Ann Allergy Asthma Immunol 1998;78:37-40

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[10] - Moneret-Vautrin DA, Morisset M, Lemerdy P, Croizier A, Kanny G. Food allergy and IgE sensitization caused by spices: CICBAA data (based on 589 cases of food allergy). Allerg Immunol (Paris) 2002;34:135-140

BACKGROUND: Spices originate in various botanical families: Apiaceae, Lamiaceae, Lauraceae, Leguminosae, Liliaceae, Myristicaceae, Myrtaceae, Piperaceae, Solanaceae, Zingiberaceae.... METHODOLOGY: Prick-tests to native spices have been carried out in patients suspected of food allergies to spices. The CICBAA data bank includes 589 cases of food allergies, a part of which has benefited from investigations for spices. Data about the rate of sensitization and food allergy are available. RESULTS: Frequent sensitization to Apiaceae is observed: coriander, caraway, fennel, celery: 32% of prick-tests in children, 23% of prick-tests in adults. Sensitization to Liliaceae: garlic, onion, chive, is observed in 4.6% of prick-tests in children, 7.7% of prick-tests in adults. Rare cases of sensitization to paprika and saffron are recorded. Prick-tests to nutmeg, ginger and clove are currently negative. 10 food allergies related to the mugwort-celery-spices syndrome are reported: coriander: 1, caraway: 2, fennel: 3, garlic: 3, onion: 1. Food allergy to spices is unfrequent: 2% of the totality of food allergies. However, only adults are allergic to spices and allergy to spices accounts for 6.4% of food allergies in adults. Tiny amount of proteins are usually ingested. Patients at risk of spice allergy are young adults sensitized to mugwort and birch allergens, sharing cross-sensitization with various food vegetal allergens. The clinical suspicion raises from frequent post-prandial systemic reactions. Other allergens of vegetal origin have to be cleared. Diagnosis can be established by DBPCFC using powdered spices in capsules. [References: 22]

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[11] - Manzanedo L, Blanco J, Fuentes M, Caballero ML, Moneo I. Anaphylactic reaction in a patient sensitized to coriander seed. Allergy 2004;59:362-363

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[12] - Freeman GL. Allergy to fresh dill. Allergy 1999;54:531-532

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[13] - Ebo DG, Bridts CH, Mertens MH, Stevens WJ. Coriander anaphylaxis in a spice grinder with undetected occupational allergy. Acta Clin Belg 2006;61:152-156

BACKGROUND: Anaphylaxis after ingestion of spices as a result from occupational sensitization remains anecdotal. We describe a patient, working in a spice factory, with anaphylaxis from coriander in a meal. He also demonstrated urticaria, angio-edema, rhinoconjunctivitis and bronchospasm during handling coriander and fenugreek. OBJECTIVE: To determine the mechanism of the anaphylactic reaction and to evaluate cross-reactivity between both botanically unrelated spices. METHODS: Investigations comprised quantification of total and specific IgE by Immuno-CAP FEIA, skin testing, basophil activation experiments by flow-assisted determination of CD63 expression in the patient and 3 healthy controls. Immuno-CAP inhibition experiments were applied to investigate cross-reactivity. RESULTS: Specific IgE, skin tests and basophil activation tests were clearly positive in the patient, whereas they remained negative in controls. No cross-reactivity between fenugreek and coriander was demonstrable by inhibition experiments. CONCLUSIONS: The clinical manifestations in temporal relationship to ingestion of coriander and handling of coriander and/or fenugreek, the positive specific IgE results, skin tests and basophil activation assays support the diagnosis of allergy to both spices. History suggests sensitization by occupational exposure.

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[14] - Schöll I, Jensen-Jarolim E. Allergenic Potency of Spices: Hot, Medium Hot, or Very Hot. Int Arch Allergy Immunol 2004;135:247-261

Spices are the most attractive ingredients to confer an authentic taste to food. As they are derived from plants, they harbour allergenic potency and can induce symptoms ranging from mild local to severe systemic reactions. Due to the content of pharmacologically active substances of spices, the diagnosis of allergy and the differentiation from intolerance reactions may be difficult. Association with inhalative allergies via IgE cross-reactivity, but also direct gastrointestinal sensitization plays a role. This article is a botanical and allergological overview of the most important spices and molecules responsible for eliciting IgE-mediated reactions or cross-reactions. As no curative treatments are known at present, strict avoidance is recommended and, therefore, accurate labelling of pre-packed food is necessary.

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[15] - 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.

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[16] - 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.

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[17] - Cordobés-Durán C, García-Menaya JM, Lombardero M, Ledesma A, Bobadilla P. Detection of a 12-Kilodalton Lipid Transfer Protein Allergen in Parsley. J Investig Allergol Clin Immunol 2007;17:282-283

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[18] - de Lacoste-Delaval A, Guérin L, Leduc V. Allergie alimentaire à la salade. Rev Fr Allergol Immunol Clin 2006;46:374

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[19] - Pauli G, Bessot JC, Braun PA, Dietemann-Molard A, Kopferschmitt-Kubler MC, Thierry R. Celery allergy: clinical and biological study of 20 cases. Ann Allergy 1988;60:243-246

In 20 patients, the ingestion of celery was responsible for mucocutaneous symptoms (generalized urticaria and angioedema) (18/20) and respiratory disorders (7/20). Four cases of systemic anaphylaxis were observed. The main associated allergic disorder was pollinosis (16/20). Food allergy to other vegetable products, mainly other Umbelliferae and apples, coexisted with celery allergy in 12 cases. It was found that celery allergy is mediated by IgE antibodies: it is easily diagnosed by skin tests (fresh extracts of celery may be used) and by adequate RAST (17 positive results). Cosensitization with mugwort pollen (14 cases) and birch pollen (9 cases) was found. Celery allergens responsible for clinical sensitization originate chiefly in the tuber and are at least partly thermally labile. The frequent association with pollen sensitization suggests the existence of common antigenic epitopes in celery extracts and mugwort and birch pollens. The immunologic investigations carried out so far (RAST inhibition and immunoprint) seem to support this hypothesis.

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[20] - Wüthrich B, Hofer T. [Food allergy: the celery-mugwort-spice syndrome. Association with mango allergy ?]. Dtsch Med Wochenschr 1984;109:981-986

Celery allergies are commonly observed in mugwort pollen-allergic patients. This situation is frequently associated with an allergy to spices of the umbelliferous family. Thus a "celery-mugwort-spice syndrome" has been established. In 31 patients (27 females, 4 males) allergy to celery was diagnosed between 1978 and 1982. They were followed up in 1983 and complementary tests (skin and RAST) with various raw vegetables, spices and mugwort pollen were done. For specific diagnosis of allergy to celery skin testing (scratch or prick) with fresh celery bulb was reliable. In contrast, RAST was not sufficiently sensitive, 87% of patients allergic to celery had pollinosis in the form of mugwort pollen sensitisation established by skin testing and RAST. Allergy to carrots was clinically and by testing currently associated in 52%. In contrast, a clinically relevant hypersensitivity to caraway (26%), parsley (16%), fennel (13%), green pepper (10%) and aniseed (3%) was found less frequently although these allergens showed a much larger positivity in skin testing. This signifies group sensitisation within the umbelliferous plants. The association of celery-mugwort allergy with allergy to mango fruit described here for the first time cannot be explained as group sensitisation within a botanic relation, as mango, and pistachio-nut, belong to the family of anacardiaceae.

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[21] - Schiappoli M, Senna G, Dama A, Bonadonna P, Crivellaro M, Passalacqua G. Anaphylaxis due to carrot as hidden food allergen. Allergol Immunopathol (Madr) 2002;30:243-244

Carrot is frequently involved in food allergies and oral allergy syndromes, usually in association with other foods. Nevertheless, carrot alone is rarely responsible for severe systemic reactions. We report a case of anaphylactic shock due to the inadvertent ingestion of carrot as a hidden allergen contained in an ice-cream. The etiological role of carrot in provoking the symptoms was thoroughly ascertained through appropriate in vivo and in vitro assays and by excluding, through double-blind placebo-controlled challenge, the involvement of other ingredients of the food.We highlight once again the harm and risks due to hidden food allergens contained in commercial preparations and the importance of thorough patient education and information.

