Les Amaranthacées

[1] - Astier C, Moneret-Vautrin DA, Puillandre E, Bihain BE. First case report of anaphylaxis to quinoa, a novel food in France. Allergy 2009;64:815-821

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[2] - de la Hoz B, Fernandez-Rivas M, Quirce S, Cuevas M, Fraj J, Davila I, et al. Swiss chard hypersensitivity: clinical and immunologic study. Ann Allergy 1991;67:487-492

Allergy to vegetables and fruits seems to be more prevalent in atopics, especially in birch pollen-sensitized individuals. We report a case of a grass pollen-sensitized woman, in whom the inhalation of vapor from boiling Swiss chard precipitated rhinoconjunctivitis and asthma. Type I hypersensitivity to Swiss chard was demonstrated by means of immediate skin test reactivity, specific IgE determination by RAST, basophil degranulation, histamine release test, and an immediate bronchial provocation test response to Swiss chard extract. The controls did not react to any of these tests. RAST inhibition assays suggest the presence of some cross-reactivity among Swiss chard and grass pollen antigens, as well as cross-reactivity between vegetables and weed pollens of the chenopod family.

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[3] - Minciullo PL, Mistrello G, Patafi M, Zanoni D, Gangemi S. Cross-reactivity between Parietaria pollen and beet. Allergol Immunopathol (Madr) 2007;35:74-75

Allergy to beet is very rare. Until now, only a few reports about asthma induced by inhaling the vapor of cooked beet have been published. We describe two patients with allergic rhinitis and positive skin prick tests to Parietaria and beet only. To investigate possible cross-reactivity between Parietaria pollen and beet, we performed laboratory assays that showed beet-specific IgE in the sera of both patients and possible cross-reactivity between Parietaria and beet in one patient.

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[4] - Sanchez I, Rodriguez F, Garcia-Abujeta JL, Fernandez L, Quiñones D, Martin-Gil D. Oral allergy syndrome induced by spinach. Allergy 1997;53:1245-1246

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[5] - Drouet M, Le Sellin J, Gay G, el Founini M, Sabbah A. Allergie aux chénopodiacées (betterave, épinard) associée à l'allergie au latex. Allerg Immunol (Paris) 1994;26:113-114

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[6] - Fowler MR, Gartland J, Norton W, Slater A, Elliott MC, Scott NW. RS2: a sugar beet gene related to the latex allergen Hev b 5 family. J Exp Bot 2000;51:2125-2126

A novel gene (RS2) has been isolated from a Beta vulgaris (cv. Regina) cDNA library. The expression of this gene was enhanced in the mature storage organ as compared to leaf tissue. The protein encoded by this gene was found to be alanine- and glutamic acid-rich and it resembles members of the latex allergen Hev b 5 family.

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[7] - Herrera I, Moneo I, Caballero ML, de Paz S, Perez Pimiento A, Rebollo S. Food allergy to spinach and mushroom. Allergy 2002;57:261-262

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[8] - Wiker HG, Stensby BA, Løvik M. Analysis of sera submitted to the Norwegian national register of severe allergic reactions to food using a macroarray for food allergy. EAACI 21th Congress, Naples, 1-5 June, 2002, Poster n°1018

Background: There is a potential for improving current serological laboratory tests for detection of specific IgE to allergens. In conjunction with the Norwegian national reporting system and register of severe allergic reactions to food, we are developing a method that aims at detecting specific IgE against a number of food allergens in one single assay. Methods: A dot blot method with 150 different food extracts immobilized on a nitrocellulose membrane was used to determine the reactivity of 52 sera referred to the Norwegian national register of severe allergic reactions to food. Patient sera were allowed to react with the food allergens and bound IgE was detected with anti-IgE antiserum conjugated to horse radish peroxidase and using Enhanced Chemiluminiscens as substrate. The results were recorded from developed films using a computer fitted with a video camera and analyzed using a computer program. Blood donor sera were used to determine reference values for each food extract. Results: Serum IgE responses in patients experiencing severe allergic reactions to food showed a broad spectrum of reactivities. Sixtythree percent (33/52) of the sera were positive for at least one food extract. Nine sera responded to one single extract, 7 sera responded to 2 different extracts, and 2 and 3 sera responded to 3 and 4 extracts respectively. Twelve sera responded to more than 4 extracts. Some sera were highly polyreactive and responded to 15, 18, 31, 35 and 56 different extracts. Positive responses were recorded for 95 of the 150 extracts in the macroarray. Fourteen sera responded to kiwi and 9 sera to lentil. Seven positive responses were recorded to almond, haddock and peanut. There were six positive responses to crab, lobster, shrimp and sugar beet. Conclusion: The results were in general agreement with the assumption that food allergies are often directed to peanuts, treenuts, shellfish and fish, but some of the frequently observed responses need further investigation to determine their clinical relevance (f. ex. kiwi, lentil and sugar beet). One of the great advantages of the method is the simultanous detection of reactivity to crossreactive allergens, thereby adding strength to the diagnosis; i.e. sera of patients with allergy to shellfish often respond to lobster, shrimp, crab and craw fish; or in the case of allergy to cereals, there may be responses to rye, barley and wheat.

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[9] - Belmonte J, Alarcon M, Avila A, Izquierdo R, Cuevas E. Foreign allergenic pollen. Some cases observed in Catalonia (NE Spain) and Tenerife (Canary islands). Allergy 2008;63(suppl. 88):104-105

Background: Pollen dispersal, a crucial process in the life cycle of wind-pollinated plants, depends on the atmospheric dynamics, and can vary from a few meters to thousands of kilometres. Several studies have shown the occurrence of long-range pollen transport; e.g. the transcontinental transport from Africa to North America or to Europe, or from the boreal/austral forests to the poles. Pollen transport and dispersion can have important consequences, such as the expansion of the bio-geographical range of different organisms, the transport of pathogens and effects on the human health due to allergenic pollen. Methods: Singular aerobiological Hirst pollen records (appearance of unexpected pollen types or detection of simultaneous pollen peaks at different sites) were analyzed together with the back trajectories and the meteorological setting during the peak days. Dust transport model maps, satellite images and source-receptor models were also used for the analysis. Results: In Catalonia, NE of Spain, significant concentrations of Ambrosia pollen (species scarcely present in Catalonia) were collected in September1996 and its appearance were attributed to long range transport from the Lyon region in France, where the species is abundant. In the same way, Fagus pollen peaks appeared simultaneously at different sites across the Catalan geography in 2004 and 2006, indicating a broad-scale transport phenomenon. The analysis of back trajectories and the meteorological setting during the peak days indicated a consistent European provenance. The application of a source-receptor model showed regions in Central Europe as the probable areas of Fagus pollen emissions. Also, Chenopodiaceae/Amaranthaceae, Artemisia and Casuarina pollen collected in Santa Cruz de Tenerife and Izan˘ a (Canary Islands) has been clearly related to an African provenance and associated with dust storms. Conclusion: The aerobiological pollen spectra are occasionally influenced by pollen and spores of far provenance transported by synoptic movements of the air masses. Sporadic episodes of allergy may be associated with these situations.

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

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[11] - Bousquet J, Cour P, Guerin B, Michel FB. Allergy in the Mediterranean area I. Pollen counts and pollinosis of Montpellier. Clin Allergy 1984;14:249-258

The climatic conditions of the Mediterranean area result in vegetation and pollen very different from that of the other parts of Europe. The pollen content of the atmosphere of Montpellier, southern France, was examined using a filter sampler which was shown to be more efficient than most of the current devices for air sampling. Pollen counts were subsequently compared with pollinosis of patients born and living in and around Montpellier. The mean annual pollen counts showed that grass pollens and Cupressaceae pollens (cypress and juniper) are the highest. Some Mediterranean pollens (Oleaceae, London plane, Parietaria) are also important. Plantain and oak pollens are also present in relatively large amounts. Grass pollen allergy was found to be present in 86.5% of pollen-allergic patients. It was followed by plantain, Parietaria, Oleaceae, London plane and Cupressaceae pollens which were allergenic in 13-36% of pollen-allergic patients. Oak and pine pollens were present in large quantities in the counts but few persons were sensitive to oak and none to pine. By contrast, some patients had positive skin tests to alfalfa, red clover, acacia and lime tree pollens though these pollens were almost absent from the counts. In a few cases local sources of these pollens could account for the positive skin tests but cross-sensitivities could also occur. In summary, pollinosis of the Northern Mediterranean area is intermediate between the southern part of the area and the other parts of Europe.

