La carotte

[1] - 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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[2] - Augustin-Aubide MP, Martinez-Cocera C, Alonso-Llamazares A, Robledo T, Lombardero M, Dominguez J, et al. Diagnostic approach to anaphylaxis by carrot, related vegetables and horsetail (Equisetum arvense) in a homemaker. Allergy 2004;59:786-791

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[3] - 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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[4] - Moreno-Ancillo A, Gil-Adrados AC, Cosmes PM, Dominguez-Noche C, Pineda F. Role of Dau c 1 in three different patterns of carrot-induced asthma. Allergol Immunopathol (Madr) 2006;34:116-120

OBJECTIVE: To assess the role of Dau c 1 in three patients with carrot induced asthma. MATERIAL AND METHODS: Patient 1 had asthma when handling raw carrots. Sensitization to pollens wasn't detected. Patient 2 had rhinoconjunctivitis due to grass and olive pollen allergy. She had asthma when handling raw carrots. Patient 3 was diagnosed of rhinoconjunctivitis and asthma due to allergic sensitization to mites, several pollens and cat. She had asthma due to raw carrot ingestion and inhalation. IgE immunobot analysis and ELISA inhibition assay were used to investigate the allergens and specific antibodies. RESULTS: IgE Immunoblot Analysis: Dau c 1 from carrot extract and the recombinant rDau c 1 were recognized by IgE from patients 1 and 2. Band of Bet v 1 in birch pollen extract wasn't recognized. Patient 3 didn't recognize any of these allergens. Specific IgE to rDau c 1 was measured by ELISA. Specific IgE ELISA-inhibition with carrot as solid phase showed an intermediate inhibition (30 %) between carrot and rDau c 1 in patient 1; and a considerable inhibition (nearly 100 %) between carrot and rDau c 1 in patient 2. No inhibition was found in patient 3. Specific IgE ELISA inhibition between rDau c 1 and rBet v 1, employing rDau c 1 as solid phase was made in patients 1 and 2. Bet v 1 showed less than 40 % of inhibition of rDau c 1 in patient 1; and an intermediate inhibition (> 40 %) between rBet v 1 and rDau c 1 in patient 2. CONCLUSIONS: Airborne carrot allergens are able to sensitize without the implication of a previous pollen allergy. Dau c 1 was the main allergen in patient 2. In patient 1, there was a band of 30 kd that looks like the predominant allergen. Patients 1 and 2 were sensitized directly from carrot allergens. In patient 3, Dau c 1 isn't related to the carrot allergy. Allergy to carrot in patient 3 seems to be related to her allergy to different pollens; however, it wasn't related to birch pollen. Mediterranean countries didn't show the same patterns of food-related pollen allergy than Nordic countries.

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[5] - Quirce S, Blanco R, Diez-Gomez ML, Cuevas M, Eiras P, Losada E. Carrot-induced asthma: immunodetection of allergens. J Allergy Clin Immunol 1997;99:718-719

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[6] - 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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[7] - 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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[8] - Wüthrich B. Lethal or life-threatening allergic reactions to food. J Investig Allergol Clin Immunol 2000;10:59-65

Fatal or life-threatening anaphylactic reactions to food occur in infants, children and adults. Atopic individuals with bronchial asthma and prior allergic reactions to the same food are at a particularly high risk, whereby even the mere inhalation of the allergenic food can be fatal. Not only peanuts, seafood and milk can induce severe, potentially lethal anaphylaxis, but indeed a wide spectrum of foods, according to the different patterns of food sensitivity in different countries. Foods with "hidden" allergens and meals at restaurants are particularly dangerous for patients with food allergies. Similarly, schools, public places and restaurants are the major places of risk. However, the main factor contributing to a fatal outcome is the fact that the victims did not carry their emergency kit with adrenaline (epinephrine) with them. In cases of death where food anaphylaxis is suspected, it is important for forensic reasons to preserve uneaten portions of the food in order to identify (hidden) allergens. It is also important to determine postmortem specific serum IgE, tryptase and histamine levels to document the anaphylaxis. There is a need to raise awareness of the diagnosis and treatment of anaphylaxis among doctors, those called upon to administer emergency medical care, and the public, and also to provide increased support for those with potentially fatal food allergies through the help of patients' organizations, and national and international medical societies. The food industry should ensure a policy of comprehensive labelling of ingredients so that even the smallest amount of potentially lethal foodstuffs can be clearly identified. Finally, the pharmaceutical industry should be persuaded to reintroduce an adrenaline inhaler onto the market.

