Les Brassicacées : généralités

Genre	Espèce	Sous-espèce	Français	Anglais
Brassica	oleracea	italica	brocoli	broccoli
capitata	chou commun	cabbage
gemmifera	chou de Bruxelles	Brussels sprouts
botrytis	chou romanesco	romanesco broccoli
cauliflora	chou-fleur	cauliflower
gongyloides	kohlrabi	kohlrabi
campestris	chinensis	pak-choi	chinese cabbage
rapa	oleifera	navette	turnip rape
rapa	navet	turnip
napus	oleifera	colza	rape
napobrassica	rutabaga	swede
juncea		moutarde de Chine	oriental mustard
nigra		moutarde noire	black mustard
Lepidium	sativum		cresson alénois	garden cress
Nasturtium	officinale		cresson de fontaine	watercress
Sinapsis	alba		moutarde jaune	yellow mustard
Raphanus	sativus		radis	radish
Armoracia	rusticana		raifort	horseradish
Eruca	sativa		roquette	rocketsalad
Capparis	spinosa		câpre	caper

[1] - Chardin H, Mayer C, Sénéchal H, Wal JM, Poncet P, Desvaux FX, et al. Lipid Transfer Protein 1 Is a Possible Allergen in Arabidopsis thaliana. Int Arch Allergy Immunol 2003;131:85-90

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[2] - Sirvent S, Palomares O, Vereda A, Villalba M, Cuesta-Herranz J, Rodríguez R. nsLTP and profilin are allergens in mustard seeds: cloning, sequencing and recombinant production of Sin a 3 and Sin a 4. Clin Exp Allergy 2009;39:1929-1936

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[3] - Nebiolo F, Castiglione G, Bommarito L, Bergia R, Heffler E, Carosso A, et al. Anaphylaxis after ingestion of Pantelleria’s caper (Capparis spinosa’s flower bud). Allergy 2008;63(suppl. 88):658

Background: Capparis spinosa is a small bush belonging to Capperaceae family, Brassicales order, and it is widely cultivated in Mediterranean countries for its flower bud, called Pantelleria‚s caper, which is used as an aromatizing ingredient for a great variety of dishes: from meat, to fish, to pasta recipes. In literature no cases of allergic reactions to Pantelleria‚s caper were described and its allergens have not been identified. Methods: A 33-year-old woman referred to our Allergy Outpatients‚ Clinic reporting an episode of diffuse pruritus, face erythema, eyelids oedema and abdominal pain about 45 min after having ingested several capers. The patient referred to an emergency unit where fexofenadine, chlorophenamine and metil-prednisolon were administred with improvement of itching and erythema in few hours, while eyelids oedema persisted 2 days more. The patient has not drunk any alcoholic or taken any drug before the above mentioned reaction. In her clinical history she referred seasonal oculorhinitis since the age of eight years. Skin Prick Tests (SPT) for may food and inhalant allergens, prick-byprick with Pantelleria‚s caper and specific IgE dosage (CAP-Pharmacia) for food allergens were performed. Results: Allergometric tests were positive for bean (2 mm), peach (4 mm), peanut (4 mm), hazelnut (3 mm), soybean (2 mm), apple (2 mm), grass pollen (8 mm), cynodon pollen (2mm) and Pantelleria‚s caper (8 mm), with an histamine wheal of 7 mm. During prick-by-prick with caper the patient manifested eyelids oedema resolved with a tablet of ebastine. IgE dosage was high for peach (2.41 kU/L), peanut (0.92 kU/L), soybean (0.76 kU/L) and walnut (0.21 kU/L). The patient usually eats peach, peanuts, soybean and walnuts without any allergic manifestation. Double Blind Placebo Controlled Food Challenge with caper was not performed because the patient had a mild systemic reaction after prick-by-prick with caper. Conclusions: In our knowledge this is the fist case of anaphylaxis after ingestion of Pantelleria‚s caper. About the allergenic molecules causing the reaction, the positive tests to peach, peanut and soybean suggest a possible involvement of lipid transfer protein. We are carrying on SDS/PAGE, bidimensional electrophoresis and immunoblotting with the patient serum to characterize food allergens involved in the allergic reaction.

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[4] - Pereira C, Tavares B, Loureiro G, Lundberg M, Chieira C. Turnip and zucchini: new foods in the latex-fruit syndrome. Allergy 2007;62:452-453

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[5] - Hemmer W. The health effects of oilseed rape: myth or reality? No clear evidence that it has adverse effects on health. BMJ 1998;316:1327-1328

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[6] - Hermanides HK, Laheÿ-de Boer AM, Zuidmeer L, Guikers C, van Ree R, Knulst AC. Brassica oleracea pollen, a new source of occupational allergens. Allergy 2006;61:498-502

BACKGROUND: Vegetable pollen is a rare source of occupational allergens. Occupational allergy has only been described in the case of paprika pollen and tomato pollen. We describe a new source of occupational pollen allergy. AIM: To study the incidence and the impact of broccoli and cauliflower pollen allergy in employees involved in classical plant breeding . METHODS: Fifty-four employees of five companies working with cauliflower (Brassica oleracea botrytis) and broccoli (B. oleracea italica/cymosa) pollen were eligible for complete evaluation. Allergy to cauliflower and broccoli pollen was evaluated by questionnaire and determination of sensitization by radioallergosorbent test (RAST) and skin-prick tests (SPT). SPT and RAST were performed with a panel of commercial and homemade extracts from cauliflower and broccoli pollen . RESULTS: Work-related symptoms such as rhinitis, conjunctivitis, asthma and urticaria caused by B. oleracea pollen were reported by 44% of the participants (24/54), of whom all but one had positive SPT for cauliflower- and/or broccoli-pollen/flower extracts and 58% (14/24) had positive RAST results. Symptoms had developed within the first 2 years in 33% of the patients. Six patients had to stop or change work . CONCLUSIONS: Brassica oleracea pollen is a new source of occupational allergen with strong allergenic potential leading to symptoms in almost half of the exposed employees.

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[7] - Anguita JL, Palacios L, Ruiz-Valenzuela L, Bartolomé B, Lopez-Urbano MJ, Saenz de San Pedro B, et al. An occupational respiratory allergy caused by Sinapis alba pollen in olive farmers. Allergy 2007;62:447-450

BACKGROUND: Sinapis alba (white mustard) is a entomophilic species included in the Brassicaceae family. To date it has not been related to allergic sensitization or clinical respiratory disease . METHODS: Twelve olive orchard workers had a history of rhinitis and/or bronchial asthma that occurred during control weed management and/or harvest, from January to March. They underwent skin prick tests (SPT) with S. alba pollen extract and a standard battery of aeroallergens. Sinapis alba pollen extract was prepared for performing quantitative skin tests, enzyme allergosorbent test and nasal challenge test (NCT). A portable monitoring station and an urban volumetric Hirst-type spore trap were used for the aerobiological study . RESULTS: Eleven patients suffered from rhinitis and bronchial asthma and one had only from rhinitis. All patients were sensitized to S. alba pollen extract, and they showed a positive NCT response. In the urban aerobiologic monitoring station the amount of S. alba pollen only exceptionally reached peaks of 21 grains/m(3), whereas in the work environment peaks of 1801 grains/m(3) were detected between 15 February and 7 April . CONCLUSIONS: We demonstrate the existence of a new occupational allergen for olive farmers: S. alba pollen. We point out the importance of perform aerobiological sampling within the occupational environment for the detection and quantification of the allergenic source.

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[8] - Yates B, De Soyza A, Harkawat R, Stenton C. Occupational asthma caused by Arabidopsis thaliana: a case of laboratory plant allergy. Eur Respir J 2008;32:1111-1112

A 36-yr-old male never-smoker with an 8-yr history of hay fever but no past history of asthma undertook a 3-yr research project involving the plant Arabidopsis thaliana. The subject was based in a small laboratory with an attached growing room. After 30 months of research, he began to develop breathlessness within 5-10 min of entering the laboratory. Initial investigations confirmed asthma with airflow obstruction (forced expiratory volume in one second (FEV(1))/forced vital capacity was 3.01/4.75 L; predicted values were 3.67/4.43 L) and increased airway responsiveness. Serial peak expiratory flow measurements showed a work-related pattern. A supervised workplace challenge test led to a fall in FEV(1) from the baseline value of 3.10 L to 1.95 L within 20 min of entering the growing room. Skin-prick solutions were prepared from Arabidopsis leaves and flower heads; positive 4-mm responses were obtained to the flower heads (i.e. to the pollen). Arabidopsis is a member of the Brassicaceae family. It is used extensively in plant biology research as its genome is small, has been fully sequenced and is easily manipulated. The present article represents the first reported case of occupational asthma due to Arabidopsis thaliana.