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[12] - Ferrer A, Huertas J, de Larramendi CH, Pagan J, Garcia-Abujeta J, Lavin J et al. Cross-reactivity Between Tomato And Pollen Extracts. J Allergy Clin Immunol 2009;123:S30

RATIONALE: Cross reactivity among fruits and different pollen species have been extensively described. The objective of this study was to analyze cross-reactivity between tomato and other common pollen extracts. METHODS: A serum pool from 18 individuals (9 males and 9 females; age 25.4 ± 8.4) with positive skin prick test to peel tomato was prepared. Extracts from Olea europaea, Platanus acerifolia, Plantago lanceolata, Chenopodium album, Salsola kali, Betula alba, Artemisia vulgaris, Phoenix dactyliphera and grasses (Avena sativa and Poa pratensis) were prepared. Specific IgE was determined by direct ELISA. Allergenic profile of different extracts was evaluated by Immunoblot. Percentage of cross-reactivity between extracts was analyzed by CAP inhibition and the allergens implicated by immunoblot inhibition. RESULTS: Eleven individuals (61.1%) reported symptoms after ingestion and/or contact with tomato. Twelve (66.7%) individuals were sensitized to Chenopodiaceae, 11 (61.1%) to A. vulgaris, 10 (55.5%) to O. europaea, 3 (16.7%) to P. acerofila and 6 (33.3%) to grasses. The highest specific IgE levels were obtained to grasses, P. acerifolia, O. europaea and Chenopodiaceae. Most pollen extracts showed a similar capacity of inhibition (about 35%). A. sativa was capable to inhibit 53% and P. pratensis 47%. P. dactyliphera showed the less capacity of inhibition with 23.4%. The most strongly inhibited allergens corresponded to bands of 32 and 45 kDa. The band of 10 kDa was poorly inhibited by all extracts except by P. acerifolia. CONCLUSIONS: Cross-reactivity between tomato and pollen extracts have been demonstrated, especially in grasses. Allergens with high molecular weight range seem to be the responsible.

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[13] - Luoto S, Lambert W, Blomqvist A, Emanuelsson C The identification of allergen proteins in sugar beet (Beta vulgaris) pollen causing occupational allergy in greenhouses. Clin Mol Allergy 2008;6(7):1-27

Abstract Background. During production of sugar beet (Beta vulgaris) seeds in greenhouses, workers frequently develop allergic symptoms. The aim of this study was to identify and characterize possible allergens in sugar beet pollen. Methods. Sera from individuals at a local sugar beet seed producing company, having positive SPT and specific IgE to sugar beet pollen extract, were used for immunoblotting. Proteins in sugar beet pollen extracts were separated by 1- and 2-dimensional electrophoresis, and IgE-reactive proteins analyzed by liquid chromatography tandem mass spectrometry. Results. A 14 kDa protein was identified as an allergen, since IgE-binding was inhibited by the well-characterized allergen Che a 2, profilin, from the related species Chenopodium album. The presence of 17 kDa and 14 kDa protein homologues to both the allergens Che a 1 and Che a 2 were detected in an extract from sugar beet pollen, and partial amino acid sequences were determined, using inclusion lists for tandem mass spectrometry based on homologous sequences. Conclusions. Two occupational allergens were identified in sugar beet pollen showing sequence similarity with Chenopodium allergens. Sequence data were obtained by mass spectrometry (70 and 25%, respectively for Beta v 1 and Beta v 2), and can be used for cloning and recombinant expression of the allergens. As for treatment of Chenopodium pollinosis, immunotherapy with sugar beet pollen extracts may be feasible.

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[14] - Cosmes Martin PM, Moreno Ancillo A, Dominguez Noche C, Gutierrez Vivas A, Belmonte Soler J, Roure Nolla JM. [Sensitization to Castanea sativa pollen and pollinosis in northern Extremadura (Spain)]. Allergol Immunopathol (Madr) 2005;33:145-150

BACKGROUND: Castanea sativa pollen allergy has generally been considered to be uncommon and clinically insignificant. In our geographical area (Plasencia, Caceres, Spain) Castanea sativa pollen is a major pollen. OBJECTIVE: To determine the atmospheric fluctuations and prevalence of patients sensitized to Castanea pollen in our region and to compare this sensitization with sensitizations to other pollens. METHODS: Patients with respiratory symptoms attending our outpatient clinic for the first time in 2003 were studied. The patients underwent skin prick tests with commercial extracts of a battery of inhalants including Castanea sativa pollen. Serologic specific IgE to Castanea sativa pollen was determined using the CAP system (Pharmacia and Upjohn, Uppsala, Sweden). Airborne pollen counts in our city were obtained using Cour collection apparatus over a 4-year period (2000 to 2003). RESULTS: The most predominant pollens detected were (mean of the maximal weekly concentrations over 4 years in pollen grains/m3): Quercus 968, Poacea 660, Olea 325, Platanus 229, Pinus 126, Cupresaceae 117, Plantago 109, Alnus 41, Populus 40, Castanea 32. We studied 346 patients (mean age: 24.1 years). In 210 patients with a diagnosis of pollinosis, the percentages of sensitization were: Dactylis glomerata 80.4%, Olea europea 71.9%, Fraxinus excelsior 68%, Plantago lanceolata 62.8%, Chenopodium album 60.9%, Robinia pseudoacacia 49%, Artemisia vulgaris 43.8%, Platanus acerifolia 36.6%, Parietaria judaica 36.1%, Populus nigra 32.3%, Betula alba 27.6%, Quercus ilex 21.4%, Alnus glutinosa 20.9%, Cupressus arizonica 7.6% and Castanea sativa 7.1%. Fifteen patients were sensitized to Castanea sativa and 14 had seasonal rhinoconjunctivitis and asthma. Ten patients had serum specific IgE to Castanea pollen (maximum value: 17.4 Ku/l). Castanea pollen is present in our area in large amounts from the 23rd to the 28th weeks of the year, with a peak pollen count in the 25th week. CONCLUSIONS: The most important allergenic pollens in northern Extremadura were Poaceae, Olea europaea and Plantago sp. The prevalence of sensitization to Castanea sativa pollen was very low (7.1%). Most sensitized patients had asthma and polysensitization. Castanea sativa pollen is not a major cause of pollinosis in our area.

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[15] - Lombardero M, Garcia-Sellés FJ, Polo F, Jimeno L, Chamorro MJ, Garcia-Casado G, et al. Prevalence of sensitization to Artemisia allergens Art v 1, Art v 3 and Art v 60 kDa. Cross-reactivity among Art v 3 and other relevant lipid-transfer protein allergens. Clin Exp Allergy 2004;34:1415-1421

BACKGROUND: Artemisia vulgaris is a widespread weed in the Mediterranean area and several allergens have been detected in its pollen. One of them, Art v 3, belongs to the lipid-transfer protein (LTP) family and its prevalence in Artemisia-sensitized patients or its relationship with other LTP allergens is not clear . OBJECTIVE: To assess the pattern of sensitization to an array of mugwort allergens in a Mediterranean population, and to study the cross-reactivity of Art v 3 with Pru p 3 and Par j 1, relevant LTP allergens in the area . METHODS: Skin prick test was performed with whole extracts (A. vulgaris, Parietaria judaica and peach) and pure natural allergens Art v 1, Art v 3, Art v 60 kDa and Par j 1 in 24 mugwort-allergic patients from a Mediterranean area. In vitro assays included measurement of specific IgE and ELISA inhibition among LTP allergens . RESULTS: The three Artemisia allergens elicited a positive skin response in 70-80% of the patients. Seven patients were clearly sensitized to Par j 1 and 11 to Pru p 3. There was no correlation between Par j 1 and Pru p 3 sensitization, but a highly significant correlation was found between peach extract and Art v 3 as regards the skin response. No IgE cross-reactivity was observed between Art v 3/Par j 1 or Pru p 3/Par j 1. In contrast, Art v 3 significantly inhibited the binding to Pru p 3 of IgE from three patients' sera out of six studied, but Pru p 3 was not able to inhibit the IgE binding to Art v 3 . CONCLUSION: Art v 3 is a major mugwort allergen and in some patients with IgE to both Art v 3 and Pru p 3, Art v 3 behaves as the primary sensitizing agent.

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

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

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[17] - Belchi-Hernandez J, Moreno-Grau S, Sanchez-Gascon F, Bayo J, Elvira Rendueles B, Bartolomé B, et al. Sensitization to Zygophyllum fabago pollen: a clinical and immunologic study. Allergy 1998;53:241-248

Zygophyllum fabago is a herbaceous plant found widely in the Mediterranean area. There are no previous reports of its allergenicity. An aerobiologic and clinical survey was conducted in Murcia, southern Spain, to determine the quantity of airborne pollen and establish the possible role of this pollen as a cause of allergic symptoms. With a Hirst volumetric trap, we determined the atmospheric concentrations of this pollen in 1993, 1994, 1995, and 1996. Of 1180 patients tested, 181 (15.34%) had a positive skin test. To determine its allergenicity, we divided 47 patients into three groups: in group 1, all the patients had symptoms of rhinoconjunctivitis plus asthma; in groups 2 and 3, rhinoconjunctivitis. In group 1, we performed a bronchial provocation test (BPT); in groups 2 and 3, we performed nasal provocation (NPT) and conjunctival provocation (CPT) tests, respectively. SDS-PAGE was used to characterize the antigenic fractions and RAST inhibition to determine cross-reactivity with other pollens. The pollen dispersion period is from May to September (445 grains/m3). BPT was positive in 13 of 15 patients, NPT in 14 of 16 patients, and CPT in 13 of 16 patients. RAST inhibition revealed cross-reactivity with Mercurialis, Ricinus, Olea, and Betula. SDS-PAGE identified 25 IgE antibody-binding components, five of which (60, 65, 41, 38, and 15.5/14.7 kDa) were recognized by 40% of the sera. By SDS-PAGE immunoblotting with sunflower antiprofilin rabbit serum and affinity chromatography we established that the Z. fabago extract has profilin. This study shows that this pollen becomes airborne and elicits an IgE response which triggers respiratory symptoms in allergic subjects.