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[9] - Hoffmann-Sommergruber K, O'Riordain G, Ahorn H, Ebner C, Dacamara Machado ML, Puhringer H, et al. Molecular characterization of Dau c1, the Bet v1 homolous protein from carrot and its cross reactivity with Bet v1 and Api g1. Clin Exp Allergy 1999;29:840-847

BACKGROUND: Up to 70% of patients with birch pollen allergy exhibit the so-called oral allergy syndrome, an IgE-mediated food allergy. The most frequent and therefore best characterized pollen-fruit syndrome is apple allergy in patients suffering from tree pollen-induced pollinosis. The occurrence of adverse reactions to proteins present in vegetables such as celery and carrots in patients suffering from pollen allergy has also been reported. cDNAs for Bet v 1 homologous proteins have been cloned from celery, apple and cherry. Objective The aim of the study was to identify Bet v 1 homologues from carrot (Daucus carota), to isolate the respective cDNA, to compare the IgE-binding capacity of the natural protein to the recombinant allergen and determine the cross-reactivity to Api g 1 and Bet v 1. METHODS: Molecular characterization of the carrot allergen was performed using IgE-immunoblotting, cross-inhibition assays, N-terminal sequencing, PCR-based cDNA cloning and expression of the recombinant protein in Escherichia coli. RESULTS: A 16-kDa protein from carrot was identified as a major IgE-binding component and designated Dau c 1. Sequencing corresponding cDNAs revealed three extremely similar sequences (Dau c 1.1, 1.2 and 1.3) with an open reading frame of 462 bp coding for 154 amino acid residues. CONCLUSIONS: Purified recombinant Dau c 1.2 was tested in immunoblots displaying IgE-binding capacity comparable to its natural counterpart. Cross-inhibition assays verified the existence of common B-cell epitopes present on Dau c 1, Api g 1 as well as on Bet v 1.

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

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

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[11] - Asero R, Mistrello G, Roncarolo D, Amato S, Van Ree R. A case of allergy to beer showing cross-reactivity between lipid transfer proteins. Ann Allergy Asthma Immunol 2001;87:65-67

BACKGROUND: Lipid transfer proteins (LTPs) are highly conserved proteins present in a broad spectrum of fruits and vegetables that might represent a novel plant panallergen . OBJECTIVE: To demonstrate that LTP is an important allergen in beer and that beer LTP cross-reacts with LTP from botanically unrelated plant-derived foods . METHODS: Serum from a patient with clinical allergy to both beer and Rosaceae was studied for IgE reactivity to LTP to several vegetable foods by RAST, ELISA, immunoblot, and inhibition studies . RESULTS: Patient's serum showed a strong IgE reactivity to LTP purified from peach peel, carrot, and broccoli, and to a 10 kD protein in both apple and peach immunoblots, whereas no reactivity to birch cross-reactive allergens such as Bet v 1, profilin, or carbohydrates was found. In inhibition studies, preabsorption of serum with apple, walnut, hazelnut, peanut, corn, and rice caused a fall of 97%, 20%, 66%, 91%, 94%, and 93%, respectively, of its reactivity to peach LTP. Beer RAST fell from 1.8 IU/mL to <0.1 IU/mL when a patient's serum was preabsorbed with recombinant carrot LTP . CONCLUSIONS: LTP is a relevant allergen in beer. Beer LTP may cross-react with LTP from several other plant-derived foods.