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[9] - Hemmer W, Focke M, Wantke F, Jager S, Gotz M, Jarisch R. Oilseed rape pollen is a potentially relevant allergen. Clin Exp Allergy 1997;27:156-161

Oilseed rape (Brassica napus) (OSR) is a partly wind-pollinated crop which has been increasingly cultivated both in Europe and overseas. Allergological data about OSR is scarce and controversial. We evaluated the frequency of sensitization to OSR pollen by skin prick test and RAST over a period of 1 yr. Airborne OSR pollen load and the agricultural role of this crop were analysed. Furthermore, six patients were investigated by immunoblot. In 4468 patients with suspect inhalant allergy investigated between June 1994 and May 1995, routine skin prick testing revealed OSR sensitivity in 7.1% of pollen-allergic patients. In all, monovalent sensitization was detected in nine patients. Routine pollen counts showed daily maxima not exceeding 50 grains/m3/24 h, but airborne OSR pollen has continuously increased during the last decade correlating with the increasing acreage. Characterization of OSR allergens by immunoblot revealed major allergens of 6/8 kD, 12/14 kD and in the high molecular weight range at 33, 42, 51, 58/61 and 70 kD. Some OSR proteins may cross-react with birch pollen allergens. In summary, the results suggest that OSR pollen is a moderate but true source of allergy and may sensitize despite low pollen exposure.

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[10] - Muehlmeier G, Kuehn M, Maier H. Increasing danger of allergic rhinitis by oilseed rape pollen. Allergy 2008;63(suppl. 88):597

The increasing cultivation of oilseed rape (OSR, Brassica napus) in Middle Europe in order to gain renewable energy sources promotes the exposition to volatile rape components. OSR is a partially wind pollinating plant with several different allergic proteins. That rises the question of significance of allergic diseases, especially allergic rhinitis, caused by OSR pollen. In a two year period from January 2006 to December 2007 1036 patients showed positive skin prick test reactions on different pollen. In 91 patients an at least one fold positive reaction was found on OSR pollen. These patients were monitored on allergen profiles, cross allergies and prevalence of panallergens.Relevant amounts of IgE antibodies were measured in 37% of the patients. Those had significantly higher total IgE levels, in 79% further pollen allergies with the highest affinity to grass pollen and in 56% IgE antibodies against profilin. 55% showed a positive result in nasal provocation testing with an OSR pollen extract. The distribution of concomitant allergens in OSR pollen allergics indicates the high rate of cross allergies to different pollen species. Solitary OSR pollen allergies were discovered in 6 patients. The great amount of mucosa irritating volatile components seems to play an additional role in the expression of OSR pollen allergic rhinitis.In conclusion the therapy of the cross-linked allergies is the first choice in most of the OSR pollen allergics, but in selected cases a specific immune therapy against OSR pollen is indicated.

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[11] - Parisot L. Sensibilisation aux pollens de maïs et de colza en France. Alim'Inter 2005;10:198-199

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[12] - Muehlmeier G, Kuehn M, Maier H. Increasing danger of allergic rhinitis by oilseed rape pollen. Allergy 2008;63(suppl. 88):597

The increasing cultivation of oilseed rape (OSR, Brassica napus) in Middle Europe in order to gain renewable energy sources promotes the exposition to volatile rape components. OSR is a partially wind pollinating plant with several different allergic proteins. That rises the question of significance of allergic diseases, especially allergic rhinitis, caused by OSR pollen. In a two year period from January 2006 to December 2007 1036 patients showed positive skin prick test reactions on different pollen. In 91 patients an at least one fold positive reaction was found on OSR pollen. These patients were monitored on allergen profiles, cross allergies and prevalence of panallergens.Relevant amounts of IgE antibodies were measured in 37% of the patients. Those had significantly higher total IgE levels, in 79% further pollen allergies with the highest affinity to grass pollen and in 56% IgE antibodies against profilin. 55% showed a positive result in nasal provocation testing with an OSR pollen extract. The distribution of concomitant allergens in OSR pollen allergics indicates the high rate of cross allergies to different pollen species. Solitary OSR pollen allergies were discovered in 6 patients. The great amount of mucosa irritating volatile components seems to play an additional role in the expression of OSR pollen allergic rhinitis.In conclusion the therapy of the cross-linked allergies is the first choice in most of the OSR pollen allergics, but in selected cases a specific immune therapy against OSR pollen is indicated.

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[13] - Hemmer W. The health effects of oilseed rape: myth or reality? No clear evidence that it has adverse effects on health. BMJ 1998;316:1327-1328

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[14] - Welch J, Jones MG, Cullinan P, Coates OA, Newman Taylor AJ. Sensitization to oilseed rape is not due to cross-reactivity with grass pollen. Clin Exp Allergy 2000;30:370-375

Oilseed rape is an important crop grown in the UK which can cause specific immunological sensitization with clinical symptoms in a relatively small number of the general population. Individuals with immunoglobulin (Ig) E-mediated allergy to oilseed rape have also been found to be sensitized to other pollen allergens, most frequently being grass pollen. Cross-reactivity between common grass and oilseed rape would have important implications, especially as their flowering period coincides. OBJECTIVE: We have investigated whether the cosensitization found in individuals sensitized to both oilseed rape and grass pollen is due to cross-reactivity. METHODS: Cross-reactivity between oilseed rape and grass pollen was determined using RAST, RAST inhibition, Western blotting and inhibition studies with Western blotting. RESULTS: Competitive RAST inhibition studies between pollen of oilseed rape and grass failed to show any cross-reactivity between the pollen types. Self-inhibition with oilseed rape resulted in 90% inhibition, whereas there was less than 10% inhibition with grass pollen. Western blotting revealed allergens of similar molecular weight in both oilseed rape and grass pollen. Despite allergens of similar molecular weights being present in both pollen types, inhibition immunoblot studies confirmed that the allergens in the two allergens were immunologically distinct. CONCLUSION: The allergens of oilseed rape and grass pollen, although similar in molecular weights, are immunologically distinct and there is no evidence of cross-reactivity between them. Individuals allergic to grass pollen will not necessarily develop a specific nasal or airway response to inhaled oilseed rape pollens.

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[15] - Chardin H, Mayer C, Sénéchal H, Tepfer M, Desvaux FX, Peltre G. Characterization of high-molecular-mass allergens in oilseed rape pollen. Int Arch Allergy Immunol 2001;125:128-134

BACKGROUND: Oilseed rape pollen allergies have been previously described as the result of cross-sensitization with various pollens. Recently, several proteins have been identified as oilseed rape allergens. The aim of the present work was the characterization of oilseed rape pollen allergens by two-dimensional (2-D) gel analysis and amino acid microsequencing. METHODS: Water extractable proteins from oilseed rape pollen were separated by isoelectrofocusing and then transferred onto a nitrocellulose sheet. Twenty-one human sera from pollen- or mustard-allergic individuals were screened for their reactivity to oilseed rape proteins. Eleven sera possessed IgE which recognized oilseed rape pollen proteins and one serum was selected for further 2-D characterization and amino acid microsequencing of the allergens. RESULTS: The results showed that three molecules from oilseed rape pollen were identified as oilseed rape allergens which have not yet been described. These three proteins were molecules of 70 kD with a pI >8, 40 kD with a pI around 10 and 80 kD with a pI around 5. These proteins displayed identities with the berberine bridge protein, a receptor-like protein kinase and the cobalamin-independent methionine synthetase from Arabidopsis thaliana, respectively. The genes encoding the putative Arabidopsis molecules are located on chromosome 1 (berberine bridge protein) and chromosomes 3 and 4 (receptor-like protein kinases). CONCLUSION: These results show that certain high-molecular-mass proteins from oilseed rape pollen are allergens

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[16] - Chardin H, Mayer C, Sénéchal H, Poncet P, Clément G, Wal JM, et al. Polygalacturonase (pectinase), a new oilseed rape allergen. Allergy 2003;58:407-411

BACKGROUND: Type I hypersensitivity to rapeseed pollen allergens was described as the result of a cross-sensitization with various pollens that could constitute an aggravating factor in birch or grass pollen allergies. Recently, a few rapeseed pollen allergens were described. The aim of the present work was to identify new rapeseed pollen allergens by using two-dimensional gel analysis, microsequencing, and mass spectrometry . METHODS: Water extractable proteins from oilseed rape pollen or stamen were separated by two-dimensional gel electrophoresis. The proteins were then electroblotted onto a nitrocellulose (NC) sheet. The NC sheets were successively incubated with (1) individual human sera pre-selected for their immunoglobulin E (IgE) reactivity to rapeseed pollen proteins, (2) alkaline phosphatase (AP)-conjugated goat anti-human IgE and (3) AP substrate. The allergens localized by this method were then identified by microsequencing and MALDI-TOF mass spectrometry analysis . RESULTS: Of the 18 sera studied, five recognized a wide multispot zone with a molecular mass around 43 kD and pIs between 6.5 and 8.5. The results obtained with two representative sera are shown. From this zone, two isoforms of the polygalacturonase enzyme were identified by microsequencing. Confirmation was obtained through MALDI-TOF mass spectrometry analysis . CONCLUSION: The present results allow the identification of a new rapeseed allergen that can be the main allergen for some patients.

