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Sample records for major buckwheat allergen

  1. Synthesis of hypoallergenic derivatives of the major allergen Fag t 1 from tartary buckwheat via sequence restructuring.

    PubMed

    Yang, Zhenhuang; Li, Yuying; Li, Chen; Wang, Zhuanhua

    2012-08-01

    Fag t 1, a legumin-type protein, is the major allergen in tartary buckwheat. In the current study, three recombinant derivatives of Fag t 1, designated as Fag t 1-rs1, Fag t 1-rs2, and Fag t 1-rs3, were constructed via rational design and genetic engineering. However, because of the loss of their native-like folds, the Fag t 1 derivatives failed to bind IgE, and their allergenic activities were reduced. The recombinant hypoallergenic variants are promising vaccine candidates for specific immunotherapy of buckwheat allergy. The unfolding of the Fag t 1 structure reduced its high resistance to gastrointestinal proteolysis and strongly reduced its IgE reactivity. The derivatives showed a more than 90% reduction in allergenic activity compared with rFag t 1. These results suggest that the structure-dependent stability of 11S seed storage proteins is directly related to digestive stability and allergenic potential. Therefore, the destruction of the native conformation is the appropriate strategy to reduce the allergenicity of the cupin family food allergens.

  2. Buckwheat anaphylaxis: an unusual allergen in Taiwan.

    PubMed

    Wang, Tsung-Chi; Shyur, Shyh-Dar; Wen, Da-Chin; Kao, Yu-Hsuan; Huang, Li-Hsin

    2006-01-01

    IgE-mediated hypersensitivity to buckwheat is common in Korea, Japan, and some other Asian countries. However, buckwheat is not a common allergen in Taiwan. We report a woman with asthma who had anaphylactic shock, generalized urticaria, and an acute exacerbation of asthma five minutes after ingesting buckwheat. The patient underwent skin prick and Pharmacia CAP testing (Uppsala, Sweden) for specific IgE to buckwheat, white sesame and soybean as well as other common allergens in Taiwan including Dermatophagoides pteronyssinus (Dp), D. farinae (Df), cat and dog dander, cockroach, egg white, cow milk and codfish. The patient had a strongly positive skin prick test response to buckwheat and positive reactions to Dp and latex. Specific IgE results were class 6 for buckwheat, class 4 for Dp and Df, and class 2 for dog dander, wheat, sesame and soybean. Results of an open food challenge with white sesame and soybean were negative. Although buckwheat is a rare allergen in Taiwan, it can cause extremely serious reactions and should be considered in patients presenting with anaphylaxis after exposure to buckwheat. PMID:17136883

  3. Effects of enzymatic hydrolysis of buckwheat protein on antigenicity and allergenicity

    PubMed Central

    Sung, Dong-Eun; Lee, Jeongok; Han, Youngshin; Shon, Dong-Hwa; Ahn, Kangmo

    2014-01-01

    BACKGROUND/OBJECTIVES Due to its beneficial health effects, use of buckwheat has shown a continuous increase, and concerns regarding the allergic property of buckwheat have also increased. This study was conducted for evaluation of the hydrolytic effects of seven commercial proteases on buckwheat allergens and its allergenicity. MATERIALS/METHODS Extracted buckwheat protein was hydrolyzed by seven proteolytic enzymes at individual optimum temperature and pH for four hours. Analysis was then performed using SDS-PAGE, immunoblotting, and competitive inhibition ELISA (ciELISA) with rabbit antiserum to buckwheat protein, and direct ELISA with pooled serum of 21 buckwheat-sensitive patients. RESULTS Alkaline protease, classified as serine peptidase, was most effective in reducing allergenicity of buckwheat protein. It caused decomposition of the whole buckwheat protein, as shown on SDS-PAGE, and results of immunoblotting showed that the rabbit antiserum to buckwheat protein no longer recognized it as an antigen. Allergenicity showed a decrease of more than 50% when pooled serum of patients was used in ELISA. Two proteolytic enzymes from Aspergillus sp. could not hydrolyze buckwheat allergens effectively, and the allergenicity even appeared to increase. CONCLUSIONS Serine-type peptidases appeared to show a relatively effective reduction of buckwheat allergenicity. However, the antigenicity measured using rabbit antiserum did not correspond to the allergenicity measured using sera from human patients. Production of less allergenic buckwheat protein may be possible using enzymatic hydrolysis. PMID:24944772

  4. Redefining the major peanut allergens.

    PubMed

    Zhuang, Yonghua; Dreskin, Stephen C

    2013-03-01

    Food allergy has become a major public health concern in westernized countries, and allergic reactions to peanuts are particularly common and severe. Allergens are defined as antigens that elicit an IgE response, and most allergenic materials (e.g., pollens, danders, and foods) contain multiple allergenic proteins. This has led to the concept that there are "major" allergens and allergens of less importance. "Major allergens" have been defined as allergens that bind a large amount of IgE from the majority of patients and have biologic activity. However, the ability of an allergen to cross-link complexes of IgE and its high-affinity receptor FcεRI (IgE/FcεRI), which we have termed its allergic effector activity, does not correlate well with assays of IgE binding. To identify the proteins that are the most active allergens in peanuts, we and others have employed in vitro model assays of allergen-mediated cross-linking of IgE/FcεRI complexes and have demonstrated that the most potent allergens are not necessarily those that bind the most IgE. The importance of a specific allergen can be determined by measuring the allergic effector activity of that allergen following purification under non-denaturing conditions and by specifically removing the allergen from a complex allergenic extract either by chromatography or by specific immunodepletion. In our studies of peanut allergens, our laboratory has found that two related allergens, Ara h 2 and Ara h 6, together account for the majority of the effector activity in a crude peanut extract. Furthermore, murine studies demonstrated that Ara h 2 and Ara h 6 are not only the major elicitors of anaphylaxis in this system, but also can effectively desensitize peanut-allergic mice. As a result of these observations, we propose that the definition of a major allergen should be based on the potency of that allergen in assays of allergic effector activity and demonstration that removal of that allergen from an extract results in

  5. Anaphylaxis after eating Italian pizza containing buckwheat as the hidden food allergen.

    PubMed

    Heffler, E; Guida, G; Badiu, I; Nebiolo, F; Rolla, G

    2007-01-01

    A 20-year-old woman developed anaphylaxis after eating pizza on 4 different occasions in 2 restaurants. Both restaurants made their pizza dough with a mixture of wheat and buckwheat flours. A prick-to-prick test with buckwheat flour was positive. Skin prick tests and specific immunoglobulin E responses to soybean and peanut were weakly positive while the response to buckwheat was negative. We ruled out a pathogenic role for peanut and soybean because the patient usually eats both with no signs of allergic reaction. Double-blind, placebo-controlled food challenges with buckwheat flour were positive after the administration of a cumulative dose of 2.3 g of the culprit flour. To our knowledge, our report describes the first case of anaphylaxis after intake of buckwheat flour as the hidden allergen in pizza dough.

  6. Endotoxin and House Dust Mite Allergen Levels on Synthetic and Buckwheat Pillows

    PubMed Central

    Nam, Hae-Seon; Park, Choon-Sik; Crane, Julian

    2004-01-01

    Pillows are known to contain significant levels of indoor allergens and endotoxin, that are of importance to house dust mite sensitized asthmatics. Buckwheat pillows are commonly used in Korea. We studied the levels of the house dust mite allergen, Der f 1, and endotoxin on new synthetic and buckwheat pillows and their accumulation over three months. Endotoxin levels were significantly higher on new buckwheat pillows compared to synthetic pillows; geometric mean levels (95% CI) were 60,950 EU/g (30,270-122,700) and 4,887 EU/g (2,570-9,311) respectively (p<0.001). No Der f 1 was detected on the new pillows. After three months Der f 1 levels were similar on buckwheat and synthetic pillows, geometric mean levels (95% CI) were 1.16 µg/g (0.02-8.13) and 1.08 µg/g (0.19-1.68) respectively. These results indicate that buckwheat pillows are a source of very high endotoxin levels that may be of relevance to asthma severity of atopic asthmatics. PMID:15308838

  7. Parvalbumin--the major tropical fish allergen.

    PubMed

    Lim, Dawn Li-Chern; Neo, Keng Hwee; Yi, Fong Cheng; Chua, Kaw Yan; Goh, Denise Li-Meng; Shek, Lynette Pei-Chi; Giam, Yoke Chin; Van Bever, Hugo P S; Lee, Bee Wah

    2008-08-01

    Fish allergy is common in countries where consumption is high. Asian nations are amongst the world's largest consumers of fish but the allergen profiles of tropical fish are unknown. This study sought to evaluate the allergenicity of four commonly consumed tropical fish, the threadfin (Polynemus indicus), Indian anchovy (Stolephorus indicus), pomfret (Pampus chinensis) and tengirri (Scomberomorus guttatus). Immunoglobulin E (IgE) cross-reactivity with parvalbumin of cod fish (Gad c 1), the major fish allergen, was also studied. Detection of tropical fish and cod specific-IgE was performed by UniCap assay, and skin prick tests were also carried out. The IgE-binding components of tropical fish were identified using IgE immunoblot techniques, and cross-reactivity with Gad c 1 was assessed by ELISA inhibition and IgE immunoblot inhibition. Clinically, nine of 10 patients studied were allergic to multiple fish. All patients exhibited detectable specific-IgE to cod fish (10 of 10 skin prick test positive, eight of 10 UniCap assay positive) despite lack of previous exposure. The major allergen of the four tropical fish was the 12-kDa parvalbumin. IgE cross-reactivity of these allergens to Gad c 1 was observed to be moderate to high in the tropical fish studied. Parvalbumins are the major allergens in commonly consumed tropical fish. They are cross-reactive with each other as well as with Gad c 1. Commercial tests for cod fish appear to be sufficient for the detection of tropical fish specific-IgE.

  8. Fish Allergens at a Glance: Variable Allergenicity of Parvalbumins, the Major Fish Allergens

    PubMed Central

    Kuehn, Annette; Swoboda, Ines; Arumugam, Karthik; Hilger, Christiane; Hentges, François

    2014-01-01

    Fish is a common trigger of severe, food-allergic reactions. Only a limited number of proteins induce specific IgE-mediated immune reactions. The major fish allergens are the parvalbumins. They are members of the calcium-binding EF-hand protein family characterized by a conserved protein structure. They represent highly cross-reactive allergens for patients with specific IgE to conserved epitopes. These patients might experience clinical reactions with various fish species. On the other hand, some individuals have IgE antibodies directed against unique, species-specific parvalbumin epitopes, and these patients show clinical symptoms only with certain fish species. Furthermore, different parvalbumin isoforms and isoallergens are present in the same fish and might display variable allergenicity. This was shown for salmon homologs, where only a single parvalbumin (beta-1) isoform was identified as allergen in specific patients. In addition to the parvalbumins, several other fish proteins, enolases, aldolases, and fish gelatin, seem to be important allergens. New clinical and molecular insights advanced the knowledge and understanding of fish allergy in the last years. These findings were useful for the advancement of the IgE-based diagnosis and also for the management of fish allergies consisting of advice and treatment of fish-allergic patients. PMID:24795722

  9. IgE-binding epitopic peptide mapping on a three-dimensional model built for the 13S globulin allergen of buckwheat (Fagopyrum esculentum).

    PubMed

    Sordet, Camille; Culerrier, Raphaël; Granier, Claude; Didier, Alain; Rougé, Pierre

    2009-06-01

    The three-dimensional model built for the 13S globulin allergen of buckwheat (Fagopyrum esculentum) consists of three protomers exhibiting the cupin motif, arranged in a homotrimer around a three-fold symmetry axis. Using the SPOT technique, 11 continuous IgE-binding epitopic peptides were characterized on the molecular surface of the 13S globulin allergen of buckwheat. Except for one of them, they all correspond to well exposed regions containing electropositiveley and/or electronegatively charged residues, which cover up to 40% of the molecular surface of the allergen. Some of these epitopes come in close contact to probably create more extended discontinuous epitopes, especially those located on the edge of the 13S globulin homotrimer. Half of the identified epitope peptides remain unaltered in a core structure protected against hydrolysis by digestive proteases and are thus assumed to promote the allergenicity of the 13S globulin. In addition, a few of these epitopes coincide with sequential IgE-binding epitopes previously characterized in soybean 11S globulins, that could account for the IgE-binding cross-reactions observed between soybean and buckwheat in Western blot experiments.

  10. Comparative and Evolutionary Analysis of Major Peanut Allergen Gene Families

    PubMed Central

    Ratnaparkhe, Milind B.; Lee, Tae-Ho; Tan, Xu; Wang, Xiyin; Li, Jingping; Kim, Changsoo; Rainville, Lisa K.; Lemke, Cornelia; Compton, Rosana O.; Robertson, Jon; Gallo, Maria; Bertioli, David J.; Paterson, Andrew H.

    2014-01-01

    Peanut (Arachis hypogaea L.) causes one of the most serious food allergies. Peanut seed proteins, Arah1, Arah2, and Arah3, are considered to be among the most important peanut allergens. To gain insights into genome organization and evolution of allergen-encoding genes, approximately 617 kb from the genome of cultivated peanut and 215 kb from a wild relative were sequenced including three Arah1, one Arah2, eight Arah3, and two Arah6 gene family members. To assign polarity to differences between homoeologous regions in peanut, we used as outgroups the single orthologous regions in Medicago, Lotus, common bean, chickpea, and pigeonpea, which diverged from peanut about 50 Ma and have not undergone subsequent polyploidy. These regions were also compared with orthologs in many additional dicot plant species to help clarify the timing of evolutionary events. The lack of conservation of allergenic epitopes between species, and the fact that many different proteins can be allergenic, makes the identification of allergens across species by comparative studies difficult. The peanut allergen genes are interspersed with low-copy genes and transposable elements. Phylogenetic analyses revealed lineage-specific expansion and loss of low-copy genes between species and homoeologs. Arah1 syntenic regions are conserved in soybean, pigeonpea, tomato, grape, Lotus, and Arabidopsis, whereas Arah3 syntenic regions show genome rearrangements. We infer that tandem and segmental duplications led to the establishment of the Arah3 gene family. Our analysis indicates differences in conserved motifs in allergen proteins and in the promoter regions of the allergen-encoding genes. Phylogenetic analysis and genomic organization studies provide new insights into the evolution of the major peanut allergen-encoding genes. PMID:25193311

  11. Comparative and evolutionary analysis of major peanut allergen gene families.

    PubMed

    Ratnaparkhe, Milind B; Lee, Tae-Ho; Tan, Xu; Wang, Xiyin; Li, Jingping; Kim, Changsoo; Rainville, Lisa K; Lemke, Cornelia; Compton, Rosana O; Robertson, Jon; Gallo, Maria; Bertioli, David J; Paterson, Andrew H

    2014-09-01

    Peanut (Arachis hypogaea L.) causes one of the most serious food allergies. Peanut seed proteins, Arah1, Arah2, and Arah3, are considered to be among the most important peanut allergens. To gain insights into genome organization and evolution of allergen-encoding genes, approximately 617 kb from the genome of cultivated peanut and 215 kb from a wild relative were sequenced including three Arah1, one Arah2, eight Arah3, and two Arah6 gene family members. To assign polarity to differences between homoeologous regions in peanut, we used as outgroups the single orthologous regions in Medicago, Lotus, common bean, chickpea, and pigeonpea, which diverged from peanut about 50 Ma and have not undergone subsequent polyploidy. These regions were also compared with orthologs in many additional dicot plant species to help clarify the timing of evolutionary events. The lack of conservation of allergenic epitopes between species, and the fact that many different proteins can be allergenic, makes the identification of allergens across species by comparative studies difficult. The peanut allergen genes are interspersed with low-copy genes and transposable elements. Phylogenetic analyses revealed lineage-specific expansion and loss of low-copy genes between species and homoeologs. Arah1 syntenic regions are conserved in soybean, pigeonpea, tomato, grape, Lotus, and Arabidopsis, whereas Arah3 syntenic regions show genome rearrangements. We infer that tandem and segmental duplications led to the establishment of the Arah3 gene family. Our analysis indicates differences in conserved motifs in allergen proteins and in the promoter regions of the allergen-encoding genes. Phylogenetic analysis and genomic organization studies provide new insights into the evolution of the major peanut allergen-encoding genes. PMID:25193311

  12. Comparative and evolutionary analysis of major peanut allergen gene families.

    PubMed

    Ratnaparkhe, Milind B; Lee, Tae-Ho; Tan, Xu; Wang, Xiyin; Li, Jingping; Kim, Changsoo; Rainville, Lisa K; Lemke, Cornelia; Compton, Rosana O; Robertson, Jon; Gallo, Maria; Bertioli, David J; Paterson, Andrew H

    2014-09-04

    Peanut (Arachis hypogaea L.) causes one of the most serious food allergies. Peanut seed proteins, Arah1, Arah2, and Arah3, are considered to be among the most important peanut allergens. To gain insights into genome organization and evolution of allergen-encoding genes, approximately 617 kb from the genome of cultivated peanut and 215 kb from a wild relative were sequenced including three Arah1, one Arah2, eight Arah3, and two Arah6 gene family members. To assign polarity to differences between homoeologous regions in peanut, we used as outgroups the single orthologous regions in Medicago, Lotus, common bean, chickpea, and pigeonpea, which diverged from peanut about 50 Ma and have not undergone subsequent polyploidy. These regions were also compared with orthologs in many additional dicot plant species to help clarify the timing of evolutionary events. The lack of conservation of allergenic epitopes between species, and the fact that many different proteins can be allergenic, makes the identification of allergens across species by comparative studies difficult. The peanut allergen genes are interspersed with low-copy genes and transposable elements. Phylogenetic analyses revealed lineage-specific expansion and loss of low-copy genes between species and homoeologs. Arah1 syntenic regions are conserved in soybean, pigeonpea, tomato, grape, Lotus, and Arabidopsis, whereas Arah3 syntenic regions show genome rearrangements. We infer that tandem and segmental duplications led to the establishment of the Arah3 gene family. Our analysis indicates differences in conserved motifs in allergen proteins and in the promoter regions of the allergen-encoding genes. Phylogenetic analysis and genomic organization studies provide new insights into the evolution of the major peanut allergen-encoding genes.

  13. Identification of the major allergens of Indian scad (Decapterus russelli).

    PubMed

    Misnan, Rosmilah; Murad, Shahnaz; Jones, Meinir; Taylor, Graham; Rahman, Dinah; Arip, Masita; Abdullah, Noormalin; Mohamed, Jamaluddin

    2008-12-01

    The purpose of this study was to characterize major allergens of Indian scad (Decapterus russelli) which is among the most commonly consumed fish in Malaysia. Raw and cooked extracts of the fish were prepared. Protein profiles and IgE binding patterns were produced by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting using sera from subjects with fish allergy. The major allergens of the fish were then identified by two-dimensional electrophoresis (2-DE), followed by mass spectrometry of the peptide digests. The SDS-PAGE of the raw extract revealed 27 protein fractions over a wide molecular weight range, while the cooked extract demonstrated only six protein fractions. The 1-DE immunoblotting detected 14 IgE-binding proteins, with a molecular weight range from 90 to < 6.5 kDa. Three protein fractions with molecular weights of approximately 51, 46 and 12 kDa were identified as the major allergens of this fish. The approximately 12 kDa band was a heat-resistant protein while the approximately 51 and 46 kDa proteins were sensitive to heat. The 2-DE gel profile of the raw extract demonstrated > 100 distinct protein spots and immunoblotting detected at least 10 different major IgE reactive spots with molecular masses as expected and isoelectric point (pI) values ranging from 4.0 to 7.0. A comparison of the major allergenic spot sequences of the 12 kDa proteins with known protein sequences in databases revealed extensive similarity with fish parvalbumin. In conclusion, this study demonstrated that a parvalbumin which is similar to Gad c 1 is the major allergen of Indian scad. Interestingly, we also detected heat-sensitive proteins as major allergenic components in our fish allergy patients. PMID:19317337

  14. Pine nut allergy: clinical features and major allergens characterization

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pine nuts, the seeds of pine trees, are widely used for human consumption in Europe, America, and Asia. The aims of this study were to evaluate IgE-mediated hypersensitivity to pine nut in a large number of patients with details of clinical reactions, and to characterize major pine nut allergens. Th...

  15. Characterization of major allergens of royal jelly Apis mellifera.

    PubMed

    Rosmilah, M; Shahnaz, M; Patel, G; Lock, J; Rahman, D; Masita, A; Noormalin, A

    2008-12-01

    Royal jelly is widely consumed in the community and has perceived benefits ranging from promoting growth in children and improvement of general health status to enhancement of longevity for the elderly. However, royal jelly consumption has been linked to contact dermatitis, acute asthma, anaphylaxis and death. High prevalence of positive skin tests to royal jelly have been reported among atopic populations in countries with a high rate of royal jelly consumption. The present study is aimed to identify the major allergens of royal jelly. Royal jelly extract was separated by sodium dodecyl polyacrylamide gel electrophoresis (SDS-PAGE) and 2-dimensional electrophoresis (2-D). Immunoblotting of the SDS-PAGE and 2-D profiles were performed to identify the allergenic spots. Spots were then excised from the 2-D gel, digested with trypsin and analyzed by mass spectrometry. The SDS-PAGE of royal jelly extract revealed 18 bands between 10 to 167 kD. Western blot of the fractionated proteins detected 15 IgE-binding bands between 14 to 127 kD with seven major allergens of 32, 40, 42, 49, 55, 60 and 67 kD using serum from 53 subjects with royal jelly allergy. The 2-D gel fractionated the royal jelly proteins to more than 50 different protein spots. Out of these, 30 spots demonstrated specific IgE affinity to the sera tested. Eight spots of the major royal jelly allergens were selected for mass-spectrometry analysis. Digested tryptic peptides of the spots were compared to the amino acid sequence search in protein databases which identified the fragments of royal jelly homologus to major royal jelly protein 1 (MRJ1) and major royal jelly protein 2 (MRJ2). In conclusion, the major allergens of royal jelly are MRJ1 and MRJ2 in our patients' population.

  16. Reduction of the Number of Major Representative Allergens: From Clinical Testing to 3-Dimensional Structures

    PubMed Central

    He, Ying; Liu, Xueting; Huang, Yuyi; Zou, Zehong; Chen, Huifang; Lai, He; Wu, Qiurong; Zhang, Junyan; Wang, Shan; Zhang, Jianguo; Tao, Ailin; Sun, Baoqing

    2014-01-01

    Vast amounts of allergen sequence data have been accumulated, thus complicating the identification of specific allergenic proteins when performing diagnostic allergy tests and immunotherapy. This study aims to rank the importance/potency of the allergens so as to logically reduce the number of allergens and/or allergenic sources. Meta-analysis of 62 allergenic sources used for intradermal testing on 3,335 allergic patients demonstrated that in southern China, mite, sesame, spiny amaranth, Pseudomonas aeruginosa, and house dust account for 88.0% to 100% of the observed positive reactions to the 62 types of allergenic sources tested. The Kolmogorov-Smironov Test results of the website-obtained allergen data and allergen family featured peptides suggested that allergen research in laboratories worldwide has been conducted in parallel on many of the same species. The major allergens were reduced to 21 representative allergens, which were further divided into seven structural classes, each of which contains similar structural components. This study therefore has condensed numerous allergenic sources and major allergens into fewer major representative ones, thus allowing for the use of a smaller number of allergens when conducting comprehensive allergen testing and immunotherapy treatments. PMID:24778467

  17. Wild buckwheat is unlikely to pose a risk to buckwheat-allergic individuals.

    PubMed

    Nordlee, Julie A; Panda, Rakhi; Baumert, Joseph L; Goodman, Richard E; Taylor, Steve L

    2011-10-01

    Buckwheat (Fagopyrum esculentum) is a commonly allergenic food especially in Asia where buckwheat is more commonly consumed. Wild buckwheat (Polygonum convolvulus, recently changed to Fallopia convolvulus) is an annual weed prevalent in grain-growing areas of the United States. Wild buckwheat is not closely related to edible buckwheat although the seeds do have some physical resemblance. A large shipment of wheat into Japan was halted by the discovery of the adventitious presence of wild buckwheat seeds over possible concerns for buckwheat-allergic consumers. However, IgE-binding was not observed to an extract of wild buckwheat using sera from 3 buckwheat-allergic individuals either by radio-allergosorbent test inhibition or by immunoblotting after protein separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Furthermore, the extract of wild buckwheat was not detected in a buckwheat enzyme-linked immunosorbent assay developed with antisera against common buckwheat. Thus, wild buckwheat is highly unlikely to pose any risk to buckwheat-allergic individuals. The common names of plants should not be a factor in the risk assessment for possible cross-allergenicity.

  18. Crystal structure of the major peanut allergen Ara h 1.

    PubMed

    Cabanos, Cerrone; Urabe, Hiroyuki; Tandang-Silvas, Mary Rose; Utsumi, Shigeru; Mikami, Bunzo; Maruyama, Nobuyuki

    2011-10-01

    Ara h 1, a 7S globulin, is one of the three major peanut allergens. We previously reported the crystallization of the core region of recombinant Ara h 1. Here, we present the crystal structure of the Ara h 1 core at a resolution of 2.43 Å. We also assayed the Ara h 1 core thermal stability and compared its final structure against other 7S globulins. The Ara h 1 core has a thermal denaturation temperature of 88.3°C and a structure that is very similar to other 7S globulins. Previously identified linear IgE epitopes were also mapped on the three-dimensional structure. Most linear epitopes were found in the extended loop domains and the coils between the N- and C-terminal modules, while others were found in the less accessible β-sheets of the C-terminal core β-barrel domain of each monomer. Most of these epitopes become either slightly or significantly buried upon trimer formation, implying that allergen digestion in the gut is required for these epitopes to be accessible to immunoglobulins. Our findings also suggest that both intact and partially degraded allergens should be employed in future diagnostic and immunotherapeutic strategies. PMID:21903274

  19. PCR method for detecting trace amounts of buckwheat (Fagopyrum spp.) in food.

    PubMed

    Hirao, Takashi; Imai, Shinsuke; Sawada, Hiroshi; Shiomi, Nobuo; Hachimura, Satoshi; Kato, Hisanori

    2005-04-01

    Buckwheat often causes severe allergic reactions, even when its ingestion level is extremely low. Therefore, buckwheat is listed in several countries as a common food allergen. In addition to common buckwheat and Tartarian buckwheat that are cultivated and consumed widely, wild buckwheat may be potentially allergenic. Food containing undeclared buckwheat poses a risk to patients with the buckwheat allergy. We describe in this report a PCR method to detect buckwheat DNA by using primers corresponding to the internal transcribed spacer region and the 5.8S rRNA gene. The method is buckwheat-specific and compatible with both cultivated and wild buckwheat of the Fagopyrum spp. Its sensitivity was sufficient to detect 1 ppm (w/w) of buckwheat DNA spiked in wheat DNA. This method should benefit food manufacturers, clinical doctors, and allergic patients by providing information on the presence of buckwheat contamination in food.

  20. Allergy vaccines: a need for standardisation in mass units of major allergen.

    PubMed

    van Ree, R; Dorpema, J W; Vieths, S

    2005-09-01

    Treatment of respiratory allergies can be performed with allergen-specific immunotherapy using allergen extracts. These products are biologicals with an extremely complex and variable composition. Only a few components are of major importance for the disease, the so-called major allergens. At present, standardisation of allergen extracts is dominated by techniques that aim at establishing their overall IgE-binding potencies using pooled sera of allergic patients. Each company in the market uses its own type of units to express potencies, thus hampering comparability. Another disadvantage is that the major allergen composition is not determined. Most companies have introduced assays for the measurement of major allergens in their quality control systems, but these data are not yet used for labelling purposes. The need to include major allergen content in standardisation protocols is now widely accepted. To support future labelling on the basis of major allergen content the European Union has funded the multidisciplinary multicentre project CREATE. This project aims at developing international certified references for the most important major respiratory allergens and at evaluating the performance of available ELISA for their measurement. The project will facilitate expression of potencies by active ingredient (major allergen) content and will allow direct comparison of competitor products.