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[12] - 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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[13] - 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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[14] - Meijer EA, de Vries SC, Sterk P, Gadella DW Jr, Wirtz KW, Hendriks T. Characterization of the non-specific lipid transfer protein EP2 from carrot (Daucus carota L.). Mol Cell Biochem 1993;123:159-166

The extracellular protein EP2 was previously identified as non-specific lipid transfer protein based on its cDNA-derived amino acid sequence. Here, the purification of the EP2 protein from the medium of somatic embryo cultures is described. After two cycles of ion-exchange and gel permeation chromatography, a single silver-stained protein band with an apparent molecular mass of 10 kDa was observed on SDS-PAGE. This protein band was recognized by the antiserum raised against a EP2-beta-galactosidase fusion-protein. Employing a fluorescent phospholipid analog, it was shown that the purified EP2 protein is capable of binding phospholipids and is able to enhance their transfer between artificial membranes. Employing a gel permeation assay, it could be demonstrated that the EP2 protein is also capable of binding palmitic and oleic acid as well as oleyl-CoA. Because in plants these fatty acids are used as precursor molecules for cutin, these results are in support of the proposed role of the EP2 protein to transport cutin monomers from their site of synthesis through the cell wall of epidermal cells to sites of cutin polymerization.

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[15] - 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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[16] - Fujita C, Moriyama T, Ogawa T. Identification of cyclophilin as an IgE-binding protein from carrots. Int Arch Allergy Immunol 2001;125:44-50

BACKGROUND: Plant food allergies have been associated with pollenosis, although most of the causative allergens are as yet undefined. It is important to elucidate the properties of plant food allergens in order to minimize a patient's risks in food selection. The purpose of the present study was to examine and characterize the IgE-binding proteins in carrots as possible allergens by using patients' sera. METHOD: IgE-binding proteins in carrot extract were screened by an immunoblot technique using sera of patients with atopic dermatitis (selected based upon a case history of food allergies). An allergenic protein was purified from carrot extract by chromatographic procedures. The N-terminal amino acid sequence of allergenic protein was determined and subjected to a computer homology search. Cross-reactivity between carrot and birch allergens was examined by immunoblotting. RESULTS AND CONCLUSION: A unique, approximately 20-kDa allergenic protein that reacted with about 14% of patients' sera was isolated and characterized. The N-terminal amino acid sequence of the purified protein was found to be homologous with those of plant cyclophilins. This allergen exhibited a peptidyl-prolyl cistrans isomerase activity, which was inhibited by the conjugation of cyclosporin A. These properties of the allergenic protein isolated from carrot identified it as a cyclophilin, a possible plant food allergen. No cross-reactivity between this 20-kDa carrot allergen and Bet v 7, a birch pollen cylcophilin, was observed.

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[17] - 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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[18] - 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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[19] - 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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[20] - Wüthrich B, Dietschi R. [The celery-carrot-mugwort-condiment syndrome: skin test and RAST results]. Schweiz Med Wochenschr 1985;115:258-264

Celery is a frequent food allergen: not only raw, but also cooked and as a spice it can produce various reactions of immediate type, from oral contact urticaria to anaphylactic shock. Most celery-allergic patients suffer from hay fever and show a skin sensitization to mugwort. An associated allergy to several spices is quite common, and therefore the term "celery-mugwort-spice-syndrome" has been proposed. The authors have investigated 35 patients, 85% of them women, in whom a current celery allergy of varying severity was diagnosed during the first 8 months of 1984. A positive skin test to celery was seen in 16 patients who denied any symptoms from it. Thorough skin testing and RAST screening with different celery preparations, other foodstuffs and mugwort-pollens were performed on all patients. The modified prick test with native celery-root proved to be the best method for detecting celery sensitization, showing a positive result in 88.6%. The scratch test with celery-salt was positive in 70.5%, intracutaneous testing with commercial extract in 63.5% and the RAST with celery-sticks in 66% of the patients. Sensitization to mugwort was absent in 8 patients only. The mugwort-sensitive group frequently showed a positive test to ragweed-pollens, a common plant in North America, and this appears to indicate cross-reactivity amongst the family of the Asteraceae. Cross-reactivity amongst the Apiaceae is the cause of the many positive results obtained with carrot, parsely, anise, fennel and caraway, the carrot allergy being of clinical importance in 50% of cases, including one with a history of anaphylactic shock after ingestion of raw carrots.