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[22] - Moreno-Ancillo A, Gil-Adrados A, Dominguez-Noche C, Cosmes P, Pineda F. Occupational asthma due to carrot in a cook. Allergol Immunopathol (Madr) 2005;33:288-290

Background: Few previous reports of carrot-induced asthma have been confirmed by objective tests. Hypersensitivity to carrot is frequently associated with allergy to Apiaceae spices and sensitization to birch and mugwort pollens. Clinical case: A 40-year-old cook woman was seen with sneezing, rhinorrhea, contact urticaria and wheezing within few minutes of handling or cutting raw carrots. She needed to leave out of the kitchen while the other cooks cut raw carrots. Methods and results: Skin tests were positive to carrot, celery, aniseed and fennel. Rubbing test with fresh carrot was positive. Specific IgE to carrot was 4.44 kU/L. Determinations of specific IgE to mugwort, grass and birch pollens were negative. Inhalative provocation test, performed as a handling test, was positive. The IgE-immunoblotting showed two bands in carrot extract: a band with apparent molecular weight of 30 kd and other band of 18 kd. This band of 18 kd was Dau c 1. The band of 30 kd could correspond a phenylcoumaran benzylic ether reductase. Dau c 1 did not appear to be the unique allergen in this case. Additional allergens may induce the sensitization. Primary sensitization due to airborne allergens of foods and the lack of pollen allergy in this patient are notorious events.

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[23] - Gonzalez-Gutierrez ML, Sanchez-Fernandez C, Esteban-Lopez MI, Sempere-Ortells JM, Diaz-Alperi P. Allergy to anis. Allergy 2000;55:195-196

This report describes the case of an allergic reaction in a 32 yr old aniseed liqueur factory worker. Symptoms appeared after the handling of several plants used in the manufacture of anisette (liquorice (Glycyrrhiza glabra), mace (Myristica fragrans), coriander (Coriandrum sativum), iris root, cinnamon (Canellal cinnamomum), orange blossom flower (Citrus sinensis) and 2 types of aniseed, matalahuga aniseed (Pimpinella anisum) and star-shaped aniseed (Anisum stellatum)). The worker was able to drink anisette and eat liquorice stick without any clinical symptoms. Skin prick tests (SPT) were positive for liquorice, mace, matalahuga aniseed, coriander and iris root extracts. The patient suffered from rhinitis and asthma at his workplace due to inhalation of dust particles from these plants. However, the symptoms appeared mainly while he was handling liquorice, and SPT and RAST values for liquorice were significantly higher than for the other spices, suggesting that liquorice was the main source of the allergic reaction.

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[24] - Garcia-Gonzalez JJ, Bartolomé-Zavala B, Fernandez-Meléndez S, Barcelo-Muñoz JM, Miranda Paez A, Carmona-Bueno MJ, et al. Occupational rhinoconjunctivitis and food allergy because of aniseed sensitization. Ann Allergy Asthma Immunol 2002;88:518-522

BACKGROUND: Aniseed is a spice frequently used in Mediterranean cooking and, as with other Umbelliferae, it has been involved in clinical allergy. OBJECTIVE: This investigation was undertaken to study the allergens implicated in a case of occupational allergy to aniseed associated with rhinoconjunctivitis and gastrointestinal symptoms. METHODS: Skin prick tests were performed to inhalant allergens, spices used in the patient's workplace (aniseed and cinnamon), and 12 other Umbelliferae spices, birch, and mugwort. A nasal challenge test to aniseed and cinnamon and a double-blind placebo-controlled oral food challenge test to aniseed were also performed. The molecular weights of the allergens were studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis immunoblotting and cross-reactivity among Umbelliferae species by enzyme immunoassay inhibition. RESULTS: Skin prick tests showed a positive immediate response to aniseed, asparagus, caraway, coriander, cumin, dill, and fennel extracts, and an intense late response to aniseed. Skin prick tests to celery, carrot, birch pollen, and mugwort pollen extracts were negative. Results of a nasal challenge test were positive to aniseed and negative to cinnamon; an aniseed oral food challenge test yielded a positive response. The molecular weights of the main immunoglobulin (Ig)E-binding proteins in aniseed extracts were approximately 48, 42, 39, 37, 34, 33, and 20 kD. Caraway, fennel, cumin, and coriander extracts showed similar IgE-binding patterns. Enzyme immunoassay inhibition studies with the patient's serum revealed cross-reactivity among the IgE components from aniseed, caraway, coriander, fennel, and dill extracts. CONCLUSIONS: We demonstrate the presence of aniseed allergens in a case of occupational rhinoconjunctivitis and food allergy, with molecular weights for this spice that differed from those previously reported.

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[25] - Schwartz HJ, Jones RT, Rojas AR, Squillace DL, Yunginger JW. Occupational allergic rhinoconjunctivitis and asthma due to fennel seed. Ann Allergy Asthma Immunol 1998;78:37-40

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[26] - Garcia-Gonzalez JJ, Bartolomé-Zavala B, Fernandez-Meléndez S, Barcelo-Muñoz JM, Miranda Paez A, Carmona-Bueno MJ, et al. Occupational rhinoconjunctivitis and food allergy because of aniseed sensitization. Ann Allergy Asthma Immunol 2002;88:518-522

BACKGROUND: Aniseed is a spice frequently used in Mediterranean cooking and, as with other Umbelliferae, it has been involved in clinical allergy. OBJECTIVE: This investigation was undertaken to study the allergens implicated in a case of occupational allergy to aniseed associated with rhinoconjunctivitis and gastrointestinal symptoms. METHODS: Skin prick tests were performed to inhalant allergens, spices used in the patient's workplace (aniseed and cinnamon), and 12 other Umbelliferae spices, birch, and mugwort. A nasal challenge test to aniseed and cinnamon and a double-blind placebo-controlled oral food challenge test to aniseed were also performed. The molecular weights of the allergens were studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis immunoblotting and cross-reactivity among Umbelliferae species by enzyme immunoassay inhibition. RESULTS: Skin prick tests showed a positive immediate response to aniseed, asparagus, caraway, coriander, cumin, dill, and fennel extracts, and an intense late response to aniseed. Skin prick tests to celery, carrot, birch pollen, and mugwort pollen extracts were negative. Results of a nasal challenge test were positive to aniseed and negative to cinnamon; an aniseed oral food challenge test yielded a positive response. The molecular weights of the main immunoglobulin (Ig)E-binding proteins in aniseed extracts were approximately 48, 42, 39, 37, 34, 33, and 20 kD. Caraway, fennel, cumin, and coriander extracts showed similar IgE-binding patterns. Enzyme immunoassay inhibition studies with the patient's serum revealed cross-reactivity among the IgE components from aniseed, caraway, coriander, fennel, and dill extracts. CONCLUSIONS: We demonstrate the presence of aniseed allergens in a case of occupational rhinoconjunctivitis and food allergy, with molecular weights for this spice that differed from those previously reported.

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[27] - Moreno-Ancillo A, Gil-Adrados A, Dominguez-Noche C, Cosmes P, Pineda F. Occupational asthma due to carrot in a cook. Allergol Immunopathol (Madr) 2005;33:288-290

Background: Few previous reports of carrot-induced asthma have been confirmed by objective tests. Hypersensitivity to carrot is frequently associated with allergy to Apiaceae spices and sensitization to birch and mugwort pollens. Clinical case: A 40-year-old cook woman was seen with sneezing, rhinorrhea, contact urticaria and wheezing within few minutes of handling or cutting raw carrots. She needed to leave out of the kitchen while the other cooks cut raw carrots. Methods and results: Skin tests were positive to carrot, celery, aniseed and fennel. Rubbing test with fresh carrot was positive. Specific IgE to carrot was 4.44 kU/L. Determinations of specific IgE to mugwort, grass and birch pollens were negative. Inhalative provocation test, performed as a handling test, was positive. The IgE-immunoblotting showed two bands in carrot extract: a band with apparent molecular weight of 30 kd and other band of 18 kd. This band of 18 kd was Dau c 1. The band of 30 kd could correspond a phenylcoumaran benzylic ether reductase. Dau c 1 did not appear to be the unique allergen in this case. Additional allergens may induce the sensitization. Primary sensitization due to airborne allergens of foods and the lack of pollen allergy in this patient are notorious events.