  1. Improvement of Surface Functionalities, Including Allergenicity Attenuation, of Whole Buckwheat Protein Fraction by Maillard-Type Glycation with Dextran

    PubMed Central

    Tazawa, Shigeru; Katayama, Shigeru; Hirabayashi, Masahiro; Yamaguchi, Daiki; Nakamura, Soichiro

    2014-01-01

    The purpose of the current study was to determine the effects of the introduction of polysaccharide chains onto the molecular surface of buckwheat proteins on buckwheat protein surface functionality. The whole buckwheat protein fraction (WBP) was prepared using 50 mM phosphate buffer (pH 7.5) containing 0.5 M NaCl and covalently linked with 6 kDa, 17.5 kDa, 40 kDa, 70 kDa, or 200 kDa dextran by Maillard-type glycation through controlled dry-heating at 60°C and 79% relative humidity for two weeks. Conjugation with 40 kDa dextran improved the water solubility and emulsifying properties of WBP without causing a serious loss of available lysine; 84.9% of the free amino groups were conserved. In addition, we found that the introduction of dextran chains onto the molecular surfaces of WBP attenuated the antigenicity of WBP. PMID:25580398

  2. Molecular and immunological characterization of the first allergenic lipocalin in hamster: the major allergen from Siberian hamster (Phodopus sungorus).

    PubMed

    Torres, José Alberto; de Las Heras, Manuel; Maroto, Aroa Sanz; Vivanco, Fernando; Sastre, Joaquín; Pastor-Vargas, Carlos

    2014-08-22

    The most frequent pet allergy is to cat and dog, but in recent years, it has become increasingly popular to have other pets, and the risk of exposure to new allergens is more prevalent. The list of new pets includes hamsters, and one of the most popular hamsters is the Siberian hamster (Phodopus sungorus). The aim of this study was the characterization and cloning of the major allergen from this hamster. The study of its allergenicity and cross-reactivity could improve the specific diagnosis and treatment for hamster-allergic patients. Thirteen Siberian hamster-allergic patients were recruited at the outpatient clinic. Protein extracts were prepared from the hair, urine, and salivary glands of four hamster species (European, golden, Siberian, and Roborovski). IgE-binding proteins were detected by immunoblotting and identified by mass spectrometry. The recombinant protein was produced in Escherichia coli and then purified by metal chelate affinity chromatography. The allergenic properties of the recombinant protein were tested by ELISA and immunoblotting, and biological activity was tested according to capacity for basophil activation. Three IgE-binding proteins were identified in extracts obtained from Siberian hamster hair, urine, and salivary glands. All proteins corresponded to the same protein, which was identified as a lipocalin. This lipocalin had no cross-reactivity with common and golden hamsters. The recombinant allergen was cloned and purified, showing similar IgE reactivity in vitro to Siberian hamster protein extracts. Also, the recombinant allergen was capable of producing biological activation in vivo. The major Siberian hamster allergen was cloned, and allergenic properties were characterized, providing a new tool for specific diagnosis of allergy to Siberian hamster.

  3. Haemoglobin, a new major allergen of Anisakis simplex.

    PubMed

    González-Fernández, Juan; Daschner, Alvaro; Nieuwenhuizen, Natalie E; Lopata, Andreas L; Frutos, Consolación De; Valls, Ana; Cuéllar, Carmen

    2015-05-01

    Gastro-allergic anisakiasis and Anisakis sensitisation associated chronic urticaria are diseases which differ in their IgE and IgG4 responses against both crude extract and specific allergens. Anisakis and Ascaris are closely related nematodes that usually cause problems with specificity in immunodiagnostics. In this study we measured IgE and IgG4 antibodies against Anisakis simplex sensu lato (s. l.) and Ascaris suum haemoglobins in sera of 21 gastro-allergic anisakiasis and 23 chronic urticaria patients. We used a capture ELISA with the anti-Anisakis haemoglobin monoclonal antibody 4E8g, which also recognises Ascaris haemoglobin. In addition, we determined specific IgE and IgG4 to both nematodes by indirect ELISA and immunoblotting. Anti-A. simplex s. l. haemoglobin IgE and IgG4 levels were higher in gastro-allergic anisakiasis than in chronic urticaria patients (P=0.002 and 0.026, respectively). Surprisingly, no patient had detectable IgE levels against A. suum haemoglobin. Finally, we carried out an in silico study of the B-cell epitopes of both haemoglobin molecules. Five epitopes were predicted in Anisakis pegreffii and four in A. suum haemoglobin. The epitope propensity values of Anisakis haemoglobin in the equivalent IgE binding region of the allergenic haemoglobin Chi t 1 from Chironomus thummi, were higher those of the Ascaris haemoglobin. In conclusion, we describe A. simplex haemoglobin as a new major allergen (Ani s 13), being recognised by a large number (64.3%) of sensitised patients and up to 80.9% in patients with gastro-allergic anisakiasis. The presence of a specific epitope and the different values of epitope propensity between Anisakis and Ascaris haemoglobin could explain the lack of cross-reactivity between the two molecules. The absence of IgE reactivity to Ascaris haemoglobin in Anisakis patients makes Anisakis haemoglobin (Ani s 13) a potential candidate for developing more specific diagnosis tools. PMID:25683373

  4. 27 CFR 4.32b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... from major food allergen labeling. 4.32b Section 4.32b Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF WINE Labeling Requirements for Wine § 4.32b Petitions for exemption from major food allergen labeling....

  5. 27 CFR 4.32b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... from major food allergen labeling. 4.32b Section 4.32b Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF WINE Labeling Requirements for Wine § 4.32b Petitions for exemption from major food allergen labeling....

  6. 27 CFR 4.32b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... from major food allergen labeling. 4.32b Section 4.32b Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY ALCOHOL LABELING AND ADVERTISING OF WINE Labeling Requirements for Wine § 4.32b Petitions for exemption from major food allergen labeling....

  7. 27 CFR 4.32b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... from major food allergen labeling. 4.32b Section 4.32b Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF WINE Labeling Requirements for Wine § 4.32b Petitions for exemption from major food allergen labeling....

  8. 27 CFR 4.32b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... from major food allergen labeling. 4.32b Section 4.32b Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY ALCOHOL LABELING AND ADVERTISING OF WINE Labeling Requirements for Wine § 4.32b Petitions for exemption from major food allergen labeling....

  9. 27 CFR 5.32b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... from major food allergen labeling. 5.32b Section 5.32b Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF... food allergen labeling. (a) Submission of petition. Any person may petition the appropriate TTB...

  10. 27 CFR 5.32b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... from major food allergen labeling. 5.32b Section 5.32b Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY ALCOHOL LABELING AND ADVERTISING OF... food allergen labeling. (a) Submission of petition. Any person may petition the appropriate TTB...

  11. 27 CFR 5.32b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... from major food allergen labeling. 5.32b Section 5.32b Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY ALCOHOL LABELING AND ADVERTISING OF... food allergen labeling. (a) Submission of petition. Any person may petition the appropriate TTB...

  12. 27 CFR 5.32b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... from major food allergen labeling. 5.32b Section 5.32b Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF... food allergen labeling. (a) Submission of petition. Any person may petition the appropriate TTB...

  13. 27 CFR 5.32b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... from major food allergen labeling. 5.32b Section 5.32b Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF... food allergen labeling. (a) Submission of petition. Any person may petition the appropriate TTB...

  14. Genetic engineering of the major timothy grass pollen allergen, Phl p 6, to reduce allergenic activity and preserve immunogenicity.

    PubMed

    Vrtala, Susanne; Focke, Margarete; Kopec, Jolanta; Verdino, Petra; Hartl, Arnulf; Sperr, Wolfgang R; Fedorov, Alexander A; Ball, Tanja; Almo, Steve; Valent, Peter; Thalhamer, Josef; Keller, Walter; Valenta, Rudolf

    2007-08-01

    On the basis of IgE epitope mapping data, we have produced three allergen fragments comprising aa 1-33, 1-57, and 31-110 of the major timothy grass pollen allergen Phl p 6 aa 1-110 by expression in Escherichia coli and chemical synthesis. Circular dichroism analysis showed that the purified fragments lack the typical alpha-helical fold of the complete allergen. Superposition of the sequences of the fragments onto the three-dimensional allergen structure indicated that the removal of only one of the four helices had led to the destabilization of the alpha helical structure of Phl p 6. The lack of structural fold was accompanied by a strong reduction of IgE reactivity and allergenic activity of the three fragments as determined by basophil histamine release in allergic patients. Each of the three Phl p 6 fragments adsorbed to CFA induced Phl p 6-specific IgG Abs in rabbits. However, immunization of mice with fragments adsorbed to an adjuvant allowed for human use (AluGel-S) showed that only the Phl p 6 aa 31-110 induced Phl p 6-specific IgG Abs. Anti-Phl p 6 IgG Abs induced by vaccination with Phl p 6 aa 31-110 inhibited patients' IgE reactivity to the wild-type allergen as well as Phl p 6-induced basophil degranulation. Our results are of importance for the design of hypoallergenic allergy vaccines. They show that it has to be demonstrated that the hypoallergenic derivative induces a robust IgG response in a formulation that can be used in allergic patients.

  15. Identification of tropomyosin and arginine kinase as major allergens of Portunus pelagicus (blue swimming crab).

    PubMed

    Rosmilah, M; Shahnaz, M; Zailatul, H M Y; Noormalin, A; Normilah, I

    2012-09-01

    Crab is an important source of food allergen. Tropomyosin represents the main crab allergen and is responsible for IgE cross-reactivity between various species of crustaceans. Recently, other new crab allergens including arginine kinase have been identified. However, information on allergens of the local Portunidcrab is not available. Thus, the aim of this study was to identify the major allergens of Portunus pelagicus (blue swimming crab) using the allergenomics approach. Raw and cooked extracts of the crab were prepared from the crab meat. Protein profile and IgE binding pattern were demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting using sera from 30 patients with crab allergy. The major allergens of the crab were then identified by two-dimensional electrophoresis (2-DE), followed by mass spectrometry analysis of the peptide digests. The SDS-PAGE of raw extract revealed approximately 20 protein fractions over a wide molecular weight range, while cooked extract demonstrated fewer protein bands. The raw extract also demonstrated a higher number of IgE reactive bands than the cooked extract. A heat-resistant protein of 36 kDa has been identified as the major allergen in both raw and cooked extracts. In addition, a heat-sensitive protein of 41 kDa was also recognized as a major allergen in raw crab. The 2-DE gel profile of the raw extract demonstrated about >100 distinct proteins spots and immunoblotting of the 2-DE profile demonstrated at least 12 different major IgE reactive spots with molecular masses between 13 to 250 kDa and isoelectric point (pI) values ranging from 4.0 to 7.0. The 36 and 41 kDa proteins were identified as the crab tropomyosin and arginine kinase, respectively by mass spectrometry. Therefore, this study confirmed that tropomyosin and arginine kinase are the major allergens of the local Portunid crab, P. pelagicus.

  16. Maillard reaction and enzymatic browning affect the allergenicity of Pru av 1, the major allergen from cherry (Prunus avium).

    PubMed

    Gruber, Patrick; Vieths, Stefan; Wangorsch, Andrea; Nerkamp, Jörg; Hofmann, Thomas

    2004-06-16

    The influence of thermal processing and nonenymatic as well as polyphenoloxidase-catalyzed browning reaction on the allergenicity of the major cherry allergen Pru av 1 was investigated. After thermal treatment of the recombinant protein rPru av 1 in the absence or presence of carbohydrates, SDS-PAGE, enzyme allergosorbent tests, and inhibition assays revealed that thermal treatment of rPru av 1 alone did not show any influence on the IgE-binding activity of the protein at least for 30 min, thus correlating well with the refolding of the allergen in buffer solution as demonstrated by CD spectroscopic experiments. Incubation of the protein with starch and maltose also showed no effect on IgE-binding activity, whereas reaction with glucose and ribose and, even more pronounced, with the carbohydrate breakdown products glyceraldehyde and glyoxal induced a strong decrease of the IgE-binding capacity of rPru av 1. In the second part of the study, the effect of polyphenoloxidase-catalyzed oxidation of polyphenols on food allergen activity was investigated. Incubation of rPru av 1 with epicatechin in the presence of tyrosinase led to a drastic decrease in IgE-binding activity of the protein. Variations of the phenolic compound revealed caffeic acid and epicatechin as the most active inhibitors of the IgE-binding activity of rPru av 1, followed by catechin and gallic acid, and, finally, by quercetin and rutin, showing significantly lower activity. On the basis of these data, reactive intermediates formed during thermal carbohydrate degradation as well as during enzymatic polyphenol oxidation are suggested as the active chemical species responsible for modifying nucleophilic amino acid side chains of proteins, thus inducing an irreversible change in the tertiary structure of the protein and resulting in a loss of conformational epitopes of the allergen.

  17. Maillard reaction and enzymatic browning affect the allergenicity of Pru av 1, the major allergen from cherry (Prunus avium).

    PubMed

    Gruber, Patrick; Vieths, Stefan; Wangorsch, Andrea; Nerkamp, Jörg; Hofmann, Thomas

    2004-06-16

    The influence of thermal processing and nonenymatic as well as polyphenoloxidase-catalyzed browning reaction on the allergenicity of the major cherry allergen Pru av 1 was investigated. After thermal treatment of the recombinant protein rPru av 1 in the absence or presence of carbohydrates, SDS-PAGE, enzyme allergosorbent tests, and inhibition assays revealed that thermal treatment of rPru av 1 alone did not show any influence on the IgE-binding activity of the protein at least for 30 min, thus correlating well with the refolding of the allergen in buffer solution as demonstrated by CD spectroscopic experiments. Incubation of the protein with starch and maltose also showed no effect on IgE-binding activity, whereas reaction with glucose and ribose and, even more pronounced, with the carbohydrate breakdown products glyceraldehyde and glyoxal induced a strong decrease of the IgE-binding capacity of rPru av 1. In the second part of the study, the effect of polyphenoloxidase-catalyzed oxidation of polyphenols on food allergen activity was investigated. Incubation of rPru av 1 with epicatechin in the presence of tyrosinase led to a drastic decrease in IgE-binding activity of the protein. Variations of the phenolic compound revealed caffeic acid and epicatechin as the most active inhibitors of the IgE-binding activity of rPru av 1, followed by catechin and gallic acid, and, finally, by quercetin and rutin, showing significantly lower activity. On the basis of these data, reactive intermediates formed during thermal carbohydrate degradation as well as during enzymatic polyphenol oxidation are suggested as the active chemical species responsible for modifying nucleophilic amino acid side chains of proteins, thus inducing an irreversible change in the tertiary structure of the protein and resulting in a loss of conformational epitopes of the allergen. PMID:15186129

  18. Identification and characterization of major cat allergen Fel d 1 mimotopes on filamentous phage carriers.

    PubMed

    Luzar, Jernej; Molek, Peter; Šilar, Mira; Korošec, Peter; Košnik, Mitja; Štrukelj, Borut; Lunder, Mojca

    2016-03-01

    Cat allergy is one of the most prevalent allergies worldwide and can lead to the development of rhinitis and asthma. Thus far, only allergen extracts from natural sources have been used for allergen-specific immunotherapy. However, extracts and whole allergens in immunotherapy present an anaphylaxis risk. Identification of allergen epitopes or mimotopes has an important role in development of safe and effective allergen-specific immunotherapy. Moreover, with a suitable immunogenic carrier, the absence of sufficient immune response elicited by short peptides could be surmounted. In this study, we identified five structural mimotopes of the major cat allergen Fel d 1 by immunoscreening with random peptide phage libraries. The mimotopes were computationally mapped to the allergen surface, and their IgE reactivity was confirmed using sera from cat-allergic patients. Importantly, the mimotopes showed no basophil activation of the corresponding cat-allergic patients, which makes them good candidates for the development of hypoallergenic vaccine. As bacteriophage particles are becoming increasingly recognized as immunogenic carriers, we constructed bacteriophage particles displaying multiple copies of each selected mimotope on major phage coat protein. These constructed phages elicited T cell-mediated immune response, which was predominated by the type 1 T cell response. Mimotopes alone contributed to the type 1 T cell response by promoting IL-2 production. Fel d 1 mimotopes, as well as their filamentous phage immunogenic carriers, represent promising candidates in the development of hypoallergenic vaccine against cat allergy.

  19. Identification and characterization of major cat allergen Fel d 1 mimotopes on filamentous phage carriers.

    PubMed

    Luzar, Jernej; Molek, Peter; Šilar, Mira; Korošec, Peter; Košnik, Mitja; Štrukelj, Borut; Lunder, Mojca

    2016-03-01

    Cat allergy is one of the most prevalent allergies worldwide and can lead to the development of rhinitis and asthma. Thus far, only allergen extracts from natural sources have been used for allergen-specific immunotherapy. However, extracts and whole allergens in immunotherapy present an anaphylaxis risk. Identification of allergen epitopes or mimotopes has an important role in development of safe and effective allergen-specific immunotherapy. Moreover, with a suitable immunogenic carrier, the absence of sufficient immune response elicited by short peptides could be surmounted. In this study, we identified five structural mimotopes of the major cat allergen Fel d 1 by immunoscreening with random peptide phage libraries. The mimotopes were computationally mapped to the allergen surface, and their IgE reactivity was confirmed using sera from cat-allergic patients. Importantly, the mimotopes showed no basophil activation of the corresponding cat-allergic patients, which makes them good candidates for the development of hypoallergenic vaccine. As bacteriophage particles are becoming increasingly recognized as immunogenic carriers, we constructed bacteriophage particles displaying multiple copies of each selected mimotope on major phage coat protein. These constructed phages elicited T cell-mediated immune response, which was predominated by the type 1 T cell response. Mimotopes alone contributed to the type 1 T cell response by promoting IL-2 production. Fel d 1 mimotopes, as well as their filamentous phage immunogenic carriers, represent promising candidates in the development of hypoallergenic vaccine against cat allergy. PMID:26908079

  20. Orthologous Allergens and Diagnostic Utility of Major Allergen Alt a 1

    PubMed Central

    Moreno, Antonio; Alcover, Javier; Rodríguez, David; Palacios, Ricardo; Martínez-Naves, Eduardo

    2016-01-01

    Purpose Hypersensitivity to fungi is associated with rhinoconjunctivitis and asthma. For some fungi, such as Alternaria alternata (A. alternata), the symptoms of asthma are persistent, increasing disease severity and the risk of fatal outcomes. There are a large number of species of fungi but knowledge of them remains limited. This, together with the difficulties in obtaining adequate standardized extracts, means that there remain significant challenges in the diagnosis and immunotherapy of allergy associated with fungi. The type of indoor fungi related to asthma/allergy varies according to geographic, climatic, and seasonal factors, making their study difficult. The aim of this study was to determine hypersensitivity to indoor fungi in a population from Cuenca, Spain. Methods Thirty-five patients with symptoms compatible with rhinitis or asthma who showed clear worsening of their symptoms in their homes or workplace were included. In vivo and in vitro tests were made with a battery of fungal allergens, including the species isolated in the home or workplace. Results Ulocladium botrytis (U. botrytis) and A. alternata were the most representative species as a source of home sensitization. These species showed very high concordance in skin tests, specific IgE, and histamine release. The allergen Alt a 1, which was recognized in all patients, was detected in A. alternata, U. botrytis, and Stemphylium botryosum (S. botryosum). Conclusions U. botrytis and A. alternata were the most representative species as a source of home sensitization. Alt a 1 was recognized in all patients and may be considered a non-species-specific allergen that could be used as a diagnostic source of sensitization to some species of the Pleosporaceae family. PMID:27334781

  1. Glycoproteomic analysis of seven major allergenic proteins reveals novel post-translational modifications.

    PubMed

    Halim, Adnan; Carlsson, Michael C; Madsen, Caroline Benedicte; Brand, Stephanie; Møller, Svenning Rune; Olsen, Carl Erik; Vakhrushev, Sergey Y; Brimnes, Jens; Wurtzen, Peter Adler; Ipsen, Henrik; Petersen, Bent L; Wandall, Hans H

    2015-01-01

    Allergenic proteins such as grass pollen and house dust mite (HDM) proteins are known to trigger hypersensitivity reactions of the immune system, leading to what is commonly known as allergy. Key allergenic proteins including sequence variants have been identified but characterization of their post-translational modifications (PTMs) is still limited. Here, we present a detailed PTM(1) characterization of a series of the main and clinically relevant allergens used in allergy tests and vaccines. We employ Orbitrap-based mass spectrometry with complementary fragmentation techniques (HCD/ETD) for site-specific PTM characterization by bottom-up analysis. In addition, top-down mass spectrometry is utilized for targeted analysis of individual proteins, revealing hitherto unknown PTMs of HDM allergens. We demonstrate the presence of lysine-linked polyhexose glycans and asparagine-linked N-acetylhexosamine glycans on HDM allergens. Moreover, we identified more complex glycan structures than previously reported on the major grass pollen group 1 and 5 allergens, implicating important roles for carbohydrates in allergen recognition and response by the immune system. The new findings are important for understanding basic disease-causing mechanisms at the cellular level, which ultimately may pave the way for instigating novel approaches for targeted desensitization strategies and improved allergy vaccines.

  2. Glycoproteomic Analysis of Seven Major Allergenic Proteins Reveals Novel Post-translational Modifications*

    PubMed Central

    Halim, Adnan; Carlsson, Michael C.; Madsen, Caroline Benedicte; Brand, Stephanie; Møller, Svenning Rune; Olsen, Carl Erik; Vakhrushev, Sergey Y.; Brimnes, Jens; Wurtzen, Peter Adler; Ipsen, Henrik; Petersen, Bent L.; Wandall, Hans H.

    2015-01-01

    Allergenic proteins such as grass pollen and house dust mite (HDM) proteins are known to trigger hypersensitivity reactions of the immune system, leading to what is commonly known as allergy. Key allergenic proteins including sequence variants have been identified but characterization of their post-translational modifications (PTMs) is still limited. Here, we present a detailed PTM1 characterization of a series of the main and clinically relevant allergens used in allergy tests and vaccines. We employ Orbitrap-based mass spectrometry with complementary fragmentation techniques (HCD/ETD) for site-specific PTM characterization by bottom-up analysis. In addition, top-down mass spectrometry is utilized for targeted analysis of individual proteins, revealing hitherto unknown PTMs of HDM allergens. We demonstrate the presence of lysine-linked polyhexose glycans and asparagine-linked N-acetylhexosamine glycans on HDM allergens. Moreover, we identified more complex glycan structures than previously reported on the major grass pollen group 1 and 5 allergens, implicating important roles for carbohydrates in allergen recognition and response by the immune system. The new findings are important for understanding basic disease-causing mechanisms at the cellular level, which ultimately may pave the way for instigating novel approaches for targeted desensitization strategies and improved allergy vaccines. PMID:25389185

  3. Purification, characterization and allergenicity assessment of 26kDa protein, a major allergen from Cicer arietinum.

    PubMed

    Verma, Alok Kumar; Sharma, Akanksha; Kumar, Sandeep; Gupta, Rinkesh Kumar; Kumar, Dinesh; Gupta, Kriti; Giridhar, B H; Das, Mukul; Dwivedi, Premendra D

    2016-06-01

    Chickpea (CP), a legume of the family Fabaceae, is an important nutrient-rich food providing protein, essential amino acids, vitamins, dietary fibre, and minerals. Unfortunately, several IgE-binding proteins in CP have been detected that are responsible for allergic manifestations in sensitized population. Therefore, the prevalence of CP induced allergy prompted us towards purification, characterization and allergenicity assessment of a major ∼26kDa protein from chickpea crude protein extract (CP-CPE). Purification of CP 26kDa protein was done using a combination of fractionation and anion exchange chromatography. This protein was further characterized as "Chain A, crystal structure of a plant albumin" from Cicer arietinum with Mol wt 25.8kDa by Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Further, allergenic potential of purified 25.8kDa protein was assessed using in vivo and in vitro model. Purified protein showed IgE-binding capacity with sensitized BALB/c mice and CP allergic patient's sera. Enhanced levels of specific and total IgE, MCP-1, MCPT-1, myeloperoxidase, histamine, prostaglandin D2, and cysteinyl leukotriene were found in sera of mice treated with CP ∼26kDa protein. Further, expressions of Th2 cytokines (i.e. IL-4, IL-5, IL-13), transcription factors (i.e. GATA-3, STAT-6, SOCS-3) and mast cell signaling proteins (Lyn, cFgr, Syk, PLC-γ2, PI-3K, PKC) were also found increased at mRNA and protein levels in the intestines of mice treated with CP ∼26kDa protein. In addition, enhanced release of β-hexosaminidase, histamine, cysteinyl leukotriene and prostaglandin D2 were observed in RBL2H3 cell line when treated (125μg) with CP 26kDa protein. Conclusively, in vivo and in vitro studies revealed the allergenic potential of purified CP 26kDa protein. Being a potential allergen, plant albumin may play a pivotal role in CP induced allergenicity. Current study will be helpful for better development of therapeutic approaches to

  4. IgE cross-reactivity between the major peanut allergen Ara h 2 and the non-homologous allergens Ara h 1 and Ara h 3

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ara h 1, a vicilin, Ara h 2, a 2S albumin, and Ara h 3, a legumin, are major33 peanut allergens. Ara h 2 is an important predictor of clinical reactivity to peanut, but co-sensitization to all three allergens is correlated with the severity of patients’ symptoms. We investigated whether co-sensitiza...