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[17] - Godfrin D, Senechal H, Desvaux F-X, Mayer C, Hennion M-C, Peltre G. Allergomes of rapeseed pollen and seed (Brassica napus). Allergy Clin Immunol Int 2005;17(Suppl. 1):17

Background: Rapeseed is the main oleaginous seed cultivated in Europe and mainly used to produce seed rape oil for human consumption. Seeds and cattle-cake, left over from the oil production, are also used for animal consumption. Some Brassica napus seed components are found in shampoo, beauty care products and washing powder. Methods: Among the patients allergic to both birch and grass pollen some of them are also allergic to rapeseed pollen (which is emitted between the pollen seasons of birch and grasses). Some sera from mustard allergic patient were used to visualize allergens from Brassica napus seeds. Water soluble proteins were extracted by incubation of pollen or ground seeds in distilled water. All remaining water-soluble proteins were removed by extensive washes in distilled water and the pellet was dried. Then, non-water soluble proteins were extracted with a combination of Thiourea, Urea and a detergent, CHAPS. The watersoluble proteins were immobilized on ELISA plates, incubated with allergic patient sera and the immunodetection was performed with a second antibody anti- IgE alkaline phosphatase labelled. More than 400 patient sera were screened for rapeseed pollen or/and seed IgE, IgG4 and total IgG antibodies. IgE positive ones were used to reveal allergens on blot obtained after 1D isoelectrofocusing and 2D gel separations. Results: IgE from 50 patient sera out of 400 recognised water soluble proteins from seed and 31 of these positive sera were used to study water and nonwater soluble proteins from pollen and seed. 10 out of 31 sera were positive with water soluble pollen allergens and 9 out of 31 with non-water soluble allergens. 12 out of 31 sera were positive with water soluble seed allergens and among them 6 were positive with non-water soluble seed allergens. In water pollen extract 7 allergens were detected compared with around twenty allergens evidenced in non-water soluble pollen extract. Likewise, in water seed extract 6 allergens were detected compared with around twenty allergens visualised in non water soluble extract. Amino acid sequencing or mass spectrometry were performed allowing identification of these proteins. Conclusion: Results show that major allergens from Brassica napus pollen and seed have different molecular characteristics using isoelectric point and molecular mass criteria. Likewise hydrophilic and hydrophobic allergen patterns seem to be different.

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[18] - Toriyama K, Hanaoka K, Okada T, Watanabe M. Molecular cloning of a cDNA encoding a pollen extracellular protein as a potential source of a pollen allergen in Brassica rapa. FEBS Lett 1998;424:234-238

A polyclonal antiserum was raised against the extracellular pollen proteins of Brassica rapa and used for screening the expression cDNA libraries made from immature anthers. We obtained five groups of cDNA clones, including cDNAs similar to PCP1, thioredoxin, and lipid transfer protein (LTP). Recombinant protein of the cDNA clone showing sequence similarity to LTP was demonstrated to bind IgE of a patient allergic to Brassica pollen. The cDNA clone reported here, therefore, represents a novel pollen allergen of Brassica rapa.

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[19] - Focke M, Hemmer W, Hayek B, Gotz M, Jarisch R. Identification of allergens in oilseed rape (Brassica napus) pollen. Int Arch Allergy Immunol 1998;117:105-112

BACKGROUND: Pollen from oilseed rape (OSR), Brassica napus, an increasingly cultivated oilplant from the Brassicaceae, has been recognized as a potential cause of allergic sensitization. Allergens have been hardly investigated. METHODS: We characterized IgE binding proteins in OSR pollen by immunoblot, immunoblot inhibition and specific monoclonal antibodies using sera from 89 patients sensitized to OSR. RESULTS: Two low-molecular-weight allergens of 6/8 kD and 14 kD as well as a high molecular-weight cluster (27-69 kD) comprising six cross-reactive peptides could be identified. The three allergens were recognized by 50, 34 and 80% of patients, respectively. Immunoblot IgE binding to OSR could be totally inhibited by rye pollen and moderately by birch pollen (6/8 and 14 kD) while mugwort had little effect. An anti-profilin-specific monoclonal antibody bound specifically to a 14-kD protein in OSR. Binding to the 6/8-kD rape allergen could be effectively inhibited by rAln g 2, a calcium-binding protein from alder. Periodate treatment led to a significant reduction in IgE binding to the 27 to 69-kD OSR allergens indicating that carbohydrate determinants are involved in IgE binding. OSR proteins were capable to quench IgE binding to timothy grass pollen proteins of >/=60 kD suggesting that grass pollen group 4 allergens cross-react with the 27 to 69-kD cluster in OSR. CONCLUSIONS: The data demonstrate that OSR pollen is allergenic and indicate that the identified allergens represent cross-reacting homologues of well-known pollen allergens, i.e. calcium-binding proteins, profilins, and high-molecular-weight glycoproteins. Via cross-reactivity, exposure to OSR pollen may be a prolonging and aggravating factor in underlying birch and grass pollen allergy.

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[20] - Hemmer W, Focke M, Wantke F, Jager S, Gotz M, Jarisch R. Oilseed rape pollen is a potentially relevant allergen. Clin Exp Allergy 1997;27:156-161

Oilseed rape (Brassica napus) (OSR) is a partly wind-pollinated crop which has been increasingly cultivated both in Europe and overseas. Allergological data about OSR is scarce and controversial. We evaluated the frequency of sensitization to OSR pollen by skin prick test and RAST over a period of 1 yr. Airborne OSR pollen load and the agricultural role of this crop were analysed. Furthermore, six patients were investigated by immunoblot. In 4468 patients with suspect inhalant allergy investigated between June 1994 and May 1995, routine skin prick testing revealed OSR sensitivity in 7.1% of pollen-allergic patients. In all, monovalent sensitization was detected in nine patients. Routine pollen counts showed daily maxima not exceeding 50 grains/m3/24 h, but airborne OSR pollen has continuously increased during the last decade correlating with the increasing acreage. Characterization of OSR allergens by immunoblot revealed major allergens of 6/8 kD, 12/14 kD and in the high molecular weight range at 33, 42, 51, 58/61 and 70 kD. Some OSR proteins may cross-react with birch pollen allergens. In summary, the results suggest that OSR pollen is a moderate but true source of allergy and may sensitize despite low pollen exposure.

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[21] - Focke M, Hemmer W, Hayek B, Gotz M, Jarisch R. Identification of allergens in oilseed rape (Brassica napus) pollen. Int Arch Allergy Immunol 1998;117:105-112

BACKGROUND: Pollen from oilseed rape (OSR), Brassica napus, an increasingly cultivated oilplant from the Brassicaceae, has been recognized as a potential cause of allergic sensitization. Allergens have been hardly investigated. METHODS: We characterized IgE binding proteins in OSR pollen by immunoblot, immunoblot inhibition and specific monoclonal antibodies using sera from 89 patients sensitized to OSR. RESULTS: Two low-molecular-weight allergens of 6/8 kD and 14 kD as well as a high molecular-weight cluster (27-69 kD) comprising six cross-reactive peptides could be identified. The three allergens were recognized by 50, 34 and 80% of patients, respectively. Immunoblot IgE binding to OSR could be totally inhibited by rye pollen and moderately by birch pollen (6/8 and 14 kD) while mugwort had little effect. An anti-profilin-specific monoclonal antibody bound specifically to a 14-kD protein in OSR. Binding to the 6/8-kD rape allergen could be effectively inhibited by rAln g 2, a calcium-binding protein from alder. Periodate treatment led to a significant reduction in IgE binding to the 27 to 69-kD OSR allergens indicating that carbohydrate determinants are involved in IgE binding. OSR proteins were capable to quench IgE binding to timothy grass pollen proteins of >/=60 kD suggesting that grass pollen group 4 allergens cross-react with the 27 to 69-kD cluster in OSR. CONCLUSIONS: The data demonstrate that OSR pollen is allergenic and indicate that the identified allergens represent cross-reacting homologues of well-known pollen allergens, i.e. calcium-binding proteins, profilins, and high-molecular-weight glycoproteins. Via cross-reactivity, exposure to OSR pollen may be a prolonging and aggravating factor in underlying birch and grass pollen allergy.

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[22] - Godfrin D, Sénéchal H, Haddad I, Sutra JP, Chardin H, Desvaux FX, et al. Characterization of allergens from rapeseed pollen and seed (Brassica napus). Allergy 2007;62(suppl. 83):293

Background: Rapeseed is the main oleaginous seed cultivated in Europe and used to produce seed rape oil for human consumption. Cattle-cake is used for animal consumption and some Brassica napus seed components are found in beauty care products and washing powder. Methods: Among the patients allergic to both birch and grass pollen some of them are also allergic to rapeseed pollen, which is emitted in between the birch and grasses pollen seasons. Two extraction types were performed: first using water then a combination of thiourea, urea and a detergent, CHAPS. The water-soluble proteins were immobilized on ELISA plates, incubated with allergic patient sera and the immunodetection was performed with labelled second antibodies anti-IgE, IgG4 and total IgG. After IgE screening of more than 400 allergic patient sera, rapeseed positive ones were used to detect allergens on blots from isoelectrofocusing and 2D gel separations. Detected allergens were submitted to mass spectrometry by Maldi-ToF and LC-MS/MS. Results: Among the 400 human sera tested, 31 of the positive sera were used to study water and non-water soluble proteins from pollen and seed separated by IEF. 10 out of 31 sera were positive with water soluble pollen allergens and 9 out of 31 with non-water soluble allergens. 12 out of 31 sera were positive with water soluble seed allergens and among them 6 were positive with non-water soluble seed allergens. After a 2D separation of water pollen extract, 7 allergens were detected compared with around twenty allergens evidenced in non-water soluble extract. Likewise, in water seed extract 6 allergens were detected compared with around twenty allergens visualised in non water soluble extract. Mass spectrometry was performed allowing identification of a few proteins. Some water soluble allergens from rape seed pollen are known since the publications of Chardin et al. (1-2). We identified four new allergens in non water soluble seed extracts corresponding to myrosinase, myrosinase-associated protein, cruciferin and napin. Conclusion: The major allergens from Brassica napus pollen and seeds, water soluble or not, are different, based on their isoelectric point and molecular mass. We have identified 4 new allergens among the non water soluble proteins from Brassica napus seeds. References: 1 Chardin et al. Allergy 2003; 58:407-411 2 Chardin et al. Int. Arch. Allergy & Immunol 2001; 125:128-134