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[28] - 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.

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[29] - Scheurer S, Pastorello EA, Wangorsch A, Kästner M, Haustein D, Vieths S. Recombinant allergens Pru av 1 and Pru av 4 and a newly identified lipid transfer protein in the in vitro diagnosis of cherry allergy. J Allergy Clin Immunol 2001;107:724-731

BACKGROUND: In central and northern Europe food allergy to fruits of the Rosaceae family is strongly associated with birch pollinosis because of the existence of IgE cross-reactive homologous allergens in birch pollen and food. By contrast, in the Mediterranean population allergic reactions to these fruits frequently are not related to birch pollen allergy and are predominantly elicited by lipid transfer proteins (LTPs). OBJECTIVE: We sought to determine the prevalence of IgE sensitization to the recombinant cherry allergens Pru av 1 and Pru av 4 in comparison with cherry extract within a representative group of patients who were allergic to cherries recruited in Germany and to compare the relevance of IgE to cherry LTPs in Italian patients. METHODS: Recombinant Pru av 1 and rPru av 4 were available from earlier studies. The cDNA of the cherry LTPs was obtained by using a PCR-cloning strategy. The protein was expressed in Escherichia coli and purified by means of metal chelate affinity chromatography. Sera from 101 German patients with birch pollinosis and oral allergy syndrome to cherry and sera from 7 Italian patients with cherry allergy were investigated by using enzyme allergosorbent tests for IgE reactivity with cherry extract, rPru av 1, rPru av 4, and the recombinant cherry LTP. Inhibition experiments were performed to compare the IgE reactivity of natural and recombinant cherry LTPs and to investigate potential cross-reactivity with birch pollen allergens. RESULTS: The LTP from cherry comprises 91 amino acids and a 26 amino acid signal peptide. The mature cherry LTP shows high amino acid sequence identity with allergenic LTPs from peach (Pru p 3, 88%), apricot (Pru ar 3, 86%), and maize (Zea m 14, 59%) and displays no IgE cross-reactivity with birch pollen. The IgE prevalences in the German patients were as follows: LTP, 3 of 101 (3%); rPru av 1, 97 of 101 (96.0%); rPru av 4, 16 of 101 (16.2%); and cherry extract, 98 of 101 (97%). All 7 Italian patients had IgE against the cherry LTP. CONCLUSIONS: Recombinant allergens are useful tools for a more accurate in vitro IgE-based diagnosis of cherry allergy. Taken together, they mimic the allergenic activity of cherry extract, having slightly higher biologic activity. Sensitization to the cherry LTP is relevant for a minority of patients recruited in Germany, but our data indicate that it may be a major allergen in Italy.

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[30] - Möller M, Paschke A, Vieluf D, Kayma M, Vieths S, Steinhart H. Characterization of allergens in kiwi fruit and detection of cross-reactivities with allergens of birch pollen and related fruit allergens. Food Agric Immunol 1997;9:107-121

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[31] - Fötisch K, Altmann F, Haustein D, Vieths S. Involvement of carbohydrate epitopes in the IgE response of celery-allergic patients. Int Arch Allergy Immunol 1999;120:30-42

This study was performed to get further insights into antibody responses to cross-reactive carbohydrate determinants (CCD), including initial experiments to prove the biological activity of anti-CCD IgE. Earlier studies have shown that IgE specific for CCD occurs in about 25% of celery-allergic patients. The clinical significance of these antibody specificities is doubtful. METHODS: Patient sera were selected on the basis of a positive case history of celery allergy and multiple binding to high molecular weight celery allergens on immunoblots. Specific IgE to native and heated celery tuber was determined by the enzyme allergosorbent test (EAST). N-glycans were purified after extensive digestion of specific glycoproteins, such as pineapple stem bromelain, bovine fibrin, and human IgG, and used as antigens in an IgE ELISA as well as in EAST and immunoblotting inhibition experiments. Dose-related histamine release was performed with BSA neoglycoproteins containing 3-4 units of the purified glycopeptides. RESULTS: Seven celery-allergic patients were identified who clearly presented IgE against the N-glycan purified from bromelain which is a common structure within the plant kingdom. Chemical defucosylation showed that alpha1, 3-fucose is a key structure for IgE binding. In patients with anti-CCD IgE, the maximal inhibition of celery EAST by the bromelain glycan ranged from 22 to 100%. Inhibition of celery immunoblots by preincubation of patient serum with this glycan led to a quenching of multiple bands at masses >40 kD. After linking the bromelain glycopeptide to BSA, a strong dose-related histamine release was obtained in a celery-allergic patient, occurring at lower concentrations than with the recombinant major protein allergen from celery, Api g 1. CONCLUSIONS: Our results demonstrate that IgE specific for CCD is common in celery-allergic patients, and can represent the major proportion of IgE against this food. alpha1, 3-fucose was confirmed to be an essential part of the IgE epitope. Immunoblotting inhibition indicated the presence of this carbohydrate determinant on multiple glycoproteins in celery extract. Although histamine release was only performed in 1 patient, our data show that proteins carrying multiple glycan units can be biologically active in patients sensitized to CCD.

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

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

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

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[34] - Ballmer-Weber BK, Vieths S, Lüttkopf D, Heuschmann P, Wüthrich B. Celery allergy confirmed by double-blind, placebo-controlled food challenge: a clinical study in 32 subjects with a history of adverse reactions to celery. J Allergy Clin Immunol 2000;106:373-378

BACKGROUND: Celery root is a frequent cause of food allergy in pollen-sensitized patients. Because of problems in blinding challenges with fresh vegetables and the risk of anaphylactic reactions, no double-blind, placebo-controlled, food challenges (DBPCFCs) with celery have been published so far . OBJECTIVE: The aim of the study was to confirm the clinical relevance of celery as a food allergen by DBPCFCs and to evaluate current diagnostic procedures in patients with true allergy . METHODS: DBPCFCs were performed in 32 patients with a history of an allergic reaction to celery. The patients underwent skin prick tests (SPTs) with celery extracts, crude celery, and different pollen extracts. Specific IgE for celery was determined by using the CAP method . RESULTS: Twenty-two of 32 patients had a positive DBPCFC result. Two patients reacted to placebo, and 8 patients did not respond to the challenge. Of the nonresponders, 4 reacted to an open provocation with celery. The sensitivity of CAP determination for specific IgE (> or =0.7 kU/L) to celery in patients with a positive DBPCFC result was 73%, 48% to 86% for SPTs (> or =3 mm) with commercial extracts, and 96% for prick-to-prick tests with crude celery. The positive predictive value of the SPT and CAP tests was between 87% and 96%, whereas the specificity and negative predictive values were poor . CONCLUSION: This study confirms the importance of celery as a food allergen for use in DBPCFCs. The SPT and CAP methods proved to be reliable for the diagnosis of a relevant allergy to celery in regard to sensitivity and positive predictive value but not to specificity and negative predictive value.

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[35] - Vieths S, Lüttkopf D, Reindl J, Anliker MD, Wüthrich B, Ballmer-Weber BK. Allergens in celery and zucchini. Allergy 2002;57(Suppl. 72):100-105

The aim of this study was to confirm allergy to celery tuber and to zucchini, for the first time, by DBPCFC, and to identify the allergens recognized by IgE from DBPCFC-positive patients. Therefore, raw vegetables were hidden in a broccoli drink, and a DBPCFC-procedure was developed that consisted of a spit and swallow protocol, making sure that the procedure was safe for the patients and that reactions strictly localized to the oral cavity as well as systemic reactions could be reproduced by DBPCFC. The allergens in celery and zucchini extract were identified by immunoblot inhibition using allergen extracts, recombinant allergens and purified N-glycans as inhibitors. Celery allergy was confirmed in 69% (22/32) of subjects with a positive case history. Four subjects with a history of allergic reactions to zucchini had a positive DBPCFC to this vegetable. During DBPCFC, systemic reactions were provoked in 50% (11/22) of the patients to celery, and in 3/4 of the zucchini-allergic patients. The Bet v 1-related major celery allergen was detected by IgE of 59% (13/22) of the patients. Cross-reactive carbohydrate epitopes (CCD) bound IgE of 55% (12/22) of the celery-allergic patients and in 2/4 of the subjects with zucchini allergy. Profilin was a food allergen in celery in 23% (5/22) and in zucchini in 2/4 of the cases. A zucchini-specific allergen was detected by IgE from one patient. We conclude that ubiquitous cross-reactive structures are important in allergy to both, celery and zucchini, and that a specific association to birch pollen allergy exists in allergy to celery (mediated by Api g 1), but not in zucchini allergy.