  5. Molecular cloning, characterization, and expression of Cuc m 2, a major allergen in Cucumis melo

    PubMed Central

    Sankian, Mojtaba; Mahmoudi, Mahmoud; Varasteh, Abdol-Reza

    2013-01-01

    Background: Several studies reported the clinical features of IgE-mediated hypersensitivity after ingestion of melon. Melon allergy is a common IgE-mediated fruit allergy in Iran. This prompted us to investigate immunochemical and molecular properties of the major allergen in melon fruit, to compare the IgE-binding capacity of the natural protein with the recombinant allergen, and to determine cross-reactivity of the major allergen with closely-related allergens from other plants displaying clinical cross-reactivity with melon. Methods: Identification and molecular characterization of the major melon allergen were performed using IgE immunoblotting, allergen-specific ELISA, affinity-based purifications, cross-inhibition assays, cloning, and expression of the allergen in Escherichia coli. Results: Melon profilin was identified and isolated as a major IgE-binding component and designated as Cuc m 2. Sequencing corresponding cDNA revealed an open reading frame of 363 bp coding for 131 amino acid residues and two fragments of 171 bp and 383 bps for the 5’and 3’ UTRs, respectively. Significant cross-reactivity was found between melon profilin and Cynodon dactylon, tomato, peach, and grape profilins in cross-inhibition assays. Although the highest degree of amino acid identity was revealed with watermelon profilin, there was no significant cross-reactivity between melon and watermelon profilins. Conclusion: Melon profilin is the major IgE-binding component in melon extract, and the recombinant and natural forms exhibited similar IgE-binding capacities. A part of the fruit-fruit and pollen-fruit cross-reactions could be explained by the presence of this conserved protein; however, sequence homology provides insufficient information to predict IgE cross-reactivity of profilins. PMID:26989709

  6. A novel IgE-binding epitope of cat major allergen, Fel d 1.

    PubMed

    Tasaniyananda, Natt; Tungtrongchitr, Anchalee; Seesuay, Watee; Sakolvaree, Yuwaporn; Indrawattana, Nitaya; Chaicumpa, Wanpen; Sookrung, Nitat

    2016-02-12

    Information on the antigenic repertoire, especially the IgE-binding epitopes of an allergen is important for understanding the allergen induced immune response and cross-reactivity, as well as for generating the hypoallergenic variants for specific component resolved immunotherapy/diagnosis (CRIT and CRD). Data on the IgE-binding epitopes of cat allergens are scarce. In this study, a novel IgE-binding epitope of the cat major allergen, Fel d 1, was identified. Mouse monoclonal antibody (MAb) specific to the Fel d 1 was produced. Computerized intermolecular docking was used for determining the residues of the Fel d 1 bound by the specific MAb. The presumptive surface exposed residues of the Fel d 1 intrigued by the MAb are located on the chain 1. They are: L34 and T37 (helix 1); T39 (between helices 1 and 2); P40, E42 and E45 (helix 2); R61, K64, N65 and D68 (helix 3); and E73 and K76 (helix 4). The MAb competed efficiently with the cat allergic patients' serum IgE for Fel d 1 binding in the competitive IgE binding assay, indicating allergenicity of the MAb epitope. The newly identified allergenic epitope of the Fel d 1 is useful in a design of the CRIT and CRD for cat allergy. PMID:26797272

  7. A novel IgE-binding epitope of cat major allergen, Fel d 1.

    PubMed

    Tasaniyananda, Natt; Tungtrongchitr, Anchalee; Seesuay, Watee; Sakolvaree, Yuwaporn; Indrawattana, Nitaya; Chaicumpa, Wanpen; Sookrung, Nitat

    2016-02-12

    Information on the antigenic repertoire, especially the IgE-binding epitopes of an allergen is important for understanding the allergen induced immune response and cross-reactivity, as well as for generating the hypoallergenic variants for specific component resolved immunotherapy/diagnosis (CRIT and CRD). Data on the IgE-binding epitopes of cat allergens are scarce. In this study, a novel IgE-binding epitope of the cat major allergen, Fel d 1, was identified. Mouse monoclonal antibody (MAb) specific to the Fel d 1 was produced. Computerized intermolecular docking was used for determining the residues of the Fel d 1 bound by the specific MAb. The presumptive surface exposed residues of the Fel d 1 intrigued by the MAb are located on the chain 1. They are: L34 and T37 (helix 1); T39 (between helices 1 and 2); P40, E42 and E45 (helix 2); R61, K64, N65 and D68 (helix 3); and E73 and K76 (helix 4). The MAb competed efficiently with the cat allergic patients' serum IgE for Fel d 1 binding in the competitive IgE binding assay, indicating allergenicity of the MAb epitope. The newly identified allergenic epitope of the Fel d 1 is useful in a design of the CRIT and CRD for cat allergy.

  8. 27 CFR 7.22b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... from major food allergen labeling. 7.22b Section 7.22b Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY ALCOHOL LABELING AND ADVERTISING OF MALT BEVERAGES Labeling Requirements for Malt Beverages § 7.22b Petitions for exemption from major food...

  9. 27 CFR 7.22b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... from major food allergen labeling. 7.22b Section 7.22b Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF MALT BEVERAGES Labeling Requirements for Malt Beverages § 7.22b Petitions for exemption from major food...

  10. 27 CFR 7.22b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... from major food allergen labeling. 7.22b Section 7.22b Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY ALCOHOL LABELING AND ADVERTISING OF MALT BEVERAGES Labeling Requirements for Malt Beverages § 7.22b Petitions for exemption from major food...

  11. 27 CFR 7.22b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... from major food allergen labeling. 7.22b Section 7.22b Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF MALT BEVERAGES Labeling Requirements for Malt Beverages § 7.22b Petitions for exemption from major food...

  12. 27 CFR 7.22b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... from major food allergen labeling. 7.22b Section 7.22b Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF MALT BEVERAGES Labeling Requirements for Malt Beverages § 7.22b Petitions for exemption from major food...

  13. Structural and immunologic characterization of Ara h 1 – a major peanut allergen

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Allergic reactions to peanut and tree nuts is the major cause of anaphylaxis in the U.S. In this report; structural, immunologic, and bioinformatics analysis of natural, and a recombinant variant of the major peanut allergen Ara h 1 is presented. Small angle X-ray scattering studies show that natura...

  14. NADP-dependent mannitol dehydrogenase, a major allergen of Cladosporium herbarum.

    PubMed

    Simon-Nobbe, Birgit; Denk, Ursula; Schneider, Peter Bernhard; Radauer, Christian; Teige, Markus; Crameri, Reto; Hawranek, Thomas; Lang, Roland; Richter, Klaus; Schmid-Grendelmeier, Peter; Nobbe, Stephan; Hartl, Arnulf; Breitenbach, Michael

    2006-06-16

    Cladosporium herbarum is an important allergenic fungal species that has been reported to cause allergic diseases in nearly all climatic zones. 5-30% of the allergic population displays IgE antibodies against molds. Sensitization to Cladosporium has often been associated with severe asthma and less frequently with chronic urticaria and atopic eczema. However, no dominant major allergen of this species has been found so far. We present cloning, production, and characterization of NADP-dependent mannitol dehydrogenase of C. herbarum (Cla h 8) and show that this protein is a major allergen that is recognized by IgE antibodies of approximately 57% of all Cladosporium allergic patients. This is the highest percentage of patients reacting with any Cladosporium allergen characterized so far. Cla h 8 was purified to homogeneity by standard chromatographic methods, and both N-terminal and internal amino acid sequences of protein fragments were determined. Enzymatic analysis of the purified natural protein revealed that this allergen represents a NADP-dependent mannitol dehydrogenase that interconverts mannitol and d-fructose. It is a soluble, non-glycosylated cytoplasmic protein. Two-dimensional protein analysis indicated that mannitol dehydrogenase is present as a single isoform. The cDNA encoding Cla h 8 was cloned from a lambda-ZAP library constructed from hyphae and spores. The recombinant non-fusion protein was expressed in Escherichia coli and purified to homogeneity. Its immunological and biochemical identity with the natural protein was shown by enzyme activity tests, CD spectroscopy, IgE immunoblots with sera of patients, and by skin prick testing of Cladosporium allergic patients. This protein therefore is a new major allergen of C. herbarum.

  15. In vitro cross-allergenicity of major aeroallergenic pollens by the radioallergosorbent technique.

    PubMed

    Bernstein, I L; Perera, M; Gallagher, J; Michael, J G; Johansson, S G

    1976-02-01

    Cross-reactivity between major classes of aeroallergenic pollens has been estimated by comparing the inhibitory effects of selected allergens upon the reaginic activity of other native and non-native varieties, as measured by the radioallergosorbent technique (RAST). Common allergenic determinants were demonstrated in indigenous and nonindigenous ragween species. Although patterns of inhibitory capacity were less uniform among grass pollens, endemic species tended to have more common allergenic properties than nonendemic species. Tree pollens exhibited the greatest degree of heterogeneity, confirming the previous view that these pollens tend to be less cross-reactive. Concomitant skin test threshold studies utilizing the same groups of pollens were in general agreement with in vitro results. Because, threshold skin testing was affected by a number of uncontrollable variables, however, the current modification of RAST in estimating cross-allergenicity was more accurate and reliable than data based upon cutaneous tests. It was concluded that reagin neutralization by the RAST method is the best currently available method of assessing cross-allergenic properties of pollens.

  16. Antisense-mediated silencing of a gene encoding a major ryegrass pollen allergen.

    PubMed

    Bhalla, P L; Swoboda, I; Singh, M B

    1999-09-28

    Type 1 allergic reactions, such as hay fever and allergic asthma, triggered by grass pollen allergens are a global health problem that affects approximately 20% of the population in cool, temperate climates. Ryegrass is the dominant source of allergens because of its prodigious production of airborne pollen. Lol p 5 is the major allergenic protein of ryegrass pollen, judging from the fact that almost all of the individuals allergic to grass pollen show presence of serum IgE antibodies against this protein. Moreover, nearly two-thirds of the IgE reactivity of ryegrass pollen has been attributed to this protein. Therefore, it can be expected that down-regulation of Lol p 5 production can significantly reduce the allergic potential of ryegrass pollen. Here, we report down-regulation of Lol p 5 with an antisense construct targeted to the Lol p 5 gene in ryegrass. The expression of antisense RNA was regulated by a pollen-specific promoter. Immunoblot analysis of proteins with allergen-specific antibodies did not detect Lol p 5 in the transgenic pollen. The transgenic pollen showed remarkably reduced allergenicity as reflected by low IgE-binding capacity of pollen extract as compared with that of control pollen. The transgenic ryegrass plants in which Lol p 5 gene expression is perturbed showed normal fertile pollen development, indicating that genetic engineering of hypoallergenic grass plants is possible.

  17. Tropomyosin and Actin Identified as Major Allergens of the Carpet Clam (Paphia textile) and the Effect of Cooking on Their Allergenicity

    PubMed Central

    Mohamad Yadzir, Zailatul Hani; Misnan, Rosmilah; Bakhtiar, Faizal; Abdullah, Noormalin; Murad, Shahnaz

    2015-01-01

    Objectives. To identify the major allergenic proteins of clam (Paphia textile) and to investigate the effect of different cooking methods on the allergenicity of these identified proteins. Methods. Clam protein extracts were separated by denaturing polyacrylamide gel electrophoresis. IgE reactive proteins were then analyzed by immunoblotting with sera from patients with positive skin prick tests (SPT) to the raw clam extract. Mass spectrometry was used to identify the major allergenic proteins of this clam. Results. Raw extract showed 12 protein bands (18–150 kDa). In contrast, fewer protein bands were seen in the boiled extract; those ranging from 40 to 150 kDa were denatured. The protein profiles were similarly altered by frying or roasting. The immunoblots of raw and boiled extracts yielded 10 and 2 IgE-binding proteins, respectively. The fried and roasted extracts showed only a single IgE-binding protein at 37 kDa. Mass spectrometry analysis of the 37 and 42 kDa major allergens indicated that these spots were tropomyosin and actin, respectively. Conclusion. The two major allergens of Paphia textile were identified as the thermostable tropomyosin and a new thermolabile allergen actin. PMID:26413512

  18. Identification of IgE-binding proteins from Lepidoglyphus destructor and production of monoclonal antibodies to a major allergen.

    PubMed

    Ventas, P; Carreira, J; Polo, F

    1991-08-01

    The allergen composition of one of the most important storage mites, Lepidoglyphus destructor, has been studied by immunodetection after SDS-PAGE with individual patient sera. An allergenic polypeptide of 14 kDa was identified with 95% of the sera. This major allergen was isolated in the supernatant of 60% ammonium sulfate salt precipitation of the whole extract, which was subsequently used to immunize BALB/c mice so as to produce monoclonal antibodies. Four mAbs recognizing molecules with IgE-binding ability were obtained. The specificity of the mAbs was assayed against different allergenic extracts, and the molecules recognized by them were characterized by immunoblotting. Two mAbs (Le5B5 and Le9E4) were directed to the 14-kDa allergen; the other two to several proteins of lesser allergenic significance. PMID:1722776

  19. Qualitative polymerase chain reaction methods for detecting major food allergens (peanut, soybean, and wheat) by using internal transcribed spacer region.

    PubMed

    Hirao, Takashi; Watanabe, Satoshi; Temmei, Yusuke; Hiramoto, Masayuki; Kato, Hisanori

    2009-01-01

    Allergen detection methods for peanut, soybean, and wheat were developed by designing PCR primer pairs for specific amplification of a fragment of the internal transcribed spacer (ITS) region reported for Arachis spp. for peanut, Glycine spp. for soybean, and Triticum and Aegilops spp. for wheat. The target species for detection included not only cultivated, but also wild and ancestor species, which were thought to be potentially allergenic. The ability of the resultant primer pairs to detect the target species was verified using genomic DNA extracted from A. hypogaea for peanut and G max for soybean; T. aestivum, T. turgidum, T. durum, T. aestivum-rye amphidiploid, T. monococcum, T. timopheevi, Ae. speltoides, and Ae. squarrosa for wheat. The LODs were 50-500 fg of target DNA, which were comparable to those of the most sensitive PCR methods previously reported. The results from the present work, as well as those from our previous work on buckwheat and kiwifruit, prove that the ITS region, for its high copy number and interspecific diversity, is particularly useful as the target of allergen detection methods.

  20. Advances in the development of functional foods from buckwheat.

    PubMed

    Li, S Q; Zhang, Q H

    2001-09-01

    Buckwheat originated in North or East Asia and is widely adapted in North America. It has been grown since at least 1000 BC in China. It has very strong adaptability to adverse environments with a very short growing span. Many varieties are growing around the world, but mainly in the north hemisphere. Currently the most common buckwheat spice is Fagopyrum esculentum Moench (common buckwheat or sweet buckwheat), while Fagopyrum tartaricum is also available in some mountainous regions. Many nutraceutical compounds exist in buckwheat seeds and other tissues. Buckwheat has been used and will be better used as an important raw material for functional food production. In this review we focus on works related to the development of functional foods from common buckwheat, Fagopyrum esculentum Moench. A lot of research has be conducted in the functionalities and properties of buckwheat proteins, flavonoids, flavones, phytosterols, thiamin-binding proteins, and other rare compounds in buckwheat seeds. Buckwheat proteins have unique amino acid composition with special biological activities of cholesterol-lowering effects, antihypertensition effects, and improving the constipation and obisity conditions by acting similar as to dietary fiber and interrupting the in vivo metabolisms. The trypsin inhibitors isolated from buckwheat seeds are heat stable and can cause poor digestion if they are not suitably cooked before consumption. The allergenic proteins existing in the buckwheat seeds and their derivatives were reviewed with respect to their chemical and biochemical characteristics as well as the physiological reactions after digestion. Some possible mechanisms involved in these effects are discussed in this review. Experiments, both with animal models and with human beings, revealed that buckwheat flour can improve diabetes, obesity, hypertension, hypercholesterolemia and constipation. Methods to exploit buckwheat seeds and flour to produce highly effective nutraceuticals are

  1. Comparison of the digestibility of the major peanut allergens in thermally processed peanuts and in pure form

    Technology Transfer Automated Retrieval System (TEKTRAN)

    It has been suggested that boiling or frying of peanuts lead to less allergenic products than roasting. Here, we have compared the digestibility of the major peanut allergens in the context of peanuts subjected to boiling, frying, or roasting, and in purified form. The soluble peanut extracts and ...

  2. Crystal Structure of Prunin-1, a Major Component of the Almond (Prunus dulcis) Allergen Amandin

    SciTech Connect

    Jin, Tengchuan; Albillos, Silvia M.; Guo, Feng; Howard, Andrew; Fu, Tong-Jen; Kothary, Mahendra H.; Zhang, Yu-Zhu

    2010-10-28

    Seed storage proteins are accumulated during seed development and act as a reserve of nutrition for seed germination and young sprout growth. Plant seeds play an important role in human nutrition by providing a relatively inexpensive source of protein. However, many plant foods contain allergenic proteins, and the number of people suffering from food allergies has increased rapidly in recent years. The 11S globulins are the most widespread seed storage proteins, present in monocotyledonous and dicotyledonous seeds as well as in gymnosperms (conifers) and other spermatophytes. This family of proteins accounts for a number of known major food allergens. They are of interest to both the public and industry due to food safety concerns. Because of the interests in the structural basis of the allergenicity of food allergens, we sought to determine the crystal structure of Pru1, the major component of the 11 S storage protein from almonds. The structure was refined to 2.4 {angstrom}, and the R/Rfree for the final refined structure is 17.2/22.9. Pru1 is a hexamer made of two trimers. Most of the back-to-back trimer-trimer association was contributed by monomer-monomer interactions. An {alpha} helix (helix 6) at the C-terminal end of the acidic domain of one of the interacting monomers lies at the cleft of the two protomers. The residues in this helix correspond to a flexible region in the peanut allergen Ara h 3 that encompasses a previously defined linear IgE epitope.

  3. Crystal structure of prunin-1, a major component of the almond (Prunus dulcis) allergen amandin.

    PubMed

    Jin, Tengchuan; Albillos, Silvia M; Guo, Feng; Howard, Andrew; Fu, Tong-Jen; Kothary, Mahendra H; Zhang, Yu-Zhu

    2009-09-23

    Seed storage proteins are accumulated during seed development and act as a reserve of nutrition for seed germination and young sprout growth. Plant seeds play an important role in human nutrition by providing a relatively inexpensive source of protein. However, many plant foods contain allergenic proteins, and the number of people suffering from food allergies has increased rapidly in recent years. The 11S globulins are the most widespread seed storage proteins, present in monocotyledonous and dicotyledonous seeds as well as in gymnosperms (conifers) and other spermatophytes. This family of proteins accounts for a number of known major food allergens. They are of interest to both the public and industry due to food safety concerns. Because of the interests in the structural basis of the allergenicity of food allergens, we sought to determine the crystal structure of Pru1, the major component of the 11 S storage protein from almonds. The structure was refined to 2.4 A, and the R/Rfree for the final refined structure is 17.2/22.9. Pru1 is a hexamer made of two trimers. Most of the back-to-back trimer-trimer association was contributed by monomer-monomer interactions. An alpha helix (helix 6) at the C-terminal end of the acidic domain of one of the interacting monomers lies at the cleft of the two protomers. The residues in this helix correspond to a flexible region in the peanut allergen Ara h 3 that encompasses a previously defined linear IgE epitope.

  4. Molecular Cloning and Expression of a New Major Allergen, Ani s 14, from Anisakis simplex.

    PubMed

    Kobayashi, Yukihiro; Kakemoto, Seiko; Shimakura, Kuniyoshi; Shiomi, Kazuo

    2015-01-01

    The nematode Anisakis simplex is a representative parasite infecting marine animals. When third stage larvae of A. simplex infecting fish and squids are ingested by humans, individuals previously sensitized by this parasite may experience IgE-mediated allergic reactions. So far, as many as 13 kinds of proteins (Ani s 1-13) have been identified as A. simplex allergens but several more unknown allergens are suggested to exist. In this study, therefore, chemiluminescent immunoscreening of an expression cDNA library constructed from the third stage larvae was conducted to identify a new allergen. As a result, an IgE-positive clone coding for a 23.5 kDa protein (named Ani s 14) composed of 217 amino acid residues was isolated. The regions 4-147 and 34-123 of Ani s 14 share 31% identity with the region 796-940 of Ani s 7 and 32% identity with the region 2-91 of Ani s 12, respectively. Recombinant Ani s 14 was successfully expressed in Escherichia coli as a His-tagged protein and shown to be IgE reactive to 14 (54%) of 26 sera from Anisakis-allergic patients. In conclusion, Ani s 14 is a new major allergen of A. simplex that is specific to Anisakis-allergic patients.

  5. NMR resonance assignments of the major apple allergen Mal d 1.

    PubMed

    Ahammer, Linda; Grutsch, Sarina; Tollinger, Martin

    2016-10-01

    The major apple allergen Mal d 1 is the predominant cause of apple (Malus domestica) allergies in large parts of Europe and Northern America. Allergic reactions against this 17.5 kDa protein are the consequence of initial sensitization to the structurally homologous major allergen from birch pollen, Bet v 1. Consumption of apples can subsequently provoke immunologic cross-reactivity of Bet v 1-specific antibodies with Mal d 1 and trigger severe oral allergic syndroms, affecting more than 70 % of all individuals that are sensitized to birch pollen. While the accumulated immunological data suggest that Mal d 1 has a three-dimensional fold that is similar to Bet v 1, experimental structural data for this protein are not available to date. In a first step towards structural characterization of Mal d 1, backbone and side chain (1)H, (13)C and (15)N chemical shifts of the isoform Mal d 1.0101 were assigned. The NMR-chemical shift data show that this protein is composed of seven β-strands and three α-helices, which is in accordance with the reported secondary structure of the major birch pollen allergen, indicating that Mal d 1 and Bet v 1 indeed have similar three-dimensional folds. The next stage in the characterization of Mal d 1 will be to utilize these resonance assignments in solving the solution structure of this protein. PMID:27165578

  6. Molecular basis of IgE-recognition of Lol p 5, a major allergen of rye-grass pollen.

    PubMed

    Suphioglu, C; Blaher, B; Rolland, J M; McCluskey, J; Schäppi, G; Kenrick, J; Singh, M B; Knox, R B

    1998-04-01

    Grass pollen, especially of rye-grass (Lolium perenne). represents an important cause of type I allergy. Identification of IgE-binding (allergenic) epitopes of major grass pollen allergens is essential for understanding the molecular basis of interaction between allergens and human IgE antibodies and therefore facilitates the devising of safer and more effective diagnostic and immunotherapy reagents. The aim of this study was to identify the allergenic epitopes of Lol p 5, a major allergen of rye-grass pollen, immunodissect these epitopes further so that the amino acid residues critical for antibody binding can be determined and investigate the conservation and nature of these epitopes within the context of the natural grass pollen allergens. Peptides, 12-13 amino acid residues long and overlapping each other by 4 amino acid residues, based on the entire deduced amino acid sequence of the coding region of Lol p 5, were synthesised and assayed for IgE-binding. Two strong IgE-binding epitopes (Lol p 5 (49-60) and (265-276), referred to as peptides 7 and 34, respectively) were identified. These epitopes were further resolved by truncated peptides and amino acid replacement studies and the amino acid residues critical for IgE-binding determined (Lol p 5 (49-60) residue Lys57 and (265-276) residue Lys275). Sequences of these epitopes were conserved in related allergens and may form the conserved allergenic domains responsible for the cross-reactivity observed between pollen allergens of taxonomically related grasses. Furthermore, due to its strong IgE-reactivity, synthetic peptide Lol p 5 (265-276) was used to affinity-purify specific IgE antibodies which recognised proteins of other clinically important grass pollens. further indicating presence of allergenic cross-reactivity at the level of allergenic epitope. Moreover, Lol p 5 (265 276) demonstrated a strong capacity to inhibit IgE-binding to natural rye-grass pollen proteins highlighting the antibody accessibility

  7. Influence of the maillard reaction on the allergenicity of rAra h 2, a recombinant major allergen from peanut (Arachis hypogaea), its major epitopes, and peanut agglutinin.

    PubMed

    Gruber, Patrick; Becker, Wolf-Meinhard; Hofmann, Thomas

    2005-03-23

    The influence of thermal processing and nonenzymatic browning reactions on the IgE-binding activity of rAra h 2 was studied and compared to findings recently reported for the allergen's natural counterpart. ELISA experiments as well as inhibition assays revealed that thermal treatment of rAra h 2 in the presence of reactive carbohydrates and carbohydrate breakdown products induces a strong increase of the IgE-binding activity, thus collaborating with the data reported for the natural protein isolated from peanuts. To localize the Ara h 2 sequences responsible for the formation of highly IgE-affine glycation sites, model peptides have been synthesized mimicking sequences which contain possible targets for glycation as well as the immunodominant epitopes. Immunological evaluation of these peptides heated in the absence or presence of reducing sugars and carbonyls, respectively, revealed that neither the two lysine residues of Ara h 2 nor its N-terminus are involved in the formation of IgE-affine structures by Maillard reaction. Also, the cysteine-containing major epitope 3 (aa 27-36) was found to lose its IgE-binding capacity upon heating. By contrast, the overlapping major epitopes 6 and 7, which do not contain any lysine or arginine moieties, showed a distinct higher level of IgE binding when subjected to Maillard reaction, thus giving the first evidence that nonbasic amino acids might be accessible for nonenzymatic glycation reactions and that these posttranslational modifications might induce increased IgE binding of the glycated Ara h 2. Analogous experiments were performed with peanut agglutinin, considered in the literature as a minor allergen. ELISA experiments revealed that the majority of tested sera samples from peanut-sensitive patients showed a high level of IgE binding to the lectin even after heat treatment. In contradiction to published data, nonenzymatic browning reactions seem to deteriorate the IgE affinity of the lectin. PMID:15769170

  8. Purification and characterization of Lep d I, a major allergen from the mite Lepidoglyphus destructor.

    PubMed

    Ventas, P; Carreira, J; Polo, F

    1992-04-01

    A major allergen of the storage mite Lepidoglyphus destructor (Lep d I) has been purified by affinity chromatography using an anti-Lep d I monoclonal antibody. The purity of the protein obtained by this procedure was assessed by reverse-phase HPLC. Lep d I displayed a molecular weight of 14 kD on SDS-PAGE under non-reducing conditions, and 16 kD in the presence of a reducing agent. Analytical IEF revealed a little charge microheterogeneity, showing three bands with pIs 7.6-7.8. Purified Lep d I retained IgE-binding ability, as proved by immunoblotting experiments after SDS-PAGE and RAST with individual sera from L. destructor-sensitive patients. Results from the latter technique demonstrated that 87% of L. destructor-allergic patients had specific IgE to Lep d I, and a good correlation between IgE reactivity with L. destructor extract and Lep d I was found. In addition, RAST inhibition experiments showed that IgE-binding sites on Lep d I are major L. destructor-allergenic determinants, since Lep d I could inhibit up to 75% the binding of specific IgE to L. destructor extract; on the other hand, Lep d I did not cross-react with D. pteronyssinus allergens. PMID:1611545

  9. Quantification of major allergen parvalbumin in 22 species of fish by SDS-PAGE.

    PubMed

    Kobayashi, Yukihiro; Yang, Tao; Yu, Cheng-Tao; Ume, Chiaki; Kubota, Hiroyuki; Shimakura, Kuniyoshi; Shiomi, Kazuo; Hamada-Sato, Naoko

    2016-03-01

    Fish is an important causative material of food allergy. Although the allergenicity of fish is considered to correlate with the content of parvalbumin, the major fish allergen, available information about the parvalbumin content in fish is limited. In this study, a simple and reliable quantification method for fish parvalbumin by SDS-PAGE was first established. Application of the SDS-PAGE method to 22 species of fish revealed a marked variation in parvalbumin content among fish. Furthermore, the parvalbumin content was found to be higher in dorsal white muscle than in ventral white muscle, in rostral part of white muscle than in caudal part of white muscle and in white muscle than in dark muscle. IgE reactivity of fish was roughly proportional to parvalbumin content. Interestingly, large-sized migratory fish, such as salmon, swordfish and tuna, were commonly very low in both parvalbumin content and IgE reactivity. PMID:26471564

  10. Identification of major rice allergen and their clinical significance in children

    PubMed Central

    Jeon, You Hoon; Oh, Se Jo; Yang, Hyeon Jong; Lee, Soo Young

    2011-01-01

    Purpose Recently, an increase in the number of patients sensitized to rice allergen with or without clinical symptoms has been reported. This study was designed to determine the major allergens in rice and their clinical significance. Methods Twenty-four children (15 boys and 9 girls; mean age, 16.3 months) with allergic disease, who were sensitized to rice antigen (by UniCAP) in the Pediatric Allergy Respiratory Center at Soonchunhyang University Hospital, were enrolled in this study. The allergenicity of various types of rice (raw, cooked, and heat-treated, simulated gastric fluid [SGF], and simulated intestinal fluid [SIF]) was investigated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoglobulin E (IgE) immunoblots. The patients' medical records, including laboratory data and allergy symptoms after ingestion of rice were reviewed. Results Patients were sensitized to an average of 13.5 food antigens and their mean total IgE was 6,888.7 kU/L. In SDS-PAGE, more than 16 protein bands were observed in the raw rice, whereas only 14-16 kDa and 31-35 kDa protein bands were observed in cooked rice. The common SDS-PAGE protein bands observed in SGF-, SIF-, and heat-treated rice were 9, 14, and 31 kDa. In a heated-rice IgE immunoblot, protein bands of 9, 14, and 31-33 kDa were found in 27.8%, 38.9%, and 38.9% of all sera, respectively, and in 50%, 50%, and 75%, of ser a from the 4 symptomatic patients, respectively. Conclusion The 9-, 14-, and 31-kDa protein bands appeared to be the major allergens responsible for rice allergy symptoms. PMID:22232624

  11. Structural and functional alterations in major peanut allergens caused by thermal processing.

    PubMed

    Maleki, Soheila J; Hurlburt, Barry K

    2004-01-01

    The majority of foods that we eat are subjected to some type of processing either at home or by the manufacturer. The biochemical reactions that occur in foods as a result of thermal processing can be both beneficial and harmful. Here, we briefly review the effects of thermal processing and some of the effects of the Maillard reaction on the allergenicity of food proteins. Specifically, we focus on the known effects of roasting on the allergenic properties of peanut proteins and the contribution of Maillard reaction products or advanced glycation end products to these observed effects. The most thorough understanding of the effects of thermal processing on allergenicity involves the peanut proteins. Thermal processing alters specific biophysical and immunological properties of peanut proteins, such as structure, function, solubility, digestibility, immunoglobulin E (IgE) binding, and T-cell responses. A better understanding of the effects of thermal processing-induced biochemical and immunological alterations is of utmost importance for proper risk assessment of existing and newly introduced proteins in the food source, as well as development of effective diagnostic tools and therapeutic treatments for food allergy.