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[23] - Chardin H, Sénéchal H, Wal JM, Desvaux FX, Godfrin D, Peltre G. Characterization of peptidic and carbohydrate cross-reactive determinants in pollen polysensitization. Clin Exp Allergy 2008;38:680-685

BACKGROUND: Cross-reactivity may be due to protein sequence or domain homologies and/or the existence of cross-reactive carbohydrate determinants (CCDs). The clinical relevance of peptidic cross-reactivities is well known, whereas that of CCDs is still a question of debate. The aim of this study is to characterize the IgE specificity of various patients suffering from pollen polysensitization to identify both peptidic and carbohydrate cross-reactive determinants. MATERIAL AND METHODS: Rapeseed, grass and Arabidopsis proteins were separated by isoelectric focusing, followed by SDS-PAGE, and transferred to a nitrocellulose sheet. The sheets were incubated either with an individual serum from a birch+grass-sensitive patient, followed by anti-human IgE, or with labelled Concanavalin A (ConA). Binding inhibition was tested by incubation of the sera with a mixture of sugar residues . RESULTS: The results showed two different patterns of cross-reacting sera: a pattern that implies few proteins, not always glycosylated and known as allergens, and a pattern that implies numerous proteins with molecular masses over 30 kDa. This second pattern was very close to the ConA -binding pattern. The IgE binding was abolished by pre-incubation with sugar residues only in the case of the second pattern. DISCUSSION: This study shows that multiple pollen sensitizations could result from multiple sensitizations to specific proteins or from a cross-sensitization to a wide range of glycoproteins. Two-D blots allow to characterize a cross-sensitization due to carbohydrate determinants, and thus to improve the diagnosis of allergy and its medical treatment.

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[24] - Focke M, Hemmer W, Hayek B, Gotz M, Jarisch R. Identification of allergens in oilseed rape (Brassica napus) pollen. Int Arch Allergy Immunol 1998;117:105-112

BACKGROUND: Pollen from oilseed rape (OSR), Brassica napus, an increasingly cultivated oilplant from the Brassicaceae, has been recognized as a potential cause of allergic sensitization. Allergens have been hardly investigated. METHODS: We characterized IgE binding proteins in OSR pollen by immunoblot, immunoblot inhibition and specific monoclonal antibodies using sera from 89 patients sensitized to OSR. RESULTS: Two low-molecular-weight allergens of 6/8 kD and 14 kD as well as a high molecular-weight cluster (27-69 kD) comprising six cross-reactive peptides could be identified. The three allergens were recognized by 50, 34 and 80% of patients, respectively. Immunoblot IgE binding to OSR could be totally inhibited by rye pollen and moderately by birch pollen (6/8 and 14 kD) while mugwort had little effect. An anti-profilin-specific monoclonal antibody bound specifically to a 14-kD protein in OSR. Binding to the 6/8-kD rape allergen could be effectively inhibited by rAln g 2, a calcium-binding protein from alder. Periodate treatment led to a significant reduction in IgE binding to the 27 to 69-kD OSR allergens indicating that carbohydrate determinants are involved in IgE binding. OSR proteins were capable to quench IgE binding to timothy grass pollen proteins of >/=60 kD suggesting that grass pollen group 4 allergens cross-react with the 27 to 69-kD cluster in OSR. CONCLUSIONS: The data demonstrate that OSR pollen is allergenic and indicate that the identified allergens represent cross-reacting homologues of well-known pollen allergens, i.e. calcium-binding proteins, profilins, and high-molecular-weight glycoproteins. Via cross-reactivity, exposure to OSR pollen may be a prolonging and aggravating factor in underlying birch and grass pollen allergy.

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[25] - Hemmer W, Focke M, Kolarich D, Wilson IBH, Altmann F, Wöhrl S, et al. Antibody binding to venom carbohydrates is a frequent cause for double positivity to honeybee and yellow jacket venom in patients with stinging-insect allergy. J Allergy Clin Immunol 2001;108:1045-1052

Background: Up to 50% of patients with stinging-insect allergy have double-positive RAST results to honeybee and yellow jacket (YJ) venom. True double sensitization and crossreactivity through venom hyaluronidases are considered main reasons for this multiple reactivity. OBJECTIVE: We investigated the role of antibodies against cross-reactive carbohydrate determinants in venom double positivity. METHODS: CAP inhibition experiments were performed with crude oilseed rape (OSR) and timothy grass pollen extracts and a neoglycoprotein construct displaying a MUXF glycan, as present in pineapple-stem bromelain (MUXF-BSA). CAP to OSR was used as a rough measure for carbohydrate-specific IgE in individual sera. RESULTS: CAP results to OSR pollen were positive in 2 of 14 single-positive honeybee venom sera, 2 of 16 single-positive YJ venom sera, and 33 (80.5%) of 41 double-positive sera (P < .00001, 2 test). CAP inhibition was performed in 16 selected patients with a CAP class of 3 or higher to both venoms. In 9 of 11 patients with a highly positive CAP result to OSR (CAP score to OSR > CAP score to second venom), pollen extracts, MUXF-BSA, or both were able to completely inhibit IgE binding to one of the venoms, whereas this was not the case in 5 patients with a negative or weakly positive CAP result to OSR (CAP score to OSR < CAP score to second venom). CONCLUSIONS: The data suggest that carbohydrate-specific IgE is a major cause for the double positivity to honeybee and YJ venom seen in patients with Hymenoptera allergy. Because these antibodies may have low clinical relevance, they may severely impede the correct diagnosis of Hymenoptera venom allergy

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[26] - Jappe U, Raulf-Heimsoth M, Hoffmann M, Burow G, Hübsch-Müller C, Enk A. In vitro hymenoptera venom allergy diagnosis: improved by screening for cross-reactive carbohydrate determinants and reciprocal inhibition. Allergy 2006;61:1220-1229

BACKGROUND: Immunoglobulin (Ig) E-double positivity for honeybee (HB) and yellow jacket (YJ) venom causes diagnostic difficulties concerning therapeutical strategies. The aim of this study was to clarify the cause and relation of the cross-reactivity in patients with insect venom allergy . METHODS: For this purpose, 147 patients with suspected stinging insect allergy and CAP-FEIA-double positivity were investigated for specific sIgE to additional cross-reactive carbohydrate determinant (CCD)-containing allergens: timothy grass pollen, rape pollen, natural rubber latex (NRL), bromelain, and horseradish peroxidase (HRP). Sera with sIgE to NRL were further investigated with the commercially available recombinant latex allergens. Reciprocal inhibition assays with both venoms and HRP were performed . RESULTS: About 36 of 147 (24.5%) patients had sIgE to both venoms only. However, 111 of 147 (75.5%) additionally reacted to CCD-carrying allergens. 89 of 111 CCD-reactive sera had NRL-sIgE. In cases where inhibition experiments were performed, the NRL-sIgE binding was completely abolished in the presence of HRP. Only nine of 61 sera were positive for at least one recombinant latex allergen; all of them were negative in history and NRL-skin prick test. In 43 sera containing sIgE to CCD, HRP inhibition revealed unequivocal results: In 28 of 43 (65%) an HRP-inhibition >70% of sIgE to one venom occurred, pointing out the relevant venom. In three of 43 sIgE proved to be entirely CCD-specific . CONCLUSIONS: Our data indicate that in cases of IgE positivity to both insect venoms supplementary screening tests with at least one CCD-containing allergen should be performed; HRP being a suitable tool for this test. In addition, subsequent reciprocal inhibition is an essential diagnostic method to specify cross-reacting sIgE results.

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[27] - Focke M, Hemmer W, Hayek B, Gotz M, Jarisch R. Identification of allergens in oilseed rape (Brassica napus) pollen. Int Arch Allergy Immunol 1998;117:105-112

BACKGROUND: Pollen from oilseed rape (OSR), Brassica napus, an increasingly cultivated oilplant from the Brassicaceae, has been recognized as a potential cause of allergic sensitization. Allergens have been hardly investigated. METHODS: We characterized IgE binding proteins in OSR pollen by immunoblot, immunoblot inhibition and specific monoclonal antibodies using sera from 89 patients sensitized to OSR. RESULTS: Two low-molecular-weight allergens of 6/8 kD and 14 kD as well as a high molecular-weight cluster (27-69 kD) comprising six cross-reactive peptides could be identified. The three allergens were recognized by 50, 34 and 80% of patients, respectively. Immunoblot IgE binding to OSR could be totally inhibited by rye pollen and moderately by birch pollen (6/8 and 14 kD) while mugwort had little effect. An anti-profilin-specific monoclonal antibody bound specifically to a 14-kD protein in OSR. Binding to the 6/8-kD rape allergen could be effectively inhibited by rAln g 2, a calcium-binding protein from alder. Periodate treatment led to a significant reduction in IgE binding to the 27 to 69-kD OSR allergens indicating that carbohydrate determinants are involved in IgE binding. OSR proteins were capable to quench IgE binding to timothy grass pollen proteins of >/=60 kD suggesting that grass pollen group 4 allergens cross-react with the 27 to 69-kD cluster in OSR. CONCLUSIONS: The data demonstrate that OSR pollen is allergenic and indicate that the identified allergens represent cross-reacting homologues of well-known pollen allergens, i.e. calcium-binding proteins, profilins, and high-molecular-weight glycoproteins. Via cross-reactivity, exposure to OSR pollen may be a prolonging and aggravating factor in underlying birch and grass pollen allergy.