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[36] - Paschke A, Kinder H, Zunker K, Wigotzki M, Wessbecher R, Vieluf D, et al. Characterization of allergens in mango fruit and ripening dependence of the allergenic potency. Food Agric Immunol 2001;13:51-61

Characterization of IgE-related mango allergens in human sera, and the occurrence of allergens during ripening of mangoes were investigated. Mol. wt. of IgE-binding allergens were determined by SDS-PAGE/immunoblotting. Of 52 sera from patients with IgE-mediated sensitization to mangoes, 46 contained 2 allergens with mol. wt. of approx. 40 and 30 kDa; these were described as the major allergens. Other IgE-binding patterns were detected in <50% of assigned sera. 2D-electrophoresis proved that the 40 kDa major allergen exists as an isoallergen with isoelectric points of 4.6 and 4.8. The 30 kDa allergen has an isoelectric point of 4.9. According to international rules, the 40 kDa allergen was named Mangifera indica 1 (Man i 1) and the 30 kDa allergen, Mangifera indica 2 (Man i 2). SDS-PAGE/immunoblotting, immunoblot inhibition and EAST-inhibition analyses showed that there was no significant difference in allergenic potency during fruit ripening

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[37] - 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.

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

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

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

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

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[40] - Hoffmann-Sommergruber K, Demoly P, Crameri R, Breiteneder H, Ebner C, Laimer Da Camara Machado M, et al. IgE reactivity to Api g 1, a major celery allergen, in a Central European population is based on primary sensitization by Bet v 1. J Allergy Clin Immunol 1999;104:478-484

BACKGROUND: Up to 70% of patients with tree pollen allergy display allergic symptoms when eating certain fruits and vegetables. Homologous proteins with allergenic features are present in a wide range of plant species and can cause allergic reactions. OBJECTIVE: The aim of this study was to evaluate recombinant Api g 1, a major celery allergen, for in vivo and in vitro diagnosis of celery allergy in populations from Davos, Switzerland, and Montpellier, France. METHODS: A group of patients with celery and birch pollen allergy from Davos was tested, and the results from those tests were compared with results from a group of patients allergic to celery from Montpellier. Skin prick tests were performed with a commercial celery extract, crude celery, and purified recombinant Api g 1. Quantitative and qualitative serology was done with natural and recombinant allergens by means of RASTs and immunoblotting. RESULTS: Recombinant Api g 1 allowed accurate in vivo diagnosis of celery allergy in all patients from the Swiss group. RAST results with celery extract were negative in 8 of 24 patients; results of immunoblotting with celery extract were negative in 4 of 24 patients, and results of immunoblotting with recombinant (r)Api g 1 were negative in 8 of 24 patients. In the French group 11 of 12 patients had a positive skin reaction with crude celery extract, but only 2 patients reacted with rApi g 1. RAST results for celery were positive in 8 of 12 patients. In immunoblotting experiments 8 patient sera displayed IgE directed against various celery allergens, whereas no patients sera had rApi g 1-specific IgE. CONCLUSION: Our results document that rApi g 1 allows accurate in vivo diagnosis only in areas where birch trees are common. In areas where no birch trees grow, primary sensitization takes place through different pollen allergens (eg, mugwort pollen). Moreover, it became evident that birch pollen and celery allergy are highly related in Central Europe, whereas in Southern Europe the mugwort-celery type is predominant.

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[41] - Wüthrich B, Hofer T. [Food allergy: the celery-mugwort-spice syndrome. Association with mango allergy ?]. Dtsch Med Wochenschr 1984;109:981-986

Celery allergies are commonly observed in mugwort pollen-allergic patients. This situation is frequently associated with an allergy to spices of the umbelliferous family. Thus a "celery-mugwort-spice syndrome" has been established. In 31 patients (27 females, 4 males) allergy to celery was diagnosed between 1978 and 1982. They were followed up in 1983 and complementary tests (skin and RAST) with various raw vegetables, spices and mugwort pollen were done. For specific diagnosis of allergy to celery skin testing (scratch or prick) with fresh celery bulb was reliable. In contrast, RAST was not sufficiently sensitive, 87% of patients allergic to celery had pollinosis in the form of mugwort pollen sensitisation established by skin testing and RAST. Allergy to carrots was clinically and by testing currently associated in 52%. In contrast, a clinically relevant hypersensitivity to caraway (26%), parsley (16%), fennel (13%), green pepper (10%) and aniseed (3%) was found less frequently although these allergens showed a much larger positivity in skin testing. This signifies group sensitisation within the umbelliferous plants. The association of celery-mugwort allergy with allergy to mango fruit described here for the first time cannot be explained as group sensitisation within a botanic relation, as mango, and pistachio-nut, belong to the family of anacardiaceae.

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[42] - Wüthrich B, Stäger J, Johansson SG. Celery allergy associated with birch and mugwort pollinosis. Allergy 1990;45:566-571

Skin prick tests (SPT) with various celery, carrot and potato preparations (raw, cooked, cooking water of each vegetable and allergen extracts) as well as specific IgE determinations by RAST to celery mix, celeriac (or root celery), stick celery and heated celery extracts were performed in 70 patients with positive prick or intracutaneous tests to birch and/or mugwort pollens and celery (extract and/or raw). 94% of the patients showed positive prick tests to raw celeriac, 36% to cooked celeriac and 8/13 to cooking water. Celery-birch positive patients (n = 13) showed negative or low RASTs to heated celery extracts and to stick celery. By contrast, in the celery-mugwort sensitive patients (n = 6) the celery RASTs with heated celery extracts remained clearly positive and high RAST values to stick celery could be found. Celery-birch-mugwort-association (n = 22) favoured more positive results with relatively high values of RAST to celeriac. The results of homologous and heterologous RAST inhibition experiments with birch, mugwort, unheated and heated celery (100 degrees C) carried out in nine celery-RAST positive sera are also discussed.

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[43] - Ballmer-Weber BK, Wüthrich B, Wangorsch A, Fötisch K, Altmann F, Vieths S. Carrot allergy: double-blind, placebo-controlled food challenge and identification of allergens. J Allergy Clin Immunol 2001;108:301-307

BACKGROUND: Allergic reactions to carrot affect up to 25% of food-allergic subjects. Clinical manifestations of carrot allergy and IgE responses to carrot proteins, however, have never been studied in subjects with carrot allergy confirmed by means of double-blinded, placebo-controlled food challenge (DBPCFC) . OBJECTIVE: The purposes of this investigation were to confirm clinically relevant sensitizations to carrot by means of DBPCFC, to validate current diagnostic methods, and to identify IgE-reactive carrot proteins in patients with true allergy . METHODS: DBPCFCs were performed in 26 subjects with histories of allergic reactions to carrot. Patients underwent skin prick tests with carrot extract, fresh carrot, and various pollen extracts. Specific IgE to carrot, celery, birch, and mugwort pollen and to rBet v 1, rBet v 2, and rBet v 6 were measured through use of the CAP method. Carrot allergens were identified by means of immunoblotting and blotting inhibition . RESULTS: Twenty of 26 patients had positive DBPCFC results. The sensitivity of the determination of carrot-specific IgE antibodies through use of the CAP method (> or =0.7 kU/L) was 90%, the sensitivity for skin prick testing with commercial extracts was 26%, and the sensitivity for prick-to-prick tests with raw carrot was 100%. The Bet v 1--related major carrot allergen Dau c 1 was recognized by IgE from 85% of patients; 45% were sensitized to cross-reactive carbohydrate determinants and 20% to carrot profilin. In 1 subject, a Bet v 6--related carrot allergen was recognized. In 4 patients, IgE binding to Dau c 1 was not inhibited or was weakly inhibited by rBet v 1 or birch pollen extract . CONCLUSION: This study confirmed the allergenicity of carrot by means of DBPCFC. DBPCFC-positive patients had exclusively specific IgE antibodies to birch pollen--related carrot allergens, Dau c 1 being the major allergen. The lack of inhibition of IgE binding to Dau c 1 by birch allergens in a subgroup of patients might indicate an secondary immune response to new epitopes on the food allergen that are not cross-reactive with Bet v 1.