  12. Ligand Recognition of the Major Birch Pollen Allergen Bet v 1 is Isoform Dependent

    PubMed Central

    Seutter von Loetzen, Christian; Jacob, Thessa; Hartl-Spiegelhauer, Olivia; Vogel, Lothar; Schiller, Dirk; Spörlein-Güttler, Cornelia; Schobert, Rainer; Vieths, Stefan; Hartl, Maximilian Johannes; Rösch, Paul

    2015-01-01

    Each spring millions of patients suffer from allergies when birch pollen is released into the air. In most cases, the major pollen allergen Bet v 1 is the elicitor of the allergy symptoms. Bet v 1 comes in a variety of isoforms that share virtually identical conformations, but their relative concentrations are plant-specific. Glycosylated flavonoids, such as quercetin-3-O-sophoroside, are the physiological ligands of Bet v 1, and here we found that three isoforms differing in their allergenic potential also show an individual, highly specific binding behaviour for the different ligands. This specificity is driven by the sugar moieties of the ligands rather than the flavonols. While the influence of the ligands on the allergenicity of the Bet v 1 isoforms may be limited, the isoform and ligand mixtures add up to a complex and thus individual fingerprint of the pollen. We suggest that this mixture is not only acting as an effective chemical sunscreen for pollen DNA, but may also play an important role in recognition processes during pollination. PMID:26042900

  13. Carbohydrate moieties can induce mediator release: a detailed characterization of two major timothy grass pollen allergens.

    PubMed

    Wicklein, Daniel; Lindner, Buko; Moll, Hermann; Kolarich, Daniel; Altmann, Friedrich; Becker, Wolf-Meinhard; Petersen, Arnd

    2004-05-01

    Specific IgE binding to carbohydrate moieties of glycosylated allergens has been known for years, but the importance of these structures for the elicitation of allergic reactions is still a matter of debate. Because of their conserved carbohydrate structures, especially N-glycans have always been prime candidates for IgE cross-reactivity between allergens from unrelated species. The aim of our study was to determine whether carbohydrate structures on glycoproteins can by themselves elucidate allergic reactions. We characterized in detail the carbohydrate moieties of the major allergens Phl p 1 and Phl p 13 of timothy grass pollen (Phleum pratense L.) by performing tryptic digests followed by HPLC, N-terminal sequencing, sugar analysis, MALDI-TOF- and ESI-ICRFT-MS. Phl p 1 contains one N-glycan with one of the two glycoforms MMXF3 and M0XF3 and a single furanosidic arabinose, which is bound to a hydroxyproline residue in direct vicinity to the N-glycan. This O-glycosylation is probably due to an arabinosylation consensus sequence found in the N-terminal part of Phl p 1 and other group 1 allergens, but displayed no IgE-reactivity. Thus, Phl p 1 is monovalent with respect to its IgE-binding carbohydrate epitopes and showed no mediator release. In contrast, the carbohydrate moiety of Phl p 13, which carries four of the same N-glycans (like Phl p 1), can cross-link IgE-receptors via carbohydrate chains and elicits IL-4 release from basophils.

  14. Development of sandwich ELISA for detection and quantification of invertebrate major allergen tropomyosin by a monoclonal antibody.

    PubMed

    Zhang, Hong; Lu, Ying; Ushio, Hideki; Shiomi, Kazuo

    2014-05-01

    Muscle protein tropomyosins of invertebrates are major allergens responsible for wide spread allergic reactions against invertebrates such as shellfish and insects. In order to develop a sandwich enzyme-linked immunoadsorbent assay (ELISA) for detection and quantification of the invertebrate pan-allergen tropomyosin, a specific monoclonal antibody (MAb), CE7B2, was produced. We have successfully established a sandwich ELISA for measuring invertebrate tropomyosin concentrations in food and food materials. The sandwich ELISA system using the MAb CE7B2 is a useful tool to detect and quantify levels of tropomyosin in food. The method is also helpful to detect mite and cockroach tropomyosins, the important indoor allergens.

  15. Homology modelling of the major peanut allergen Ara h 2 and surface mapping of IgE-binding epitopes.

    PubMed

    Barre, Annick; Borges, Jean-Philippe; Culerrier, Raphaël; Rougé, Pierre

    2005-09-15

    Three-dimensional models built for the peanut Ara h 2 allergen and other structurally-related 2S albumin allergens of dietary nuts exhibited an overall three-dimensional fold stabilized by disulphide bridges well conserved among all the members of the 2S albumin superfamily. Conformational analysis of the linear IgE-binding epitopes mapped on the molecular surface of Ara h 2 showed no structural homology with the corresponding regions of the walnut Jug r 1, the pecan nut Car i 1 or the Brazil nut Ber e 1 allergens. The absence of epitopic community does not support the allergenic cross-reactivity observed between peanut and walnut or Brazil nut, which presumably depends on other ubiquitous seed storage protein allergens, namely the vicilins. However, the major IgE-binding epitope identified on the molecular surface of the walnut Jug r 1 allergen shared a pronounced structural homology with the corresponding region of the pecan nut Car i 1 allergen. With the exception of peanut, 2S albumins could thus account for the IgE-binding cross-reactivity observed between some other dietary nuts, e.g. walnut and pecan nut. PMID:15899521

  16. Changes in Major Peanut Allergens Under Different pH Conditions.

    PubMed

    Kim, Jihyun; Lee, Jeongok; Seo, Won Hee; Han, Youngshin; Ahn, Kangmo; Lee, Sang-Il

    2012-05-01

    Regional dietary habits and cooking methods affect the prevalence of specific food allergies; therefore, we determined the effects of various pH conditions on major peanut allergens. Peanut kernels were soaked overnight in commercial vinegar (pH 2.3) or acetic acid solutions at pH 1.0, 3.0, or 5.0. Protein extracts from the sera of seven patients with peanut-specific IgE levels >15 kU(A)/L were analyzed by SDS-PAGE and immunolabeling. A densitometer was used to quantify and compare the allergenicity of each protein. The density of Ara h 1 was reduced by treatment with pH 1.0, 3.0, or 5.0 acetic acid, or commercial vinegar. Ara h 2 remained largely unchanged after treatment with pH 5.0 acetic acid, and was decreased following treatment with pH 1.0, 2.3, or 3.0 acetic acid. Ara h 3 and Ara h 6 appeared as a thick band after treatment with pH 1.0 acetic acid and commercial vinegar. IgE-binding intensities to Ara h 1, Ara h 2, and Ara h 3 were significantly reduced after treatment with pH 1.0 acetic acid or commercial vinegar. These data suggest that treatment with acetic acid at various pH values affects peanut allergenicity and may explain the low prevalence of peanut allergy in Korea. PMID:22548209

  17. Search for Allergens from the Pollen Proteome of Sunflower (Helianthus annuus L.): A Major Sensitizer for Respiratory Allergy Patients

    PubMed Central

    Saha, Bodhisattwa; Pandey, Naren; Gupta Bhattacharya, Swati

    2015-01-01

    Background Respiratory allergy triggered by pollen allergens is increasing at an alarming rate worldwide. Sunflower pollen is thought to be an important source of inhalant allergens. Present study aims to identify the prevalence of sunflower pollinosis among the Indian allergic population and characterizes the pollen allergens using immuno-proteomic tools. Methodology Clinico-immunological tests were performed to understand the prevalence of sensitivity towards sunflower pollen among the atopic population. Sera from selected sunflower positive patients were used as probe to detect the IgE-reactive proteins from the one and two dimensional electrophoretic separated proteome of sunflower pollen. The antigenic nature of the sugar moiety of the glycoallergens was studied by meta-periodate modification of IgE-immunoblot. Finally, these allergens were identified by mass-spectrometry. Results Prevalence of sunflower pollen sensitization was observed among 21% of the pollen allergic population and associated with elevated level of specific IgE and histamine in the sera of these patients. Immunoscreening of sunflower pollen proteome with patient sera detected seven IgE-reactive proteins with varying molecular weight and pI. Hierarchical clustering of 2D-immunoblot data highlighted three allergens characterized by a more frequent immuno-reactivity and increased levels of IgE antibodies in the sera of susceptible patients. These allergens were considered as the major allergens of sunflower pollen and were found to have their glycan moiety critical for inducing IgE response. Homology driven search of MS/MS data of these IgE-reactive proteins identified seven previously unreported allergens from sunflower pollen. Three major allergenic proteins were identified as two pectate lyases and a cysteine protease. Conclusion Novelty of the present report is the identification of a panel of seven sunflower pollen allergens for the first time at immuno-biochemical and proteomic level

  18. Vacuolar Serine Protease Is a Major Allergen of Fusarium proliferatum and an IgE-Cross Reactive Pan-Fungal Allergen

    PubMed Central

    Yeh, Chang-Ching; Tai, Hsiao-Yun; Chou, Hong; Wu, Keh-Gong

    2016-01-01

    Purpose Fusarium species are among prevalent airborne fungi and causative agents of human respiratory atopic disorders. We previously identified a 36.5-kDa F. proliferatum component recognized by IgE antibodies in 9 (53%) of the 17 F. proliferatum-sensitized atopic serum samples. The purpose of this study is to characterize the 36.5-kDa allergen of F. proliferatum. Methods Characterization of allergens and determination of IgE cross-reactivity were performed by cDNA cloning/expression and immunoblot inhibition studies. Results Based on the finding that the 36.5-kDa IgE-binding component reacted with the mouse monoclonal antibody FUM20 against fungal vacuolar serine protease allergens, the cDNA of F. proliferatum vacuolar serine protease (Fus p 9.0101) was subsequently cloned. Nine serum samples from respiratory atopic patients with IgE binding to the vacuolar serine protease allergen of Penicillium chrysogenum (Pen ch 18) also showed IgE-immunoblot reactivity to rFus p 9.0101. The purified rFus p 9.0101 can inhibit IgE and FUM20 binding to the 36.5-kDa component of F. proliferatum. Thus, a novel and important Fus p 9.0101 was identified. The rPen ch 18 can inhibit IgE binding to Fus p 9.0101. It indicates that IgE cross-reactivity between Fus p 9.0101 and Pen ch 18 also exists. Furthermore, neither rFus p 9.0101 K88A nor rPen ch 18 K89A mutants inhibited IgE binding to rFus p 9.0101. Lys88 was considered a critical core amino acid in IgE binding to r Fus p 9.0101 and a residue responsible for IgE cross-reactivity between Fus p 9.0101 and Pen ch 18 allergens. Conclusions Results obtained from this study indicate that vacuolar serine protease may be a major allergen of F. proliferatum and an important IgE cross-reactive pan-fungal allergen, and provide important bases for clinical diagnosis of fungal allergy. PMID:27334782

  19. Der p 11 Is a Major Allergen for House Dust Mite-Allergic Patients Suffering from Atopic Dermatitis

    PubMed Central

    Banerjee, Srinita; Resch, Yvonne; Chen, Kuan-Wei; Swoboda, Ines; Focke-Tejkl, Margit; Blatt, Katharina; Novak, Natalija; Wickman, Magnus; van Hage, Marianne; Ferrara, Rosetta; Mari, Adriano; Purohit, Ashok; Pauli, Gabrielle; Sibanda, Elopy N.; Ndlovu, Portia; Thomas, Wayne R.; Krzyzanek, Vladislav; Tacke, Sebastian; Malkus, Ursula; Valent, Peter; Valenta, Rudolf; Vrtala, Susanne

    2015-01-01

    House dust mites (HDMs) belong to the most potent indoor allergen sources worldwide and are associated with allergic manifestations in the respiratory tract and the skin. Here we studied the importance of the high-molecular-weight group 11 allergen from Dermatophagoides pteronyssinus (Der p 11) in HDM allergy. Sequence analysis showed that Der p 11 has high homology to paramyosins from mites, ticks, and other invertebrates. A synthetic gene coding for Der p 11 was expressed in Escherichia coli and rDer p 11 purified to homogeneity as folded, alpha-helical protein as determined by circular dichroism spectroscopy. Using antibodies raised against rDer p 11 and immunogold electron microscopy, the allergen was localized in the muscle beneath the skin of mite bodies but not in feces. IgE reactivity of rDer p 11 was tested with sera from HDM-allergic patients from Europe and Africa in radioallergosorbent test–based dot-blot assays. Interestingly, we found that Der p 11 is a major allergen for patients suffering from atopic dermatitis (AD), whereas it is only a minor allergen for patients suffering from respiratory forms of HDM allergy. Thus, rDer p 11 might be a useful serological marker allergen for the identification of a subgroup of HDM-allergic patients suffering from HDM-associated AD. PMID:24999597

  20. Targeting allergenic fungi in agricultural environments aids the identification of major sources and potential risks for human health.

    PubMed

    Weikl, F; Radl, V; Munch, J C; Pritsch, K

    2015-10-01

    Fungi are, after pollen, the second most important producers of outdoor airborne allergens. To identify sources of airborne fungal allergens, a workflow for qPCR quantification from environmental samples was developed, thoroughly tested, and finally applied. We concentrated on determining the levels of allergenic fungi belonging to Alternaria, Cladosporium, Fusarium, and Trichoderma in plant and soil samples from agricultural fields in which cereals were grown. Our aims were to identify the major sources of allergenic fungi and factors potentially influencing their occurrence. Plant materials were the main source of the tested fungi at and after harvest. Amounts of A. alternata and C. cladosporioides varied significantly in fields under different management conditions, but absolute levels were very high in all cases. This finding suggests that high numbers of allergenic fungi may be an inevitable side effect of farming in several crops. Applied in large-scale studies, the concept described here may help to explain the high number of sensitization to airborne fungal allergens.

  1. Creation of an engineered APC system to explore and optimize the presentation of immunodominant peptides of major allergens

    PubMed Central

    Rosskopf, Sandra; Jutz, Sabrina; Neunkirchner, Alina; Candia, Martín R.; Jahn-Schmid, Beatrice; Bohle, Barbara; Pickl, Winfried F.; Steinberger, Peter

    2016-01-01

    We have generated engineered APC to present immunodominant peptides derived from the major aero-allergens of birch and mugwort pollen, Bet v 1142–153 and Art v 125–36, respectively. Jurkat-based T cell reporter lines expressing the cognate allergen-specific T cell receptors were used to read out the presentation of allergenic peptides on the engineered APC. Different modalities of peptide loading and presentation on MHC class II molecules were compared. Upon exogenous loading with allergenic peptides, the engineered APC elicited a dose-dependent response in the reporter T cells and the presence of chemical loading enhancers strongly increased reporter activation. Invariant chain-based MHC class II targeting strategies of endogenously expressed peptides resulted in stronger activation of the reporters than exogenous loading. Moreover, we used Bet v 1 as model allergen to study the ability of K562 cells to present antigenic peptides derived from whole proteins either taken up or endogenously expressed as LAMP-1 fusion protein. In both cases the ability of these cells to process and present peptides derived from whole proteins critically depended on the expression of HLA-DM. We have identified strategies to achieve efficient presentation of allergenic peptides on engineered APC and demonstrate their use to stimulate T cells from allergic individuals. PMID:27539532

  2. Targeting allergenic fungi in agricultural environments aids the identification of major sources and potential risks for human health.

    PubMed

    Weikl, F; Radl, V; Munch, J C; Pritsch, K

    2015-10-01

    Fungi are, after pollen, the second most important producers of outdoor airborne allergens. To identify sources of airborne fungal allergens, a workflow for qPCR quantification from environmental samples was developed, thoroughly tested, and finally applied. We concentrated on determining the levels of allergenic fungi belonging to Alternaria, Cladosporium, Fusarium, and Trichoderma in plant and soil samples from agricultural fields in which cereals were grown. Our aims were to identify the major sources of allergenic fungi and factors potentially influencing their occurrence. Plant materials were the main source of the tested fungi at and after harvest. Amounts of A. alternata and C. cladosporioides varied significantly in fields under different management conditions, but absolute levels were very high in all cases. This finding suggests that high numbers of allergenic fungi may be an inevitable side effect of farming in several crops. Applied in large-scale studies, the concept described here may help to explain the high number of sensitization to airborne fungal allergens. PMID:26022406

  3. Creation of an engineered APC system to explore and optimize the presentation of immunodominant peptides of major allergens.

    PubMed

    Rosskopf, Sandra; Jutz, Sabrina; Neunkirchner, Alina; Candia, Martín R; Jahn-Schmid, Beatrice; Bohle, Barbara; Pickl, Winfried F; Steinberger, Peter

    2016-01-01

    We have generated engineered APC to present immunodominant peptides derived from the major aero-allergens of birch and mugwort pollen, Bet v 1142-153 and Art v 125-36, respectively. Jurkat-based T cell reporter lines expressing the cognate allergen-specific T cell receptors were used to read out the presentation of allergenic peptides on the engineered APC. Different modalities of peptide loading and presentation on MHC class II molecules were compared. Upon exogenous loading with allergenic peptides, the engineered APC elicited a dose-dependent response in the reporter T cells and the presence of chemical loading enhancers strongly increased reporter activation. Invariant chain-based MHC class II targeting strategies of endogenously expressed peptides resulted in stronger activation of the reporters than exogenous loading. Moreover, we used Bet v 1 as model allergen to study the ability of K562 cells to present antigenic peptides derived from whole proteins either taken up or endogenously expressed as LAMP-1 fusion protein. In both cases the ability of these cells to process and present peptides derived from whole proteins critically depended on the expression of HLA-DM. We have identified strategies to achieve efficient presentation of allergenic peptides on engineered APC and demonstrate their use to stimulate T cells from allergic individuals. PMID:27539532

  4. Purification and recombinant expression of major peanut allergen Ara h 1.

    PubMed

    Wu, Zhihua; Yan, Fei; Wei, Xiaonan; Li, Xin; Tong, Ping; Yang, Anshu; Tang, Ronghua; Chen, Hongbing

    2015-01-01

    Reaction to peanut, as one of the major food allergens, has become an increasingly common life-threatening disorder. Although peanut allergens have been extensively identified, Ara h 1 is still too expensive to be applied in food safety or clinical utility. In this study, the purification, expression, and immunological analyses of Ara h 1 are investigated. It was shown that a high purity (>95%) of Ara h 1 could be prepared by either purification or expression. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), Western blot, and mass spectroscopy were used to identify the Ara h 1, and it was found that natural Ara h 1 (nAra h 1) and expressed Ara h 1 (rAra h 1) have the same properties, including amino acid sequence. In particular, rAra h 1 reacted positively with anti-nAra h 1 serum, showing their similar immunological property. Thus, by either purification or expression, Ara h 1 could be prepared with low cost, as performed in the present work. SDS-PAGE, mag trix-assisted laser desorption/ionization time-of-flight mass spectroscopy (MALDI-TOF MS), and immunological analysis confirmed that both forms of Ara h 1 had the same properties. PMID:25036339

  5. Purification and recombinant expression of major peanut allergen Ara h 1.

    PubMed

    Wu, Zhihua; Yan, Fei; Wei, Xiaonan; Li, Xin; Tong, Ping; Yang, Anshu; Tang, Ronghua; Chen, Hongbing

    2015-01-01

    Reaction to peanut, as one of the major food allergens, has become an increasingly common life-threatening disorder. Although peanut allergens have been extensively identified, Ara h 1 is still too expensive to be applied in food safety or clinical utility. In this study, the purification, expression, and immunological analyses of Ara h 1 are investigated. It was shown that a high purity (>95%) of Ara h 1 could be prepared by either purification or expression. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), Western blot, and mass spectroscopy were used to identify the Ara h 1, and it was found that natural Ara h 1 (nAra h 1) and expressed Ara h 1 (rAra h 1) have the same properties, including amino acid sequence. In particular, rAra h 1 reacted positively with anti-nAra h 1 serum, showing their similar immunological property. Thus, by either purification or expression, Ara h 1 could be prepared with low cost, as performed in the present work. SDS-PAGE, mag trix-assisted laser desorption/ionization time-of-flight mass spectroscopy (MALDI-TOF MS), and immunological analysis confirmed that both forms of Ara h 1 had the same properties.

  6. Identification of tropomyosin as major allergen of white squid (Loligo edulis) by two-dimensional immunoblotting and mass spectrometry.

    PubMed

    Yadzir, Zailatul Hani Mohamad; Misnan, Rosmilah; Murad, Shahnaz

    2012-01-01

    IgE-mediated allergic reaction to squid is one of the most frequent molluscan shellfish allergies. Previously, we have detected a 36 kDa protein as the major allergen of Loligo edulis (white squid) by immunoblotting using sera from patients with squid allergy. The aim of this present study was to further identify this major allergen using a proteomics approach. The major allergen was identified by a combination of two-dimensional electrophoresis (2-DE), immunoblotting, mass spectrometry and bioinformatics tools. The 2-DE gel fractionated the cooked white squid proteins to more than 50 different protein spots between 10 to 38 kDa and isoelectric point (pI) from 3.0 to 10.0. A highly reactive protein spot of a molecular mass of 36 kDa and pI of 4.55 was observed in all of the patients' serum samples tested. Mass spectrometry analysis led to identification of this allergen as tropomyosin. This finding can contribute to advancement in component-based diagnosis, management of squid allergic patients, to the development of immunotherapy and to the standardization of allergenic test products as tools in molecular allergology.

  7. Assessment of the allergenic potential of transgenic wheat (Triticum aestivum) with reduced levels of omega-5 gliadins, the major sensitizing allergen in wheat-dependent exercise-induced anaphylaxis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The omega-5 gliadins are the major sensitizing allergens in wheat-dependent exercise-induced anaphylaxis (WDEIA). In this study, two-dimensional immunoblot analysis was used to assess the allergenic potential of two transgenic wheat lines in which omega-5 gliadin genes were silenced by RNA interfe...

  8. Identification of tropomyosins as major allergens in antarctic krill and mantis shrimp and their amino acid sequence characteristics.

    PubMed

    Motoyama, Kanna; Suma, Yota; Ishizaki, Shoichiro; Nagashima, Yuji; Lu, Ying; Ushio, Hideki; Shiomi, Kazuo

    2008-01-01

    Tropomyosin represents a major allergen of decapod crustaceans such as shrimps and crabs, and its highly conserved amino acid sequence (>90% identity) is a molecular basis of the immunoglobulin E (IgE) cross-reactivity among decapods. At present, however, little information is available about allergens in edible crustaceans other than decapods. In this study, the major allergen in two species of edible crustaceans, Antarctic krill Euphausia superba and mantis shrimp Oratosquilla oratoria that are taxonomically distinct from decapods, was demonstrated to be tropomyosin by IgE-immunoblotting using patient sera. The cross-reactivity of the tropomyosins from both species with decapod tropomyosins was also confirmed by inhibition IgE immunoblotting. Sequences of the tropomyosins from both species were determined by complementary deoxyribonucleic acid cloning. The mantis shrimp tropomyosin has high sequence identity (>90% identity) with decapod tropomyosins, especially with fast-type tropomyosins. On the other hand, the Antarctic krill tropomyosin is characterized by diverse alterations in region 13-42, the amino acid sequence of which is highly conserved for decapod tropomyosins, and hence, it shares somewhat lower sequence identity (82.4-89.8% identity) with decapod tropomyosins than the mantis shrimp tropomyosin. Quantification by enzyme-linked immunosorbent assay revealed that Antarctic krill contains tropomyosin at almost the same level as decapods, suggesting that its allergenicity is equivalent to decapods. However, mantis shrimp was assumed to be substantially not allergenic because of the extremely low content of tropomyosin. PMID:18521668

  9. Expression of the major mugwort pollen allergen Art v 1 in tobacco plants and cell cultures: problems and perspectives for allergen production in plants.

    PubMed

    Siegert, Marc; Pertl-Obermeyer, Heidi; Gadermaier, Gabriele; Ferreira, Fatima; Obermeyer, Gerhard

    2012-03-01

    An economic and cheap production of large amounts of recombinant allergenic proteins might become a prerequisite for the common use of microarray-based diagnostic allergy assays which allow a component-specific diagnosis. A molecular pharming strategy was applied to express the major allergen of Artemisia vulgaris pollen, Art v 1, in tobacco plants and tobacco cell cultures. The original Art v 1 with its endogenous signal peptide which directs Art v 1 to the secretory pathway, was expressed in transiently transformed tobacco leaves but was lost in stable transformed tobacco plants during the alternation of generations. Using a light-regulated promoter and "hiding" the recombinant Art v 1 in the ER succeeded in expression of Art v 1 over three generations of tobacco plants and in cell cultures generated from stable transformed plants. However, the amounts of the recombinant allergen were sufficient for analysis but not high enough to allow an economic production. Although molecular pharming has been shown to work well for the production of non-plant therapeutic proteins, it might be less efficient for closely related plant proteins.

  10. Expression of the major mugwort pollen allergen Art v 1 in tobacco plants and cell cultures: problems and perspectives for allergen production in plants.