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[28] - Focke M, Hemmer W, Valenta R, Götz M, Jarisch R. Identification of Oilseed Rape (Brassica napus) Pollen Profilin as a Cross-Reactive Allergen. Int Arch Allergy Immunol 2003;132:116-123

BACKGROUND: Major allergens of oilseed rape (OSR) pollen with molecular weights of 6/8, 14 and between 27 and 69 kD have been described. The aim of the present study was to further characterize the 14-kD allergen . METHODS: The 14-kD protein was purified from OSR pollen extracts by poly-(L-proline) (PLP)-Sepharose affinity chromatography and characterized immunologically by means of allergic patients' IgE antibodies, profilin-specific rabbit antisera, Western blot and ELISA inhibition using recombinant birch profilin (rBet v 2), and skin prick testing . RESULTS: By PLP affinity chromatography, OSR pollen profilin was purified as a single protein of 14.5 kD and further identified as a profilin by three polyclonal rabbit antisera raised against ragweed and tobacco pollen profilin and the C-terminus of birch profilin. IgE binding of a human serum pool (n = 15) and four profilin-reactive sera to nitrocellulose-blotted OSR profilin was completely inhibited by 1 microg/ml rBet v 2 (birch profilin). Reciprocal ELISA inhibition using increasing concentrations of rBet v 2 and purified OSR profilin, respectively, showed that rBet v 2 strongly inhibits antibody binding to OSR profilin, whereas almost 100 times the amount of OSR profilin was needed to inhibit IgE binding to rBet v 2. Skin prick tests were positive (wheal >/=3 mm) with 5 microg/ml rBet v 2 in all three patients tested, and with OSR profilin in two patients at a concentration of 50 microg/ml . CONCLUSIONS: OSR pollen profilin shares IgE and IgG epitopes with Bet v 2 and other plant profilins and may represent a potentially relevant allergen for profilin-sensitized patients.

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[29] - Smith PM, Xu H, Swoboda I, Singh MB. Identification of a Ca2+ binding protein as a new Bermuda grass pollen allergen Cyn d7: IgE cross-reactivity with oilseed rape pollen allergen Bra r1. Int Arch Allergy Immunol 1997;114:265-271

cDNA clones encoding two isoforms of an allergen from pollen of Bermuda grass (Cynodon dactylon) have been isolated using IgE from allergic patients. Homologous transcripts are present in pollen of 15 other grasses tested. This allergen, tentatively designated as Cyn d 7, contains two calcium binding domains and shows significant sequence similarity with other Ca2+ binding pollen allergens, namely Bet v 4 from birch and Bra r 1 from oilseed rape. Approximately 10% of allergic sera tested showed IgE reactivity to this allergen. IgE cross-reactivity was observed between this allergen and Bra r 1 of oilseed rape. IgE reactivity of this allergen requires protein-bound Ca2+. Using IgE affinity-purified from the recombinant allergen to probe Western blots of pollen extracts Cyn d 7 has been identified as a 12 kDA protein.

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[30] - Welch J, Jones MG, Cullinan P, Coates OA, Newman Taylor AJ. Sensitization to oilseed rape is not due to cross-reactivity with grass pollen. Clin Exp Allergy 2000;30:370-375

Oilseed rape is an important crop grown in the UK which can cause specific immunological sensitization with clinical symptoms in a relatively small number of the general population. Individuals with immunoglobulin (Ig) E-mediated allergy to oilseed rape have also been found to be sensitized to other pollen allergens, most frequently being grass pollen. Cross-reactivity between common grass and oilseed rape would have important implications, especially as their flowering period coincides. OBJECTIVE: We have investigated whether the cosensitization found in individuals sensitized to both oilseed rape and grass pollen is due to cross-reactivity. METHODS: Cross-reactivity between oilseed rape and grass pollen was determined using RAST, RAST inhibition, Western blotting and inhibition studies with Western blotting. RESULTS: Competitive RAST inhibition studies between pollen of oilseed rape and grass failed to show any cross-reactivity between the pollen types. Self-inhibition with oilseed rape resulted in 90% inhibition, whereas there was less than 10% inhibition with grass pollen. Western blotting revealed allergens of similar molecular weight in both oilseed rape and grass pollen. Despite allergens of similar molecular weights being present in both pollen types, inhibition immunoblot studies confirmed that the allergens in the two allergens were immunologically distinct. CONCLUSION: The allergens of oilseed rape and grass pollen, although similar in molecular weights, are immunologically distinct and there is no evidence of cross-reactivity between them. Individuals allergic to grass pollen will not necessarily develop a specific nasal or airway response to inhaled oilseed rape pollens.

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[31] - Poikonen S, Rancé F, Puumalainen T, Le Manach G, Reunala T, Turjanmaa K. Sensitization and allergy to turnip rape: a comparison between the Finnish and French children with atopic dermatitis. Acta Paediatr 2009;98:310-315

Aim: Finnish children with atopic dermatitis (AD) are frequently sensitized and show positive food challenge to turnip rape. We examined whether French children are also allergic to this oilseed plant and whether mustard could be the cross-reacting allergen. Methods: Turnip rape and mustard challenge was performed to 14 Finnish and 14 French children with atopic dermatitis and positive skin prick test to turnip rape. Specific IgE antibodies were measured by ImmunoCAP and enzyme-linked immunosorbent assay (ELISA). Results: Open labial or oral challenge to turnip rape was positive in 14 (100%) Finnish and five (36%) French children and mustard challenge in five Finnish and five French children. IgE antibodies to oilseed rape and mustard were slightly more frequent in the Finnish (100% and 93%) than in the French (93% and 71%) children but rare (4%) in the 28 matched controls. The same findings were true for IgE antibodies to purified 2S albumin allergens, which showed similar cross-wise IgE inhibition patterns. Conclusion: French children with atopic dermatitis show IgE antibodies to turnip rape, oilseed rape and mustard similarly to the Finnish children. 2S albumin allergens in the seeds of these plants are highly cross-reactive and therefore, they all could be important sensitizers in children with atopic dermatitis.

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[32] - Alvarez MJ, Estrada JL, Gozalo F, Fernandez-Rojo F, Barber D. Oilseed rape flour: another allergen causing occupational asthma among farmers. Allergy 2001;56:185-188

BACKGROUND: Farmers are exposed to a wide variety of sensitizers. Since occupational asthma (OA) can lead to permanent disability, exposure discontinuation is the preferred treatment. When this is not possible, the identification of the causative allergen may allow an alternative therapy. METHODS: We present three farmers diagnosed with OA as a consequence of handling fodder. We carried out skin tests with common and occupational allergens and with oilseed rape (OSR) extract. Total and specific serum IgE levels were measured. The patients underwent the OSR-bronchial provocation test (OSR-BPT). The day before and 24 h after the OSR-BPT, the methacholine (M)-BPT and induced sputum were performed. Eosinophil percentages and ECP levels were measured in the sputum samples. RESULTS: OSR sensitization (skin tests and specific serum IgE) was detected in all the patients. The OSR-BPT elicited early responses in two subjects. Methacholine sensitivity, sputum eosinophils, and sputum ECP levels increased 24 h after the OSR-BPT in all the patients. CONCLUSIONS: We have demonstrated that inhalation of OSR flour causes bronchoconstriction, induces an eosinophilic inflammatory bronchial response, and increases bronchial hyperresponsiveness in sensitized asthmatics. OSR flour contained in animal fodder should be considered another potential cause of OA among farmers.

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[33] - Monsalve RI, Delapena MAG, Lopezotiin C, Fiandor A, Fernandez C, Villalba M, et al. Detection, isolation and complete amino acid sequence of an aeroallergenic protein from rapeseed flour. Clin Exp Allergy 1997;27:833-841

Seed proteins have been found to cause hypersensitivity by ingestion or inhalation. Rapeseed flour was responsible for allergic symptoms in a patient, who develops into allergy to mustard spice. Objective To determine the presence of allergenic proteins in rapeseed flour, and analyse the structure of the main component and its crossreactivity with the mustard allergen. Methods SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and subsequent immunoblotting with a serum from a rapeseed allergic patient were performed to detect IgE-binding proteins. Proteolytic digestions and high performance liquid chromatography were used to obtain the peptides from the allergenic BnIII napin from rapeseed flour. Automatic Edman degradations were carried out to determine their amino acid sequences, which were compared with other sequences in nucleotide and amino acid sequence databases. Crossreactivity assays were carried out by ELISA inhibition using sera from a rapeseed allergic patient and from patients allergic to mustard. Results The 2S albumins of rapeseed were recognized by the serum from a patient allergic to this seed. The most abundant isoform of the allergenic napins, BnIII, was used for structural and immunological analysis. The protein consists of two different chains of 9.5 and 4.5 kDa. Their complete amino acid sequences were determined. The protein exhibited structural relationships with other napin-like storage proteins from seeds. IgE and IgG crossreactivity between rapeseed and mustard allergens was also demonstrated. Considering the structural and immunological data, certain polypeptide regions are suggested to be involved in the allergenicity of these proteins. Conclusions Rapeseed contains 2S storage proteins which may cause allergy in hypersensitive individuals. These proteins exhibit great sequence similarity with 2S albumins from different seeds. Crossreactivity between mustard and rapeseed flours can be explained by sequence homology