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[18] - Colás C, Monzon S, Venturini M, Lezaun A, Laclaustra M, Lara S, et al. Correlation between Chenopodiacea/Amaranthacea pollen counts and allergic symptoms in Salsola kali monosensitized patients. J Investig Allergol Clin Immunol 2005;15:254-258

We performed a prospective observational study to establish a relationship between pollen counts of Chenopodiacea/Amaranthacea and clinical symptoms of rhinoconjunctivitis and asthma in a group of monosensitised patients. MATERIAL AND METHODS: A total of 60 patients (19 with asthma) were included in the study. All patients collected daily symptom scores during the summer months of 1999, 2000 and 2001. The questionnaire included ocular, nasal and pulmonary symptoms. Pollen counts were expressed as pollen grains/m3. Symptom scores and pollen counts were correlated using correlation coefficients and Log transformed variables. RESULTS: In the 3 seasons studied we identified a peak of pollen and clinical symptoms in the second half of August and first half of September. In 1999, there was a significant positive correlation between total symptoms and daily pollen grains/m3 (p<0.005, r = 0.347). This correlation was not significant for the summers of 2000 and 2001. After further analysis, and by displacing one of both variables between 11 to 17 days, the correlation coefficients for total symptoms, improved for 1999 (r = 0. 744; p < 0.0001) and became significant for 2000 (r = 0. 521; p < 0.0001) and 2001 (r = 0.635; p < 0.0001). CONCLUSION: We identified a significant time lag between pollen counts and symptom scores in S. kali monosensitized patients.

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[19] - Mohapatra SS, Lockey RF, Polo F. Weed pollen allergens. In Lockey RF. and Ledford DK, (eds). Allergens and Allergen Immunotherapy for Allergic Diseases. 4th ed. 2008, New York: Informa Healthcare USA, Inc., p. 127-39

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[20] - Barderas R, Villalba M, Rodriguez R. Recombinant expression, purification and cross-reactivity of chenopod profilin: rChe a 2 as a good marker for profilin sensitization. Biol Chem 2004;385:731-737

Chenopod pollen is one of the major sources of allergens in some locations in the US, southern Europe and desert countries, and pollen profilin (Che a 2) is a major allergen. Recombinant Che a 2 (rChe a 2) has been produced in Escherichia coil cells with a final yield of 25 mg/l of cell culture. The expressed protein was isolated and structurally characterized by means of mass spectrometry, Edman degradation and circular dichroism. rChe a 2 displayed a molecular mass of 13 959 Da, which agrees with that of the amino acid sequence. The N-terminal amino acid sequence indicated the correct processing of the recombinant product. The immunological analysis of rChe a 2 showed IgG- and IgE-binding capabilities equivalent to those of its natural counterpart, Che a 2, isolated from the pollen. Inhibition experiments showed high cross-reactivity degrees with different allergenic sources. Inhibition degrees of >95% and >80% were obtained for chenopod profilin and, respectively, latex and pollen extracts, whereas 10-95% of inhibition was observed for different plant-derived foods. Due to its close relation to other allergenic profilins from pollens, plant-derived foods and latex, rChe a 2 could be a useful tool in clinical trials to detect profilin-allergic patients and perhaps, depending on its clinical relevance, in specific immunotherapy of these hypersensitive individuals.

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[21] - Luoto S, Lambert W, Blomqvist A, Emanuelsson C The identification of allergen proteins in sugar beet (Beta vulgaris) pollen causing occupational allergy in greenhouses. Clin Mol Allergy 2008;6(7):1-27

Abstract Background. During production of sugar beet (Beta vulgaris) seeds in greenhouses, workers frequently develop allergic symptoms. The aim of this study was to identify and characterize possible allergens in sugar beet pollen. Methods. Sera from individuals at a local sugar beet seed producing company, having positive SPT and specific IgE to sugar beet pollen extract, were used for immunoblotting. Proteins in sugar beet pollen extracts were separated by 1- and 2-dimensional electrophoresis, and IgE-reactive proteins analyzed by liquid chromatography tandem mass spectrometry. Results. A 14 kDa protein was identified as an allergen, since IgE-binding was inhibited by the well-characterized allergen Che a 2, profilin, from the related species Chenopodium album. The presence of 17 kDa and 14 kDa protein homologues to both the allergens Che a 1 and Che a 2 were detected in an extract from sugar beet pollen, and partial amino acid sequences were determined, using inclusion lists for tandem mass spectrometry based on homologous sequences. Conclusions. Two occupational allergens were identified in sugar beet pollen showing sequence similarity with Chenopodium allergens. Sequence data were obtained by mass spectrometry (70 and 25%, respectively for Beta v 1 and Beta v 2), and can be used for cloning and recombinant expression of the allergens. As for treatment of Chenopodium pollinosis, immunotherapy with sugar beet pollen extracts may be feasible.

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[22] - Barderas R, Villalba M, Pascual CY, Batenero E, Rodriguez E. Profilin (Che a 2) and polcalcin (Che a 3) are relevant allergens of Chenopodium album pollen: Isolation, amino acid sequences, and immunologic properties. J Allergy Clin Immunol 2004;113:1192-1198

Background Little is known about the molecular properties of chenopod allergens. Recently, profilin and 2 EF-hand calcium-binding protein (polcalcin) have been shown to play a role in chenopod pollinosis. Objective : We sought to analyze these panallergens in chenopod pollen and to evaluate their involvement in the allergy to this biologic source. Method s : Profilin and polcalcin were purified to homogeneity and characterized by using spectrometric and chemical methods. Immunologic analyses were performed by means of immunoblotting, ELISA, and competitive inhibition assays with olive profilinˆ and polcalcin-specific rabbit polyclonal antibodies and sera from patients with chenopod allergy. cDNAs encoding these proteins were cloned by means of PCR and sequenced. Result s : Purified Che a 2 (profilin) and Che a 3 (polcalcin) exhibited prevalences of 55% and 46%, respectively, in patients (n=104) hypersensitive to chenopod pollen. Both purified allergens individually inhibited the IgE binding to the whole pollen extract and showed strong cross-reactivity with the corresponding olive pollen profilin (Ole e 2) and polcalcin (Ole e 3). Chenopod profilin consists of a 131-amino-acid chain that displays identities of approximately 75% and 82% with pollen and food profilins, respectively. Che a 3 (86 amino acids) displays similarity (65% to 82% identity) with polcalcins from pollens of olive, birch, alder, rapeseed, and timothy. Conclusion : Profilin and polcalcin are relevant panallergens in chenopod pollen and good candidates to be involved in IgE cross-reactivity with other pollen sources, thus explaining the highly frequent polysensitization of patients allergic to chenopod.

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[23] - Barderas R, Villalba M, Rodriguez R. Recombinant expression, purification and cross-reactivity of chenopod profilin: rChe a 2 as a good marker for profilin sensitization. Biol Chem 2004;385:731-737

Chenopod pollen is one of the major sources of allergens in some locations in the US, southern Europe and desert countries, and pollen profilin (Che a 2) is a major allergen. Recombinant Che a 2 (rChe a 2) has been produced in Escherichia coil cells with a final yield of 25 mg/l of cell culture. The expressed protein was isolated and structurally characterized by means of mass spectrometry, Edman degradation and circular dichroism. rChe a 2 displayed a molecular mass of 13 959 Da, which agrees with that of the amino acid sequence. The N-terminal amino acid sequence indicated the correct processing of the recombinant product. The immunological analysis of rChe a 2 showed IgG- and IgE-binding capabilities equivalent to those of its natural counterpart, Che a 2, isolated from the pollen. Inhibition experiments showed high cross-reactivity degrees with different allergenic sources. Inhibition degrees of >95% and >80% were obtained for chenopod profilin and, respectively, latex and pollen extracts, whereas 10-95% of inhibition was observed for different plant-derived foods. Due to its close relation to other allergenic profilins from pollens, plant-derived foods and latex, rChe a 2 could be a useful tool in clinical trials to detect profilin-allergic patients and perhaps, depending on its clinical relevance, in specific immunotherapy of these hypersensitive individuals.