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[44] - Ballmer-Weber BK, Wangorsch A, Bohle B, Kaul S, Kündig T, Fötisch K, et al. Component-resolved in vitro diagnosis in carrot allergy: Does the use of recombinant carrot allergens improve the reliability of the diagnostic procedure ? Clin Exp Allergy 2005;35:970-978

BACKGROUND: In Europe, pollen-related food allergy is the most frequent form of food allergy in adults. Reliability of current diagnostic procedures, however, is poor and therapeutic options are not available . OBJECTIVES: In the present study, we created a panel of recombinant allergens from carrot and evaluated its potential in component-resolved in vitro diagnosis of carrot allergy . METHODS: Recombinant (r) Dau c 1.0104, Dau c 1.0201 and Dau c 4 were cloned by a polymerase chain reaction strategy, expressed in Escherichia coli and purified. Carrot lipid transfer protein (LTP) was expressed in the yeast Pichia pastoris. Sera from 40 carrot-allergic patients were investigated. Twenty-one birch pollen-allergic subjects with negative open provocation to carrot and 20 non-allergic subjects were included as controls. IgE binding to recombinant allergens as well as to cross-reactive carbohydrate determinants (CCD) was measured by ELISA. Cross-reactivity between Dau c 1 isoforms and Bet v 1 was assayed by ELISA inhibition. Biological activity of the recombinant carrot allergens was assessed by histamine release assay and peripheral blood mononuclear cells stimulation . RESULTS: Ninety-eight percent of the carrot-allergic patients were positive to at least one recombinant allergen; 98% reacted to rDau c 1.0104, 65% to rDau c 1.0201, 38% to rDau c 4 and 20% had IgE against CCD. Specificity using the recombinant allergens was high when compared with non-allergic controls, but low compared with birch-sensitized subjects without carrot allergy. Sensitization to Dau c 1.0201, however, proved to be highly specific for clinically relevant sensitization. Inhibition assays indicated the absence of LTP in carrot root extract, and epitope diversity between Dau c 1.0104, Dau c 1.0201 and Bet v 1 . CONCLUSIONS: Our panel of recombinant allergens from carrot can provide a standardized tool for in vitro diagnosis of carrot allergy, and for epitope studies.

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[45] - Bauer L, Ebner C, Hirschwehr R, Wüthrich B, Pichler C, Fritsch R, et al. IgE cross-reactivity between birch pollen, mugwort pollen and celery is due to at least three distinct cross-reacting allergens: immunoblot investigation of the birch-mugwort-celery syndrome. Clin Exp Allergy 1996;26:1161-1170

Allergy to celery is often associated with sensitization to birch and/or mugwort pollen. OBJECTIVE and METHODS: In a multi-centre study, sera from 23 patients suffering from type 1 allergy to celery and 15 patients with positive celery RAST but no clinical sensitization were compared. To examine whether cross-reactivity between celery and mugwort pollen includes cross-sensitization to birch pollen allergens, we determined cross-reacting structures in birch pollen, mugwort pollen and celery by means of immunoblotting. Inhibition studies were performed by preincubation of sera with extracts of birch pollen, mugwort pollen, and celery. RESULTS: We identified three groups of proteins--homologues of Bet v 1 and birch profilin (Bet v 2) as well as a group of proteins with a molecular range of 46 to 60 kD--displaying IgE-cross-reactivity, which were shared by birch pollen and celery. Two of these groups of allergens (profilin and the 46 to 60 kD proteins) were also present in mugwort pollen. In this paper we demonstrate that most cross-reacting allergens present in mugwort pollen and celery can also be detected in birch pollen extract. CONCLUSION: Therefore we propose, from a serological point of view, to extend the mugwort-celery syndrome to the birch-mugwort-celery syndrome.

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

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

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[47] - Pauli G, Bessot JC, Braun PA, Dietemann-Molard A, Kopferschmitt-Kubler MC, Thierry R. Celery allergy: clinical and biological study of 20 cases. Ann Allergy 1988;60:243-246

In 20 patients, the ingestion of celery was responsible for mucocutaneous symptoms (generalized urticaria and angioedema) (18/20) and respiratory disorders (7/20). Four cases of systemic anaphylaxis were observed. The main associated allergic disorder was pollinosis (16/20). Food allergy to other vegetable products, mainly other Umbelliferae and apples, coexisted with celery allergy in 12 cases. It was found that celery allergy is mediated by IgE antibodies: it is easily diagnosed by skin tests (fresh extracts of celery may be used) and by adequate RAST (17 positive results). Cosensitization with mugwort pollen (14 cases) and birch pollen (9 cases) was found. Celery allergens responsible for clinical sensitization originate chiefly in the tuber and are at least partly thermally labile. The frequent association with pollen sensitization suggests the existence of common antigenic epitopes in celery extracts and mugwort and birch pollens. The immunologic investigations carried out so far (RAST inhibition and immunoprint) seem to support this hypothesis.

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[48] - Pauli G, Bessot JC, Dietemann-Molard A, Braun PA, Thierry R. Celery sensitivity: clinical and immunological correlations with pollen allergy. Clin Allergy 1985;15:273-279

The authors studied twenty patients with celery allergy and concomitant hypersensitivity to certain pollens (mugwort, birch). The specific symptoms induced by eating celery were attacks of urticaria and angio oedema (seventeen out of twenty) respiratory complaints (eight out of twenty), systemic anaphylaxis with vascular collapse (three out of twenty). A strong association between clinical reactions to celery and mugwort sensitization, and to a lesser degree between celery allergy and birch pollen sensitization was established. Celery allergy is mediated by IgE antibodies and can be easily diagnosed by cutaneous tests using fresh material and/or by adequate RAST test. RAST inhibitions performed on individual sera suggest the existence of common antigens in celery and mugwort, and in celery and birch pollen. However, the exact nature of these common antigens has not yet been determined.

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

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

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[50] - Hoffmann-Sommergruber K, Demoly P, Crameri R, Breiteneder H, Ebner C, Laimer Da Camara Machado M, et al. IgE reactivity to Api g 1, a major celery allergen, in a Central European population is based on primary sensitization by Bet v 1. J Allergy Clin Immunol 1999;104:478-484

BACKGROUND: Up to 70% of patients with tree pollen allergy display allergic symptoms when eating certain fruits and vegetables. Homologous proteins with allergenic features are present in a wide range of plant species and can cause allergic reactions. OBJECTIVE: The aim of this study was to evaluate recombinant Api g 1, a major celery allergen, for in vivo and in vitro diagnosis of celery allergy in populations from Davos, Switzerland, and Montpellier, France. METHODS: A group of patients with celery and birch pollen allergy from Davos was tested, and the results from those tests were compared with results from a group of patients allergic to celery from Montpellier. Skin prick tests were performed with a commercial celery extract, crude celery, and purified recombinant Api g 1. Quantitative and qualitative serology was done with natural and recombinant allergens by means of RASTs and immunoblotting. RESULTS: Recombinant Api g 1 allowed accurate in vivo diagnosis of celery allergy in all patients from the Swiss group. RAST results with celery extract were negative in 8 of 24 patients; results of immunoblotting with celery extract were negative in 4 of 24 patients, and results of immunoblotting with recombinant (r)Api g 1 were negative in 8 of 24 patients. In the French group 11 of 12 patients had a positive skin reaction with crude celery extract, but only 2 patients reacted with rApi g 1. RAST results for celery were positive in 8 of 12 patients. In immunoblotting experiments 8 patient sera displayed IgE directed against various celery allergens, whereas no patients sera had rApi g 1-specific IgE. CONCLUSION: Our results document that rApi g 1 allows accurate in vivo diagnosis only in areas where birch trees are common. In areas where no birch trees grow, primary sensitization takes place through different pollen allergens (eg, mugwort pollen). Moreover, it became evident that birch pollen and celery allergy are highly related in Central Europe, whereas in Southern Europe the mugwort-celery type is predominant.