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[34] - Monsalve RI, Delapena MAG, Lopezotiin C, Fiandor A, Fernandez C, Villalba M, et al. Detection, isolation and complete amino acid sequence of an aeroallergenic protein from rapeseed flour. Clin Exp Allergy 1997;27:833-841

Seed proteins have been found to cause hypersensitivity by ingestion or inhalation. Rapeseed flour was responsible for allergic symptoms in a patient, who develops into allergy to mustard spice. Objective To determine the presence of allergenic proteins in rapeseed flour, and analyse the structure of the main component and its crossreactivity with the mustard allergen. Methods SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and subsequent immunoblotting with a serum from a rapeseed allergic patient were performed to detect IgE-binding proteins. Proteolytic digestions and high performance liquid chromatography were used to obtain the peptides from the allergenic BnIII napin from rapeseed flour. Automatic Edman degradations were carried out to determine their amino acid sequences, which were compared with other sequences in nucleotide and amino acid sequence databases. Crossreactivity assays were carried out by ELISA inhibition using sera from a rapeseed allergic patient and from patients allergic to mustard. Results The 2S albumins of rapeseed were recognized by the serum from a patient allergic to this seed. The most abundant isoform of the allergenic napins, BnIII, was used for structural and immunological analysis. The protein consists of two different chains of 9.5 and 4.5 kDa. Their complete amino acid sequences were determined. The protein exhibited structural relationships with other napin-like storage proteins from seeds. IgE and IgG crossreactivity between rapeseed and mustard allergens was also demonstrated. Considering the structural and immunological data, certain polypeptide regions are suggested to be involved in the allergenicity of these proteins. Conclusions Rapeseed contains 2S storage proteins which may cause allergy in hypersensitive individuals. These proteins exhibit great sequence similarity with 2S albumins from different seeds. Crossreactivity between mustard and rapeseed flours can be explained by sequence homology

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[35] - Le Manach G, Breinig S, Rancé F. Allergie alimentaire associée à la moutarde et au colza. Rev Fr Allergol Immunol Clin 2007;47:284

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[36] - Acero S, Blanco R, Lombardero M. Rapeseed: an indoor aeroallergen ? EAACI 21th Congress, Naples, 1-5 June, 2002, Poster n°891

Volatile compounds emitted during the rape flowering season have been reported to cause bronchial and conjunctival irritation. Rape flour has also been previously described to cause allergic symptoms. A 36-year-old atopic man reported intermittent nasal and mild ocular symptoms. He noticed that his symptoms worsened when feeding his canary birds or cleaning their cage. Results of skin prick tests (SPT) were positive for house dust mites, rapeseed and birdseed. SPTs were negative for other common aeroallergens, other components of the bird food and canary feathers. He experienced quick improvement when he got ride of the birds. A 39-year-old atopic woman (suffering from rhinoconjunctivitis and asthma due to grass pollen) developed nasal and conjunctival symptoms, out of the grass pollen season, which seemed to be related to the acquisition of some canary birds. SPTs with the components of the bird food and canary feathers were positive for rapeseed. She became nearly asymptomatic, out of grass pollen season, when exposure to the birds ceased. Several extracts prepared with the seeds provided by one of our patients were used for SDS-PAGE-Immunoblotting. Patients' sera contained specific IgE for different proteins of the rapeseed extract with different molecular weights, the main ones with about 16, 45, 75, 100 and >100 kDa, as shown by immunoblotting. Both cases are suggestive of rhinoconjunctivitis induced by rapeseed, likely due to an IgE mechanism. Rapeseed should be considered as a trigger of allergic symptoms in exposed patients. Rapeseed can be an indoor allergen in homes where there are birds.

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[37] - Compés E, Palomares O, Fernandez-Nieto M, Escudero M, Cuesta-Herranz J. Allergy to turnip seeds in a bird fancier. Allergy 2007;62:1472-1473

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[38] - Martin-Hernandez C, Bénet S, Obert L. Determination of Proteins in Refined and Nonrefined Oils. J Agric Food Chem 2008;56:4348-4351

Five methods using aqueous/organic solvents for the separation of proteins from oils were compared. The extraction with acetone-hexane followed by amino acid analysis was found to be the most suitable method for isolation and quantification of proteins from oils. The detection limit of the method was 0.18 mg protein/kg oil, and the quantification limit was 0.6 mg protein/kg. The relative repeatability limit for samples containing 1-5 mg protein/kg sample was 27%. The protein recovery ranged between 68 and 133%. Using this method, the protein content of 14 refined and nonrefined oils was determined. In none of the refined oils were proteins detected, whereas the protein content of the unrefined oils ranged between undetectable in extra virgin olive oil to 11 mg/kg in rapeseed oil. With sodium dodecyl sulfate-polyacrylamide gel electrophoresis in combination with silver staining, many protein bands were visible in the unrefined soy, olive, peanut, and rapeseed oil samples. Proteins bands were not obtained from the refined fish oil. In the other refined oil samples, a few proteins bands could be visualized. Two protein bands with apparent molecular molecular masses of 58 and 64 kDa were always observed in these oils.

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[39] - Poikonen S, Puumalainen TJ, Kautiainen H, Burri P, Palosuo T, Reunala T, et al. Turnip rape and oilseed rape are new potential food allergens in children with atopic dermatitis. Allergy 2006;61:124-127

BACKGROUND: When skin prick testing (SPT) young children with atopic dermatitis (AD) for suspected food allergy, we frequently found positive reactions with turnip rape (Brassica rapa) and oilseed rape (Brassica napus). We performed food challenge to examine whether these children react clinically to turnip rape . METHODS: A total of 1887 children were screened with SPTs for sensitization to turnip rape and oilseed rape. Twenty-eight children with clearly positive SPT (> or =5 mm) were first subjected to labial challenge with turnip rape seeds followed, if negative, by open oral challenge for up to 7 days. Twenty-five children with AD but negative SPT to turnip rape and oilseed rape served as controls . RESULTS: Two-hundred and six (10.9%) children had positive SPT to turnip rape and/or oilseed rape. Twenty-five (89%) of 28 children showed a positive challenge reaction to turnip rape. Seventeen reacted with labial whealing, and eight in oral challenge with facial urticaria, flare-up of AD or abdominal symptoms. All 25 control children remained negative in the labial challenge . CONCLUSIONS: Turnip rape and oilseed rape seem to be new important food allergens in young children with AD. The modes of exposure to these allergens and the possible routes of sensitization remain to be established.

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[40] - Poikonen S, Rancé F, Puumalainen T, Le Manach G, Reunala T, Turjanmaa K. Sensitization and allergy to turnip rape: a comparison between the Finnish and French children with atopic dermatitis. Acta Paediatr 2009;98:310-315

Aim: Finnish children with atopic dermatitis (AD) are frequently sensitized and show positive food challenge to turnip rape. We examined whether French children are also allergic to this oilseed plant and whether mustard could be the cross-reacting allergen. Methods: Turnip rape and mustard challenge was performed to 14 Finnish and 14 French children with atopic dermatitis and positive skin prick test to turnip rape. Specific IgE antibodies were measured by ImmunoCAP and enzyme-linked immunosorbent assay (ELISA). Results: Open labial or oral challenge to turnip rape was positive in 14 (100%) Finnish and five (36%) French children and mustard challenge in five Finnish and five French children. IgE antibodies to oilseed rape and mustard were slightly more frequent in the Finnish (100% and 93%) than in the French (93% and 71%) children but rare (4%) in the 28 matched controls. The same findings were true for IgE antibodies to purified 2S albumin allergens, which showed similar cross-wise IgE inhibition patterns. Conclusion: French children with atopic dermatitis show IgE antibodies to turnip rape, oilseed rape and mustard similarly to the Finnish children. 2S albumin allergens in the seeds of these plants are highly cross-reactive and therefore, they all could be important sensitizers in children with atopic dermatitis.

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[41] - Puumalainen TJ, Poikonen S, Kotovuori A, Vaali K, Kalkkinen N, Reunala T, et al. Napins, 2S albumins, are major allergens in oilseed rape and turnip rape. J Allergy Clin Immunol 2006;117:426-432

BACKGROUND: Children with IgE-mediated allergy to foods frequently react to seeds of oilseed rape (Brassica napus ssp. oleifera) and turnip rape (Brassica rapa ssp. oleifera) in skin prick tests (SPTs). Sensitization pathways are not known . OBJECTIVE: We identified possible major allergens in oilseed rape and turnip rape using sera from 72 atopic children (mean age, 3.3 years) with positive SPT responses to oilseed rape and turnip rape . METHODS: Allergens from oilseed rape and turnip rape seed extracts were purified by using gel filtration and cation exchange chromatography and characterized by means of reversed-phase chromatography, N-terminal amino acid sequencing, and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. IgE binding of sera from 72 children with positive SPT reactions to oilseed rape and turnip rape and 72 age- and sex-matched atopic control subjects with negative SPT responses were analyzed by means of IgE ELISA and immunoblotting. In vivo reactivity of the purified allergens was tested with SPTs in 6 children . RESULTS: In IgE immunoblotting and IgE ELISA major reactivity was to a group of homologous, approximately 9.5- to 14.5-kd proteins. These allergens were identified as 2S albumins, also known as napins, by means of N-terminal amino acid sequencing. In ELISA approximately 80% of the patients had IgE to purified napins from both plants. In SPTs purified napins caused positive reactions in all 6 children tested . CONCLUSIONS: This study shows that 2S albumins in oilseed rape and turnip rape are new potential food allergens. Further studies are needed to clarify the routes of exposure and mechanisms of sensitization.