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

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[25] - Barderas R, Villalba M, Lombardero M, Rodríguez R. Identification and Characterization of Che a 1 Allergen from Chenopodium album Pollen. Int Arch Allergy Immunol 2002;127:47-54

Background: Pollinosis to Chenopodium album has been reported, but no data are available on its allergenic proteins. Methods: An allergen from C. album pollen has been isolated by means of gel permeation and reverse-phase high-performance liquid chromatography. Molecular characterization was achieved by concanavalin A reaction, mass spectrometry, Edman degradation and cDNA sequence. Antigenic analyses were performed by immunoblotting, ELISA, and ELISA inhibition, using sera from allergic patients, two Ole e 1-specific monoclonal antibodies and an Ole e 1-specific polyclonal antiserum. Results: The isolated allergen, Che a 1, is a glycoprotein of molecular mass 17.088 kD and 143 amino acid residues, whose sequence exhibits 27-45% identity with known members of the Ole e 1-like protein family. 77% of sera from patients allergic to chenopod pollen were reactive to Che a 1. No correlation was found between the IgE reactivities to Che a 1 and Ole e 1, the major allergens from olive pollen, and both allergens display low, although detectable, IgE and IgG cross-reactivities. Conclusions: Che a 1, a relevant allergen from chenopod pollen, is structurally related to the Ole e 1-like protein family, but exhibits significant differences on its polypeptide sequence that could explain its different antigenic behavior and limited cross-reactivity.

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[26] - Luoto S, Lambert W, Blomqvist A, Emanuelsson C The identification of allergen proteins in sugar beet (Beta vulgaris) pollen causing occupational allergy in greenhouses. Clin Mol Allergy 2008;6(7):1-27

Abstract Background. During production of sugar beet (Beta vulgaris) seeds in greenhouses, workers frequently develop allergic symptoms. The aim of this study was to identify and characterize possible allergens in sugar beet pollen. Methods. Sera from individuals at a local sugar beet seed producing company, having positive SPT and specific IgE to sugar beet pollen extract, were used for immunoblotting. Proteins in sugar beet pollen extracts were separated by 1- and 2-dimensional electrophoresis, and IgE-reactive proteins analyzed by liquid chromatography tandem mass spectrometry. Results. A 14 kDa protein was identified as an allergen, since IgE-binding was inhibited by the well-characterized allergen Che a 2, profilin, from the related species Chenopodium album. The presence of 17 kDa and 14 kDa protein homologues to both the allergens Che a 1 and Che a 2 were detected in an extract from sugar beet pollen, and partial amino acid sequences were determined, using inclusion lists for tandem mass spectrometry based on homologous sequences. Conclusions. Two occupational allergens were identified in sugar beet pollen showing sequence similarity with Chenopodium allergens. Sequence data were obtained by mass spectrometry (70 and 25%, respectively for Beta v 1 and Beta v 2), and can be used for cloning and recombinant expression of the allergens. As for treatment of Chenopodium pollinosis, immunotherapy with sugar beet pollen extracts may be feasible.

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[27] - Castro L, Villalba M, Abian S, Prado N, Sirvent S, Palomares O, et al. Isolation, characterisation, and cloning of Sal k 4, an Ole e 1-like protein from Salsola kali. Allergy 2008;63(suppl. 88):545

Background: Soil desertification of the Mediterranean area is enhancing the incidence of certain weeds as Salsola kali, which was until just recently a pollinosis typical of desert countries as Kuwait. To date only Sal k 1, a pectin-methylesterase, has been isolated and characterized. Other protein bands of 32, 20, 15 kDa, were recognized by the IgE of patients‚ sera. In this work, we present the isolation, characterisation, and cloning of the 20 kDa protein showing a preliminary clinical analysis of the relevance of this allergen in a S. kali-allergic population. Methods: Three chromatographic steps: Sephacryl S-100, Phenyl-Sepharose, and reverse- phase HPLC were carried out to purify Sal k 4. 2D-dimensional electrophoresis and fingerprint analysis was used to determine peptide sequences in order to clone and sequencing by PCR this allergen. Immunoblotting and ELISA were used to analyze IgE and IgG binding by using polyclonal and monoclonal antibodies specific to Ole e 1 and sera of patients allergic to S. kali. Results: This S. kali allergen belongs to the Ole e 1-like protein family and has been named as Sal k 4. It is a polypeptide chain of 151 amino acids and exhibits an identity degree of 68% to Che a 1, its homologous counterpart in Chenopodium album pollen. The immunologic similarity to Ole e 1 is not significant as it is proven after immunoblotting inhibition experiments and can be explained by the low identity % to Ole e 1. The prevalence data vary with geographical regions reaching values up to 64% in some populations. Conclusion: A new allergen from Salsola kali has been identified. The protein belongs to the Ole e 1-like protein family exhibiting a high degree of identity with members of the same Chenopodiaceae family, but lower with other members of the Ole e 1-protein family. Its significant prevalence makes this allergen necessary in the component-based diagnosis and specific immunotherapy.

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[28] - Barderas R, Villalba M, Lombardero M, Rodríguez R. Identification and Characterization of Che a 1 Allergen from Chenopodium album Pollen. Int Arch Allergy Immunol 2002;127:47-54

Background: Pollinosis to Chenopodium album has been reported, but no data are available on its allergenic proteins. Methods: An allergen from C. album pollen has been isolated by means of gel permeation and reverse-phase high-performance liquid chromatography. Molecular characterization was achieved by concanavalin A reaction, mass spectrometry, Edman degradation and cDNA sequence. Antigenic analyses were performed by immunoblotting, ELISA, and ELISA inhibition, using sera from allergic patients, two Ole e 1-specific monoclonal antibodies and an Ole e 1-specific polyclonal antiserum. Results: The isolated allergen, Che a 1, is a glycoprotein of molecular mass 17.088 kD and 143 amino acid residues, whose sequence exhibits 27-45% identity with known members of the Ole e 1-like protein family. 77% of sera from patients allergic to chenopod pollen were reactive to Che a 1. No correlation was found between the IgE reactivities to Che a 1 and Ole e 1, the major allergens from olive pollen, and both allergens display low, although detectable, IgE and IgG cross-reactivities. Conclusions: Che a 1, a relevant allergen from chenopod pollen, is structurally related to the Ole e 1-like protein family, but exhibits significant differences on its polypeptide sequence that could explain its different antigenic behavior and limited cross-reactivity.

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[29] - Barderas R, Villalba M, Rodriguez R. Che a 1: Recombinant Expression, Purification and Correspondence to the Natural Form. Int Arch Allergy Immunol 2004;135:284-292

BACKGROUND: Pollinosis to chenopods is one of the main causes of allergy in desertic regions and it is increasing in the South of Europe and Western USA. Che a 1 is a major allergen for chenopod-allergic subjects and belongs to the Ole-e-1-like family of proteins . METHODS: Pichia pastoris yeast has been used as expression system to produce the recombinant form of Che a 1 (rChe a 1). The allergen was isolated using a gel permeation column and reverse-phase/high-performance liquid chromatography. Molecular characterization was performed using Edman degradation, mass spectrometry and concanavalin A staining. Sera from patients allergic to chenopod pollen, as well as polyclonal and monoclonal antibodies raised against Ole e 1, were used in immunoblotting, ELISA and inhibition assays for immunological characterization of rChe a 1 . RESULTS: The allergen was purified to homogeneity with a final yield of 15 mg/l of cell culture and showed a glycosylated character. N-terminal amino acid sequence of rChe a 1 and molecular mass were according to those of the protein isolated from chenopod pollen. The recombinant allergen maintained the IgG and IgE epitopes of the natural allergen deduced from the immunological assays . CONCLUSIONS: Structural and in vitro immunological properties of rChe a 1 produced in P. pastoris were equivalent to those of the natural form of the allergen and, thus, it could be used in testing patients allergic to chenopods.

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[30] - Barderas R, Villalba M, Pascual CY, Batenero E, Rodriguez E. Profilin (Che a 2) and polcalcin (Che a 3) are relevant allergens of Chenopodium album pollen: Isolation, amino acid sequences, and immunologic properties. J Allergy Clin Immunol 2004;113:1192-1198

Background Little is known about the molecular properties of chenopod allergens. Recently, profilin and 2 EF-hand calcium-binding protein (polcalcin) have been shown to play a role in chenopod pollinosis. Objective : We sought to analyze these panallergens in chenopod pollen and to evaluate their involvement in the allergy to this biologic source. Method s : Profilin and polcalcin were purified to homogeneity and characterized by using spectrometric and chemical methods. Immunologic analyses were performed by means of immunoblotting, ELISA, and competitive inhibition assays with olive profilinˆ and polcalcin-specific rabbit polyclonal antibodies and sera from patients with chenopod allergy. cDNAs encoding these proteins were cloned by means of PCR and sequenced. Result s : Purified Che a 2 (profilin) and Che a 3 (polcalcin) exhibited prevalences of 55% and 46%, respectively, in patients (n=104) hypersensitive to chenopod pollen. Both purified allergens individually inhibited the IgE binding to the whole pollen extract and showed strong cross-reactivity with the corresponding olive pollen profilin (Ole e 2) and polcalcin (Ole e 3). Chenopod profilin consists of a 131-amino-acid chain that displays identities of approximately 75% and 82% with pollen and food profilins, respectively. Che a 3 (86 amino acids) displays similarity (65% to 82% identity) with polcalcins from pollens of olive, birch, alder, rapeseed, and timothy. Conclusion : Profilin and polcalcin are relevant panallergens in chenopod pollen and good candidates to be involved in IgE cross-reactivity with other pollen sources, thus explaining the highly frequent polysensitization of patients allergic to chenopod.