    PubMed

    Siegert, Marc; Pertl-Obermeyer, Heidi; Gadermaier, Gabriele; Ferreira, Fatima; Obermeyer, Gerhard

    2012-03-01

    An economic and cheap production of large amounts of recombinant allergenic proteins might become a prerequisite for the common use of microarray-based diagnostic allergy assays which allow a component-specific diagnosis. A molecular pharming strategy was applied to express the major allergen of Artemisia vulgaris pollen, Art v 1, in tobacco plants and tobacco cell cultures. The original Art v 1 with its endogenous signal peptide which directs Art v 1 to the secretory pathway, was expressed in transiently transformed tobacco leaves but was lost in stable transformed tobacco plants during the alternation of generations. Using a light-regulated promoter and "hiding" the recombinant Art v 1 in the ER succeeded in expression of Art v 1 over three generations of tobacco plants and in cell cultures generated from stable transformed plants. However, the amounts of the recombinant allergen were sufficient for analysis but not high enough to allow an economic production. Although molecular pharming has been shown to work well for the production of non-plant therapeutic proteins, it might be less efficient for closely related plant proteins. PMID:22159963

  11. Conversion of Der p 23, a New Major House Dust Mite Allergen, into a Hypoallergenic Vaccine

    PubMed Central

    Banerjee, Srinita; Weber, Milena; Blatt, Katharina; Swoboda, Ines; Focke-Tejkl, Margit; Valent, Peter; Valenta, Rudolf; Vrtala, Susanne

    2015-01-01

    Der p 23, a new, major house dust mite (HDM) allergen that is recognized by >70% of HDM-allergic patients, has high allergenic activity and, therefore, must be considered an important component for HDM-specific immunotherapy. We constructed and characterized a hypoallergenic Der p 23 vaccine for HDM immunotherapy. Three nonallergenic peptides from the C-terminal IgE epitope-containing part of Der p 23 (P4, P5) and P6, a mutant peptide containing serines instead of cysteines, were identified. Peptides were fused to the hepatitis B virus–derived PreS domain as recombinant fusion proteins (i.e., PreS-2XP4P5 and PreS-4XP6) that were expressed in Escherichia coli and purified to homogeneity. Compared with Der p 23, PreS-2XP4P5 and PreS-4XP6 showed no relevant IgE reactivity and exhibited considerably reduced allergenic activity in basophil activation tests using blood from HDM-allergic patients. Upon immunization of rabbits, only PreS-2XP4P5 induced high levels of Der p 23–specific IgG Abs that inhibited binding of patients’ IgE to Der p 23, comparable to IgG Abs induced with Der p 23, whereas Abs induced with PreS-4XP6 had only low blocking capacity. Additionally, IgG Abs induced with PreS-2XP4P5 inhibited Der p 23–induced basophil activation comparable to IgG Abs induced with Der p 23. Compared with Der p 23, PreS-2XP4P5 induced lower T cell proliferation but higher levels of the tolerogenic cytokine IL-10 and the Th1 cytokine IFN-γ in PBMCs from HDM-allergic patients, indicating an immunomodulatory capacity of the fusion protein. Therefore, PreS-2XP4P5 represents a promising candidate for immunotherapy of HDM-allergic patients. PMID:24733847

  12. Molecular characterization of major allergens Ara h 1, 2, 3 in peanut seed.

    PubMed

    Jiang, Shengjuan; Wang, Songhua; Sun, Yujun; Zhou, Zhengyi; Wang, Guiqin

    2011-06-01

    Peanut is among the most commonly used dietary seeds, but peanut allergens, especially Ara h 1 (Arachis hypogaea allergy 1), 2 and 3, can cause severe IgE-mediated reactions. In this study, the molecular characterization and expression pattern of three allergens in peanut LUHUA 8, the representative of the cultivated lines in China, are reported. In situ hybridization and real time PCR analysis revealed high expression levels and different tissue expression patterns of the three allergens, which might be connected with many aspects, such as the strong conservation of intron phase of the allergen genes, the low energy of the mRNA's regions, and the complicated post-translational modifications. Furthermore, the different sequences between the cloned allergens and the reported sequences previously involved the charged amino acids especially in IgE epitopes, which might alter specific physicochemical and physiological properties, and thus influence the immunity of the allergens. The identification of the specific features of the allergen genes would be of considerable importance to the basic understanding of the specific characteristics of peanut seed allergens.

  13. Standardization of RP-HPLC methods for the detection of the major peanut allergens Ara h 1, Ara h 2 and Ara h 3.

    PubMed

    Singh, Harmit; Cantoria, Mary Jo; Malave, Poonam; Saputra, Denny; Maleki, Soheila

    2016-03-01

    Crude peanut extract (CPE) was analyzed for three major allergens (Ara h 1, h 2, and h 3) using a C12 and a C18 column at two wavelengths (280 and 220nm) and under different solvent conditions. HPLC profiles were compared for retention time, resolution, and peak heights. CPE samples were spiked with pure allergens to identify the peaks corresponding to allergens. The HPLC fractions of corresponding allergens were collected and freeze-dried in order to perform SDS-PAGE and immunoblotting tests. The best method was identified the one with a shorter retention time, better resolution, and greater peak height as compared with the other methods. In general, the peak heights were greater at 220nm than at 280nm. The major disadvantage of the C12 column was the need for two sets of conditions to identify the allergens as compared to the C18 column where all three allergens could be identified in one run. PMID:26471570

  14. Standardization of RP-HPLC methods for the detection of the major peanut allergens Ara h 1, Ara h 2 and Ara h 3.

    PubMed

    Singh, Harmit; Cantoria, Mary Jo; Malave, Poonam; Saputra, Denny; Maleki, Soheila

    2016-03-01

    Crude peanut extract (CPE) was analyzed for three major allergens (Ara h 1, h 2, and h 3) using a C12 and a C18 column at two wavelengths (280 and 220nm) and under different solvent conditions. HPLC profiles were compared for retention time, resolution, and peak heights. CPE samples were spiked with pure allergens to identify the peaks corresponding to allergens. The HPLC fractions of corresponding allergens were collected and freeze-dried in order to perform SDS-PAGE and immunoblotting tests. The best method was identified the one with a shorter retention time, better resolution, and greater peak height as compared with the other methods. In general, the peak heights were greater at 220nm than at 280nm. The major disadvantage of the C12 column was the need for two sets of conditions to identify the allergens as compared to the C18 column where all three allergens could be identified in one run.

  15. Changes in phenols contents from buckwheat sprouts during growth stage.

    PubMed

    Koyama, Masahiro; Nakamura, Chiho; Nakamura, Kozo

    2013-02-01

    Germinated buckwheat is buckwheat seeds soaked in water just until it begins to bud. Buckwheat sprouts are seedling plants of buckwheat grown up to 10-15 cm. The purpose of this study was to determine the optimal growth period for accumulating the most abundant functional phenol(s) in germinated buckwheat that had been soaked in darkness and buckwheat sprouts cultivated by hydroponic culture. The rutin contained in germinated buckwheat was analyzed by CE (capillary electrophoresis). Phenols, including isoorientin, orientin, isovitexin, vitexin, and rutin were separated from buckwheat sprouts by HPLC and identified by LC-MS. The highest rutin content in germinated buckwheat was found to be 15.8 mg/100 g DW at 20 h after germination. Buckwheat sprouts contained five kinds of major phenols. The highest amounts of isoorientin, orientin, isovitexin, and vitexin were measured at day 3, with the exception of rutin, and then a gradual decrease was observed as the sprouts grew. The quantities of isoorientin, orientin, isovitexin, and vitexin at day 3 were 5.8, 11.7, 26.2, and 28.9 mg/100 g FW, respectively. The rutin content rapidly increased to 109.0 mg/100 g FW until day 6. The highest total phenols in buckwheat sprouts were 162.9 mg/100 g FW at day 6. Germinated buckwheat soaked for 20 h and buckwheat sprouts cultivated for 6 days were rich in dietary phenol(s), which makes these plants a valuable functional food for human consumption. PMID:24425891

  16. Comparative identification of protein profiles and major allergens of saliva, salivary gland and whole body extracts of mosquito species in Thailand.

    PubMed

    Wongkamchai, Sirichit; Khongtak, Pacharee; Leemingsawat, Somjai; Komalamisra, Narumon; Junsong, Nujorn; Kulthanan, Kanokvalai; Wisuthsarewong, Wanee; Boitano, John J

    2010-01-01

    Allergic reactions to mosquito bites, such as generalized urticaria or severe local reactions are common problems worldwide. The diverse sources of allergen prepared from different mosquito body parts usage are a major obstacle to obtaining safe and effective tests and immunotherapy for mosquito bite allergy. Thus, the reactions are often not recognized and allergen immunotherapy is seldom used for severe reaction to mosquito bites. In a search for appropriate allergen sources, the protein profiles of saliva, salivary glands and whole body extracts were comparatively analyzed from 4 common mosquito species of Thailand and/or South East Asia; viz. Culex quinquefasciatus, Aedes aegypti, Aedes albopictus and a zoophilic strain, Anopheles minimus. The major allergens in the extracts which elicited specific IgE responses in the pooled sera of subjects allergic to mosquito bites were identified. It was concluded that mosquito saliva was the best source of allergens. Additionally, both species-specific and species-shared allergens of the 4 mosquito species were identified. The major saliva allergens having MWs of 36, 32 and 22 kDa were identified. The identificstion of major allergens should facilitate the production of specific recombinant allergens and contribute to improvement in the diagnosis and specific immunotherapy of Thai mosquito bite allergy patients.

  17. Identification of major allergens of Malassezia pachydermatis in dogs with atopic dermatitis and Malassezia overgrowth.

    PubMed

    Chen, Tai-An; Halliwell, Richard E W; Pemberton, Alan D; Hill, Peter B

    2002-06-01

    We have previously shown that both atopic and normal dogs generate an IgG response to antigens of Malassezia pachydermatis. The aim of this study was to compare IgE responses to separated proteins of M. pachydermatis in 28 atopic dogs with Malassezia dermatitis and 22 clinically normal dogs using Western immunoblotting. Six different detection systems were evaluated in order to assess sensitivity and eliminate nonspecific binding and cross-reactivity. The protocol yielding the best results utilized a monoclonal mouse antidog IgE, an alkaline phosphatase conjugated goat antimouse IgG which had been passed through a canine IgG column 3 times, a chemiluminescent substrate and a digital imaging system. Proteins of 45, 52, 56 and 63 kDa were recognized by more than 50% of the atopic dog sera and thus represented major allergens. Only a minority of normal dogs showed faint IgE binding to these proteins. The results indicate that the majority of atopic dogs with Malassezia dermatitis have a greater IgE response than normal dogs, suggesting an IgE-mediated immune response may be clinically important in the pathogenesis of the disease. PMID:12074703

  18. Effects of adjuvants on the immune response to allergens in a murine model of allergen inhalation: cholera toxin induces a Th1-like response to Bet v 1, the major birch pollen allergen.

    PubMed

    Wiedermann, U; Jahn-Schmid, B; Fritsch, R; Bauer, L; Renz, H; Kraft, D; Ebner, C

    1998-01-01

    Based on the fact that type I allergies are frequently elicited by inhalant allergens, we have established a model of aerosol inhalation leading to allergic sensitization in BALB/c mice. Using this model we studied the effects of aluminium hydroxide (Al(OH)3), known to enhance IgE antibody responses, compared with cholera toxin (CT), a potent mucosal adjuvant, on the immune response to birch pollen (BP) and its major allergen Bet v 1. Two groups of BALB/c mice were either systemically immunized with recombinant Bet v 1 in Al(OH)3 and subsequently aerosol exposed to BP allergen, or aerosolized with BP and CT. IgE-mediated skin reactions were only elicited in the mice which had received Bet v 1/Al(OH)3. Allergen-specific serum IgE and IgG1 antibodies dominated in the Al(OH)3 group, IgG2a antibody levels to BP and rBet v 1 were markedly higher in the sera of mice exposed to CT with the allergen. IgA antibodies were only detected in the bronchial lavage of the CT-treated group. Moreover, the latter group displayed consistently higher T cell proliferative responses to BP and interferon-gamma production in vitro. Thus, the systemic immunization with rBet v 1 in Al(OH)3 before inhalation of the BP extract promoted a Th2-like immune response, while CT mixed with the aerosolized BP extract rather induced a Th1-like immune response. In an attempt to reverse these ongoing immune responses we could achieve a shift towards a Th0 response. Immunization with BP extract without adjuvant treatment led to undetectable antibody or cellular immune responses. We conclude from the present study that the induction of an immune response to BP allergen after aerosol inhalation can be directed towards a Th1- or a Th2-like response. Once established, the immune response can be modulated.

  19. 27 CFR 4.32a - Voluntary disclosure of major food allergens.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... allergen means any of the following: (i) Milk, egg, fish (for example, bass, flounder, or cod), Crustacean..., or walnuts); (ii) In the case of Crustacean shellfish, it means the name of the species of...

  20. 27 CFR 4.32a - Voluntary disclosure of major food allergens.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... allergen means any of the following: (i) Milk, egg, fish (for example, bass, flounder, or cod), Crustacean..., or walnuts); (ii) In the case of Crustacean shellfish, it means the name of the species of...

  1. 27 CFR 4.32a - Voluntary disclosure of major food allergens.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... allergen means any of the following: (i) Milk, egg, fish (for example, bass, flounder, or cod), Crustacean..., or walnuts); (ii) In the case of Crustacean shellfish, it means the name of the species of...

  2. 27 CFR 4.32a - Voluntary disclosure of major food allergens.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... allergen means any of the following: (i) Milk, egg, fish (for example, bass, flounder, or cod), Crustacean..., or walnuts); (ii) In the case of Crustacean shellfish, it means the name of the species of...

  3. 27 CFR 4.32a - Voluntary disclosure of major food allergens.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... allergen means any of the following: (i) Milk, egg, fish (for example, bass, flounder, or cod), Crustacean..., or walnuts); (ii) In the case of Crustacean shellfish, it means the name of the species of...

  4. Rubber Elongation Factor (REF), a Major Allergen Component in Hevea brasiliensis Latex Has Amyloid Properties

    PubMed Central

    Berthelot, Karine; Lecomte, Sophie; Estevez, Yannick; Coulary-Salin, Bénédicte; Bentaleb, Ahmed; Cullin, Christophe; Deffieux, Alain; Peruch, Frédéric

    2012-01-01

    REF (Hevb1) and SRPP (Hevb3) are two major components of Hevea brasiliensis latex, well known for their allergenic properties. They are obviously taking part in the biosynthesis of natural rubber, but their exact function is still unclear. They could be involved in defense/stress mechanisms after tapping or directly acting on the isoprenoid biosynthetic pathway. The structure of these two proteins is still not described. In this work, it was discovered that REF has amyloid properties, contrary to SRPP. We investigated their structure by CD, TEM, ATR-FTIR and WAXS and neatly showed the presence of β-sheet organized aggregates for REF, whereas SRPP mainly fold as a helical protein. Both proteins are highly hydrophobic but differ in their interaction with lipid monolayers used to mimic the monomembrane surrounding the rubber particles. Ellipsometry experiments showed that REF seems to penetrate deeply into the monolayer and SRPP only binds to the lipid surface. These results could therefore clarify the role of these two paralogous proteins in latex production, either in the coagulation of natural rubber or in stress-related responses. To our knowledge, this is the first report of an amyloid formed from a plant protein. This suggests also the presence of functional amyloid in the plant kingdom. PMID:23133547

  5. Rubber elongation factor (REF), a major allergen component in Hevea brasiliensis latex has amyloid properties.

    PubMed

    Berthelot, Karine; Lecomte, Sophie; Estevez, Yannick; Coulary-Salin, Bénédicte; Bentaleb, Ahmed; Cullin, Christophe; Deffieux, Alain; Peruch, Frédéric

    2012-01-01

    REF (Hevb1) and SRPP (Hevb3) are two major components of Hevea brasiliensis latex, well known for their allergenic properties. They are obviously taking part in the biosynthesis of natural rubber, but their exact function is still unclear. They could be involved in defense/stress mechanisms after tapping or directly acting on the isoprenoid biosynthetic pathway. The structure of these two proteins is still not described. In this work, it was discovered that REF has amyloid properties, contrary to SRPP. We investigated their structure by CD, TEM, ATR-FTIR and WAXS and neatly showed the presence of β-sheet organized aggregates for REF, whereas SRPP mainly fold as a helical protein. Both proteins are highly hydrophobic but differ in their interaction with lipid monolayers used to mimic the monomembrane surrounding the rubber particles. Ellipsometry experiments showed that REF seems to penetrate deeply into the monolayer and SRPP only binds to the lipid surface. These results could therefore clarify the role of these two paralogous proteins in latex production, either in the coagulation of natural rubber or in stress-related responses. To our knowledge, this is the first report of an amyloid formed from a plant protein. This suggests also the presence of functional amyloid in the plant kingdom.

  6. Monoclonal antibodies to the major Lolium perenne (rye grass) pollen allergen Lol p I (Rye I).

    PubMed

    Kahn, C R; Marsh, D G

    1986-12-01

    Thirteen monoclonal antibodies (MAbs) were produced against Lol p I (Rye I), the major Lolium perenne (rye grass) pollen allergen. Spleen cells from A/J and SJL mice immunized with highly purified Lol p I (Lol I) were allowed to fuse with cells from the non-secreting Sp2/0-Ag14 myeloma cell line. Each MAb was analyzed for antigenic specificity by radioimmunoassay (RIA) using 125I-Lol I. The epitope specificities of seven of the MAbs were examined by competitive binding against a labelled standard MAb for the Lol I antigen (Ag). The dissociation constant, Kd, of one MAb (No. 3.2) that was studied most extensively was determined by double Ab RIA to be 3.5 X 10(-6) L/M. This MAb recognized the related 27,000-30,000 Group I glycoproteins found in the pollens of nine other species of grass pollens tested, including weak binding to Bermuda grass Group I (Cyn d I), which by conventional analysis using polyclonal anti-Lol I serum shows no detectable binding. Monoclonal antibody No. 3.2 was coupled covalently to Sepharose 4B and used to prepare highly purified Lol I from a partially purified rye pollen extract. Finally, an RIA was developed which permitted the analysis of the Group I components in rye grass and nine other grass pollen species. The latter assay is likely to prove useful in the standardization of grass pollen extracts according to their Group I contents.

  7. Multiple IgE recognition on the major allergen of the Parietaria pollen Par j 2.

    PubMed

    Longo, Valeria; Costa, Maria Assunta; Cibella, Fabio; Cuttitta, Giuseppina; La Grutta, Stefania; Colombo, Paolo

    2015-02-01

    The interaction between IgE antibodies and allergens is a key event in triggering an allergic reaction. The characterization of this region provides information of paramount importance for diagnosis and therapy. Par j 2 Lipid Transfer Protein is one of the most important allergens in southern Europe and a well-established marker of sensitization in Parietaria pollen allergy. The main aim of this study was to map the IgE binding regions of this allergen and to study the pattern of reactivity of individual Parietaria-allergic patients. By means of gene fragmentation, six overlapping peptides were expressed in Escherichia coli, and their IgE binding activity was evaluated by immunoblotting in a cohort of 79 Parietaria-allergic patients. Our results showed that Pj-allergic patients display a heterogeneous pattern of IgE binding to the different recombinant fragments, and that patients reacted simultaneously against several protein domains spread all the over the molecule, even in fragments which do not contain structural features resembling the native allergen. Our results reveal the presence of a large number of linear and conformational epitopes on the Par j 2 sequence, which probably explains the high allergenic activity of this allergen.

  8. Identification of Der p 23, a peritrophin-like protein, as a new major Dermatophagoides pteronyssinus allergen associated with the peritrophic matrix of mite fecal pellets.

    PubMed

    Weghofer, Margit; Grote, Monika; Resch, Yvonne; Casset, Anne; Kneidinger, Michael; Kopec, Jolanta; Thomas, Wayne R; Fernández-Caldas, Enrique; Kabesch, Michael; Ferrara, Rosetta; Mari, Adriano; Purohit, Ashok; Pauli, Gabrielle; Horak, Friedrich; Keller, Walter; Valent, Peter; Valenta, Rudolf; Vrtala, Susanne

    2013-04-01

    The house dust mite (HDM) Dermatophagoides pteronyssinus is one of most important allergen sources and a major elicitor of allergic asthma. We screened a D. pteronyssinus expression cDNA library with IgE Abs from HDM allergic patients. A cDNA coding for a new major allergen was isolated, which showed sequence homology to peritrophins, which contain chitin-binding domains and are part of the peritrophic matrix lining the gut of arthropods. The mature Der p 23 allergen was expressed in Escherichia coli as an 8-kDa protein without its hydrophobic leader sequence and purified to homogeneity. It reacted with IgE Abs from 74% of D. pteronyssinus allergic patients (n = 347) at levels comparable to the two major HDM allergens, Der p 1 and Der p 2. Thus, Der p 23 represents a new major D. pteronyssinus allergen. Furthermore, rDer p 23 exhibited high allergenic activity as demonstrated by upregulation of CD203c expression on basophils from D. pteronyssinus allergic patients. Immunogold electron microscopy localized the allergen in the peritrophic matrix lining the midgut of D. pteronyssinus as well as on the surface of the fecal pellets. Thus, we identified a new major D. pteronyssinus allergen as peritrophin-like protein. The high allergenic activity of Der p 23 and its frequent recognition as respiratory allergen may be explained by the fact that it becomes airborne and respirable through its association with mite feces. Der p 23 may be an essential component for diagnosis and specific immunotherapy of HDM allergy. PMID:23460742

  9. Specific B-cell Epitope of Per a 1: A Major Allergen of American Cockroach (Periplaneta americana) and Anatomical Localization

    PubMed Central

    Sookrung, Nitat; Khetsuphan, Thanyathon; Chaisri, Urai; Indrawattana, Nitaya; Reamtong, Onrapak; Chaicumpa, Wanpen

    2014-01-01

    Purpose Cockroach (CR) is a common source of indoor allergens, and Per a 1 is a major American CR (Periplaneta americana) allergen; however, several attributes of this protein remain unknown. This study identifies a novel specific B cell epitope and anatomical locations of Per a 1.0105. Methods Recombinant Per a 1.0105 (rPer a 1.0105) was used as BALB/c mouse immunogen for the production of monoclonal antibodies (MAb). The MAb specific B cell epitope was identified by determining phage mimotopic peptides and pair-wise alignment of the peptides with the rPer a 1.0105 amino acid sequence. Locations of the Per a 1.0105 in P. americana were investigated by immunohistochemical staining. Results The rPer a 1.0105 (~13 kDa) had 100%, 98% and ≥90% identity to Per a 1.0105, Per a 1.0101, and Cr-PII, respectively. The B-cell epitope of the Per a 1.0105 specific-MAb was located at residues99 QDLLLQLRDKGV110 contained in all 5 Per a 1.01 isoforms and Per a 1.02. The epitope was analogous to the Bla g 1.02 epitope; however, this B-cell epitope was not an IgE inducer. Per a 1.0105 was found in the midgut and intestinal content of American CR but not in the other organs. The amount of the Per a 1 was ~544 ℃g per gram of feces. Conclusions The novel Per a 1 B-cell epitope described in this study is a useful target for allergen quantification in samples; however, the specific MAb can be used as an allergen detection reagent. The MAb based-affinity resin can be made for allergen purification, and the so-purified protein can serve as a standard and diagnostic allergen as well as a therapeutic vaccine component. The finding that the Per a 1 is contained in the midgut and feces is useful to increase yield and purity when preparing this allergen. PMID:24991456

  10. Purification, crystallization and initial crystallographic characterization of peanut major allergen Ara h 3

    SciTech Connect

    Jin, Tengchuan; Howard, Andrew; Zhang, Yu-Zhu

    2007-10-01

    The crystallization of peanut allergen Ara h 3 is reported. The peanut is a significant food source, but is responsible for many cases of anaphylaxis. The peanut 11S legumin-like seed storage protein Ara h 3 is one of the best characterized allergens. In this study, Ara h 3 was extracted from peanut kernels and purified by sequential anion-exchange, hydrophobic interaction and gel-filtration chromatography to very high purity to facilitate crystallization and structural studies. Well diffracting single crystals were obtained by the vapor-diffusion method. A molecular-replacement structural solution has been obtained and refinement of the structure is currently under way.

  11. Cloning and sequencing of Lol pI, the major allergenic protein of rye-grass pollen.

    PubMed

    Griffith, I J; Smith, P M; Pollock, J; Theerakulpisut, P; Avjioglu, A; Davies, S; Hough, T; Singh, M B; Simpson, R J; Ward, L D

    1991-02-25

    We have isolated a full length cDNA clone encoding the major glycoprotein allergen Lol pI. The clone was selected using a combination of immunological screening of a cDNA expression library and PCR amplification of Lol pI-specific transcripts. Lol pI expressed in bacteria as a fusion protein shows recognition by specific IgE antibodies present in sera of grass pollen-allergic subjects. Northern analysis has shown that the Lol pI transcripts are expressed only in pollen of rye-grass. Molecular cloning of Lol pI provides a molecular genetic approach to study the structure-function relationship of allergens.

  12. Crystallization and preliminary X-ray analysis of the major peanut allergen Ara h 1 core region

    PubMed Central

    Cabanos, Cerrone; Urabe, Hiroyuki; Masuda, Taro; Tandang-Silvas, Mary Rose; Utsumi, Shigeru; Mikami, Bunzo; Maruyama, Nobuyuki

    2010-01-01

    Peanuts contain some of the most potent food allergens known to date. Ara h 1 is one of the three major peanut allergens. As a first step towards three-dimensional structure elucidation, recombinant Ara h 1 core region was cloned, expressed in Escherichia coli and purified to homogeneity. Crystals were obtained using 0.1 M sodium citrate pH 5.6, 0.1 M NaCl, 15% PEG 400 as precipitant. The crystals diffracted to 2.25 Å resolution using synchrotron radiation and belonged to the monoclinic space group C2, with unit-cell parameters a = 156.521, b = 88.991, c = 158.971 Å, β = 107.144°. Data were collected at the BL-38B1 station of SPring-8 (Hyogo, Japan). PMID:20823529

  13. Phytochemical and Pharmacological Profiles of Three Fagopyrum Buckwheats.

    PubMed

    Jing, Rui; Li, Hua-Qiang; Hu, Chang-Ling; Jiang, Yi-Ping; Qin, Lu-Ping; Zheng, Cheng-Jian

    2016-04-19

    The genus Fagopyrum (Polygonaceae), currently comprising 15 species of plants, includes three important buckwheat species: Fagopyrum esculentum (F. esculentum) Moench. (common buckwheat), Fagopyrum tataricum (F. tataricum) (L.) Gaertn. (tartary buckwheat) and Fagopyrum dibotrys (F. dibotrys) (D. Don) Hara. (perennial buckwheat), which have been well explored due to their long tradition of both edible and medicinal use. This review aimed to present an up-to-date and comprehensive analysis of the phytochemistry and pharmacology of the three Fagopyrum buckwheats. In addition, the scope for future research was also discussed. All available references included in this paper were compiled from major databases, such as MEDLINE, Pubmed, Scholar, Elsevier, Springer, Wiley and CNKI. A total of 106 compounds isolated from three Fagopyrum buckwheats can be mainly divided into six classes: flavonoids, phenolics, fagopyritols, triterpenoids, steroids and fatty acids. Flavonoids and phenolic compounds were considered to be the major active components. Considerable pharmacological experiments both in vitro and in vivo have validated that Fagopyrum buckwheats possess antitumor, anti-oxidant, anti-inflammatory, hepatoprotective, anti-diabetic activities, etc. All reported data lead us to conclude that Fagopyrum buckwheats have convincing medicinal potential. However, further research is needed to explore its bioactive constituents, the relationship to their structural activities and the molecular mechanisms of action.