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[21] - 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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[22] - 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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[23] - Lahti A, Hannuksela M. Hypersensitivity to apple and carrot can be reliably detected with fresh material. Allergy 1978;33:143-146

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

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[24] - 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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[25] - Kazemi-Shirazi L, Pauli G, Purohit A, Spitzauer S, Fröschl R, Hoffmann-Sommergruber K, et al. Quantitative IgE inhibition with purified recombinant allergens indicate pollen-derived allergens as the sensitizing agents responsible for many forms of plant food allergy. J Allergy Clin Immunol 2000;105:116-125

Type I allergic symptoms in the oropharyngeal mucosa upon contact with plant-derived food in patients with pollen allergies have been termed oral allergy syndrome (OAS). IgE cross-reactivity between pollen and food allergens represents the molecular basis for this phenomenon. The sensitizing allergen source (pollen or plant food) in OAS is a controversial issue. OBJECTIVE: We sought to determine the primary sensitizing molecules in patients with OAS. METHODS: We used recombinant birch pollen (rBet v 1 and rBet v 2) and plant food allergens (apple, rMal d 1; celery, rApi g 1; and carrot, rDau c 1), as well as natural pollen (birch and timothy grass) and plant food (apple, peach, kiwi, hazelnut, celery, and carrot) allergens, to identify cross-reactive allergens by using qualitative immunoblot inhibitions. In addition, we determined the percentage of plant food-specific IgE that can be preadsorbed with recombinant and natural pollen allergens by quantitative RAST inhibitions by using sera from 71 patients with OAS. RESULTS: Preincubation of sera with recombinant and natural pollen allergens led to an almost complete inhibition of IgE binding to plant food allergens in Western blots, as well as in RAST inhibition experiments. In contrast, recombinant plant food allergens poorly inhibited IgE binding to Bet v 1. CONCLUSION: Most IgE epitopes in plant food recognized by patients with OAS are resembled by pollen allergens. Thus pollen allergens may be responsible for the elicitation and maintenance of OAS.

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[26] - Moneo I, Gomez M, Sanchez-Monge R, Alday E, Delasheras M, Esteban I, et al. Lack of cross-reaction with Bet v1 in patients sensitized to Dau c1, a carrot allergen. Ann Allergy Asthma Immunol 1999;83:71-75

BACKGROUND: Pollen-related food allergies to fresh fruit and vegetables are a well-known clinical phenomenon. Allergens related to Bet v 1 are responsible for these cross-reactions. OBJECTIVE: To characterize the allergen recognized by four carrot-allergic patients. METHODS: Sera from four patients showing strong immediate systemic reactions after contact or ingestion of raw carrot were studied by immunoblotting. The 18-kD allergen, named Dau c 1, was isolated by ethanol precipitation and specific extraction after SDS-PAGE and its N-terminal amino acid sequence was determined. RESULTS: All the patients had significant levels of specific IgE to carrot, but no specific IgE to birch pollen was detected in any of them. IgE immunodetection with the sera only recognized a single band of around 18 kD in raw carrot and in celery (with weaker reaction). No reactive band was found with birch pollen. These results were confirmed using a polyclonal anti-carrot antiserum. The carrot IgE-binding protein had a pl of 4.2 and its N-terminal sequence was homologous to that of Bet v 1 and to allergens previously described in celery and other foods. The four patients studied were not sensitized to birch pollen and three of them tolerated fruit ingestion. CONCLUSION: The whole study indicated that a sensitization to Dau c 1 induces IgE antibodies that do not cross-react with birch pollen allergens.