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[31] - Civantos E, del Pozo V, López E, Llanes ME, Cárdaba B, Gallardo S, et al. Characterization and molecular cloning of the major pollen allergen from Salsola kali : Sal k 1. EAACI 22th Congress, Paris, 7-11 June, 2003, Poster n°1353

Background: Salsola kali is a tumbleweed which belong to the Chenopodiacea family. It‚s the most allergenic of the family and it's widely distributed along the coast of Europe, North Africa, USA and Australia. The aim of this study is the characterization of Salsola kali major pollen antigen, and proceed with its cloning and sequencing. Methods: SDS-PAGE and immunoblot were utilized to identify allergenic molecules in a extract of Salsola kali pollen. A patient population of 67 was included in the study. Bidimensional PAGE was performed to obtain the protein pattern of Salsola kali and a pool of sera from sensitive patients were used. Cross-reactivity with Salsola kali was demonstrated by immunoblot inhibition with Chenopodium album pollen extract. Messenger RNA from Salsola kali pollen was obtained and the Sal k 1 sequence was established using 3'RACE system and primers based on the N´terminal sequence of a similar protein from Chenopodium album pollen. Recombinant Sal k 1 was cloned in pGEM-T easy and glycosylated form of protein is subcloning in pPICZaA for expression in Pichia pastoris. Results: A protein of 43 kDa was recognized by 58% of sensitive patients sera. Six isophorms from major allergen were identified by 2-D electrophoresis whose pH swing between 5.5 to 7, and all of them are recognized by a pool patients sera. A PCR fragment of 1074 pb was obtained and cloned in pGEM-T easy vector. Three clones were positive and their sequence completely identical. The sequence codifies for a 36 kDa protein with a pH estimated is 5.7. It has some putative glycosilation and phosphorilation sites and has homology with kinase from Oriza sativa, Zea mays and Arabidopsis thaliana. The nucleotide sequence was submitted to the NCBI Nucleotide Database (AF449490). Conclusions: The major allergen of Salsola kali pollen, Sal k 1, has a molecular weight of 43 kDa with six different isophorms (pI 5.5-7). The sequence does not present homology with any of allergens sequence until now known. The recombinant protein is a good tool to the standardization of the allergens used for diagnosis and immunotherapy.

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[32] - Carnés J, Fernandez-Caldas E, Marina A, Alonso C, Lahoz C, Colas C, et al. Immunochemical characterization of Russian thistle (Salsola kali) pollen extracts. Purification of the allergen Sal k 1. Allergy 2003;58:1152-1156

BACKGROUND: Salsola kali (Russian thistle) is a weed which belongs to the Chaenopodiacea family. It is widely distributed along the coasts of Europe, North Africa, USA and Australia. The objectives of this study were to study the allergenic composition of S. kali pollen and to purify an important allergen from the pollen extracts of this plant . METHODS: A population of 66 individuals with specific IgE-mediated allergic symptoms and positive skin tests to S. kali were included in the study. Specific IgE to S. kali was determined by direct enzyme-linked immunosorbent assay (ELISA). The antigenic and allergenic profile of S. kali was evaluated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focussing (IEF) and immunoblot. Allergen purification was conducted by preparative SDS-PAGE. The allergenicity of the protein was evaluated by skin testing, direct ELISA, ELISA inhibition and immunoblots . RESULTS: Specific IgE to S. kali was detected in 39 of the 66 individuals (59%). An allergen with a molecular weight of approximately 43 kDa was purified. This allergen was termed Sal k 1. A partial sequencing was obtained and no homology was found with other known proteins/allergens. The allergenicity of Sal k 1 was tested in vitro and in vivo. Of the 39 individuals with a positive specific IgE determination to S. kali, 26 (66.6%) had detectable specific IgE to Sal k 1. Twenty of these 39 individuals were skin-prick tested with the purified allergen (0.5 mg/ml) and all of them had a positive skin test to the purified allergen. Ten additional individuals, used as negative controls, had a negative response . CONCLUSIONS: Sal k 1, an important allergen of S. kali, is recognized, in vitro, by approximately 67% of the patients sensitized to S. kali. Twenty patients with a positive skin test to a standardized S. kali extract had a positive reaction to the purified allergen.

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[33] - Barderas R, García-Sellés J, Salamanca G, Colás C, Barber D, Rodríguez R, et al. A pectin methylesterase as an allergenic marker for the sensitization to Russian thistle (Salsola kali) pollen. Clin Exp Allergy 2007;37:1111-1119

Background Chenopodiaceae pollen is considered the main cause of pollen allergy in desert countries and its incidence is world-wide increasing by the desertization of extensive zones. Although the correlation between the sensitization to Chenopodium album and Salsola kali pollens of patients suffering from allergy to Chenopodiaceae pollens is high, a significant number of patients exhibited IgE sensitivity exclusively towards S. kali. Objective To analyse this differential reactivity and to purify, clone and characterize the putative responsible allergen. Methods Immunoblotting was used to analyse the IgE binding to pollen extract for S. kali and C. album. The protein was isolated by two chromatographic steps and characterized by Edman degradation, mass spectrometry, finger print analysis and Concanavalin A lectin staining. Specific cDNA was amplified by polymerase chain reaction, cloned in Escherichia coli and sequenced. Immunologic characterization was performed by immunoblotting, enzyme-linked immunoassay detection and inhibition experiments using sera from 11 patients allergic to S. kali pollen. Results cDNA codifies for a mature protein of 339 amino acids plus a putative signal peptide of 23 residues and it belongs to the plant pectin methylesterase (PME) family. It is a mildly basic and polymorphic protein and was recognized by the IgE from all the patients allergic to S. kali included in the study, and was called Sal k 1. The protein was not recognized in the C. album pollen extract using the sera of these patients. Conclusion Sal k 1 is a protein from the PME family with a high allergenic relevance. Considering this allergen as responsible for the different sensitization between S. kali and C. album pollen, it may be a useful marker to classify patients allergic to Chenopodiaceae allowing a safer and more specific immunotherapy.

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[34] - Lombardero M, Duffort O, Selles JG, Hernandez J, Carreira J. Cross-reactivity among Chenopodiaceae and Amaranthaceae. Ann Allergy 1985;54:430-436

Pollen extracts from Atriplex latifolia, Beta vulgaris, Salsola kali and Amaranthus retroflexus were compared with an extract from Chenopodium album by both in vivo and in vitro methods. Skin prick tests on 20 C. album-sensitive patients were positive with all extracts. RAST inhibition together with two-dimensional immunoelectrophoresis and two-dimensional radioimmunoelectrophoresis indicate that common allergenic determinants are present. Electrophoretic transfer for detection of IgE binding molecules from sodium dodecyl sulfate-polyacrylamide gel electrophoresis or isoelectricfocusing gels suggests that the common allergenic determinants are present in molecules with various molecular weights or isoelectric points.

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[35] - Würtzen PA, Nelson HS, Lowenstein H, Ipsen H. Characterization of Chenopodiales (Amaranthus retroflexus, Chenopodium album, Kochia scoparia, Salsola pestifer) pollen allergens. Allergy 1995;50:489-497

Pollen extracts of the four taxonomically related weeds, Amaranthus retroflexus (Ama r), Chenopodium album (Che a), Kochia scoparia (Koc s), and Salsola pestifer (S. kali) (Sal p), were characterized by various methods including crossed immunoelectrophoresis (CIE), crossed radioimmunoelectrophoresis (CRIE), and SDS-PAGE immunoblotting. The allergen profiles were determined by CRIE and SDS-PAGE IgE immunoblotting. CRIE detected from one to four important allergens, while SDS showed up to four bands that bound IgE from a number of patient sera. CRIE and SDS-PAGE immunoblotting did not recognize the same number of important allergens in the individual weeds, and the number of allergens detected by the two methods differed considerably, suggesting that IgE-binding epitopes may be denatured during SDS-PAGE. However, it was possible to correlate the identity of some of the important allergens detected by CRIE and SDS-PAGE immunoblotting in all four weeds.

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[36] - Luoto S, Lambert W, Blomqvist A, Emanuelsson C The identification of allergen proteins in sugar beet (Beta vulgaris) pollen causing occupational allergy in greenhouses. Clin Mol Allergy 2008;6(7):1-27

Abstract Background. During production of sugar beet (Beta vulgaris) seeds in greenhouses, workers frequently develop allergic symptoms. The aim of this study was to identify and characterize possible allergens in sugar beet pollen. Methods. Sera from individuals at a local sugar beet seed producing company, having positive SPT and specific IgE to sugar beet pollen extract, were used for immunoblotting. Proteins in sugar beet pollen extracts were separated by 1- and 2-dimensional electrophoresis, and IgE-reactive proteins analyzed by liquid chromatography tandem mass spectrometry. Results. A 14 kDa protein was identified as an allergen, since IgE-binding was inhibited by the well-characterized allergen Che a 2, profilin, from the related species Chenopodium album. The presence of 17 kDa and 14 kDa protein homologues to both the allergens Che a 1 and Che a 2 were detected in an extract from sugar beet pollen, and partial amino acid sequences were determined, using inclusion lists for tandem mass spectrometry based on homologous sequences. Conclusions. Two occupational allergens were identified in sugar beet pollen showing sequence similarity with Chenopodium allergens. Sequence data were obtained by mass spectrometry (70 and 25%, respectively for Beta v 1 and Beta v 2), and can be used for cloning and recombinant expression of the allergens. As for treatment of Chenopodium pollinosis, immunotherapy with sugar beet pollen extracts may be feasible.