  14. Phytochemical and Pharmacological Profiles of Three Fagopyrum Buckwheats

    PubMed Central

    Jing, Rui; Li, Hua-Qiang; Hu, Chang-Ling; Jiang, Yi-Ping; Qin, Lu-Ping; Zheng, Cheng-Jian

    2016-01-01

    The genus Fagopyrum (Polygonaceae), currently comprising 15 species of plants, includes three important buckwheat species: Fagopyrum esculentum (F. esculentum) Moench. (common buckwheat), Fagopyrum tataricum (F. tataricum) (L.) Gaertn. (tartary buckwheat) and Fagopyrum dibotrys (F. dibotrys) (D. Don) Hara. (perennial buckwheat), which have been well explored due to their long tradition of both edible and medicinal use. This review aimed to present an up-to-date and comprehensive analysis of the phytochemistry and pharmacology of the three Fagopyrum buckwheats. In addition, the scope for future research was also discussed. All available references included in this paper were compiled from major databases, such as MEDLINE, Pubmed, Scholar, Elsevier, Springer, Wiley and CNKI. A total of 106 compounds isolated from three Fagopyrum buckwheats can be mainly divided into six classes: flavonoids, phenolics, fagopyritols, triterpenoids, steroids and fatty acids. Flavonoids and phenolic compounds were considered to be the major active components. Considerable pharmacological experiments both in vitro and in vivo have validated that Fagopyrum buckwheats possess antitumor, anti-oxidant, anti-inflammatory, hepatoprotective, anti-diabetic activities, etc. All reported data lead us to conclude that Fagopyrum buckwheats have convincing medicinal potential. However, further research is needed to explore its bioactive constituents, the relationship to their structural activities and the molecular mechanisms of action. PMID:27104519

  15. Molecular and immunological characterization and IgE epitope mapping of Pen n 18, a major allergen of Penicillium notatum.

    PubMed Central

    Yu, Chia-Jung; Chen, Yen-Ming; Su, Song-Nan; Forouhar, Farhad; Lee, Shu-Hua; Chow, Lu-Ping

    2002-01-01

    The mould genus, Penicillium, is a significant source of environmental aero-allergens. A major allergen from Penicillium notatum, Pen n 18, was identified by two-dimensional immunoblotting using monoclonal antibody G11A10, raised against the vacuolar serine protease of Penicillium citrinum, followed by matrix-assisted laser-desorption ionization-time-of-flight MS analysis of the peptide digest. Pen n 18 was then cloned and the amino acid sequence deduced from the cDNA sequence. The cDNA encoded a 494 amino acid protein, considerably larger than mature Pen n 18, the differences being due to the N- and C-terminal prosequences. The deduced amino acid sequence showed extensive similarity with those of vacuolar serine proteases from various fungi. The Pen n 18 coding sequence was expressed in Escherichia coli as a His-tagged fusion protein and purified by Ni(2+)-chelate affinity chromatography. On immunoblots, the purified recombinant protein specifically bound IgE from mould-allergic patients, and cross-inhibition assays demonstrated the presence of common IgE-binding epitopes on Pen n 18 and a major allergen of P. citrinum, Pen c 18. When mapping of the allergenic epitopes was performed, at least nine different linear IgE-binding epitopes, located throughout the Pen n 18 protein, were identified. Of these, peptide C12, located in the N-terminal region of the molecule, was recognized by serum from 75% of the patients tested and therefore appears to be an immunodominant IgE-binding epitope. PMID:11964171

  16. Competitive inhibition ELISA for quantification of Ara h 1 and Ara h 2, the major allergens of peanuts.

    PubMed

    Schmitt, David A; Cheng, Hsiaopo; Maleki, Soheila J; Burks, A Wesley

    2004-01-01

    Allergies to peanuts are becoming an increasingly important health problem as a result of the persistence and severity of the reaction in allergic individuals. Because no treatment currently is available, avoidance is the only option for peanut-allergic individuals. Avoidance of an abundant and often disguised food such as peanuts, however, is very difficult; therefore, competitive inhibition ELISAs were developed to detect and quantitate each of the major peanut allergens, Ara h 1 and Ara h 2. Under optimal conditions for each assay, the sensitivity of the Ara h 1 and Ara h 2 detection assays were 12 and 0.5 ng/mL, respectively. These assays were primarily devised to effectively compare the levels of Ara h 1 and Ara h 2 in a wide variety of peanuts or peanut products but can also be used to identify cross-reactive antigens. The method is simple and rapid, requiring only one allergen-specific antibody and, therefore, could be adapted specifically to detect the presence of these individual allergens in different foods.

  17. Characterization of a mouse model of allergy to a major occupational latex glove allergen Hev b 5.

    PubMed

    Hardy, Charles L; Kenins, Linda; Drew, Alexander C; Rolland, Jennifer M; O'Hehir, Robyn E

    2003-05-15

    Allergen-specific immunotherapy is a clinically proven effective treatment for many allergic diseases, including asthma; however, it is not currently available for latex allergy because of the high risk of anaphylaxis. There is, therefore, a crucial need for an animal model of latex allergy in which to develop effective immunotherapy. Previous mouse models of latex allergy either did not characterize the allergic pulmonary immune response or used crude latex extracts, making it difficult to quantify the contribution of individual proteins and limiting their usefulness for developing specific immunotherapy. We immunized mice with recombinant Hev b 5, a defined major latex allergen, or latex glove protein extract, representing the range of occupationally encountered processed latex allergens. The immune response was compared with that seen in ovalbumin-immunized mice. Immunization with Hev b 5 or glove extract elicits hallmarks of allergic pulmonary Th2-type immune responses, comparable to those for ovalbumin, including (1) serum antigen-specific IgE, (2) an eosinophilic inflammatory infiltrate in the lung, (3) increased interleukin-5 in lung bronchoalveolar lavage fluid, and (4) mucus hypersecretion by epithelial cells in the lung airways. This mouse model will aid the development of potentially curative treatments for latex-sensitized individuals, including those with occupational asthma.

  18. Comparison of six commercial ELISA kits for their specificity and sensitivity in detecting different major peanut allergens.

    PubMed

    Jayasena, Shyamali; Smits, Mieke; Fiechter, Daniëlle; de Jong, Aard; Nordlee, Julie; Baumert, Joe; Taylor, Steve L; Pieters, Raymond H; Koppelman, Stef J

    2015-02-18

    Six commercial peanut enzyme-linked immunosorbent assay kits were assessed for their ability to recover peanut from the standard reference material 2387 peanut butter and also for their specificity in detecting four major peanut allergens, Ara h 1, Ara h 2, Ara h 3, and Ara h 6. The percentage recovery of peanut from peanut butter differed across different kits as well as at different sample concentrations. The highest recovery was observed with the Romer and R-Biopharm kits, while four other kits were found to underestimate the protein content of the reference peanut butter samples. Five of the kits were most sensitive in detecting Ara h 3 followed by Ara h 1, while hardly recognizing Ara h 2 and Ara h 6. The other kit showed the highest sensitivity to Ara h 2 and Ara h 6, while Ara h 1 and Ara h 3 were poorly recognized. Although Ara h 2 and Ara h 6 are known to be heat stable and more potent allergens, antisera specific to any of these four peanut proteins/allergens may serve as good markers for the detection of peanut residues.

  19. The Major Soybean Allergen Gly m Bd 28K Induces Hypersensitivity Reactions in Mice Sensitized to Cow's Milk Proteins.

    PubMed

    Candreva, Ángela María; Smaldini, Paola Lorena; Curciarello, Renata; Fossati, Carlos Alberto; Docena, Guillermo Horacio; Petruccelli, Silvana

    2016-02-24

    Reactions to soy have been reported in a proportion of patients with IgE-mediated cow's milk allergy (CMA). In this work, we analyzed if Gly m Bd 28K/P28, one of the major soybean allergens, is a cross-reactive allergen with cow milk proteins (CMP). We showed that P28 was recognized by IgE sera from CMA patients and activated human peripheral basophils degranulation. Moreover, IgE sera of mice exclusively sensitized to CMP recognized P28. Splenocytes from sensitized animals secreted IL-5 and IL-13 when incubated with CMP or soy proteins, but only IL-13 when treated with P28. In addition, a skin test was strongly positive for CMP and weakly positive for P28. Remarkably, milk-sensitized mice showed hypersensitivity symptoms following sublingual challenge with P28 or CMP. With the use of bioinformatics' tools seven putative cross-reactive epitopes were identified. In conclusion, using in vitro and in vivo tests we demonstrated that P28 is a novel cross-reactive allergen with CMP.

  20. Comparison of six commercial ELISA kits for their specificity and sensitivity in detecting different major peanut allergens.

    PubMed

    Jayasena, Shyamali; Smits, Mieke; Fiechter, Daniëlle; de Jong, Aard; Nordlee, Julie; Baumert, Joe; Taylor, Steve L; Pieters, Raymond H; Koppelman, Stef J

    2015-02-18

    Six commercial peanut enzyme-linked immunosorbent assay kits were assessed for their ability to recover peanut from the standard reference material 2387 peanut butter and also for their specificity in detecting four major peanut allergens, Ara h 1, Ara h 2, Ara h 3, and Ara h 6. The percentage recovery of peanut from peanut butter differed across different kits as well as at different sample concentrations. The highest recovery was observed with the Romer and R-Biopharm kits, while four other kits were found to underestimate the protein content of the reference peanut butter samples. Five of the kits were most sensitive in detecting Ara h 3 followed by Ara h 1, while hardly recognizing Ara h 2 and Ara h 6. The other kit showed the highest sensitivity to Ara h 2 and Ara h 6, while Ara h 1 and Ara h 3 were poorly recognized. Although Ara h 2 and Ara h 6 are known to be heat stable and more potent allergens, antisera specific to any of these four peanut proteins/allergens may serve as good markers for the detection of peanut residues. PMID:25651402

  1. Genetic engineering of trimers of hypoallergenic fragments of the major birch pollen allergen, Bet v 1, for allergy vaccination.

    PubMed

    Vrtala, Susanne; Fohr, Monika; Campana, Raffaela; Baumgartner, Christian; Valent, Peter; Valenta, Rudolf

    2011-03-01

    An immunotherapy trial performed in allergic patients with hypoallergenic recombinant fragments, comprising aa 1-74 and 75-160 of the major birch pollen allergen, Bet v 1, has indicated that the induction of allergen-specific IgG responses may be an important mechanism of this treatment. To investigate whether the immunogenicity of the rBet v 1 fragments can be increased, recombinant trimers of the fragments were produced. For this purpose, DNA trimers of rBet v 1 aa 1-74 as well as of rBet v 1 aa 75-160 were subcloned into expression plasmid pET 17b, expressed in Escherichia coli and purified. The fragments as well as the fragment trimers showed a reduced IgE-binding capacity and allergenic activity compared to rBet v 1 wildtype when tested in allergic patients. Both rBet v 1 aa 75-160 monomer and trimer induced high titers of allergen-specific IgG1 Abs in mice. Interestingly, rBet v 1 aa 1-74 trimer induced a much higher IgG(1) response to rBet v 1 than rBet v 1 aa 1-74 monomer. Consequently, IgG Abs induced with the rBet v 1 aa 1-74 trimer inhibited birch pollen allergic patients' IgE-binding 10-fold more efficiently than IgG Abs induced with the monomer. Our data show that the immunogenicity of allergy vaccines can be increased by oligomerization.

  2. Solution Structure, Copper Binding and Backbone Dynamics of Recombinant Ber e 1–The Major Allergen from Brazil Nut

    PubMed Central

    Rundqvist, Louise; Tengel, Tobias; Zdunek, Janusz; Björn, Erik; Schleucher, Jürgen; Alcocer, Marcos J. C.; Larsson, Göran

    2012-01-01

    Background The 2S albumin Ber e 1 is the major allergen in Brazil nuts. Previous findings indicated that the protein alone does not cause an allergenic response in mice, but the addition of components from a Brazil nut lipid fraction were required. Structural details of Ber e 1 may contribute to the understanding of the allergenic properties of the protein and its potential interaction partners. Methodology/Principal Findings The solution structure of recombinant Ber e 1 was solved using NMR spectroscopy and measurements of the protein back bone dynamics at a residue-specific level were extracted using 15N-spin relaxation. A hydrophobic cavity was identified in the structure of Ber e 1. Using the paramagnetic relaxation enhancement property of Cu2+ in conjunction with NMR, it was shown that Ber e 1 is able to specifically interact with the divalent copper ion and the binding site was modeled into the structure. The IgE binding region as well as the copper binding site show increased dynamics on both fast ps-ns timescale as well as slower µs-ms timescale. Conclusions/Significance The overall fold of Ber e 1 is similar to other 2S albumins, but the hydrophobic cavity resembles that of a homologous non-specific lipid transfer protein. Ber e 1 is the first 2S albumin shown to interact with Cu2+ ions. This Cu2+ binding has minimal effect on the electrostatic potential on the surface of the protein, but the charge distribution within the hydrophobic cavity is significantly altered. As the hydrophobic cavity is likely to be involved in a putative lipid interaction the Cu2+ can in turn affect the interaction that is essential to provoke an allergenic response. PMID:23056307

  3. Immunological cross-reactivity of the major allergen from perennial ryegrass (Lolium perenne), Lol p I, and the cysteine proteinase, bromelain.

    PubMed

    Pike, R N; Bagarozzi, D; Travis, J

    1997-04-01

    Antibodies prepared in rabbits against the major allergen from ryegrass (Lolium perenne), Lol p I, cross-reacted with the cysteine proteinase bromelain from pineapple and vice versa. Deglycosylation of the proteins showed that the cross-reaction was based on recognition of the carbohydrate moiety of the allergen, but for bromelain the cross-reaction was most likely due to a combination of factors. The results indicate that the carbohydrate residues from these allergens play an important role in cross-reactions found between them and possibly those from other species.

  4. Antioxidant activity of tartary (Fagopyrum tataricum (L.) Gaertn.) and common (Fagopyrum esculentum moench) buckwheat sprouts.

    PubMed

    Liu, Chia-Ling; Chen, Yih-Shyuan; Yang, Joan-Hwa; Chiang, Been-Huang

    2008-01-01

    This study compared the differences of two types of buckwheat sprouts, namely, common buckwheat ( Fagopyrum esculentum Moench) and tartary buckwheat ( Fagopyrum tataricum (L.) Gaertn.), in general composition, functional components, and antioxidant capacity. The ethanol extracts of tartary buckwheat sprouts (TBS) had higher reducing power, free radical scavenging activity, and superoxide anion scavenging activity than those of common buckwheat sprouts (CBS). As for chelating effects on ferrous ions, CBS had higher values than TBS. Rutin was the major flavonoid found in these two types of buckwheat sprouts, and TBS was 5 fold higher in rutin than CBS. The antioxidant effects of buckwheat sprouts on human hepatoma HepG2 cells revealed that both of TBS and CBS could decrease the production of intracellular peroxide and remove the intracellular superoxide anions in HepG2 cells, but TBS reduced the cellular oxidative stress more effectively than CBS, possibly because of its higher rutin (and quercetin) content.

  5. New tree nut allergens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The 7S vicilin and 11S legumin seed storage globulins belong to the cupin protein superfamily and are major food allergens in many of the “big eight” food allergen groups. Korean pine vicilin and pecan vicilin are thus predicted to be food allergens. Recombinant vicilins were expressed in E. coli an...

  6. Cloning and characterization of an 11S legumin, Car i 4, a major allergen in pecan.

    PubMed

    Sharma, Girdhari M; Irsigler, Andre; Dhanarajan, Pushparani; Ayuso, Rosalia; Bardina, Luda; Sampson, Hugh A; Roux, Kenneth H; Sathe, Shridhar K

    2011-09-14

    Among tree nut allergens, pecan allergens remain to be identified and characterized. The objective was to demonstrate the IgE-binding ability of pecan 11S legumin and characterize its sequential IgE-binding epitopes. The 11S legumin gene was amplified from a pecan cDNA library and expressed as a fusion protein in Escherichia coli. The native 11S legumin in pecan extract was identified by mass spectrometry/mass spectrometry (MS/MS). Sequential epitopes were determined by probing the overlapping peptides with three serum pools prepared from different patients' sera. A three-dimensional model was generated using almond legumin as a template and compared with known sequential epitopes on other allergenic tree nut homologues. Of 28 patients tested by dot blot, 16 (57%) bound to 11S legumin, designated Car i 4. MS/MS sequencing of native 11S legumin identified 33 kDa acidic and 20-22 kDa basic subunits. Both pecan and walnut seed protein extracts inhibited IgE binding to recombinant Car i 4, suggesting cross-reactivity with Jug r 4. Sequential epitope mapping results of Car i 4 revealed weak, moderate, and strong reactivity of serum pools against 10, 5, and 4 peptides, respectively. Seven peptides were recognized by all three serum pools, of which two were strongly reactive. The strongly reactive peptides were located in three discrete regions of the Car i 4 acidic subunit sequence (residues 118-132, 208-219, and 238-249). Homology modeling of Car i 4 revealed significant overlapping regions shared in common with other tree nut legumins. PMID:21718052

  7. Identification of T-cell epitopes of Lol p 9, a major allergen of ryegrass (Lolium perenne) pollen.

    PubMed

    Blaher, B; Suphioglu, C; Knox, R B; Singh, M B; McCluskey, J; Rolland, J M

    1996-07-01

    T-cell recognition of Lol p 9, a major allergen of ryegrass pollen, was investigated by using a T-cell line and T-cell clones generated from the peripheral blood of an atopic donor. The T-cell line reacted with purified Lol p 9, as well as with crude ryegrass pollen extract, but failed to cross-react with Bermuda grass pollen extract. All of six T-cell clones generated from this line proliferated in response to Lol p 9. Epitope mapping was carried out with a panel of 34 overlapping synthetic peptides, which spanned the entire sequence of the Lol p 9 12R isoform. The T-cell line responded to two of the peptides, Lol p 9 (105-116) and Lol p 9 (193-204), whereas reactivity with one or other of these peptides was shown by five T-cell clones. These two peptides contained sequences consistent with motifs previously reported for major histocompatibility complex class II-restricted peptides. HLA antibody blocking studies showed that presentation of peptide Lol p 9 (105-116) to one T-cell clone was HLA-DR-restricted; this clone expressed a T helper cell phenotype (CD3+, CD4+) and the T-cell receptor alpha beta. The identification of immunodominant T-cell epitope(s) on allergens is essential for devising safer and more effective immunotherapy strategies, which can interrupt the chain of events leading to allergic disease.

  8. Purification, characterization and N-terminal amino acid sequence of a new major allergen from European chestnut pollen--Cas s 1.

    PubMed

    Kos, T; Hoffmann-Sommergruber, K; Ferreira, F; Hirschwehr, R; Ahorn, H; Horak, F; Jager, S; Sperr, W; Kraft, D; Scheiner, O

    1993-11-15

    Pollens from trees of the order Fagales (e.g. birch, alder, hazel, and hornbeam) all contain one major allergen--the main cause for tree pollen allergy. So far the major allergens from birch (Bet v 1), alder (Aln g 1), hazel (Cor a 1), and hornbeam (Car b 1) have been characterized, showing high sequence similarity with each other (1-4). We present the molecular and immunologic characterization of Cas s 1, the major allergen from the European chestnut (Castanea sativa). From aqueous pollen extracts from European chestnut a protein was purified to homogeneity and was subjected to further investigation. The protein revealed a Mr of 22 kDa and was shown to represent the major allergen of the European chestnut (immunoblotting, histamine release) and designated Cas s 1. Despite a marked difference in Mr, Cas s 1 shows significant amino acid sequence similarity at the N-terminus and is antigenically closely related to the major birch pollen allergen Bet v 1 (17 kDa), as shown by binding to the anti-Bet v 1 monoclonal antibody BIP-1 and by IgE-inhibition tests using recombinant Bet v 1.

  9. Molecular characterization of Api g 1, the major allergen of celery (Apium graveolens), and its immunological and structural relationships to a group of 17-kDa tree pollen allergens.

    PubMed

    Breiteneder, H; Hoffmann-Sommergruber, K; O'Riordain, G; Susani, M; Ahorn, H; Ebner, C; Kraft, D; Scheiner, O

    1995-10-15

    Individuals suffering from immediate hypersensitivity (type-I allergy) to a particular pollen frequently display intolerance to several foods of plant origin. In this respect, individuals sensitized to birch pollen and/or mugwort pollen frequently display type-I allergic symptoms after ingestion of celery. In this study, we expressed the major allergenic protein of celery, Api g 1, which is responsible for the birch-celery syndrome, in the form of a non-fusion protein. The open reading frame of the cDNA of Api g 1 codes for a protein of 153 amino acids with a molecular mass of 16.2 kDa and 40% identity (60% similarity) to the major allergen of birch pollen, Bet v 1. Furthermore, Api g 1 exhibited similar characteristics to (a) two proteins in parsley induced by fungal infection, (b) the major tree pollen allergens and (c) pathogenesis-related and stress-induced proteins in other plant species. The reactivity of recombinant Api g 1 with IgE antibodies present in sera from celery intolerant patients was comparable to that of the natural celery allergen. Cross-reactivity with Bet v 1 was proven by cross-inhibition experiments, which provides further support for the existence of the birch-celery syndrome and for the suggestion that allergies to some vegetable foods are epiphenomena to allergies caused by inhalation of tree pollen.

  10. A sensitive assay for the detection of IgE bound to the major birch pollen allergen, Bet v 1, in the form of immune complexes.

    PubMed

    Pree, Ines; Reisinger, Jürgen; Bohle, Barbara; Frantal, Sophie; Valenta, Rudolf; Niederberger, Verena

    2009-06-30

    Allergen-IgE immune complexes, in which the IgE paratopes are occupied by allergen, remain undetected in standard IgE-detection assays which are based on capturing specific IgE by allergens.We describe an assay for the detection of immune complexes consisting of IgE bound to one of the most frequent environmental allergens, the major birch pollen allergen, Bet v 1. This assay is based on a Bet v 1-specific monoclonal antibody, Bip 1, which binds to an epitope on Bet v 1 that is distinct from the epitopes recognized by allergic patients' IgE. IgE-immune complexes formed with sera from birch pollen-allergic patients (n = 46) were undetectable with solid phase-bound allergen but could be captured by Bip 1 immobilized to nitrocellulose membranes or ELISA plates and traced with radioactively or enzymatically labelled anti-human IgE antibodies. The levels of IgE complexed with Bet v 1 measured in our assays were highly correlated with the amounts of non-complexed allergen-specific IgE as determined by the ImmunoCAP assay which is based on solid phase-bound IgE (r = 0.95; p < 0.01). Bet v 1-specific IgE could even be detected at serum dilutions below the cut off of the ImmunoCAP system (i.e., 0.35 kUA/L). We have thus developed a robust and sensitive assay for the detection and quantification of Bet v 1-IgE immune complexes which should be useful to measure allergen-bound IgE in human body fluids and in in vitro experiments.

  11. Crystal structure of Sol I 2: a major allergen from fire ant venom.

    PubMed

    Borer, Aline S; Wassmann, Paul; Schmidt, Margit; Hoffman, Donald R; Zhou, Jing-Jiang; Wright, Christine; Schirmer, Tilman; Marković-Housley, Zora

    2012-01-27

    Sol i 2 is a potent allergen from the venom of red imported fire ant, which contains allergens Sol i 1, Sol i 2, Sol i 3, and Sol i 4 that are known to be powerful triggers of anaphylaxis. Sol i 2 causes IgE antibody production in about one-third of individuals stung by fire ants. Baculovirus recombinant dimeric Sol i 2 was crystallized as a native and selenomethionyl-derivatized protein, and its structure has been determined by single-wavelength anomalous dispersion at 2.6 Å resolution. The overall fold of each subunit consists of five helices that enclose a central hydrophobic cavity. The structure is stabilized by three intramolecular disulfide bridges and one intermolecular disulfide bridge. The nearest structural homologue is the sequence-unrelated odorant binding protein and pheromone binding protein LUSH of the fruit fly Drosophila, which may suggest a similar biological function. To test this hypothesis, we measured the reversible binding of various pheromones, plant odorants, and other ligands to Sol i 2 by the changes in N-phenyl-1-naphthylamine fluorescence emission upon binding of ligands that compete with N-phenyl-1-naphthylamine. The highest binding affinity was observed for hydrophobic ligands such as aphid alarm pheromone (E)-β-farnesene, analogs of ant alarm pheromones, and plant volatiles decane, undecane, and β-caryophyllene. Conceivably, Sol i 2 may play a role in capturing and/or transporting small hydrophobic ligands such as pheromones, odors, fatty acids, or short-living hydrophobic primers. Molecular surface analysis, in combination with sequence alignment, can explain the serological cross-reactivity observed between some ant species.

  12. Crystal structure of Sol I 2: a major allergen from fire ant venom.

    PubMed

    Borer, Aline S; Wassmann, Paul; Schmidt, Margit; Hoffman, Donald R; Zhou, Jing-Jiang; Wright, Christine; Schirmer, Tilman; Marković-Housley, Zora

    2012-01-27

    Sol i 2 is a potent allergen from the venom of red imported fire ant, which contains allergens Sol i 1, Sol i 2, Sol i 3, and Sol i 4 that are known to be powerful triggers of anaphylaxis. Sol i 2 causes IgE antibody production in about one-third of individuals stung by fire ants. Baculovirus recombinant dimeric Sol i 2 was crystallized as a native and selenomethionyl-derivatized protein, and its structure has been determined by single-wavelength anomalous dispersion at 2.6 Å resolution. The overall fold of each subunit consists of five helices that enclose a central hydrophobic cavity. The structure is stabilized by three intramolecular disulfide bridges and one intermolecular disulfide bridge. The nearest structural homologue is the sequence-unrelated odorant binding protein and pheromone binding protein LUSH of the fruit fly Drosophila, which may suggest a similar biological function. To test this hypothesis, we measured the reversible binding of various pheromones, plant odorants, and other ligands to Sol i 2 by the changes in N-phenyl-1-naphthylamine fluorescence emission upon binding of ligands that compete with N-phenyl-1-naphthylamine. The highest binding affinity was observed for hydrophobic ligands such as aphid alarm pheromone (E)-β-farnesene, analogs of ant alarm pheromones, and plant volatiles decane, undecane, and β-caryophyllene. Conceivably, Sol i 2 may play a role in capturing and/or transporting small hydrophobic ligands such as pheromones, odors, fatty acids, or short-living hydrophobic primers. Molecular surface analysis, in combination with sequence alignment, can explain the serological cross-reactivity observed between some ant species. PMID:22100449

  13. Cloning and characterization of 2S albumin, Car i 1, a major allergen in pecan.

    PubMed

    Sharma, Girdhari M; Irsigler, Andre; Dhanarajan, Pushparani; Ayuso, Rosalia; Bardina, Luda; Sampson, Hugh A; Roux, Kenneth H; Sathe, Shridhar K

    2011-04-27

    Although pecans are associated with IgE-mediated food allergies, the allergens responsible remain to be identified and characterized. The 2S albumin gene was amplified from the pecan cDNA library. Dot-blots were used to screen the recombinant protein with pecan allergic patients' serum. The affinity purified native protein was analyzed by Edman sequencing and mass spectrometry/mass spectrometry (MS/MS) analysis. Cross-reactivity with walnut was determined by inhibition enzyme-linked immunosorbent assay (ELISA). Sequential epitopes were determined by probing the overlapping peptides with three different patients' serum pool. The 3-dimensional homology model was generated, and the locations of the pecan epitopes were compared with those of known sequential epitopes on other allergenic tree nut homologues. Of 28 patients tested by dot-blot, 22 (79%) bound to 2S albumin, designated as Car i 1. Edman sequencing and the MS/MS sequencing of native 2S albumin confirmed the identity of recombinant (r) Car i 1. Both pecan and walnut protein extracts inhibited the IgE-binding to rCar i 1. Sequential epitope mapping indicated weak, moderate, and strong reactivity against 12, 7, and 5 peptides, respectively. Of the 11 peptides recognized by all serum pools, 5 peptides were strongly reactive and located in 3 discrete regions of the Car i 1 (amino acids 43-57, 67-78, and 106-120). Three-dimensional modeling revealed IgE-reactive epitopes to be solvent accessible and share significant homology with other tree nuts providing a possible basis for previously observed cross-reactivity. PMID:21395309

  14. Reduction of major peanut allergens Ara h 1 and Ara h 2, in roasted peanuts by ultrasound assisted enzymatic treatment.