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[51] - Rancé F, Juchet-Gibon A, Fejji S, Brémont F, Dutau G. Sensibilisation aux épices chez l'enfant. Rev Fr Allergol Immunol Clin 1994;34:475-479

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[52] - Moneret-Vautrin DA, Morisset M, Lemerdy P, Croizier A, Kanny G. Food allergy and IgE sensitization caused by spices: CICBAA data (based on 589 cases of food allergy). Allerg Immunol (Paris) 2002;34:135-140

BACKGROUND: Spices originate in various botanical families: Apiaceae, Lamiaceae, Lauraceae, Leguminosae, Liliaceae, Myristicaceae, Myrtaceae, Piperaceae, Solanaceae, Zingiberaceae.... METHODOLOGY: Prick-tests to native spices have been carried out in patients suspected of food allergies to spices. The CICBAA data bank includes 589 cases of food allergies, a part of which has benefited from investigations for spices. Data about the rate of sensitization and food allergy are available. RESULTS: Frequent sensitization to Apiaceae is observed: coriander, caraway, fennel, celery: 32% of prick-tests in children, 23% of prick-tests in adults. Sensitization to Liliaceae: garlic, onion, chive, is observed in 4.6% of prick-tests in children, 7.7% of prick-tests in adults. Rare cases of sensitization to paprika and saffron are recorded. Prick-tests to nutmeg, ginger and clove are currently negative. 10 food allergies related to the mugwort-celery-spices syndrome are reported: coriander: 1, caraway: 2, fennel: 3, garlic: 3, onion: 1. Food allergy to spices is unfrequent: 2% of the totality of food allergies. However, only adults are allergic to spices and allergy to spices accounts for 6.4% of food allergies in adults. Tiny amount of proteins are usually ingested. Patients at risk of spice allergy are young adults sensitized to mugwort and birch allergens, sharing cross-sensitization with various food vegetal allergens. The clinical suspicion raises from frequent post-prandial systemic reactions. Other allergens of vegetal origin have to be cleared. Diagnosis can be established by DBPCFC using powdered spices in capsules. [References: 22]

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[53] - 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.

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[54] - Asero R. Relevance of pollen-specific IgE levels to the development of Apiaceae hypersensitivity in patients with birch pollen allergy. Allergy 1997;52:560-564

A large clinical/serologic study was carried out to determine the prevalence of Apiaceae (carrot, celery, and fennel) hypersensitivity in patients with birch pollen allergy, and to investigate its relationship with apple and hazelnut allergy and with birch pollen-specific IgE levels. A total of 196 birch pollen-hypersensitive patients with oral allergy syndrome (OAS) caused by different vegetable foods were examined in the cross-sectional part of the study. Of this total, 195 patients had apple and/or hazelnut allergy, and 103 had Apiaceae sensitivity; only one patient had Apiaceae allergy alone. Apiaceae-positive patients showed significantly higher birch pollen-specific IgE levels than negative ones (median 13 vs 7 AU/ml; P<0.0001). The prospective part of the study was performed on 103 birch pollen-hypersensitive patients who were OAS-free at the time of the first visit and were periodically followed-up for OAS. Patients who developed Apiaceae sensitivity showed much higher birch-specific IgE levels than patients who developed apple/hazelnut allergy only (median 15.5 vs 8.5 AU/ml; P<0.05), whereas those who remained OAS-free showed the lowest specific IgE levels (median 5 AU/ml). This study suggests that most Apiaceae determinants cross-react with apple or hazelnut determinants, whereas only some apple or hazelnut determinants cross-react with apiaceae-allergenic determinants; moreover, it shows that birch-specific IgE levels heavily influence the onset of OAS as a whole, and probably play a critical role in the development of allergies to distinct vegetable foods as well

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[55] - Asero R, Mistrello G, Roncarolo D, Amato S, Caldironi G, Barocci F, et al. Immunological cross-reactivity between lipid transfer proteins from botanically unrelated plant-derived foods: a clinical study. Allergy 2002;57:900-906

BACKGROUND: Lipid transfer proteins (LTP) are highly conserved and widely distributed throughout the plant kingdom. Recent studies demonstrated immunological cross-reactivity between LTP from many botanically unrelated fruits and vegetables and concluded that LTP are pan-allergens. This study aimed to evaluate the clinical relevance of such cross-reactivity in a group of subjects monosensitized to LTP . METHODS: Twenty LTP-hypersensitive patients were selected from a population of about 600 subjects with history of Rosaceae allergy by means of: 1) negative skin prick test (SPT) with a commercial birch pollen extract; 2) positive SPT with a commercial plum extract, rich in LTP but virtually lacking both Bet v 1-like proteins and profilin; 3) in-vitro IgE reactivity to the 9-10 kDa fraction of peach peel or immunoblot with peach peel showing a single band at 10 kDa; and 4) total inhibition of reactivity to whole peach extract (containing Bet v 1-related allergen, profilin, and LTP) by purified peach LTP on enzyme-linked immunoassay (ELISA). Allergy to foods other than Rosaceae was ascertained by careful interview and analysis of medical recordings. SPT with a large series of plant-derived foods were carried out as well. The cross reactivity between LTPs from botanically unrelated plant-derived foods was assessed by ELISA inhibition tests using walnut and peanut extracts as substrate, and peach LTP as inhibitor . RESULTS: All patients reported allergic reactions after the ingestion of at least one from a large number of vegetable foods other than Rosaceae, and in several cases clinical reactions were very severe (anaphylaxis, asthma, urticaria/angioedema). Nuts and peanuts were the most frequently reported causes of allergic reactions (80% and 40% of patients, respectively). All patients showed positive SPT to several non-Rosaceae food extracts. SPT with nuts, peanut, legumes, celery, rice, and corn were positive in the majority of patients. In ELISA inhibition studies, absorption of sera with peach LTP caused complete inhibition of IgE reactivity to walnut and peanut in all cases . CONCLUSION: LTP is a clinically relevant pan-allergen. Most Rosaceae-allergic, LTP-hypersensitive patients experience adverse reactions after ingestion of botanically unrelated plant-derived foods as well. In view of the high prevalence and severity of the allergic reactions induced, hazelnut, walnut, and peanut should be regarded as potentially hazardous for these patients.

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[56] - 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.

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[57] - Asero R, Mistrello G, Roncarolo D, Amato S. Detection of Some Safe Plant-Derived Foods for LTP-Allergic Patients. Int Arch Allergy Immunol 2007;144:57-63

BACKGROUND: Lipid transfer protein (LTP) is a widely cross-reacting plant pan-allergen. Adverse reactions to Rosaceae, tree nuts, peanut, beer, maize, mustard, asparagus, grapes, mulberry, cabbage, dates, orange, fig, kiwi, lupine, fennel, celery, tomato, eggplant, lettuce, chestnut and pineapple have been recorded . OBJECTIVE: To detect vegetable foods to be regarded as safe for LTP-allergic patients . METHODS: Tolerance/intolerance to a large spectrum of vegetable foods other than Rosaceae, tree nuts and peanut was assessed by interview in 49 subjects monosensitized to LTP and in three distinct groups of controls monosensitized to Bet v 1 (n = 24) or Bet v 2 (n = 18), or sensitized to both LTP and birch pollen (n = 16), all with a history of vegetable food allergy. Patients and controls underwent skin prick test (SPT) with a large spectrum of vegetable foods. The absence of IgE reactivity to foods that were negative in both clinical history and SPT was confirmed by immunoblot analysis and their clinical tolerance was finally assessed by open oral challenge (50 g per food) . RESULTS: All patients reported tolerance and showed negative SPT to carrot, potato, banana and melon; these foods scored positive in SPT and elicited clinical symptoms in a significant proportion of patients from all three control groups. All patients tolerated these four foods on oral challenge. Immunoblot analysis confirmed the lack of IgE reactivity to these foods by LTP-allergic patients . CONCLUSION: Carrot, potato, banana and melon seem safe for LTP-allergic patients. This finding may be helpful for a better management of allergy to LTP.