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[37] - Ferrer A, Larramendi CH, Huertas AJ, Pagan JA, Andreu C, Garcia-Abujeta JL et al. Cross-reactivity between different species of the Chenopodiaceae family and identification of the responsible allergens. Allergy 2009;64(Suppl. 90):507

Background: Sensitivity to Chenopodiaceae family is a frequent cause of allergic respiratory diseases. Previous studies have demonstrated that not only Salsola kali or Chenopodium album but also S. oppositifolia is capable to induce sensitization. The objective of this study was to evaluate the cross-reactivity between S. oppositifolia and the two other species and to analyze the responsible allergens. Methods: S. kali, S. oppositifolia and Ch. album pollen extracts were manufactured. Patients reporting respiratory and/or cutaneous symptoms were skin prick tested with the 3 extracts and with a battery of common aeroallergens. A total of 246 individuals with positive skin prick test to at least one of the three species were included in the study. SDS-PAGE and bidimensional studies were carried out to elucidate the protein profile of the extracts. The allergenic profile was investigated in 131 serum samples by immunoblot studies. Inhibition experiments with the 3 species were conducted with a pooll of sera, in order to establish cross-reactivity between them. Results: 246 patients (108 males and 138 females; mean age: 30.9± 13.8 years) were included. Protein profile, 2-D analysis and immunoblot demonstrated a similar pattern in all extracts, except in some low molecular weight allergens of S. oppositifolia. Immunoblot inhibition experiments demonstrated that most of the high molecular weight allergens of S. oppositifolia were inhibited by S. kali whereas low molecular weight allergens were only totally inhibited by Ch. album. Conclusions: This study confirms the allergenic importance of S. oppositifolia. We have also demonstrated that the three species share a high degree of cross-reactivity, but S. oppositifolia seems to have more allergenic similarities with Ch. album than S. kali.

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[38] - Civantos E, del Pozo V, López E, Llanes ME, Cárdaba B, Gallardo S, et al. Characterization and molecular cloning of the major pollen allergen from Salsola kali : Sal k 1. EAACI 22th Congress, Paris, 7-11 June, 2003, Poster n°1353

Background: Salsola kali is a tumbleweed which belong to the Chenopodiacea family. It‚s the most allergenic of the family and it's widely distributed along the coast of Europe, North Africa, USA and Australia. The aim of this study is the characterization of Salsola kali major pollen antigen, and proceed with its cloning and sequencing. Methods: SDS-PAGE and immunoblot were utilized to identify allergenic molecules in a extract of Salsola kali pollen. A patient population of 67 was included in the study. Bidimensional PAGE was performed to obtain the protein pattern of Salsola kali and a pool of sera from sensitive patients were used. Cross-reactivity with Salsola kali was demonstrated by immunoblot inhibition with Chenopodium album pollen extract. Messenger RNA from Salsola kali pollen was obtained and the Sal k 1 sequence was established using 3'RACE system and primers based on the N´terminal sequence of a similar protein from Chenopodium album pollen. Recombinant Sal k 1 was cloned in pGEM-T easy and glycosylated form of protein is subcloning in pPICZaA for expression in Pichia pastoris. Results: A protein of 43 kDa was recognized by 58% of sensitive patients sera. Six isophorms from major allergen were identified by 2-D electrophoresis whose pH swing between 5.5 to 7, and all of them are recognized by a pool patients sera. A PCR fragment of 1074 pb was obtained and cloned in pGEM-T easy vector. Three clones were positive and their sequence completely identical. The sequence codifies for a 36 kDa protein with a pH estimated is 5.7. It has some putative glycosilation and phosphorilation sites and has homology with kinase from Oriza sativa, Zea mays and Arabidopsis thaliana. The nucleotide sequence was submitted to the NCBI Nucleotide Database (AF449490). Conclusions: The major allergen of Salsola kali pollen, Sal k 1, has a molecular weight of 43 kDa with six different isophorms (pI 5.5-7). The sequence does not present homology with any of allergens sequence until now known. The recombinant protein is a good tool to the standardization of the allergens used for diagnosis and immunotherapy.

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[39] - Barderas R, García-Sellés J, Salamanca G, Colás C, Barber D, Rodríguez R, et al. A pectin methylesterase as an allergenic marker for the sensitization to Russian thistle (Salsola kali) pollen. Clin Exp Allergy 2007;37:1111-1119

Background Chenopodiaceae pollen is considered the main cause of pollen allergy in desert countries and its incidence is world-wide increasing by the desertization of extensive zones. Although the correlation between the sensitization to Chenopodium album and Salsola kali pollens of patients suffering from allergy to Chenopodiaceae pollens is high, a significant number of patients exhibited IgE sensitivity exclusively towards S. kali. Objective To analyse this differential reactivity and to purify, clone and characterize the putative responsible allergen. Methods Immunoblotting was used to analyse the IgE binding to pollen extract for S. kali and C. album. The protein was isolated by two chromatographic steps and characterized by Edman degradation, mass spectrometry, finger print analysis and Concanavalin A lectin staining. Specific cDNA was amplified by polymerase chain reaction, cloned in Escherichia coli and sequenced. Immunologic characterization was performed by immunoblotting, enzyme-linked immunoassay detection and inhibition experiments using sera from 11 patients allergic to S. kali pollen. Results cDNA codifies for a mature protein of 339 amino acids plus a putative signal peptide of 23 residues and it belongs to the plant pectin methylesterase (PME) family. It is a mildly basic and polymorphic protein and was recognized by the IgE from all the patients allergic to S. kali included in the study, and was called Sal k 1. The protein was not recognized in the C. album pollen extract using the sera of these patients. Conclusion Sal k 1 is a protein from the PME family with a high allergenic relevance. Considering this allergen as responsible for the different sensitization between S. kali and C. album pollen, it may be a useful marker to classify patients allergic to Chenopodiaceae allowing a safer and more specific immunotherapy.

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[40] - Lombardero M, Duffort O, Selles JG, Hernandez J, Carreira J. Cross-reactivity among Chenopodiaceae and Amaranthaceae. Ann Allergy 1985;54:430-436

Pollen extracts from Atriplex latifolia, Beta vulgaris, Salsola kali and Amaranthus retroflexus were compared with an extract from Chenopodium album by both in vivo and in vitro methods. Skin prick tests on 20 C. album-sensitive patients were positive with all extracts. RAST inhibition together with two-dimensional immunoelectrophoresis and two-dimensional radioimmunoelectrophoresis indicate that common allergenic determinants are present. Electrophoretic transfer for detection of IgE binding molecules from sodium dodecyl sulfate-polyacrylamide gel electrophoresis or isoelectricfocusing gels suggests that the common allergenic determinants are present in molecules with various molecular weights or isoelectric points.

↩

[41] - Würtzen PA, Nelson HS, Lowenstein H, Ipsen H. Characterization of Chenopodiales (Amaranthus retroflexus, Chenopodium album, Kochia scoparia, Salsola pestifer) pollen allergens. Allergy 1995;50:489-497

Pollen extracts of the four taxonomically related weeds, Amaranthus retroflexus (Ama r), Chenopodium album (Che a), Kochia scoparia (Koc s), and Salsola pestifer (S. kali) (Sal p), were characterized by various methods including crossed immunoelectrophoresis (CIE), crossed radioimmunoelectrophoresis (CRIE), and SDS-PAGE immunoblotting. The allergen profiles were determined by CRIE and SDS-PAGE IgE immunoblotting. CRIE detected from one to four important allergens, while SDS showed up to four bands that bound IgE from a number of patient sera. CRIE and SDS-PAGE immunoblotting did not recognize the same number of important allergens in the individual weeds, and the number of allergens detected by the two methods differed considerably, suggesting that IgE-binding epitopes may be denatured during SDS-PAGE. However, it was possible to correlate the identity of some of the important allergens detected by CRIE and SDS-PAGE immunoblotting in all four weeds.

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[42] - Weber RW. Cross-reactivity of Pollen Allergens. Curr Allergy Asthma Rep 2004;4:401-408

Pollen cross-allergenicity has practical implications on the management of inhalant allergy, in both evaluation and therapy, especially with allergen vaccine immunotherapy. The study of cross-reactivity among pollen allergens has expanded beyond investigation of crude extracts to the characterization and cloning of numerous pollen proteins. In this review, the interrelationships between these pollen allergens in the context of botanical systematics are examined, to provide a framework for cross-reactivity understanding. Recommendations for choices in evaluation and therapy are given.