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[27] - Ebner C, Hirschwehr R, Bauer L, Breiteneder H, Valenta R, Ebner H, et al. Identification of allergens in fruits and vegetables: IgE cross-reactivities with the important birch pollen allergens Bet v 1 and Bet v 2 (birch profilin). J Allergy Clin Immunol 1995;95:962-969

In this study serum samples collected from 20 patients with birch pollen allergy were investigated. All patients had experienced allergic symptoms after contact with or ingestion of particular fresh fruits and vegetables known as birch pollen-related foods. METHODS: Serum samples were tested by means of immunoblotting for IgE reactivities with proteins in extracts of birch pollen, apple, pear, celery, carrot, and potato. Anti-Bet v 1 and anti-Bet v 2 antibodies were used to investigate cross-reactivity. Inhibition studies were performed by preincubation of sera with recombinant Bet v 1 and Bet v 2. RESULTS: IgE binding to proteins, corresponding to the major birch pollen allergen Bet v 1 and to Bet v 2 (birch pollen profilin) could be observed. An allergen homologous to Bet v 1 could be detected in apple, pear, and celery when a Bet v 1-specific monoclonal antibody was used. Testing a polyclonal rabbit anti-Bet v 2 antibody with extracts of the respective plants revealed the presence of profilins in every source tested. Inhibition with recombinant Bet v 1 and Bet v 2 led to complete blocking or marked reduction of IgE binding to proteins of comparable molecular weights in the respective food extracts, indicating IgE cross-reactivity. CONCLUSION: Our results indicate that many plant-derived food agents contain proteins with high homology to the birch pollen allergens Bet v 1 and Bet v 2 and must therefore be considered as potentially threatening for patients with tree pollen allergy.

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[28] - 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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[29] - Cuesta J, Blanco C, Fernández-Rivas M, Villalba M. Vegetalia. J Investig Allergol Clin Immunol 2006;16(Suppl. 2):40-44

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[30] - Chalmet P, Chalmet C. Un cas d'anaphylaxie à la carotte par allergie croisée aux pollens de Composées dont l'ambroisie. Alim'Inter 2008;13:34

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[31] - Jordan-Wagner DL, Whisman BA, Goetz DW. Cross-allergenicity among celery, cucumber, carrot, and watermelon. Ann Allergy 1993;71:70-79

Cross-reactive allergens may be responsible for the clustering of food allergies seen in patients hypersensitive to fruits and vegetables. The pooled sera of six individuals were used to investigate cross-antigenicity among freshly prepared extracts of celery (Cy), cucumber (Cc), carrot (Ct), and watermelon (W). Each patient demonstrated clinical allergy to one or more study foods and, with the exception of Ct in two cases, had IgE to all four extracts by skin test or ELISA. In comparisons of each food against itself and the other three antigens, ELISA inhibition assays demonstrated allergenic similarity among Cy, Cc, Ct, and W by their similar slopes and 50% inhibition concentrations (2.0-7.3 micrograms/mL) In contrast, mountain cedar pollen (MC) produced at 50% inhibition of each food which was 10-fold higher (26.9-70.8 micrograms/mL) and had a flatter slope. Immunoblots of individual sera showed a 15-kD protein band common to all four foods. Pooled sera immunoblot inhibitions (100 and 5 micrograms/mL) demonstrated mutual inhibition of all bands in each of the four foods with the exception of a 28-kD protein of W uniquely inhibited by itself. We conclude that Cy, Cc, Ct, and W possess shared antigens that may account for clustering of these food allergies in patients.

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[32] - Quirce S, Blanco R, Diez-Gomez ML, Cuevas M, Eiras P, Losada E. Carrot-induced asthma: immunodetection of allergens. J Allergy Clin Immunol 1997;99:718-719

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[33] - Fernández-Rivas M, Gonzalez-Mancebo E, van Leeuwen WA, Leon F, van Ree R. Anaphylaxis to raw carrot not linked to pollen allergy. Allergy 2004;59:1239-1240

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[34] - Moneo I, Gomez M, Sanchez-Monge R, Alday E, Delasheras M, Esteban I, et al. Lack of cross-reaction with Bet v1 in patients sensitized to Dau c1, a carrot allergen. Ann Allergy Asthma Immunol 1999;83:71-75