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[58] - Asero R, Mistrello G, Roncarolo D, Amato S, Zanoni D, Barocci F, et al. Detection of clinical markers of sensitization to profilin in patients allergic to plant-derived foods. J Allergy Clin Immunol 2003;112:427-432

BACKGROUND: A proper classification of patients allergic to plant-derived foods is of pivotal importance because the clinical features of allergic reactions to fruits and vegetables depend on the nature and characteristics of proteins responsible for sensitization. However, in normal clinical settings this is presently impossible . OBJECTIVE: We sought to detect clinical markers of sensitization to profilin . METHODS: Seventy-one patients allergic to fruits and vegetables but not sensitized to lipid transfer protein or natural rubber latex were studied. Food allergy was ascertained on the basis of clinical history and positive skin prick test responses with fresh foods, commercial extracts, or both. Allergies to foods that had caused less than 2 adverse reactions were confirmed by means of open oral challenge. IgE reactivity to rBet v 1/rBet v 2 and to natural Phleum species profilin were detected. Moreover, IgE to the 30- to 40-kd and 60- to 90-kd birch pollen-enriched fractions, which also can be involved in cross-reactivity phenomena, were measured in sera from 52 patients by means of ELISA . RESULTS: On the basis of in vitro tests, 24, 18, and 25 patients turned out to be sensitized to Bet v 1, Bet v 2, or both, respectively. Four patients had negative test results for both allergens. Hypersensitivity to Bet v 2 was strongly associated with clinical allergy to citrus fruits (39% in patients monosensitized to Bet v 2 vs 4% in patients monosensitized to Bet v 1, P <.025), melon or watermelon (67% vs 0%, P <.001), banana (66% vs 8%, P <.001), and tomato (33% vs 0%, P <.05), whereas Bet v 1 sensitivity was associated with clinical allergy to apple (100% vs 39%, P <.001) and hazelnut (56% vs 0%, P <.001). The sensitivity of a history of allergy to gourd fruits, citrus fruits, tomato, banana, or a combination thereof as a means to detect profilin-hypersensitive patients was 85% (41/48). The specificity of an allergy to any of these fruits exceeded 85%, with positive predictive values ranging between 68% and 91% . CONCLUSION: In clinical settings in which laboratory investigations are not easily accessible, allergy to melon, watermelon, citrus fruits, tomato, and banana can be used as a marker of profilin hypersensitivity once a sensitization to natural rubber latex and lipid transfer protein is ruled out.

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[59] - Pastorello EA, Farioli L, Pravettoni V, Ortolani C, Fortunato D, Giuffrida G, et al. Identification of grape and wine allergens as an endochitinase 4, a lipid-transfer protein, and a thaumatin. J Allergy Clin Immunol 2003;111:350-359

BACKGROUND: Few allergic reactions to grape are reported in the literature. In some cases an association with peach and cherry allergy was observed. No IgE-mediated reactions to wine have been described, and no grape major allergens have yet been identified . OBJECTIVE: We describe several severe reactions to grape or wine. We characterized the grape major allergens and tried to identify the allergen in wine . METHODS: We collected documented histories of allergic reactions to grape and wine. Grape allergens were identified by means of SDS-PAGE and immunoblotting and purified by means of HPLC. Using amino acid sequencing and mass spectrometry, we identified the family of proteins to which the allergens belong. Cross-reactivity with peach and cherry was evaluated by means of cross-wise inhibition experiments . RESULTS: Eleven patients with reactions to grape and 3 with anaphylactic reactions to wine were recruited. The major allergens were an endochitinase 4A and a lipid-transfer protein (LTP) that was homologous to and cross-reactive with peach LTP. A 24-kd protein homologous to the cherry thaumatin-like allergen was a minor allergen. Endochitinase 4A is very likely the allergen in vino novello and in vino Fragolino . CONCLUSIONS: Grape and wine might cause severe allergic reactions in sensitive patients. The major allergens of grape are endochitinase 4A, which is also the allergen of wine, and an LTP cross-reacting with the peach major allergen.

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

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

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[61] - Cuesta-Herranz J, Lazaro M, Figueredo E, Igea JM, Umpierrez A, De Las Heras M. Allergy to plant-derived fresh foods in a birch- and ragweed-free area. Clin Exp Allergy 2000;30:1411-1416

BACKGROUND: Allergy to plant-derived fresh foods has often been reported in geographical areas where birch or ragweed pollens are frequent and has been attributed to cross-reactivity to pollens. OBJECTIVE: The aim of this study has been to evaluate allergy to plant-derived fresh foods among pollen-allergic patients from a birch and ragweed-free area. METHODS: Ninety-five pollen-allergic patients took part in the study. The study consisted of a questionnaire, skin prick tests and challenge tests. Pollen skin tests to five grasses, eight trees and seven weeds were performed in duplicate. Prick tests (prick by prick) and challenge tests were carried out with the fresh foods. RESULTS: Most patients allergic to pollens were sensitized to grass (Lolium and Phleum; 97.9%), followed by tree (Olea; 82.1%) and weed pollens (Plantago; 64.2%). 35 of the 95 pollen-allergic patients had positive skin test responses to some plant-derived fresh foods, the highest percentage corresponding to several fruits in the Rosaceae family (peach and pear, 26.3%), followed by Cucurbitacea fruits (melon, 13.7%). The 21. 05% of the pollen-allergic patients were allergic to some type of plant-derived fresh food. Peach was the plant-derived fresh food which most frequently elicited allergy symptoms (12.6%), followed by melon (7.36%). The cluster of positive responses to Rosaceae fruits was higher for skin testing than for challenge testing. CONCLUSION: Peach was the most important allergy provoking fruit in a birch and ragweed free-area where apples were consumed at a rate of two times more than peaches and the patients allergic to pollen were principally sensitized to grass pollens.

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[62] - Miralles JC, Caravaca F, Guillén F, Lombardero M, Negro JM. Cross-reactivity between Platanus pollen and vegetables. Allergy 2002;57:146-149

Background: Several associations have been described between tree and plant pollens and certain foods. The objective of this study is to verify whether there is cross-reactivity between Platanus pollen and vegetable origin foods. Methods: We selected 56 patients allergic to vegetable foods and subjected them to cutaneous tests with aeroallergens and vegetable foods. A statistical analysis was performed to evaluate the association of Platanus pollen with foods and with other aeroallergens. Later, a specific IgE determination was performed as well as a RAST (radioallergosorbent) inhibition experiment, to verify the existence of cross-reactivity in vitro. Results: In the cutaneous tests we found a positive correlation between Platanus pollen and hazelnut, peanut, banana and celery. The results of the RAST inhibition experiment indicate an important cross-reactivity between the pollen of Platanus acerifolia and hazelnut and banana fruit, and an intermediate cross-reactivity with celery and peanut. Conclusion: We have described an association between the pollen of the Platanus tree and some vegetable foods such as hazelnut, banana, peanut and celery. This association could be explained by the in vitro IgE cross-reactivity detected.