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[43] - Assarehzadegan MA, Sankian M, Jabbari F, Noorbakhsh R, Varasteh A. Allergy to Salsola Kali in a Salsola Incanescens-rich Area: Role of Extensive Cross Allergenicity. Allergol Int 2009;58:261-266

Background: Pollens from the Salsola spp. are an important source of respiratory allergy in tropical countries. Our aim was to characterize the IgE binding proteins of S. incanescens pollen extract and study its cross-reactivity with S. kali pollen allergens. Methods: Prick tests with S. kali and S. incanescens pollen extracts were performed on eight respiratory allergy patients from Mashhad, Northeast Iran. The antigenic profiles and IgE-binding patterns of S. kali and S. incanescens pollen extracts were compared by SDS-PAGE and Western blotting, using individual sera from the salsola pollen-sensitive patients. Cross-reactivity of proteins in the two weeds was assessed by IgE- immunoblotting inhibition. Results: S. kali and S. incanescens pollen extracts showed similar IgE-binding profiles in Western blotting. The IgE binding components of 39, 45, 66 and 85kDa were detected in both pollen extracts. Furthermore, inhibition of the immunoblots revealed extensive inhibition of IgE binding to proteins and a close relationship between these two weeds allergens. Conclusions: S. incanescens pollen is a potent allergen source with several IgE binding components that shows a close allergenic relationship with S. kali. Our results suggest that in S. incanescens-rich areas, S. kali pollen extracts could be used as a diagnostic reagent for allergic patients to S. incanescens pollen.

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[44] - Ferrer A, Larramendi CH, Huertas AJ, Pagan JA, Andreu C, Garcia-Abujeta JL et al. Cross-reactivity between different species of the Chenopodiaceae family and identification of the responsible allergens. Allergy 2009;64(Suppl. 90):507

Background: Sensitivity to Chenopodiaceae family is a frequent cause of allergic respiratory diseases. Previous studies have demonstrated that not only Salsola kali or Chenopodium album but also S. oppositifolia is capable to induce sensitization. The objective of this study was to evaluate the cross-reactivity between S. oppositifolia and the two other species and to analyze the responsible allergens. Methods: S. kali, S. oppositifolia and Ch. album pollen extracts were manufactured. Patients reporting respiratory and/or cutaneous symptoms were skin prick tested with the 3 extracts and with a battery of common aeroallergens. A total of 246 individuals with positive skin prick test to at least one of the three species were included in the study. SDS-PAGE and bidimensional studies were carried out to elucidate the protein profile of the extracts. The allergenic profile was investigated in 131 serum samples by immunoblot studies. Inhibition experiments with the 3 species were conducted with a pooll of sera, in order to establish cross-reactivity between them. Results: 246 patients (108 males and 138 females; mean age: 30.9± 13.8 years) were included. Protein profile, 2-D analysis and immunoblot demonstrated a similar pattern in all extracts, except in some low molecular weight allergens of S. oppositifolia. Immunoblot inhibition experiments demonstrated that most of the high molecular weight allergens of S. oppositifolia were inhibited by S. kali whereas low molecular weight allergens were only totally inhibited by Ch. album. Conclusions: This study confirms the allergenic importance of S. oppositifolia. We have also demonstrated that the three species share a high degree of cross-reactivity, but S. oppositifolia seems to have more allergenic similarities with Ch. album than S. kali.

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[45] - Weber RW. Cross-reactivity of pollen allergens: recommendations for immunotherapy vaccines. Curr Opin Allergy Clin Immunol 2005;5:563-569

PURPOSE OF REVIEW: This review will summarize recent research on pollen allergen and epitope cross-reactivity. Knowledge of these relationships aids in the rational formulation of allergen immunotherapy vaccines. RECENT FINDINGS: There has been further clarification of panallergens and their roles as both major and minor allergens. Recent studies have targeted non-specific lipid transfer proteins and calcium-binding proteins (polcalcins), as well as pathogenesis-related protein families and profilins. Polcalcins and non-specific lipid transfer proteins are responsible for pollen-fruit interactions as well as pollen cross-reactivity, in some cases, but not all, accounting for major allergenicity. Delineation of the enzymatic activity of certain allergens explains the ubiquitous nature of these pollen proteins. SUMMARY: Characterization of specific pollen allergens and their protein families has provided insight into the grounds for cross-reactivity. Continuing clarification of these relationships will allow the substitution and consolidation of inhalant extracts as described in the conclusion.

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[46] - Barderas R, Villalba M, Lombardero M, Rodríguez R. Identification and Characterization of Che a 1 Allergen from Chenopodium album Pollen. Int Arch Allergy Immunol 2002;127:47-54

Background: Pollinosis to Chenopodium album has been reported, but no data are available on its allergenic proteins. Methods: An allergen from C. album pollen has been isolated by means of gel permeation and reverse-phase high-performance liquid chromatography. Molecular characterization was achieved by concanavalin A reaction, mass spectrometry, Edman degradation and cDNA sequence. Antigenic analyses were performed by immunoblotting, ELISA, and ELISA inhibition, using sera from allergic patients, two Ole e 1-specific monoclonal antibodies and an Ole e 1-specific polyclonal antiserum. Results: The isolated allergen, Che a 1, is a glycoprotein of molecular mass 17.088 kD and 143 amino acid residues, whose sequence exhibits 27-45% identity with known members of the Ole e 1-like protein family. 77% of sera from patients allergic to chenopod pollen were reactive to Che a 1. No correlation was found between the IgE reactivities to Che a 1 and Ole e 1, the major allergens from olive pollen, and both allergens display low, although detectable, IgE and IgG cross-reactivities. Conclusions: Che a 1, a relevant allergen from chenopod pollen, is structurally related to the Ole e 1-like protein family, but exhibits significant differences on its polypeptide sequence that could explain its different antigenic behavior and limited cross-reactivity.

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[47] - Castro L, Villalba M, Abian S, Prado N, Sirvent S, Palomares O, et al. Isolation, characterisation, and cloning of Sal k 4, an Ole e 1-like protein from Salsola kali. Allergy 2008;63(suppl. 88):545

Background: Soil desertification of the Mediterranean area is enhancing the incidence of certain weeds as Salsola kali, which was until just recently a pollinosis typical of desert countries as Kuwait. To date only Sal k 1, a pectin-methylesterase, has been isolated and characterized. Other protein bands of 32, 20, 15 kDa, were recognized by the IgE of patients‚ sera. In this work, we present the isolation, characterisation, and cloning of the 20 kDa protein showing a preliminary clinical analysis of the relevance of this allergen in a S. kali-allergic population. Methods: Three chromatographic steps: Sephacryl S-100, Phenyl-Sepharose, and reverse- phase HPLC were carried out to purify Sal k 4. 2D-dimensional electrophoresis and fingerprint analysis was used to determine peptide sequences in order to clone and sequencing by PCR this allergen. Immunoblotting and ELISA were used to analyze IgE and IgG binding by using polyclonal and monoclonal antibodies specific to Ole e 1 and sera of patients allergic to S. kali. Results: This S. kali allergen belongs to the Ole e 1-like protein family and has been named as Sal k 4. It is a polypeptide chain of 151 amino acids and exhibits an identity degree of 68% to Che a 1, its homologous counterpart in Chenopodium album pollen. The immunologic similarity to Ole e 1 is not significant as it is proven after immunoblotting inhibition experiments and can be explained by the low identity % to Ole e 1. The prevalence data vary with geographical regions reaching values up to 64% in some populations. Conclusion: A new allergen from Salsola kali has been identified. The protein belongs to the Ole e 1-like protein family exhibiting a high degree of identity with members of the same Chenopodiaceae family, but lower with other members of the Ole e 1-protein family. Its significant prevalence makes this allergen necessary in the component-based diagnosis and specific immunotherapy.

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[48] - Barderas R, Villalba M, Pascual CY, Batenero E, Rodriguez E. Profilin (Che a 2) and polcalcin (Che a 3) are relevant allergens of Chenopodium album pollen: Isolation, amino acid sequences, and immunologic properties. J Allergy Clin Immunol 2004;113:1192-1198

Background Little is known about the molecular properties of chenopod allergens. Recently, profilin and 2 EF-hand calcium-binding protein (polcalcin) have been shown to play a role in chenopod pollinosis. Objective : We sought to analyze these panallergens in chenopod pollen and to evaluate their involvement in the allergy to this biologic source. Method s : Profilin and polcalcin were purified to homogeneity and characterized by using spectrometric and chemical methods. Immunologic analyses were performed by means of immunoblotting, ELISA, and competitive inhibition assays with olive profilinˆ and polcalcin-specific rabbit polyclonal antibodies and sera from patients with chenopod allergy. cDNAs encoding these proteins were cloned by means of PCR and sequenced. Result s : Purified Che a 2 (profilin) and Che a 3 (polcalcin) exhibited prevalences of 55% and 46%, respectively, in patients (n=104) hypersensitive to chenopod pollen. Both purified allergens individually inhibited the IgE binding to the whole pollen extract and showed strong cross-reactivity with the corresponding olive pollen profilin (Ole e 2) and polcalcin (Ole e 3). Chenopod profilin consists of a 131-amino-acid chain that displays identities of approximately 75% and 82% with pollen and food profilins, respectively. Che a 3 (86 amino acids) displays similarity (65% to 82% identity) with polcalcins from pollens of olive, birch, alder, rapeseed, and timothy. Conclusion : Profilin and polcalcin are relevant panallergens in chenopod pollen and good candidates to be involved in IgE cross-reactivity with other pollen sources, thus explaining the highly frequent polysensitization of patients allergic to chenopod.