    PubMed

    Li, Hao; Yu, Jianmei; Ahmedna, Mohamed; Goktepe, Ipek

    2013-11-15

    This study investigated the effects of ultrasound, enzyme concentration and enzyme treatment time on soluble protein and major allergenic proteins (Ara h 1 and Ara h 2) of roasted peanut kernels. A 3-factor, five-level orthogonal experimental design was implemented with various ultrasonication times, concentrations of trypsin or α-chymotrypsin and treatment times. The total soluble proteins were determined by the Bicinchoninic acid (BCA) method, Ara h 1 and Ara h 2 were evaluated by SDS-PAGE and sandwich ELISA. The IgE-binding of peanut extracts was analysed by a competitive inhibition ELISA. Results indicate that ultrasound treatment, followed by protease digestion of peanuts, significantly increased the solubility of peanut protein and decreased the concentrations of Ara h 1 and Ara h 2. The sequential treatment of peanuts by ultrasonication-trypsin-alpha chymotrypsin, resulted in maximum reductions of Ara h 1/Ara h 2, and lowest IgE-binding. This study provides an approach to significantly reduce allergenic proteins in peanut product. PMID:23790845

  15. A rapid and efficient two-step gel electrophoresis method for the purification of major rye grass pollen allergens.

    PubMed

    Levy, D; Davies, J; O'Hehir, R; Suphioglu, C

    2001-06-01

    Purified proteins are mandatory for molecular, immunological and cellular studies. However, purification of proteins from complex mixtures requires specialised chromatography methods (i.e., gel filtration, ion exchange, etc.) using fast protein liquid chromatography (FPLC) or high-performance liquid chromatography (HPLC) systems. Such systems are expensive and certain proteins require two or more different steps for sufficient purity and generally result in low recovery. The aim of this study was to develop a rapid, inexpensive and efficient gel-electrophoresis-based protein purification method using basic and readily available laboratory equipment. We have used crude rye grass pollen extract to purify the major allergens Lol p 1 and Lol p 5 as the model protein candidates. Total proteins were resolved on large primary gel and Coomassie Brilliant Blue (CBB)-stained Lol p 1/5 allergens were excised and purified on a secondary "mini"-gel. Purified proteins were extracted from unstained separating gels and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analyses. Silver-stained SDS-PAGE gels resolved pure proteins (i.e., 875 microg of Lol p 1 recovered from a 8 mg crude starting material) while immunoblot analysis confirmed immunological reactivity of the purified proteins. Such a purification method is rapid, inexpensive, and efficient in generating proteins of sufficient purity for use in monoclonal antibody (mAb) production, protein sequencing and general molecular, immunological, and cellular studies.

  16. Analysis of the allergenic proteins in black tiger prawn (Penaeus monodon) and characterization of the major allergen tropomyosin using mass spectrometry.

    PubMed

    Abdel Rahman, Anas M; Rahman, Anas M Abdel; Kamath, Sandip; Lopata, Andreas L; Helleur, Robert J

    2010-08-30

    Crustaceans are the third most prevalent cause of food-induced anaphylaxis after peanuts and tree nuts. The severity of the allergenic proteins depends mainly on the amino acid sequence that induces production of IgE antibodies. In black tiger prawn (Penaeus monodon), the crude protein extract was profiled and its allergenic potency was examined against patient's sera. Proteins having strong immunoreactivity with patient's IgE were characterized using peptide mass fingerprinting (PMF). Tropomyosin (TM) (33 kDa), myosin light chain (20 kDa), and arginine kinase (40 kDa) were identified as allergenic proteins. Tropomyosin, the most abundant and potent allergen, was purified using ion-exchange chromatography for de novo sequencing experiments. Using bottom up tandem mass spectrometry, the full amino acid sequence was achieved by a combination of matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI) tandem mass spectrometry (QqToF). Myosin light chain and arginine kinase were also characterized, and their related peptides were de novo sequenced using the same approach. The immunological reactivity of the crude prawn extracts and purified TM samples were analyzed using a large number of patients' sera. A signature peptide was assigned for the TM protein for future quantification work of black tiger prawn TM levels in different matrices (i.e. water, air, food) in the seafood industry. PMID:20658686

  17. Allergens in veterinary medicine

    PubMed Central

    Mueller, R. S.; Janda, J.; Jensen-Jarolim, E.; Rhyner, C.; Marti, E.

    2015-01-01

    Allergic diseases in animals are increasingly gaining importance in veterinary practice and as research models. For intradermal testing and allergen immunotherapy, a good knowledge of relevant allergens for the individual species is of great importance. Currently, the knowledge about relevant veterinary allergens is based on sensitization rates identified by intradermal testing or serum testing for allergen-specific IgE; crude extracts are the basis for most evaluations. Only a few studies provide evidence about the molecular structure of (particularly) dust mite, insect and mould allergens in dogs and horses, respectively. In those species, some major allergens differ from those in humans. This position paper summarizes the current knowledge about relevant allergens in dogs, cats and horses. PMID:26280544

  18. The Calcium Goes Meow: Effects of Ions and Glycosylation on Fel d 1, the Major Cat Allergen

    PubMed Central

    Pol-Fachin, Laércio; Verli, Hugo

    2015-01-01

    The major cat allergen, Fel d 1, is a structurally complex protein with two N-glycosylation sites that may be filled by different glycoforms. In addition, the protein contains three putative Ca2+ binding sites. Since the impact of these Fel d 1 structure modifications on the protein dynamics, physiology and pathology are not well established, the present work employed computational biology techniques to tackle these issues. While conformational effects brought upon by glycosylation were identified, potentially involved in cavity volume regulation, our results indicate that only the central Ca2+ ion remains coordinated to Fel d 1 in biological solutions, impairing its proposed role in modulating phospholipase A2 activity. As these results increase our understanding of Fel d 1 structural biology, they may offer new support for understanding its physiological role and impact into cat-promoted allergy. PMID:26134118

  19. Pin p 1 is a major allergen in pine nut and the first food allergen described in the plant group of gymnosperms.

    PubMed

    Cabanillas, Beatriz; Crespo, Jesus F; Maleki, Soheila J; Rodriguez, Julia; Novak, Natalija

    2016-11-01

    This study aimed to report the complete sequence of a 2S albumin purified from pine nut and to analyze its allergenic properties. Individual recognition of this protein by serum IgE from pine nut-allergic patients was assessed. IgE cross-linking capacity was analyzed in a basophil activation test. Inhibition of IgE-binding and stability to heating was also assessed. The complete nucleotide sequence was obtained and a phylogenetic study was carried out. 2S albumin from pine nut (registered as Pin p 1.0101) was recognized by IgE of 75% of sera. The allergen was heat-stable and had a robust capacity to inhibit IgE-binding to whole pine nut extract. The IgE cross-linking capacity of Pin p 1 on basophils was also demonstrated. Despite the low homology of Pin p 1 sequence with other allergenic 2S albumins from angiosperms, Pin p 1 contains the typical skeleton of 8 cysteine residues, important for its α-helixes enriched structure. PMID:27211622

  20. Purification, Crystallization and Preliminary X-ray Characterization of Prunin-1, a Major Component of the Almond (Prunus dulcis) Allergen Amandin

    SciTech Connect

    Albillos, Silvia M.; Jin, Tengchuan; Howard, Andrew; Zhang, Yuzhu; Kothary, Mahendra H.; Fu, Tong-Jen

    2008-08-04

    The 11S globulins from plant seeds account for a number of major food allergens. Because of the interest in the structural basis underlying the allergenicity of food allergens, we sought to crystallize the main 11S seed storage protein from almond (Prunus dulcis). Prunin-1 (Pru1) was purified from defatted almond flour by water extraction, cryoprecipitation, followed by sequential anion exchange, hydrophobic interaction, and size exclusion chromatography. Single crystals of Pru1 were obtained in a screening with a crystal screen kit, using the hanging-drop vapor diffusion method. Diffraction quality crystals were grown after optimization. The Pru1 crystals diffracted to at least 3.0 {angstrom} and belong to the tetragonal space group P4{sub 1}22, with unit cell parameters of a = b = 150.912 {angstrom}, c = 165.248 {angstrom}. Self-rotation functions and molecular replacement calculations showed that there are three molecules in the asymmetry unit with water content of 51.41%. The three Pru1 protomers are related by a noncrystallographic 3-fold axis and they form a doughnut-shaped trimer. Two prunin trimers form a homohexamer. Elucidation of prunin structure will allow further characterization of the allergenic features of the 11S protein allergens at the molecular level.

  1. Purification, crystallization and preliminary X-ray characterization of prunin-1, a major component of the almond (Prunus dulcis) allergen amandin.

    PubMed

    Albillos, Silvia M; Jin, Tengchuan; Howard, Andrew; Zhang, Yuzhu; Kothary, Mahendra H; Fu, Tong-Jen

    2008-07-01

    The 11S globulins from plant seeds account for a number of major food allergens. Because of the interest in the structural basis underlying the allergenicity of food allergens, we sought to crystallize the main 11S seed storage protein from almond ( Prunus dulcis). Prunin-1 (Pru1) was purified from defatted almond flour by water extraction, cryoprecipitation, followed by sequential anion exchange, hydrophobic interaction, and size exclusion chromatography. Single crystals of Pru1 were obtained in a screening with a crystal screen kit, using the hanging-drop vapor diffusion method. Diffraction quality crystals were grown after optimization. The Pru1 crystals diffracted to at least 3.0 A and belong to the tetragonal space group P4(1)22, with unit cell parameters of a = b = 150.912 A, c = 165.248 A. Self-rotation functions and molecular replacement calculations showed that there are three molecules in the asymmetry unit with water content of 51.41%. The three Pru1 protomers are related by a noncrystallographic 3-fold axis and they form a doughnut-shaped trimer. Two prunin trimers form a homohexamer. Elucidation of prunin structure will allow further characterization of the allergenic features of the 11S protein allergens at the molecular level.

  2. Correlation between airborne Olea europaea pollen concentrations and levels of the major allergen Ole e 1 in Córdoba, Spain, 2012-2014

    NASA Astrophysics Data System (ADS)

    Plaza, M. P.; Alcázar, P.; Galán, C.

    2016-04-01

    Olea europaea L. pollen is the second-largest cause of pollinosis in the southern Iberian Peninsula. Airborne-pollen monitoring networks provide essential data on pollen dynamics over a given study area. Recent research, however, has shown that airborne pollen levels alone do not always provide a clear indicator of actual exposure to aeroallergens. This study sought to evaluate correlations between airborne concentrations of olive pollen and Ole e 1 allergen levels in Córdoba (southern Spain), in order to determine whether atmospheric pollen concentrations alone are sufficient to chart changes in hay fever symptoms. The influence of major weather-related variables on local airborne pollen and allergen levels was also examined. Monitoring was carried out from 2012 to 2014. Pollen sampling was performed using a Hirst-type sampler, following the protocol recommended by the Spanish Aerobiology Network. A multi-vial cyclone sampler was used to collect aeroallergens, and allergenic particles were quantified by ELISA assay. Significant positive correlations were found between daily airborne allergen levels and atmospheric pollen concentrations, although there were occasions when allergen was detected before and after the pollen season and in the absence of airborne pollen. The correlation between the two was irregular, and pollen potency displayed year-on-year variations and did not necessarily match pollen-season-intensity.

  3. A sensitive fluorescent assay for measuring the cysteine protease activity of Der p 1, a major allergen from the dust mite Dermatophagoides pteronyssinus.

    PubMed Central

    Schulz, O; Sewell, H F; Shakib, F

    1998-01-01

    The potent allergenicity of Der p 1, a major allergen of the house dust mite Dermatophagoides pteronyssinus, is thought to be related to its cysteine protease activity. Therefore, there is considerable interest in developing a sensitive assay for measuring Der p 1 activity to screen for specific inhibitors. This study demonstrates for the first time that the activity of Der p 1 can be measured conveniently in a continuous rate assay with the fluorogenic substrate Boc-Gln-Ala-Arg-AMC (K(m) = 280 microM and kcat/K(m) = 4.6 x 10(3)/M/s). PMID:9893750

  4. Standardization of RP-HPLC methods for the detection of the major peanut allergens Ara h 1, Ara h 2 and Ara h 3

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Crude peanut extract (CPE) was analyzed for three major allergens (Ara h 1, h 2, and h 3) using a C12 and a C18 column at two wavelengths (280 and 220 nm) and under different solvent conditions. HPLC profiles were compared for retention time, resolution, and peak heights. CPE samples were spiked wit...

  5. Allergen nomenclature*

    PubMed Central

    1994-01-01

    The revised nomenclature for allergens is presented together with proposed nomenclatures for (a) allergen genes, mRNAs and cDNAs, and (b) recombinant and synthetic peptides of allergenic interest. PMID:7955031

  6. Flavonoid analysis of buckwheat sprouts.

    PubMed

    Nam, Tae-Gyu; Lee, Sun Mi; Park, Ji-Hae; Kim, Dae-Ok; Baek, Nam-in; Eom, Seok Hyun

    2015-03-01

    It is known that common buckwheat sprouts contain several flavonoids, including orientin, isoorientin, vitexin, isovitexin, rutin, and quercetrin, whereas tartary buckwheat sprouts contain only rutin. In this study, we evaluated flavonoids present in buckwheat sprouts and identified a previously unreported flavonoid. Simultaneous detection by HPLC was used to separate rutin and a compound that was not separated in previous studies. We used a novel HPLC elution gradient method to successfully separate rutin and the previously unidentified compound, for which we performed structural analysis. The identification of six flavonoids by HPLC was confirmed using HPLC-ESI-MS/MS analysis. The newly identified compound, [M+H](+) =611.17, was identified by NMR as the rutin epimer quercetin-3-O-robinobioside. Unlike common buckwheat sprout, tartary buckwheat sprout contained rutin as a main flavonoid, whereas other flavonoids appeared only in trace amounts or were not detected. Quercetin-3-O-robinobioside was not detected in tartary buckwheat sprout.

  7. Allergen composition analysis and allergenicity assessment of Chinese peanut cultivars.

    PubMed

    Wu, Zhihua; Zhou, Ningling; Xiong, Faqian; Li, Xin; Yang, Anshu; Tong, Ping; Tang, Ronghua; Chen, Hongbing

    2016-04-01

    Peanut (Arachis hypogaea) is among the eight major food allergens in the world. Several attempts have been made to decrease or eliminate the allergenicity of peanut. Systemic screening of thousands of peanut cultivars may identify peanut with low allergenicity. In this study, the allergen compositions of 53 Chinese peanut cultivars were characterized, and their allergenicity to sera IgE of Chinese patients and in a mouse model was assessed. Contents of total protein and allergens were quantified by SDS-PAGE and densitometry analysis on gel. Although the contents of allergens broadly varied among cultivars, they were related to one another. The IgE binding capacity of cultivars was tested by ELISA, and their allergenicity was further evaluated in a mouse model by oral sensitization. Results showed that the allergenicity of peanut was affected by allergen composition rather than a single allergen. Peanut cultivars with low allergenicity may contain more Ara h 3/4 (24 kDa), Ara h 2 and less Ara h 3/4 (43, 38, and 36 kDa), Ara h 6. Screening based on allergen composition would facilitate the identification of low-allergenic peanut. PMID:26593515

  8. Allergen composition analysis and allergenicity assessment of Chinese peanut cultivars.

    PubMed

    Wu, Zhihua; Zhou, Ningling; Xiong, Faqian; Li, Xin; Yang, Anshu; Tong, Ping; Tang, Ronghua; Chen, Hongbing

    2016-04-01

    Peanut (Arachis hypogaea) is among the eight major food allergens in the world. Several attempts have been made to decrease or eliminate the allergenicity of peanut. Systemic screening of thousands of peanut cultivars may identify peanut with low allergenicity. In this study, the allergen compositions of 53 Chinese peanut cultivars were characterized, and their allergenicity to sera IgE of Chinese patients and in a mouse model was assessed. Contents of total protein and allergens were quantified by SDS-PAGE and densitometry analysis on gel. Although the contents of allergens broadly varied among cultivars, they were related to one another. The IgE binding capacity of cultivars was tested by ELISA, and their allergenicity was further evaluated in a mouse model by oral sensitization. Results showed that the allergenicity of peanut was affected by allergen composition rather than a single allergen. Peanut cultivars with low allergenicity may contain more Ara h 3/4 (24 kDa), Ara h 2 and less Ara h 3/4 (43, 38, and 36 kDa), Ara h 6. Screening based on allergen composition would facilitate the identification of low-allergenic peanut.

  9. Confirmation of immuno-reactivity of the recombinant major birch pollen allergen Bet v 1a by affinity-CIEF.

    PubMed

    Dullnig, Verena; Weiss, Richard; Amon, Sabine; Rizzi, Andreas; Stutz, Hanno

    2009-07-01

    Affinity-CIEF has been applied to characterize a recombinant product of the major birch pollen allergen Betula verrucosa isoform 1a (Bet v 1a) immuno-chemically. For this purpose mAbs of the IgG-type have been produced in-lab from two murine hybridoma lines, specified as clones 2 and 5.1. Both IgG clones were characterized by SDS-PAGE, MALDI-TOF-MS and CIEF. The purified IgG solutions had to be dialysed against 10 mmol/L phosphate (pH 7.4) to prevent IgG precipitation and to ensure appropriate CIEF separation. Both tested monoclonal IgGs (mIgGs) comprised four constituents covering pI ranges of 6.98-7.09 and 6.78-7.03 for clones 2 and 5.1 with major peaks at pI 7.09 and 7.03, respectively. When increasing amounts of Bet v 1a (pI 4.95) were incubated with 2.0 mumol/L mIgG, novel peaks were progressively induced in a pI range slightly more acidic than the focusing region of mIgGs. These peaks grew on the expense of original mIgG peaks. All pI values were calculated using two pI marker compounds with a repeatability of better than 0.03 units. New peaks represent complexes between Bet v 1a and mIgG either of 1:1 or of 2:1 binding stoichiometry. At a molar ratio of 2:1, saturation of both IgG paratopes with allergen (Ag) molecules was achieved as indicated by unbound Bet v 1a. The current CIEF approach addresses the proof of single epitope integrity in the course of immuno-chemical characterization of Bet v 1a. Contrary to traditional immunoassays, affinity CIEF allows for a distinction and relative quantification of mAbs, Ag-antibody complexes and Ag variants coexisting in one sample. PMID:19621361

  10. Structures of Two Major Allergens, Bla g 4 and Per a 4, From Cockroaches and Their IgE Binding Epitopes

    SciTech Connect

    Tan, Y.; Chan, S; Ong, T; Yit, L; Tiong, Y; Chew, F; Sivaraman, J; Mok, Y

    2009-01-01

    Inhalant allergens from cockroaches are an important cause of asthma to millions of individuals worldwide. Here we report for the first time the structures of two major cockroach allergens, Bla g 4 and Per a 4, that adopt a typical lipocalin fold but with distinct structural features as compared with other known lipocalin allergens. Both Bla g 4 and Per a 4 contain two long-range disulfide bonds linking the N and C termini to a beta-barrel. The C-terminal helix of Bla g 4 is bent and greatly extended toward the N terminus. Bla g 4 is found to be a monomer, whereas Per a 4 exists as a dimer in solution with a novel dimeric interface involving residues from loops at the top and bottom of the beta-barrel. Putative ligand binding sites of both allergens are determined by docking of the juvenile hormone III inside the beta-barrel and found to interact with the ligand using non-conserved residues. Bla g 4 and Per a 4 are found to be cross-reactive in sera IgE binding, at least in the Singaporean Chinese population tested. A major IgE binding epitope unique to Per a 4 is found on the loops at the bottom of the beta-barrel that may aid the development of hypoallergens for immunotherapy.

  11. Atopic donor status does not influence the uptake of the major grass pollen allergen, Phl p 5, by dendritic cells

    PubMed Central

    Ashjaei, Kazem; Palmberger, Dieter; Bublin, Merima; Bajna, Erika; Breiteneder, Heimo; Grabherr, Reingard; Ellinger, Isabella; Hoffmann-Sommergruber, Karin

    2016-01-01

    Dendritic cells (DCs) are sentinels of the immune system for antigen recognition and uptake, as well as presentation to naïve T cells for stimulation or priming. Internalization and endocytic degradation of allergens by DCs are important steps required for T cell priming. In the current study we investigated binding and internalization of purified recombinant non-glycosylated grass pollen allergen, Phl p 5, and natural non-specific lipid transfer protein from sunflower, SF-nsLTP to human monocyte derived dendritic cells (MoDCs). Colocalization of Phl p 5 with low affinity (CD23) or high affinity receptor (FcεRI) was investigated by immunofluorescence staining. Likewise, localization of the allergens in early (EE) and late endosomes (LE) was detected by co-staining for early endosome antigen (EEA1) and lysosomal-associated membrane protein 1 (LAMP1). In our experimental setting we could demonstrate that Phl p 5 as well as SF-nsLTP bound to MoDCs from both, grass pollen allergic and non-allergic individuals. Competitive allergen uptake experiments demonstrated non-preferential and simultaneous uptake of Phl p 5 and SF-nsLTP by MoDCs. No overlap of signals from Phl p 5 and CD23 or FcεRI was detectable, excluding IgE-mediated uptake for this allergen. Both allergens, Phl p 5 and SF-nsLTP, were localized in early and late endosomes. The present study applied a set of methods to assess the allergen uptake by MoDCs in an in vitro model. No qualitative and quantitative differences in the allergen uptake of both, Phl p 5 and SF-nsLTP were detected in single and competitive assays. PMID:26055335

  12. Genomic, RNAseq, and molecular modeling evidence suggests that the major allergen domain in insects evolved from a homodimeric origin.

    PubMed

    Randall, Thomas A; Perera, Lalith; London, Robert E; Mueller, Geoffrey A

    2013-01-01

    The major allergen domain (MA) is widely distributed in insects. The crystal structure of a single Bla g 1 MA revealed a novel protein fold in which the fundamental structure was a duplex of two subsequences (monomers), which had diverged over time. This suggested that the evolutionary origin of the MA structure may have been a homodimer of this smaller subsequence. Using publicly available genomic data, the distribution of the basic unit of this class of proteins was determined to better understand its evolutionary history. The duplication and divergence is examined at three distinct levels of resolution: 1) within the orders Diptera and Hymenoptera, 2) within one genus Drosophila, and 3) within one species Aedes aegypti. Within the family Culicidae, we have found two separate occurrences of monomers as independent genes. The organization of the gene family in A. aegypti shows a common evolutionary origin for its monomer and several closely related MAs. Molecular modeling of the A. aegypti monomer with the unique Bla g 1 fold confirms the distant evolutionary relationship and supports the feasibility of homodimer formation from a single monomer. RNAseq data for A. aegypti confirms that the monomer is expressed in the mosquito similar to other A. aegypti MAs after a blood meal. Together, these data support the contention that the detected monomer shares similar functional characteristics to related MAs in other insects. An extensive search for this domain outside of Insecta confirms that the MAs are restricted to insects.

  13. Common food allergens and their IgE-binding epitopes.

    PubMed

    Matsuo, Hiroaki; Yokooji, Tomoharu; Taogoshi, Takanori

    2015-10-01

    Food allergy is an adverse immune response to certain kinds of food. Although any food can cause allergic reactions, chicken egg, cow's milk, wheat, shellfish, fruit, and buckwheat account for 75% of food allergies in Japan. Allergen-specific immunoglobulin E (IgE) antibodies play a pivotal role in the development of food allergy. Recent advances in molecular biological techniques have enabled the efficient analysis of food allergens. As a result, many food allergens have been identified, and their molecular structure and IgE-binding epitopes have also been identified. Studies of allergens have demonstrated that IgE antibodies specific to allergen components and/or the peptide epitopes are good indicators for the identification of patients with food allergy, prediction of clinical severity and development of tolerance. In this review, we summarize our current knowledge regarding the allergens and IgE epitopes in the well-researched allergies to chicken egg, cow's milk, wheat, shrimp, and peanut.

  14. Assessment of the Olea pollen and its major allergen Ole e 1 concentrations in the bioearosol of two biogeographical areas

    NASA Astrophysics Data System (ADS)

    Moreno-Grau, S.; Aira, M. J.; Elvira-Rendueles, B.; Fernández-González, M.; Fernández-González, D.; García-Sánchez, A.; Martínez-García, M. J.; Moreno, J. M.; Negral, L.; Vara, A.; Rodríguez-Rajo, F. J.

    2016-11-01

    The Olea pollen is currently an important allergy source. In some regions of Southern Spain, olive pollen is the main cause of allergic sensitization exceeding 40% of the sensitized individuals. Due to the scarce presence of olive trees in Northern Spain, limited to some cultivated fields in the South of the Galicia region where they also grow wild, only 8% of the sensitized individuals showed positive results for Olea pollen. The aim of the paper was to assess the behaviour pattern of the Olea pollen and its aeroallergens in the atmosphere, as this information could help us to improve the understanding and prevention of clinical symptoms. Airborne Olea pollen and Ole e 1 allergens were quantified in Cartagena (South-eastern Spain) and Ourense (North-western Spain). A volumetric pollen trap and a Burkard Cyclone sampler were used for pollen and allergen quantification. The Olea flowering took place in April or May in both biometeorological sampling areas. The higher concentrations were registered in the Southern area of Spain, for both pollen and Ole e 1, with values 8 times higher for pollen concentrations and 40 times higher for allergens. An alternate bearing pattern could be observed, characterized by years with high pollen values and low allergen concentrations and vice versa. Moreover, during some flowering seasons the allergen concentrations did not correspond to the atmospheric pollen values. Variations in weather conditions or Long Distance Transport (LDT) processes could explain the discordance. The back trajectory analysis shows that the most important contributions of pollen and allergens in the atmosphere are coincident with air masses passing through potential source areas. The exposure to olive pollen may not be synonym of antigen exposure.

  15. Tree nut allergens.

    PubMed

    Roux, Kenneth H; Teuber, Suzanne S; Sathe, Shridhar K

    2003-08-01

    Allergic reactions to tree nuts can be serious and life threatening. Considerable research has been conducted in recent years in an attempt to characterize those allergens that are most responsible for allergy sensitization and triggering. Both native and recombinant nut allergens have been identified and characterized and, for some, the IgE-reactive epitopes described. Some allergens, such as lipid transfer proteins, profilins, and members of the Bet v 1-related family, represent minor constituents in tree nuts. These allergens are frequently cross-reactive with other food and pollen homologues, and are considered panallergens. Others, such as legumins, vicilins, and 2S albumins, represent major seed storage protein constituents of the nuts. The allergenic tree nuts discussed in this review include those most commonly responsible for allergic reactions such as hazelnut, walnut, cashew, and almond as well as those less frequently associated with allergies including pecan, chestnut, Brazil nut, pine nut, macadamia nut, pistachio, coconut, Nangai nut, and acorn.

  16. Tree nut allergens.

    PubMed

    Roux, Kenneth H; Teuber, Suzanne S; Sathe, Shridhar K

    2003-08-01

    Allergic reactions to tree nuts can be serious and life threatening. Considerable research has been conducted in recent years in an attempt to characterize those allergens that are most responsible for allergy sensitization and triggering. Both native and recombinant nut allergens have been identified and characterized and, for some, the IgE-reactive epitopes described. Some allergens, such as lipid transfer proteins, profilins, and members of the Bet v 1-related family, represent minor constituents in tree nuts. These allergens are frequently cross-reactive with other food and pollen homologues, and are considered panallergens. Others, such as legumins, vicilins, and 2S albumins, represent major seed storage protein constituents of the nuts. The allergenic tree nuts discussed in this review include those most commonly responsible for allergic reactions such as hazelnut, walnut, cashew, and almond as well as those less frequently associated with allergies including pecan, chestnut, Brazil nut, pine nut, macadamia nut, pistachio, coconut, Nangai nut, and acorn. PMID:12915766

  17. Impact of heat processing on the detection of the major shellfish allergen tropomyosin in crustaceans and molluscs using specific monoclonal antibodies.