BACKGROUND: Pollen-related food allergies to fresh fruit and vegetables are a well-known clinical phenomenon. Allergens related to Bet v 1 are responsible for these cross-reactions. OBJECTIVE: To characterize the allergen recognized by four carrot-allergic patients. METHODS: Sera from four patients showing strong immediate systemic reactions after contact or ingestion of raw carrot were studied by immunoblotting. The 18-kD allergen, named Dau c 1, was isolated by ethanol precipitation and specific extraction after SDS-PAGE and its N-terminal amino acid sequence was determined. RESULTS: All the patients had significant levels of specific IgE to carrot, but no specific IgE to birch pollen was detected in any of them. IgE immunodetection with the sera only recognized a single band of around 18 kD in raw carrot and in celery (with weaker reaction). No reactive band was found with birch pollen. These results were confirmed using a polyclonal anti-carrot antiserum. The carrot IgE-binding protein had a pl of 4.2 and its N-terminal sequence was homologous to that of Bet v 1 and to allergens previously described in celery and other foods. The four patients studied were not sensitized to birch pollen and three of them tolerated fruit ingestion. CONCLUSION: The whole study indicated that a sensitization to Dau c 1 induces IgE antibodies that do not cross-react with birch pollen allergens.

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[35] - Bollen M, Wichers H, Helsper J, Savelkoul H, van Boekel M. Heat stability of PR-10 isoform mixtures, purified from birch, celery and carrot. 3rd International Symposium on Molecular Allergology, Salzburg, April 18-20, 2008, Poster n°32

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[36] - Fiocchi A, Bouygue GR, Sarratud T, Terracciano L, Martelli A, Restani P. Clinical tolerance of processed foods. Ann Allergy Asthma Immunol 2004;93(5 Suppl. 3):S38-S46

OBJECTIVE: To review the effects of technological processing on selected foods of relevance to childhood allergy from the viewpoints of reduced allergenicity, contamination of processed foods by allergens introduced during processing, and ad hoc technologies to produce reduced hypoallergenic products. DATA SOURCES: We searched the literature (PubMed/MEDLINE) for articles published between January 1994 and April 2004 using the following keywords: food allergy AND process* OR heat* OR cooking OR toleran*. STUDY SELECTION: We drew on our collective clinical and biological experience to restrict retrieved studies to those of more frequent relevance to a hospital allergy practice. RESULTS: Comparatively few clinical studies address the modification of allergenicity of food through cooking or processing. Dairy foods are largely unaffected by processing and may be contaminated by, or themselves become, hidden allergens. Hypoallergenic formulas based on milk, soy, or rice and homogenized beef are successful applications of allergenicity reduction via technological processing. Egg, fish, condiments, and vegetables all carry heat-resistant allergens and should also be considered contaminants. Cereals and bakery products are generally well tolerated, but their allergenicity may be enhanced by processing; the case of rice is still open. Peanut allergens are stable, and the evidence is scant that thermal processing affects the allergenicity of soybean and soy hydrolysates. The debate is ongoing about the tolerance of vegetable oils. CONCLUSIONS: It is too early to systematize clinical studies based on single procedures. Processing affects antigenicity, but this does not always translate into safety recommendations. Industrial processing is liable to contamination, and monitoring and labeling are industry priorities. Clinicians should evaluate foods by as complete a workup as possible before recommending processed foods.