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[42] - Toriyama K, Hanaoka K, Okada T, Watanabe M. Molecular cloning of a cDNA encoding a pollen extracellular protein as a potential source of a pollen allergen in Brassica rapa. FEBS Lett 1998;424:234-238

A polyclonal antiserum was raised against the extracellular pollen proteins of Brassica rapa and used for screening the expression cDNA libraries made from immature anthers. We obtained five groups of cDNA clones, including cDNAs similar to PCP1, thioredoxin, and lipid transfer protein (LTP). Recombinant protein of the cDNA clone showing sequence similarity to LTP was demonstrated to bind IgE of a patient allergic to Brassica pollen. The cDNA clone reported here, therefore, represents a novel pollen allergen of Brassica rapa.

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[43] - Godfrin D, Senechal H, Desvaux F-X, Mayer C, Hennion M-C, Peltre G. Allergomes of rapeseed pollen and seed (Brassica napus). Allergy Clin Immunol Int 2005;17(Suppl. 1):17

Background: Rapeseed is the main oleaginous seed cultivated in Europe and mainly used to produce seed rape oil for human consumption. Seeds and cattle-cake, left over from the oil production, are also used for animal consumption. Some Brassica napus seed components are found in shampoo, beauty care products and washing powder. Methods: Among the patients allergic to both birch and grass pollen some of them are also allergic to rapeseed pollen (which is emitted between the pollen seasons of birch and grasses). Some sera from mustard allergic patient were used to visualize allergens from Brassica napus seeds. Water soluble proteins were extracted by incubation of pollen or ground seeds in distilled water. All remaining water-soluble proteins were removed by extensive washes in distilled water and the pellet was dried. Then, non-water soluble proteins were extracted with a combination of Thiourea, Urea and a detergent, CHAPS. The watersoluble proteins were immobilized on ELISA plates, incubated with allergic patient sera and the immunodetection was performed with a second antibody anti- IgE alkaline phosphatase labelled. More than 400 patient sera were screened for rapeseed pollen or/and seed IgE, IgG4 and total IgG antibodies. IgE positive ones were used to reveal allergens on blot obtained after 1D isoelectrofocusing and 2D gel separations. Results: IgE from 50 patient sera out of 400 recognised water soluble proteins from seed and 31 of these positive sera were used to study water and nonwater soluble proteins from pollen and seed. 10 out of 31 sera were positive with water soluble pollen allergens and 9 out of 31 with non-water soluble allergens. 12 out of 31 sera were positive with water soluble seed allergens and among them 6 were positive with non-water soluble seed allergens. In water pollen extract 7 allergens were detected compared with around twenty allergens evidenced in non-water soluble pollen extract. Likewise, in water seed extract 6 allergens were detected compared with around twenty allergens visualised in non water soluble extract. Amino acid sequencing or mass spectrometry were performed allowing identification of these proteins. Conclusion: Results show that major allergens from Brassica napus pollen and seed have different molecular characteristics using isoelectric point and molecular mass criteria. Likewise hydrophilic and hydrophobic allergen patterns seem to be different.

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[44] - Godfrin D, Sénéchal H, Haddad I, Sutra JP, Chardin H, Desvaux FX, et al. Caractérisation par électrophorèse 2D et spectrométrie de masse des allergènes de pollen et de graines de colza (Brassica napus). Rev Fr Allergol Immunol Clin 2007;47:270

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[45] - Quirce S, Madero MF, Fernandez-Nieto M, Jimenez A, Sastre J. Occupational asthma due to the inhalation of cauliflower and cabbage vapors. Allergy 2005;60:969-970

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[46] - Pasquet-Noualhaguet C. Anaphylaxie au chou. Alim'Inter 2008;13:100-101

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[47] - Palacin A, Cumplido J, Figueroa J, Ahrazem O, Sanchez-Monge R, Carrillo T, et al. Cabbage lipid transfer protein Bra o 3 is a major allergen responsible for cross-reactivity between plant foods and pollens. J Allergy Clin Immunol 2006;117:1423-1429

BACKGROUND: Food IgE-mediated allergy to members of the Brassicaceae family has been increasingly reported . OBJECTIVE: To characterize cabbage-Brassica oleracea var capitata-allergy and its major allergens . METHODS: A prospective study was performed, recruiting 17 patients allergic to cabbage, and control subjects. Skin prick tests and double-blind placebo-controlled food challenges were performed. A major allergen was isolated from cabbage by RP-HPLC and characterized by N-terminal amino acid sequencing and matrix-assisted laser desorption/ionization mass spectrometry analysis. Specific IgE determinations, IgE immunoblots, and CAP-inhibition assays were also performed . RESULTS: Skin prick test and specific IgE were positive to cabbage in all patients. Five of them referred anaphylactic reactions when eating cabbage, and in another 5 patients, cabbage allergy was further confirmed by double-blind placebo-controlled food challenge. Most of them showed associated sensitizations to mugwort pollen, mustard, and peach. A 9-kd cabbage IgE-binding protein, Bra o 3, was identified as a lipid transfer protein (LTP) with 50% of identity to peach LTP Pru p 3. Skin prick test with Bra o 3 showed positive results in 12 of 14 cases (86%). On CAP inhibition assays, Bra o 3 managed to inhibit significantly the IgE binding to cabbage, mugwort pollen, and peach. Both Bra o 3 and Pru p 3 were recognized by IgE from the patients' sera . CONCLUSION: Bra o 3, a cabbage LTP, is a major allergen in this food, cross-reacting with mugwort pollen and with other plant foods, such as peach. CLINICAL IMPLICATIONS: Cabbage IgE-mediated allergy is a potentially severe condition that can present with other plant food and pollen allergies.

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[48] - Blaiss MS, McCants ML, Lehrer SB. Anaphylaxis to cabbage: detection of allergens. Ann Allergy 1987;58:248-250

Allergy to cabbage and other foods in the Brassica family has rarely been observed in man. We report a case of facial and throat swelling in an atopic female after she ingested coleslaw on two separate occasions. She had 4+ reactions to cabbage, mustard plant, cauliflower, and broccoli by skin testing. A RAST using cabbage extract was positive for specific IgE antibody. Analysis of cabbage extract by gel filtration showed five peaks of ultraviolet-absorbing material at 280 nm. Allergenic activity was demonstrated by RAST inhibition in two fractions of the cabbage extract. This report confirms IgE sensitivity can occur to foods in the Brassica family.

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[49] - Vovolis V, Poulios G, Koutsostathis N. IgE-mediated allergy to raw cabbage but not to cooked. Allergy 2009;64:964-965

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[50] - Yin J, Li H. Food Dependent Exercise Induced Anaphylaxis Caused by Cauliflower: First Case Report in China. AAAAI 60th Annual Meeting, San Francisco, 19-23 March 2004, Poster n°1173

Rationale Food dependent exercise induced anaphylaxis (FDEIAn) have been studied in western countries, but no case report have been published in China. Method s : We report a 16 year-old boy with no family or personal history of allergic disease experienced an anaphylactic reaction of urticaria, facial angioedema, throat swelling, dyspnea, syncope and hypotension while playing badminton 5 minutes after eating rice, cauliflower and pork. He recovered within 30 minutes after the emergency treatment. The intradermal skin tests to 40 kinds of food and inhaler allergens were performed; The serum total IgE and specific IgE were detected by using the UniCAP specific IgE method. The food challenge test was not taken because the parents of the boy refused it. Result s : Skin tests were positive to peanut, soybean, sesame, maize, artimisia pollen and negative to rice, pork, beef, chicken, wheat, shrimp, sea fish, sea crab, egg as well as milk. The total serum IgE was 814ku/l; specific IgE were: cauliflower 4.47ku/l, peanut 7.27ku/l, sesame 1.55ku/l, soybean 1.14ku/l, maize 2.54ku/l and artimisia vulgaris 7.52ku/l. Specific IgE to egg, milk, shrimp, pork, beef, mutton and chicken were negative. Under our follow-up observation of 3 years, the boy's episodes have been prevented successfully by avoidance of cauliflower ingestion in relation to exercise. Conclusion : It is useful to test both in vivo and in vitro an extensive panel of foods. Avoidance of foods associated with skin test or RAST positively for hours before exercise can prevent further episodes with specific FDEIAn. This is the first case report of FDEIAn caused by cauliflower in China.

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[51] - Blaiss MS, McCants ML, Lehrer SB. Anaphylaxis to cabbage: detection of allergens. Ann Allergy 1987;58:248-250

Allergy to cabbage and other foods in the Brassica family has rarely been observed in man. We report a case of facial and throat swelling in an atopic female after she ingested coleslaw on two separate occasions. She had 4+ reactions to cabbage, mustard plant, cauliflower, and broccoli by skin testing. A RAST using cabbage extract was positive for specific IgE antibody. Analysis of cabbage extract by gel filtration showed five peaks of ultraviolet-absorbing material at 280 nm. Allergenic activity was demonstrated by RAST inhibition in two fractions of the cabbage extract. This report confirms IgE sensitivity can occur to foods in the Brassica family.