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[49] - Verdino P, Barderas R, Villalba M, Westritschnig K, Valenta R, Rodriguez R, et al. Three-dimensional structure of the cross-reactive pollen allergen Che a 3: visualizing cross-reactivity on the molecular surfaces of weed, grass, and tree pollen allergens. J Immunol 2008;180:2313-2321

Two EF-hand calcium-binding allergens (polcalcins) occur in the pollen of a wide variety of unrelated plants as highly cross-reactive allergenic molecules. We report the expression, purification, immunological characterization, and the 1.75-A crystal structure of recombinant Che a 3 (rChe a 3), the polcalcin from the weed Chenopodium album. The three-dimensional structure of rChe a 3 resembles an alpha-helical fold that is essentially identical with that of the two EF-hand allergens from birch pollen, Bet v 4, and timothy grass pollen, Phl p 7. The extensive cross-reactivity between Che a 3 and Phl p 7 is demonstrated by competition experiments with IgE Abs from allergic patients as well as specific Ab probes. Amino acid residues that are conserved for the two EF-hand allergen family were identified in multiple sequence alignments of polcalcins from 15 different plants. Next, the three-dimensional structures of rChe a 3, rPhl p 7, and rBet v 4 were used to identify conserved amino acids with high surface exposition to visualize surface patches as potential targets for the polyclonal IgE Ab response of allergic patients. The essentially identical three-dimensional structures of rChe a 3, rPhl p 7, and rBet v 4 explain the extensive cross-reactivity of allergic patients IgE Abs with two EF-hand allergens from unrelated plants. In addition, analyzing the three-dimensional structures of cross-reactive Ags for conserved and surface exposed amino acids may be a first approach to mapping the conformational epitopes on disease-related Ags that are recognized by polyclonal patient Abs

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[50] - Vallverdu A, Garcia-Ortega P, Martinez J, Martinez A, Esteban MI, de Molina M, et al. Mercurialis annua: characterization of main allergens and cross-reactivity with other species. Int Arch Allergy Immunol 1997;112:356-364

A multicentric study was conducted to evaluate the frequency of Mercurialis annua pollen sensitization in several areas of Spain and to select a population sample to characterize the main allergenic components in M. annua pollen. Patients were recruited from six hospitals in Spain. Out of 420 patients sensitized to pollens, 195 (46.4%) showed positive skin tests to M. annua, thus evidencing the high level of sensitization to the pollen of this plant in Spain. Thirty-seven sera with RAST class values to M. annua > or = 3 were selected for SDS-PAGE immunoblotting analysis. Two main allergenic components with molecular weights of 15.8 and 14.1 kD were detected in 59 and 51% of the sera, respectively, and they were identified as profilins. Isolation of the relevant allergens was made by affinity chromatography on a poly-L-proline-Sepharose column, followed by gel filtration and anion exchange chromatography in the micropreparative SMARTs System. A significant but low antigenic cross-reactivity between M. annua and Olea europaea, Fraxinus elatior, Ricinus communis, Salsola kali, Parietaria judaica and Artemisia vulgaris was demonstrated by several in vitro techniques.

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[51] - Egger M, Mutschlechner S, Wopfner N, Gadermaier G, Briza P, Ferreira F. Pollen-food syndromes associated with weed pollinosis: an update from the molecular point of view. Allergy 2006;61:461-476

Pollinosis patients often display adverse reactions upon the ingestion of plant-derived foods as a result of immunoglobulin E (IgE) cross-reactive structures shared by pollen and food allergen sources. The symptoms of such pollen-food syndromes (PFS) or class 2 food allergies range from local oral allergy syndrome to severe systemic anaphylaxis. Two clinical syndromes, the celery-mugwort-spice syndrome and the mugwort-mustard-allergy syndrome have been described in association with weed pollinosis. However, other associations between weed pollinosis and hypersensitivity to certain kinds of food have also been observed, like the mugwort-peach, the ragweed-melon-banana, the plantain-melon, the pellitory-pistachio, the goosefoot-fruit, the Russian thistle-saffron, and the hop-celery association. The number of allergen sources involved, the allergens, and influencing factors including geography, diet, and food preparation contribute to the high clinical complexity of PFS. So far, known causative cross-reactive allergens include profilins, lipid transfer proteins, and high-molecular weight allergens and/or glycoallergens. The current usage of nonstandardized allergen extracts poses additional problems for both diagnosis and therapy of PFS patients. Further identification and characterization of involved allergens is inescapable for better understanding of PFS and vaccine development. Panels of recombinant allergens and/or hypo-allergens are promising tools to improve both PFS diagnostics and therapy.

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[52] - Callejo A, Sanchís ME, Armentia A, Moneo I, Fernández A. A new pollen-fruit cross-reactivity. Allergy 2002;57:1088-1089

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[53] - Anibarro B, Fontela JL, De la Hoz F. Occupational asthma induces by garlic dust. J Allergy Clin Immunol 1997;100:734-738

BACKGROUND: Garlic dust has not been a frequently encountered cause of IgE-mediated disease . OBJECTIVE: We report on 12 patients (all of them garlic workers) with the clinical criteria for occupational asthma . METHODS: Skin prick tests and serum-specific IgE determinations were performed with common inhalants, garlic, and other members of the Liliaceae family (onion, leek, and asparagus). Bronchial challenge test with garlic powder was performed in all patients. Garlic and onion extract proteins were separated by sodium dodecylsulfate-polyacrylamide gel electrophoresis. Immunoblot and IgE immunoblot inhibition analyses were performed with patients' sera on extracts of garlic, onion, and pollens of Phleum pratense and Chenopodium album . RESULTS: Garlic sensitization was demonstrated by bronchial challenge test in seven patients (group 1) and ruled out in the remaining five (group 2). Clinical data were similar in both groups. The patients with garlic allergy had a mean age of 27 years, and all of them had pollen allergy; sensitization to other members of the Liliaceae family was also common. Electrophoresis of garlic extract revealed two major protein bands at approximately 12 and 54 kd. During IgE immunoblotting, the pool of sera reacted with garlic proteins mainly at 54 kd. Preincubation with onion, Phleum, and Chenopodium partially abolished the IgE binding to several allergens of garlic . CONCLUSION: We report on seven patients in whom an occupational garlic allergy was demonstrated. Garlic allergy is relatively rare but seems to affect young subjects with pollen allergy, and sensitization to other members of the Liliaceae family is common. The results of this study confirm the presence of some structurally similar allergens in garlic, onion, and certain pollens.

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[54] - Feo F, Martinez J, Martinez A, Galindo PA, Cruz A, Garcia R, et al. Occupational allergy in saffron workers. Allergy 1997;52:633-641

Sensitization to the flower of saffron, a plant commonly grown in Spain for commercial purposes, and its clinical significance as an occupational allergen were studied. The prick test and RAST, with saffron pollen, stamen, and pistil extracts, were used to evaluate the cutaneous and specific antibody responses in the studied population. Provocation tests in patients with clinical findings were used to verify the implication of saffron components in these symptoms. Fifty saffron workers were evaluated. Three of them were sensitized to saffron pollen and stamen proteins, giving prick and RAST positive values. One patient presented asthma, showing a positive bronchial provocation test, and two patients rhinoconjunctivitis, showing positive conjunctival provocation tests. Of a general allergic population of 237, 10 patients also presented cutaneous test and IgE positive to saffron. Saffron allergens (from pollen and stamens) were characterized by SDS-PAGE immunoblotting. A relevant allergen of 15.5 kDa with profilinic nature was detected and further purified by high-resolution gel filtration chromatography. No allergenic components were demonstrated in pistils. Cross-reactivity of saffron extracts was evaluated by RAST inhibition with respect to other pollen species commonly causing sensitization in the same area of study. A significant degree of cross-reactivity was demonstrated between saffron and Lolium, Salsola, or Olea. The identification of the protein components involved in the cross-reactions was investigated by blot inhibition

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Les allergènes des pollens d’Amaranthacées

Réactivités croisées entre pollens d’Amaranthacées

Réactivités croisées avec d’autres pollens

Réactivités croisées avec des aliments

Rubriques

Dans la même rubrique

Les ambroisies

L’armoise

Les Astéracées (Composées)

Les plantains

Les Euphorbiacées

Les Pariétaires