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[37] - 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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[38] - 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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[39] - 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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[40] - García González MC, de la Hoz Caballer B, Cerecedo I, Carnés J, Fernández-Caldas E. Collection and identification of aerosolised proteins during the boiling process of vegetables and legumes. EAACI 23th Congress, Amsterdam, 12-16 June, 2004, Poster n°964

Background: Although most food allergic reactions are caused by ingestion, IgE mediated respiratory reactions caused by the inhalation of vapours from boiling vegetables and legumes have been described. In these cases, IgE-mediated mechanisms have been suggested, but the responsible allergens have not been fully characterised. The aims of this study were to develop a method to collect allergens from vegetable vapours and to investigate if these proteins are capable of binding specific IgE. Material and Methods: Three types of extracts (A, B and C) were prepared with potato, carrot, Swiss chard and green beans. Extracts A (raw), extracted in PBS 0.01 M (1:2 w/v); Extracts B (boiled), cooked in PBS 0.01 M (1:2 w/v) for 30 minutes; and Extract C (lyophilised vapours), collected during the boiling process. The vapours were passed through a refrigeration column and the drops collected and freeze-dried. Raw and boiled extracts were extracted for 4 hours, dialyzed against bidistilled water in a 3.5 kDa cut-off dialysis membrane, frozen and freeze-dried. Antigenic profile of the extracts was evaluated by SDS-PAGE and analysed by scanning densitometry. Results: Several bands were visualised in the raw extracts in the molecular weight range of 9 to 90 KDa in the 4 analysed raw extracts. The protein profile of the boiled extracts showed low molecular weights bands, which could correspond to denatured proteins from the raw extracts. The total yield of vapour proteins obtained in each extract was 0,64% in potato, 0,48% in carrot, 0,4% in Swiss chard and 0,352% in green bean. Vapours extracts (Extracts C) showed several prominent bands; 1 band in potatoes (35 kDa), 2 bands in carrot vapours (60 and a 66 kDa), 2 bands in the Swiss chard vapours (62 and 71 kDa) and 1 band (38 kDa) in green bean vapours. Almost all these molecules had a corresponding band in their respective raw extracts. Faint IgE binding was observed to green bean vapours. Conclusions: We have developed a reproducible method to collect vapours during the boiling process of foods. The results suggest that proteins are present in vapour drops aerosolised during the boiling process. More studies are necessary to elucidate if these proteins are capable of inducing respiratory symptoms and to what extent these proteins are denatured and capable of binding specific IgE.

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[41] - 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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[42] - Erdmann SM, Sachs B, Schmidt A, Merk HF, Scheiner O, Moll-Slodowy S, et al. In vitro Analysis of Birch-Pollen-Associated Food Allergy by Use of Recombinant Allergens in the Basophil Activation Test. Int Arch Allergy Immunol 2005;136:230-238

BACKGROUND: Basophil activation is associated with the expression of CD63. In birch-pollen-associated food allergy to celery, carrot and apple, Bet v 1, Api g 1, Dau c 1 and Mal d 1 are major allergens. Recombinant allergens have not yet been used in the CD63-based basophil activation test (BAT) . OBJECTIVE: To evaluate the feasibility of using recombinant allergens in the BAT in the diagnosis of allergy to apple, carrot and celery and to compare results with routine tests, i.e. skin prick tests (SPTs) and specific IgE . METHODS: Thirty-two patients with an oral allergy syndrome induced by apple, carrot or celery and 22 controls were studied. SPTs were performed with native foods. Specific IgE was determined by the CAP method and basophil activation by flowcytometry upon double staining with anti-IgE/anti-CD63 monoclonal antibodies after incubating with purified recombinant Bet v 1, Bet v 2, Api g 1, Dau c 1 and Mal d 1 . RESULTS: By the combined use of the BAT and the CAP method, sensitization to Bet v 1 and Bet v 2 was detected in 100 and 25% of all subjects, respectively. Sensitivity of specific IgE for apple, carrot and celery was 60, 70 and 75% with corresponding specificities of 64, 86 and 82%. Sensitivity of the BAT for Mal d 1, Dau c 1 and Api g 1 was 75, 65 and 75% with corresponding specificities of 68, 100 and 77% . CONCLUSIONS: The BAT using recombinant allergens provides a valuable new in vitro method for the detection of sensitization to foods. Although double-blind placebo-controlled food challenges remain the gold standard to confirm food allergy, the CD63-based BAT with recombinant allergens may supplement routine tests for allergy diagnosis.