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[52] - Figueroa J, Blanco C, Dumpiérrez AG, Almeida L, Ortega N, Castillo R, et al. Mustard allergy confirmed by double-blind placebo-controlled food challenges: clinical features and cross-reactivity with mugwort pollen and plant-derived foods. Allergy 2005;60:48-55

BACKGROUND: Mustard IgE-mediated allergy is supposed to be a rare cause of food allergy, and its clinical features and cross-reactivities have not been fully elucidated . METHODS: A prospective study was carried out, recruiting mustard allergic patients, and paired control subjects. A clinical questionnaire was administered, and skin-prick tests (SPT) with panels of aeroallergens and foods, serum extraction for in vitro tests and double-blind placebo-controlled food challenges (DBPCFC) were performed . RESULTS: Thirty-eight mainly adult patients, with 10.5% reporting systemic anaphylaxis, were included in the study [age (mean +/- SD): 21.9 +/- 8.6 years]. DBPCFC were performed in 24 patients, being positive in 14 cases (58.3%). Patients with positive outcome showed significantly greater mustard SPT than those with negative outcome (8.2 +/- 3.7 vs 5.3 +/- 2.4 mm, P <0.05), and the receiver-operating characteristic (ROC) curve analysis yielded a cut-off value for mustard commercial SPT of 8 mm, with a specificity of 90% (95% CI, 55.5-98.3), and a sensitivity of 50% (95% CI, 23.1-76.9). A significant association between mustard hypersensitivity and mugwort pollen sensitization was found (97.4% of patients), with partial cross-reactivity demonstrated by UniCAP System inhibition assays. All patients showed sensitization to other members of Brassicaceae family, and cross-reactivity among them was also confirmed. Moreover, significant associations with nut (97.4%), leguminous (94.7%), corn (78.9%), and Rosaceae fruit (89.5%) sensitizations were also shown. Around 40% of these food sensitizations were symptomatic, including food-dependent exercise-induced anaphylaxis in six patients . CONCLUSIONS: Mustard allergy is a not-uncommon disorder that can induce severe reactions. Significant associations with mugwort pollinosis and several plant-derived food allergies are demonstrated, suggesting a new mustard-mugwort allergy syndrome. A relationship between this syndrome and food-dependent exercise-induced anaphylaxis is also reported.

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[53] - Hernandez E, Quirce S, Villalba M, Cuesta J, Sastre J. Anaphylaxis caused by cauliflower. J Investig Allergol Clin Immunol 2005;15:158-159

BACKGROUND: Cauliflower is a vegetable belonging to the family Cruciferae, genus Brassica, var. Botrytis. METHODS: We report the case of a 70 year-old man who suffered an acute episode consisting of oropharyngeal itching, facial and hand swelling, dyspnea and severe bronchospasm within a few minutes after eating vegetable paella containing cauliflower, green beans, red and green pepper. Due to the severity of the reaction he needed treatment in the emergency room. RESULTS: Skin prick tests with common aeroallergens were positive to Cupressus, Platanus and grass pollen. A strong skin prick test response was obtained with cauliflower and peach lipid transfer protein. Skin prick test with rice, green beans and pepper were negative. Specific IgE determinations were positive to cabbage and cauliflower, and negative to mustard. CONCLUSIONS: The clinical history and the results of the allergologic work-up point out to this patient having experienced an IgE-mediated anaphylactic reaction to cauliflower. To the best of our knowledge, this is the first case report of anaphylaxis due to cauliflower.

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

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

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[55] - Quirce S, Madero MF, Fernandez-Nieto M, Jimenez A, Sastre J. Occupational asthma due to the inhalation of cauliflower and cabbage vapors. Allergy 2005;60:969-970

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[56] - Yin J, Li H. Food Dependent Exercise Induced Anaphylaxis Caused by Cauliflower: First Case Report in China. AAAAI 60th Annual Meeting, San Francisco, 19-23 March 2004, Poster n°1173

Rationale Food dependent exercise induced anaphylaxis (FDEIAn) have been studied in western countries, but no case report have been published in China. Method s : We report a 16 year-old boy with no family or personal history of allergic disease experienced an anaphylactic reaction of urticaria, facial angioedema, throat swelling, dyspnea, syncope and hypotension while playing badminton 5 minutes after eating rice, cauliflower and pork. He recovered within 30 minutes after the emergency treatment. The intradermal skin tests to 40 kinds of food and inhaler allergens were performed; The serum total IgE and specific IgE were detected by using the UniCAP specific IgE method. The food challenge test was not taken because the parents of the boy refused it. Result s : Skin tests were positive to peanut, soybean, sesame, maize, artimisia pollen and negative to rice, pork, beef, chicken, wheat, shrimp, sea fish, sea crab, egg as well as milk. The total serum IgE was 814ku/l; specific IgE were: cauliflower 4.47ku/l, peanut 7.27ku/l, sesame 1.55ku/l, soybean 1.14ku/l, maize 2.54ku/l and artimisia vulgaris 7.52ku/l. Specific IgE to egg, milk, shrimp, pork, beef, mutton and chicken were negative. Under our follow-up observation of 3 years, the boy's episodes have been prevented successfully by avoidance of cauliflower ingestion in relation to exercise. Conclusion : It is useful to test both in vivo and in vitro an extensive panel of foods. Avoidance of foods associated with skin test or RAST positively for hours before exercise can prevent further episodes with specific FDEIAn. This is the first case report of FDEIAn caused by cauliflower in China.

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[57] - Hernandez E, Quirce S, Villalba M, Cuesta J, Sastre J. Anaphylaxis caused by cauliflower. J Investig Allergol Clin Immunol 2005;15:158-159

BACKGROUND: Cauliflower is a vegetable belonging to the family Cruciferae, genus Brassica, var. Botrytis. METHODS: We report the case of a 70 year-old man who suffered an acute episode consisting of oropharyngeal itching, facial and hand swelling, dyspnea and severe bronchospasm within a few minutes after eating vegetable paella containing cauliflower, green beans, red and green pepper. Due to the severity of the reaction he needed treatment in the emergency room. RESULTS: Skin prick tests with common aeroallergens were positive to Cupressus, Platanus and grass pollen. A strong skin prick test response was obtained with cauliflower and peach lipid transfer protein. Skin prick test with rice, green beans and pepper were negative. Specific IgE determinations were positive to cabbage and cauliflower, and negative to mustard. CONCLUSIONS: The clinical history and the results of the allergologic work-up point out to this patient having experienced an IgE-mediated anaphylactic reaction to cauliflower. To the best of our knowledge, this is the first case report of anaphylaxis due to cauliflower.

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[58] - Hincha DK, Neukamm B, Sror HA, Sieg F, Weckwarth W, Ruckels M, et al. Cabbage Cryoprotectin Is a Member of the Nonspecific Plant Lipid Transfer Protein Gene Family. Plant Physiol 2001;125:835-846

We have recently purified a protein (cryoprotectin) from the leaves of cold-acclimated cabbage (Brassica oleracea) to electrophoretic homogeneity, which protects thylakoids isolated from the leaves of nonacclimated spinach (Spinacia oleracea) from freeze-thaw damage. Sequencing of cryoprotectin showed the presence of at least three isoforms of WAX9 proteins, which belong to the class of nonspecific lipid transfer proteins. Antibodies raised against two synthetic peptides derived from the WAX9 proteins recognized a band of approximately 10 kD in western blots of crude cryoprotectin preparations. This protein and the cryoprotective activity could be precipitated from solution by the antiserum. We show further that cryoprotectin is structurally and functionally different from WAX9 isolated from the surface wax of cabbage leaves. WAX9 has lipid transfer activity for phosphatidylcholine, but no cryoprotective activity. Cryoprotectin, on the other hand, has cryoprotective, but no lipid transfer activity. The cryoprotective activity of cryoprotectin was strictly dependent on Ca(2+) and Mn(2+) and could be inhibited by chelating agents, whereas the lipid transfer activity of WAX9 was higher in the presence of ethylenediaminetetraacetate than in the presence of Ca(2+) and Mn(2+). We have recently purified a protein (cryoprotectin) from the leaves of cold-acclimated cabbage (Brassica oleracea) to electrophoretic homogeneity, which protects thylakoids isolated from the leaves of nonacclimated spinach (Spinacia oleracea) from freeze-thaw damage. Sequencing of cryoprotectin showed the presence of at least three isoforms of WAX9 proteins, which belong to the class of nonspecific lipid transfer proteins. Antibodies raised against two synthetic peptides derived from the WAX9 proteins recognized a band of approximately 10 kD in western blots of crude cryoprotectin preparations. This protein and the cryoprotective activity could be precipitated from solution by the antiserum. We show further that cryoprotectin is structurally and functionally different from WAX9 isolated from the surface wax of cabbage leaves. WAX9 has lipid transfer activity for phosphatidylcholine, but no cryoprotective activity. Cryoprotectin, on the other hand, has cryoprotective, but no lipid transfer activity. The cryoprotective activity of cryoprotectin was strictly dependent on Ca(2+) and Mn(2+) and could be inhibited by chelating agents, whereas the lipid transfer activity of WAX9 was higher in the presence of ethylenediaminetetraacetate than in the presence of Ca(2+) and Mn(2+).

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Pollen de colza

Graines et huiles de colza

Choux, brocoli, chou-fleur

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