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[63] - Asero R, Mistrello G, Roncarolo D, Amato S, Van Ree R. String bean-induced anaphylaxis. Allergy 2001;56:259-260

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[64] - Asero R, Mistrello G, Roncarolo D, de Vries SC, Gautier MF, Ciurana CL, et al. Lipid transfer protein: a pan-allergen in plant-derived foods that is highly resistant to pepsin digestion. Int Arch Allergy Immunol 2000;122:20-32

Lipid transfer proteins (LTPs) are small molecules of approximately 10 kD that demonstrate high stability. They have recently been identified as allergens in the Rosaceae subfamilies of the Prunoideae (peach, apricot, plum) and of the Pomoideae (apple). They belong to a family of structurally highly conserved proteins that are also present in non-Rosaceae vegetable foods. OBJECTIVE: The aim of this study was to investigate the cross-reactivity to non-Rosaceae LTPs, and to study the role of protein stability in allergenicity. METHODS: Thirty-eight patients with a positive SPT to Rosaceae fruit extracts enriched for LTP were characterized by interview and SPT. To investigate IgE cross-reactivity between Rosaceae and non-Rosaceae LTPs, RAST and RAST inhibition as well as ELISA and ELISA inhibition were performed, using whole food extracts and purified LTPs. Both purified natural LTPs (peach, carrot and broccoli) and Pichia pastoris recombinant LTPs (carrot and wheat) were included. Pepsin digestion was used to address the role of stability in the allergenicity of LTPs. RESULTS: IgE antibodies to Rosaceae LTPs reacted to a broad range of vegetable foods, including Gramineae (cereals), Leguminosae (peanut), Juglandaceae (walnut), Anacardiaceae (pistachio), Brassicaceae (broccoli), Umbelliferae (carrot, celery), Solanaceae (tomato), Cucurbitaceae (melon), and Actinidiaceae (kiwi). Binding and inhibition studies with purified natural and recombinant LTPs confirmed their role in this cross-reactivity. Many of these cross-reactivities were accompanied by clinical food allergy, frequently including systemic reactions. Antibody binding to LTP was shown to be resistant to pepsin treatment of whole extract or purified LTP. CONCLUSION: LTP is a pan-allergen with a degree of cross-reactivity comparable to profilin. Due to its extreme resistance to pepsin digestion, LTP is a potentially severe food allergen.

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[65] - Vieths S, Wangorsch A, Fötisch K, van Ree R, Wüthrich B, Ballmer-Weber BK. Carrot allergy: use of recombinant allergens to determine sensitisation patterns and cross-reactivity. EAACI 21th Congress, Naples, 1-5 June, 2002, Poster n°61

Background: Recently, carrot allergy was confirmed by DBPCFC for the first time (JACI 2001; 108: 301-7). Dau c 1, Dau c 4 (profilin), and cross-reactive carbohydrate epitopes (CCD) were identified as IgE-binding components in carrot-allergic patients. In 4/20 patients, IgE-binding to Dau c 1 was not inhibited by Bet v 1. In the present study we created an almost complete panel of recombinant allergens (RA) from carrot and evaluated its potential in component-resolved diagnosis of carrot allergy. Methods: Recombinant (r) Dau c 1.0104 (GenBank Z81362), Dau c 1.0201 (a homologue of Api g 1.02, GenBank AF456481), and Dau c 4 (GenBank AF456482) were cloned by a PCR strategy, expressed in E. coli, and purified. Recombinant carrot lipid transfer protein (LTP) (GenBank M64746) was expressed in the yeast Pichia pastoris. Sera from 40 carrot-allergic patients including 20 subjects with positive DBPCFC were investigated. Sera from birch pollen-allergic subjects with negative open provocation to carrot and from non-allergic subjects were used as controls. IgE binding to RA as well as to CCD was measured by ELISA. Cross-reactivity between Dau c 1 isoforms and to Bet v 1 was assayed by ELISA inhibition. Results: 39/40 (98%) of the carrot-allergic patients were positive to at least one RA. 39/40 (98%) reacted to rDau c 1.0104, 22/40 (55%) to rDau c 1.0201, 7/40 (18%) to rLTP, and 12/40 (30%) to rDau c 4. IgE against CCD were detected in 8/40 (20%) of the sera. Specificity and positive predictive value using the 4 RA were high when compared to non-allergic controls but low compared to birch-sensitised subjects without carrot allergy. No correlation was observed between sensitisation profile, levels of specific IgE, and presence or severity of symptoms. Inhibition assays indicated the absence of LTP in carrot root extract, and epitope diversity between Dau c 1.0104, Dau c 1.0201, and Bet v 1. Conclusions: Our panel of RA from carrot can provide a standardised tool for in vitro diagnosis of carrot allergy, and for epitope studies. However, so far it remains impossible to establish an in vitro assay providing reliable information on clinical relevance or severity of carrot allergy.

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[66] - Ballmer-Weber BK, Wangorsch A, Bohle B, Kaul S, Kündig T, Fötisch K, et al. Component-resolved in vitro diagnosis in carrot allergy: Does the use of recombinant carrot allergens improve the reliability of the diagnostic procedure ? Clin Exp Allergy 2005;35:970-978

BACKGROUND: In Europe, pollen-related food allergy is the most frequent form of food allergy in adults. Reliability of current diagnostic procedures, however, is poor and therapeutic options are not available . OBJECTIVES: In the present study, we created a panel of recombinant allergens from carrot and evaluated its potential in component-resolved in vitro diagnosis of carrot allergy . METHODS: Recombinant (r) Dau c 1.0104, Dau c 1.0201 and Dau c 4 were cloned by a polymerase chain reaction strategy, expressed in Escherichia coli and purified. Carrot lipid transfer protein (LTP) was expressed in the yeast Pichia pastoris. Sera from 40 carrot-allergic patients were investigated. Twenty-one birch pollen-allergic subjects with negative open provocation to carrot and 20 non-allergic subjects were included as controls. IgE binding to recombinant allergens as well as to cross-reactive carbohydrate determinants (CCD) was measured by ELISA. Cross-reactivity between Dau c 1 isoforms and Bet v 1 was assayed by ELISA inhibition. Biological activity of the recombinant carrot allergens was assessed by histamine release assay and peripheral blood mononuclear cells stimulation . RESULTS: Ninety-eight percent of the carrot-allergic patients were positive to at least one recombinant allergen; 98% reacted to rDau c 1.0104, 65% to rDau c 1.0201, 38% to rDau c 4 and 20% had IgE against CCD. Specificity using the recombinant allergens was high when compared with non-allergic controls, but low compared with birch-sensitized subjects without carrot allergy. Sensitization to Dau c 1.0201, however, proved to be highly specific for clinically relevant sensitization. Inhibition assays indicated the absence of LTP in carrot root extract, and epitope diversity between Dau c 1.0104, Dau c 1.0201 and Bet v 1 . CONCLUSIONS: Our panel of recombinant allergens from carrot can provide a standardized tool for in vitro diagnosis of carrot allergy, and for epitope studies.

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[67] - Asero R, Mistrello G, Roncarolo D, Amato S. Detection of Some Safe Plant-Derived Foods for LTP-Allergic Patients. Int Arch Allergy Immunol 2007;144:57-63

BACKGROUND: Lipid transfer protein (LTP) is a widely cross-reacting plant pan-allergen. Adverse reactions to Rosaceae, tree nuts, peanut, beer, maize, mustard, asparagus, grapes, mulberry, cabbage, dates, orange, fig, kiwi, lupine, fennel, celery, tomato, eggplant, lettuce, chestnut and pineapple have been recorded . OBJECTIVE: To detect vegetable foods to be regarded as safe for LTP-allergic patients . METHODS: Tolerance/intolerance to a large spectrum of vegetable foods other than Rosaceae, tree nuts and peanut was assessed by interview in 49 subjects monosensitized to LTP and in three distinct groups of controls monosensitized to Bet v 1 (n = 24) or Bet v 2 (n = 18), or sensitized to both LTP and birch pollen (n = 16), all with a history of vegetable food allergy. Patients and controls underwent skin prick test (SPT) with a large spectrum of vegetable foods. The absence of IgE reactivity to foods that were negative in both clinical history and SPT was confirmed by immunoblot analysis and their clinical tolerance was finally assessed by open oral challenge (50 g per food) . RESULTS: All patients reported tolerance and showed negative SPT to carrot, potato, banana and melon; these foods scored positive in SPT and elicited clinical symptoms in a significant proportion of patients from all three control groups. All patients tolerated these four foods on oral challenge. Immunoblot analysis confirmed the lack of IgE reactivity to these foods by LTP-allergic patients . CONCLUSION: Carrot, potato, banana and melon seem safe for LTP-allergic patients. This finding may be helpful for a better management of allergy to LTP.

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Les allergènes des Apiacées

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La carotte

Le céleri