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[43] - Ballmer-Weber BK, Weber JM, Vieths S, Wüthrich B. Predictive value of the sulfidoleukotriene release assay in oral allergy syndrome to celery, hazelnut, and carrot. J Investig Allergol Clin Immunol 2008;18:93-99

BACKGROUND: Patients sensitized to birch pollen frequently suffer from a food allergy to plant foods such as celery, carrots, or hazelnut. One of the main manifestations of birch pollen-related food allergy is the oral allergy syndrome. Skin tests and allergen-specific immunoglobulin (Ig) E determinations are poor predictors of such reactions when assessed by double-blind placebo-controlled food challenge (DBPCFC). OBJECTIVE: To investigate whether a cellular test based on leukotriene release from basophils, the cellular antigen stimulation test in combination with enzyme-linked immunosorbent assay (CAST-ELISA), is predictive of pollen-related food allergy. METHODS: Birch pollen-sensitized patients with positive DBPCFC to celery (n=21), hazelnut (n=15), and carrot (n=7) underwent skin tests along with determination of specific IgE and CAST-ELISA for the respective allergens. The results were compared with those of 24 birch pollen-sensitized patients with negative open food challenge to celery, hazelnut, and carrot. RESULTS: While skin prick tests had a sensitivity of 85%, 80%, and 29% for commercial extracts of celery, hazelnut, and carrot, respectively, prick testing with self-prepared extracts yielded sensitivities of 100%, 80%, and 100%, respectively. For specific IgE determinations, sensitivities were 71%, 73%, and 57%, respectively, and the respective specificities were 67%, 73%, and 60%. For CAST-ELISA with various sources and doses of allergens, the sensitivity varied from 71% to 95% for celery, 73% to 80% for hazelnut, and 43% to 86% for carrot. The respective specificities were 67% to 92%, 75% to 88%, and 77% to 91%. Analysis of the predictive value of CAST-ELISA with receiver operating characteristic curves showed that the results of the tests were more predictive of pollen-related food allergy than quantitative allergen-specific IgE determinations. CONCLUSIONS: CAST-ELISA is more specific than routine diagnostic tests for the diagnosis of pollen-related food allergy to celery, hazelnut, and carrot.

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[44] - Sturm A. Heterogeneity of the complex N-linked oligosaccharides at specific glycosylation sites of two secreted carrot glycoproteins. Eur J Biochem 1991;199:169-179

The N-linked glycans from the 52/54-kDa medium protein and cell wall beta-fructosidase, two glycoproteins secreted by carrot suspension culture cells, were characterized. Carrot cells were labelled with [3H]glucosamine or [3H]fucose. The 52/54-kDa medium protein was isolated from the culture medium and beta-fructosidase from cell walls. The purified proteins were digested with trypsin and glycopeptides were isolated and sequenced. Glycans obtained from individual glycopeptides were separated by gel filtration chromatography and characterized by concanavalin A chromatography, by treatments with exoglycosidases and by sugar composition analysis. The 52/54-kDa medium protein and cell wall beta-fructosidase have one high-mannose-type glycan similar to those from yeast and animal glycoproteins. In addition, the 52/54-kDa medium protein has three complex-type and cell wall beta-fructosidase two complex-type glycans per polypeptide. The complex-type glycans isolated from individual glycosylation sites are fairly large and very heterogeneous. The smallest of these glycans has the structure [Xyl](Man)3[Fuc](GlcNAc]2Asn (square brackets indicating branching) whereas the larger ones carry additional sugars like terminal N-acetylglucosamine and possibly rhamnose and arabinose in the case of the 52/54-kDa medium protein and only arabinose in the case of cell wall beta-fructosidase. These terminal sugars are linked to the alpha-mannose residues of the glycan cores. The 52/54-kDa medium protein is secreted with large and homogeneous complex glycans, their heterogeneity originates from slow processing after secretion. The complex glycans from cell wall beta-fructosidase are processed before the enzyme is integrated into the cell wall.

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Carotte crue vs carotte cuite

Diagnostic moléculaire

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