    PubMed

    Kamath, Sandip D; Abdel Rahman, Anas M; Komoda, Toshikazu; Lopata, Andreas L

    2013-12-15

    The major heat-stable shellfish allergen, tropomyosin, demonstrates immunological cross-reactivity, making specific differentiation of crustaceans and molluscs for food labelling very difficult. The aim of this study was to evaluate the application of allergen-specific monoclonal antibodies in differential detection of shellfish-derived tropomyosin in 11 crustacean and 7 mollusc species, and to study the impact of heating on its detection. Cross-reactive tropomyosin was detected in all crustacean species, with partial detection in molluscs: mussels, scallops and snails but none in oyster, octopus and squid. Furthermore, we have demonstrated that heating of shellfish has a profound effect on tropomyosin detection. This was evident by the enhanced recognition of multiple tropomyosin variants in the analysed shellfish species. Specific monoclonal antibodies, targetting the N-terminal region of tropomyosin, must therefore be developed to differentiate tropomyosins in crustaceans and molluscs. This can help in correct food labelling practices and thus protection of consumers.

  18. Lipocalins as allergens.

    PubMed

    Mäntyjärvi, R; Rautiainen, J; Virtanen, T

    2000-10-18

    The term allergy refers to clinical conditions caused by an inappropriate immune response to innocuous proteins in genetically predisposed persons. Allergens of animal origin are responsible for a significant proportion of allergies. In recent years, it has become evident that practically all respiratory animal allergens characterized at the molecular level belong to the lipocalin family of proteins. The current list comprises the major allergens of horse, cow, dog, mouse, rat and cockroach as well as beta-lactoglobulin of cow's milk. While the molecular structure of all these allergens is known, far less information is available regarding their immunological characteristics. Knowing the way the immune system recognizes these allergens and reacts to them might, however, be the key for discovering the common denominator of the allergenicity of lipocalins. The human body contains numerous endogenous lipocalins, and the immune system has to adapt to their presence. We have proposed that under these conditions the immune response against the lipocalin allergens which are structurally related to endogenous lipocalins might be the pathway to allergy in genetically predisposed persons. The same might well apply also to other allergens with homologous endogenous counterparts. PMID:11058771

  19. Quantitation of latex allergens.

    PubMed

    Palosuo, Timo; Alenius, Harri; Turjanmaa, Kristiina

    2002-05-01

    Minimizing allergen concentration in latex goods to prevent sensitization to natural rubber latex (NRL) and thereby the development of clinical allergy is acknowledged as of mutual interest for rubber manufacturers and regulatory health authorities. However, measuring total protein, the principal currently available method, cannot be deemed a satisfactory regulatory measure to control allergen content. Specific methods based on human IgE-containing reagents, such as radioallergosorbent test (RAST) inhibition, have been available in certain laboratories for demonstrating NRL allergens in rubber products but the methods lack standardization. Currently, one commercial test has become available for measuring individual NRL allergens by capture ELISA-based assays using monoclonal antibodies and purified or recombinant allergens. Such methods are specific, they can be properly standardized, and they are of sufficient sensitivity and reproducibility. Results from medical gloves collected in two national market surveys in Finland in 1995 and 1999, respectively, show that Hev b 6.02 and Hev b 5, the two major allergens for NRL-allergic adults, are the most abundant allergens regularly detectable in high- and moderate-allergen gloves. In addition, Hev b 3 and Hev b 1, the two major allergens for children with spina bifida, are also commonly found. In general, when the sum of the four allergens exceeded 1 microg/g, most NRL-allergic patients showed positive skin prick test reactions against them. Using these new methods assessment of threshold levels that could in due course become guidelines for the rubber industry and regulatory health authorities is becoming possible. Eventually, this progress is expected to lead to a declining incidence of latex allergy. PMID:12079417

  20. The Major Birch Pollen Allergen Bet v 1 Induces Different Responses in Dendritic Cells of Birch Pollen Allergic and Healthy Individuals

    PubMed Central

    Smole, Ursula; Radauer, Christian; Lengger, Nina; Svoboda, Martin; Rigby, Neil; Bublin, Merima; Gaier, Sonja; Hoffmann-Sommergruber, Karin; Jensen-Jarolim, Erika; Mechtcheriakova, Diana; Breiteneder, Heimo

    2015-01-01

    Dendritic cells play a fundamental role in shaping the immune response to allergens. The events that lead to allergic sensitization or tolerance induction during the interaction of the major birch pollen allergen Bet v 1 and dendritic cells are not very well studied. Here, we analyzed the uptake of Bet v 1 and the cross-reactive celery allergen Api g 1 by immature monocyte-derived dendritic cells (iMoDCs) of allergic and normal donors. In addition, we characterized the allergen-triggered intracellular signaling and transcriptional events. Uptake kinetics, competitive binding, and internalization pathways of labeled allergens by iMoDCs were visualized by live-cell imaging. Surface-bound IgE was detected by immunofluorescence microscopy and flow cytometry. Allergen- and IgE-induced gene expression of early growth response genes and Th1 and Th2 related cytokines and chemokines were analyzed by real-time PCR. Phosporylation of signaling kinases was analyzed by Western blot. Internalization of Bet v 1 by iMoDCs of both donor groups, likely by receptor-mediated caveolar endocytosis, followed similar kinetics. Bet v 1 outcompeted Api g 1 in cell surface binding and uptake. MoDCs of allergic and healthy donors displayed surface-bound IgE and showed a pronounced upregulation of Th2 cytokine- and NFκB-dependent genes upon non-specific Fcε receptor cross-linking. In contrast to these IgE-mediated responses, Bet v 1-stimulation increased transcript levels of the Th2 cytokines IL-4 and IL-13 but not of NFκB-related genes in MoDCs of BP allergic donors. Cells of healthy donors were either unresponsive or showed elevated mRNA levels of Th1-promoting chemokines. Moreover, Bet v 1 was able to induce Erk1/2 and p38 MAPK activation in BP allergics but only a slight p38 activation in normal donors. In conclusion, our data indicate that Bet v 1 favors the activation of a Th2 program only in DCs of BP allergic individuals. PMID:25635684

  1. The major birch pollen allergen Bet v 1 induces different responses in dendritic cells of birch pollen allergic and healthy individuals.

    PubMed

    Smole, Ursula; Radauer, Christian; Lengger, Nina; Svoboda, Martin; Rigby, Neil; Bublin, Merima; Gaier, Sonja; Hoffmann-Sommergruber, Karin; Jensen-Jarolim, Erika; Mechtcheriakova, Diana; Breiteneder, Heimo

    2015-01-01

    Dendritic cells play a fundamental role in shaping the immune response to allergens. The events that lead to allergic sensitization or tolerance induction during the interaction of the major birch pollen allergen Bet v 1 and dendritic cells are not very well studied. Here, we analyzed the uptake of Bet v 1 and the cross-reactive celery allergen Api g 1 by immature monocyte-derived dendritic cells (iMoDCs) of allergic and normal donors. In addition, we characterized the allergen-triggered intracellular signaling and transcriptional events. Uptake kinetics, competitive binding, and internalization pathways of labeled allergens by iMoDCs were visualized by live-cell imaging. Surface-bound IgE was detected by immunofluorescence microscopy and flow cytometry. Allergen- and IgE-induced gene expression of early growth response genes and Th1 and Th2 related cytokines and chemokines were analyzed by real-time PCR. Phosporylation of signaling kinases was analyzed by Western blot. Internalization of Bet v 1 by iMoDCs of both donor groups, likely by receptor-mediated caveolar endocytosis, followed similar kinetics. Bet v 1 outcompeted Api g 1 in cell surface binding and uptake. MoDCs of allergic and healthy donors displayed surface-bound IgE and showed a pronounced upregulation of Th2 cytokine- and NFκB-dependent genes upon non-specific Fcε receptor cross-linking. In contrast to these IgE-mediated responses, Bet v 1-stimulation increased transcript levels of the Th2 cytokines IL-4 and IL-13 but not of NFκB-related genes in MoDCs of BP allergic donors. Cells of healthy donors were either unresponsive or showed elevated mRNA levels of Th1-promoting chemokines. Moreover, Bet v 1 was able to induce Erk1/2 and p38 MAPK activation in BP allergics but only a slight p38 activation in normal donors. In conclusion, our data indicate that Bet v 1 favors the activation of a Th2 program only in DCs of BP allergic individuals. PMID:25635684

  2. Use of on-line stop-flow heart-cutting two-dimensional high performance liquid chromatography for simultaneous determination of 12 major constituents in tartary buckwheat (Fagopyrum tataricum Gaertn).

    PubMed

    Ren, Qiang; Wu, Caisheng; Zhang, Jinlan

    2013-08-23

    The use of two-dimensional liquid chromatography (2D-LC) for quantification studies presents challenges with respect to repeatability, precision, and robustness. The present study used an on-line stop-flow heart-cutting 2D-LC system to determine 12 chemical constituents in tartary buckwheat. A combination of various stationary phases was developed and bridged using two switch valves as the interface. Hydrophilic interaction chromatography was chosen for separation in the first dimension ((1)D), and mixed mode stationary phases (an amide polar-embedded phase and alkyl-phenyl phase) were used in parallel for separation in the second dimension ((2)D). The mobile phase comprised acetonitrile and water containing 0.03% aqueous phosphoric acid. The sample was separated into two fractions on the (1)D column (HILIC-10 column) using 5% acetonitrile. One fraction, mainly comprising flavonoids, was directly eluted onto the head of (2)D column (Polar Advantage II column) and further separated using a linear gradient of 11-23% acetonitrile. The second fraction, containing phenylpropanoid glycosides, was trapped on the (1)D column. This retained fraction was back-flushed onto the (2)D column (Phenyl-1 column) and separated using a linear gradient of 35-43% acetonitrile. An on-line stop-flow heart-cutting 2D-LC system was successfully developed with column switching and back-flush. This 2D-LC system was validated and was able to simultaneously determine 12 major components in tartary buckwheat: seven flavonoids, four phenylpropanoid glycosides, and N-trans-feruloyltyramine. The system showed good performance with respect to linearity (r>0.996), repeatability (RSD, relative standard deviation<3.4%), intra-day and inter-day precision (RSD<4.6%), recovery (91.2-108%), limit of detection (LOD) (0.05-0.21μg/mL), and limit of quantification (LOQ) (0.10-0.41μg/mL). The on-line stop-flow heart-cutting 2D-LC system offers a potential approach to analyze compounds, which have similar

  3. Allergenic characteristics of a modified peanut allergen.

    PubMed

    King, Nina; Helm, Ricki; Stanley, J Steven; Vieths, Stefan; Lüttkopf, Dirk; Hatahet, Lina; Sampson, Hugh; Pons, Laurent; Burks, Wesley; Bannon, Gary A

    2005-10-01

    Attempts to treat peanut allergy using traditional methods of allergen desensitization are accompanied by a high risk of anaphylaxis. The aim of this study was to determine if modifications to the IgE-binding epitopes of a major peanut allergen would result in a safer immunotherapeutic agent for the treatment of peanut-allergic patients. IgE-binding epitopes on the Ara h 2 allergen were modified, and modified Ara h 2 (mAra h 2) protein was produced. Wild-type (wAra h 2) and mAra h 2 proteins were analyzed for their ability to interact with T-cells, their ability to bind IgE, and their ability to release mediators from a passively sensitized RBL-2H3 cell line. Multiple T-cell epitopes were identified on the major peanut allergen, Ara h 2. Ara h 2 amino acid regions 11-35, 86-125, and 121-155 contained the majority of peptides that interact with T-cells from most patients. The wAra h 2 and mAra h 2 proteins stimulated proliferation of T-cells from peanut-allergic patients to similar levels. In contrast, the mAra h 2 protein exhibited greatly reduced IgE-binding capacity compared to the wild-type allergen. In addition, the modified allergen released significantly lower amounts of beta-hexosaminidase, a marker for IgE-mediated RBL-2H3 degranulation, compared to the wild-type allergen.

  4. Immunogold electron microscopy of soluble proteins: localization of Bet v I major allergen in ultra-thin sections of birch pollen after anhydrous fixation techniques.

    PubMed

    Grote, M

    1991-10-01

    To localize the highly water-soluble major allergen Bet v I in ultra-thin sections of birch pollen, pollen grains were cracked, air-dried, and processed for electron microscopy using one of the following preparation techniques: fixation in aqueous p-formaldehyde + cetylpyridinium chloride; fixation in p-formaldehyde vapor; fixation in benzoquinone vapor; inert dehydration; or no fixation. Afterwards the pollen grains were embedded in Lowicryl K4M resin at low temperature. Ultra-thin sections were cut and incubated with a monoclonal antibody against Bet v I, followed by a gold-labeled secondary antibody. In some experiments, commercial rabbit IgG antibodies against birth pollen allergens were also used, followed by incubation with the protein A-gold complex. Bet v I could be localized only after vapor fixation and in the inert dehydrated specimens. Best preservation of ultrastructure and antigenicity was obtained after p-formaldehyde vapor fixation. Bet v I antibody binding sites were detected only in the cytoplasmic matrix of the pollen grain, never in the pollen wall. Commercial rabbit antibodies bound to cytoplasm and wall of all prepared specimens, even after aqueous fixation. This might be explained by the assumption that these antibodies recognize a variety of antigenic and allergenic structures, not all of which are so highly soluble as Bet v I.

  5. Assessment of the Allergenic Potential of Transgenic Wheat (Triticum aestivum) with Reduced Levels of ω5-Gliadins, the Major Sensitizing Allergen in Wheat-Dependent Exercise-Induced Anaphylaxis.

    PubMed

    Altenbach, Susan B; Tanaka, Charlene K; Pineau, Florence; Lupi, Roberta; Drouet, Martine; Beaudouin, Etienne; Morisset, Martine; Denery-Papini, Sandra

    2015-10-28

    The ω5-gliadins are the major sensitizing allergens in wheat-dependent exercise-induced anaphylaxis (WDEIA). In this study, two-dimensional immunoblot analysis was used to assess the allergenic potential of two transgenic wheat lines in which ω5-gliadin genes were silenced by RNA interference. Sera from 7 of 11 WDEIA patients showed greatly reduced levels of immunoglobulin E (IgE) reactivity to ω5-gliadins in both transgenic lines. However, these sera also showed low levels of reactivity to other gluten proteins. Sera from three patients showed the greatest reactivity to proteins other than ω5-gliadins, either high-molecular-weight glutenin subunits (HMW-GSs), α-gliadins, or non-gluten proteins. The complexity of immunological responses among these patients suggests that flour from the transgenic lines would not be suitable for individuals already diagnosed with WDEIA. However, the introduction of wheat lacking ω5-gliadins could reduce the number of people sensitized to these proteins and thereby decrease the overall incidence of this serious food allergy.

  6. Assessment of the Allergenic Potential of Transgenic Wheat (Triticum aestivum) with Reduced Levels of ω5-Gliadins, the Major Sensitizing Allergen in Wheat-Dependent Exercise-Induced Anaphylaxis.

    PubMed

    Altenbach, Susan B; Tanaka, Charlene K; Pineau, Florence; Lupi, Roberta; Drouet, Martine; Beaudouin, Etienne; Morisset, Martine; Denery-Papini, Sandra

    2015-10-28

    The ω5-gliadins are the major sensitizing allergens in wheat-dependent exercise-induced anaphylaxis (WDEIA). In this study, two-dimensional immunoblot analysis was used to assess the allergenic potential of two transgenic wheat lines in which ω5-gliadin genes were silenced by RNA interference. Sera from 7 of 11 WDEIA patients showed greatly reduced levels of immunoglobulin E (IgE) reactivity to ω5-gliadins in both transgenic lines. However, these sera also showed low levels of reactivity to other gluten proteins. Sera from three patients showed the greatest reactivity to proteins other than ω5-gliadins, either high-molecular-weight glutenin subunits (HMW-GSs), α-gliadins, or non-gluten proteins. The complexity of immunological responses among these patients suggests that flour from the transgenic lines would not be suitable for individuals already diagnosed with WDEIA. However, the introduction of wheat lacking ω5-gliadins could reduce the number of people sensitized to these proteins and thereby decrease the overall incidence of this serious food allergy. PMID:26447559

  7. Peanut oral immunotherapy modifies IgE and IgG4 responses to major peanut allergens

    PubMed Central

    Vickery, Brian P.; Lin, Jing; Kulis, Michael; Fu, Zhiyan; Steele, Pamela H.; Jones, Stacie M.; Scurlock, Amy M.; Gimenez, Gustavo; Bardina, Ludmilla; Sampson, Hugh A.; Burks, A. Wesley

    2012-01-01

    Background Peanut-allergic subjects have highly stable pathologic antibody repertoires to the immunodominant B cell epitopes of the major peanut allergens Ara h 1-3. Objective We used a peptide microarray technique to analyze the effect of treatment with peanut oral immunotherapy (OIT) on such repertoires. Methods Measurements of total peanut-specific IgE (psIgE) and psIgG4 were made with CAP-FEIA. We analyzed sera from 22 OIT subjects and 6 controls and measured serum specific IgE and IgG4 binding to epitopes of Ara h 1-3 using a high-throughput peptide microarray technique. Antibody affinity was measured using a competitive peptide microarray as previously described. Results At baseline, psIgE and psIgG4 diversity were similar between subjects and controls, and there was broad variation in epitope recognition. After a median 41 months of OIT, polyclonal psIgG4 increased from a median 0.3 mcg/mL (IQR 0.1-0.43) at baseline to 10.5 mcg/mL (3.95-45.48) (p<0.0001) and included de novo specificities. PsIgE was reduced from a median baseline of 85.45 kUA/L (23.05-101.0) to 7.75 kUA/L (2.58-30.55) (p<0.0001). Affinity was unaffected. Although the psIgE repertoire contracted in most OIT-treated subjects, several subjects generated new IgE specificities even as the total psIgE decreased. Global epitope-specific shifts from IgE to IgG4 binding occurred, including at an informative epitope of Ara h 2. Conclusion OIT differentially alters Ara h 1-3 binding patterns. These changes are variable between subjects, not observed in controls, and include a progressive polyclonal increase in IgG4, with concurrent reduction in IgE amount and diversity. PMID:23199605

  8. Solution structure of the C-terminal domain of Ole e 9, a major allergen of olive pollen

    PubMed Central

    Treviño, Miguel Á.; Palomares, Oscar; Castrillo, Inés; Villalba, Mayte; Rodríguez, Rosalía; Rico, Manuel; Santoro, Jorge; Bruix, Marta

    2008-01-01

    Ole e 9 is an olive pollen allergen belonging to group 2 of pathogenesis-related proteins. The protein is composed of two immunological independent domains: an N-terminal domain (NtD) with 1,3-β-glucanase activity, and a C-terminal domain (CtD) that binds 1,3-β-glucans. We have determined the three-dimensional structure of CtD-Ole e 9 (101 amino acids), which consists of two parallel α-helices forming an angle of ∼55°, a small antiparallel β-sheet with two short strands, and a 3–10 helix turn, all connected by long coil segments, resembling a novel type of folding among allergens. Two regions surrounded by aromatic residues (F49, Y60, F96, Y91 and Y31, H68, Y65, F78) have been localized on the protein surface, and a role for sugar binding is suggested. The epitope mapping of CtD-Ole e 9 shows that B-cell epitopes are mainly located on loops, although some of them are contained in secondary structural elements. Interestingly, the IgG and IgE epitopes are contiguous or overlapped, rather than coincident. The three-dimensional structure of CtD-Ole e 9 might help to understand the underlying mechanism of its biochemical function and to determine possible structure–allergenicity relationships. PMID:18096638

  9. Structural stability of Amandin, a major allergen from almond (Prunus dulcis), and its acidic and basic polypeptides.

    PubMed

    Albillos, Silvia M; Menhart, Nicholas; Fu, Tong-Jen

    2009-06-10

    Information relating to the resistance of food allergens to thermal and/or chemical denaturation is critical if a reduction in protein allergenicity is to be achieved through food-processing means. This study examined the changes in the secondary structure of an almond allergen, amandin, and its acidic and basic polypeptides as a result of thermal and chemical denaturation. Amandin ( approximately 370 kDa) was purified by cryoprecipitation followed by gel filtration chromatography and subjected to thermal (13-96 degrees C) and chemical (urea and dithiothreitol) treatments. Changes in the secondary structure of the protein were followed using circular dichroism spectroscopy. The secondary structure of the hexameric amandin did not undergo remarkable changes at temperatures up to 90 degrees C, although protein aggregation was observed. In the presence of a reducing agent, irreversible denaturation occurred with the following experimental values: T(m) = 72.53 degrees C (transition temperature), DeltaH = 87.40 kcal/mol (unfolding enthalpy), and C(p) = 2.48 kcal/(mol degrees C) (heat capacity). The concentration of urea needed to achieve 50% denaturation was 2.59 M, and the Gibbs free energy of chemical denaturation was calculated to be DeltaG = 3.82 kcal/mol. The basic and acidic polypeptides of amandin had lower thermal stabilities than the multimeric protein.

  10. Ancient whole grain gluten-free buckwheat snacks

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Four kinds of whole grain Buckwheat snacks (gluten-free, low in fat and salt) were evaluated. The snacks were Buckwheat, Buckwheat-Turmeric, Buckwheat-Pepper and Buckwheat-Ginger. Turmeric and Ginger are very common ingredients of Asian cuisines. Turmeric and ginger have been reported to have heal...

  11. Japan food allergen labeling regulation--history and evaluation.

    PubMed

    Akiyama, Hiroshi; Imai, Takanori; Ebisawa, Motohiro

    2011-01-01

    According to a national survey of food allergy cases, the food-labeling system for specific allergenic ingredients (i.e., egg, milk, wheat, buckwheat, and peanut) in Japan was mandated under law on April 1, 2002. By Japanese law, labeling of allergens is designated as mandatory or recommended based on the number of cases of actual illness and the degree of seriousness. Mandatory labeling is enforced by the ministerial ordinance, and the ministerial notification recommends that foods containing walnut and soybean be labeled with subspecific allergenic ingredients. Additional labeling of shrimp/prawn and crab has also become mandatory since 2008. To monitor the validity of the labeling system, the Japanese government announced the official methods for detection of allergens in a November 2002 ministry notification. These official methods, including two kinds of enzyme-linked immunosorbent assay kits for screening, Western blotting analyses for egg and milk, and polymerase chain reaction analyses for wheat, buckwheat, peanut, shrimp/prawn and crab as confirmation tests, have provided a means to monitor the labeling system. To standardize the official methods, the Japanese government described the validation protocol criteria in the 2006 official guidelines. The guidelines stipulate that any food containing allergen proteins at greater than 10mg/kg must be labeled under the Law. This review covers the selection of the specific allergenic ingredients by the Japanese government, the implementation of regulatory action levels and the detection methods to support them, and the assessment of the effectiveness of this approach.

  12. [Food allergens].

    PubMed

    Bonneau, J C

    1997-07-01

    Perhaps more than any other kind of allergen, search for a food allergen seems to be difficult. There should be no bias about the usual allergens found in our food, that are a source of pathology that is less spectacular than shocks or giant urticaria that are provoked by easily recognised causes. Crossed allergies must be recognised in their overall features. This may give decisive aid in the etiological approach by facilitating understanding of the symptoms and the discovery of potential triggering allergens which are systematically sought.

  13. Identification of the major allergenic epitopes of Eriocheir sinensis roe hemocyanin: A novel tool for food allergy diagnoses.

    PubMed

    Zhang, Yingying; Zhu, Lina; Li, Shaoshen; Zhang, Jiayi; She, Tiantian; Yan, Juanjuan; Bian, Ying; Li, Huiqiang

    2016-06-01

    Crab meat and roe are highly nutritious delicacies in China. While extensive research has been conducted for allergens derived from crab-meat, data relevant to the allergenic potential of crab roe derived proteins, of which hemocyanin is a principal contender, are almost entirely absent. Using bioinformatics prediction and IgE-binding assays, the three principal immunodominant epitopes of hemocyanin were identified and then combined as a single recombinant fusion protein (rHc). This together with the full-length recombinant protein (Hc) were expressed in Escherichia coli and subsequently identified by SDS-PAGE and immunoblotting. Ninety-five percent of our patients were found to carry rHc-specific IgE antibodies by ELISA. Dot-blot inhibition, together with ELISA inhibition studies, showed that pre-incubation of patient sera with the recombinant epitope protein could inhibit26% to 63% (mean: 50%) of IgE binding to immobilized, full-length Hc and the dose-response curve represents as a sigmoid shape. The recombinant protein (rHc) represents a versatile biologic tool with which to diagnose and investigate therapies for E. sinensis allergy. PMID:27208437

  14. Rabbit IgG directed to a synthetic C-terminal peptide of the major grass pollen allergen Lol p I inhibits human basophil histamine release induced by natural Lol p I.

    PubMed

    van Ree, R; Aalberse, R C

    1995-03-01

    The potential role of allergen-specific IgG antibodies as 'blocking' antibodies in allergen-induced human basophil histamine release was investigated. This was studied in a model with the major grass pollen allergen Lol p I and polyclonal rabbit antisera directed against this allergen and against a synthetic peptide of its C terminus. When allergen and antibodies were allowed to preincubate, Lol p I induced histamine release was inhibited up to 85% by the antiserum against Lol p I. By omitting preincubation, and thereby more closely mimicking an in vivo situation, up to 55% inhibition was realized. This indicates that allergen-specific IgG can act as 'blocking' antibody without preincubation. Immunization of rabbits with a synthetic C-terminal peptide of Lol p I resulted in antibodies reactive with natural Lol p I. Despite their 100-fold lower avidity for Lol p I (as compared with antinatural Lol p I), these antibodies had the capacity to inhibit Lol p I induced histamine release for > 90% (up to 50% without preincubation). This indicates that it is possible to block histamine release induced by a major allergen with low-avidity IgG antibodies directed against a minor proportion of the allergen (25 amino acids). IgE antibodies from the donors studied were unreactive with this synthetic peptide, indicating that for blocking activity identical epitope specificity of IgE and IgG is not essential. This opens interesting perspectives for application of synthetic peptides in immunotherapy, distinct from their effects on T cell reactivity.

  15. Identification of a B-cell epitope of hyaluronidase, a major bee venom allergen, from its crystal structure in complex with a specific Fab.

    PubMed

    Padavattan, Sivaraman; Schirmer, Tilman; Schmidt, Margit; Akdis, Cezmi; Valenta, Rudolf; Mittermann, Irene; Soldatova, Lyudmila; Slater, Jay; Mueller, Ulrich; Markovic-Housley, Zora

    2007-05-01

    The major allergens of honeybee venom, hyaluronidase (Hyal) and phospholipase A2, can induce life-threatening IgE-mediated allergic reactions in humans. Although conventional immunotherapy is effective, up to 40% of patients develop allergic side effects including anaphylaxis and thus, there is a need for an improved immunotherapy. A murine monoclonal anti-Hyal IgG1 antibody (mAb 21E11), that competed for Hyal binding with IgEs from sera of bee venom allergic patients, was raised. The fragment of these IgG antibodies which bind to antigen (Fab) was produced and complexed (1:1) with Hyal. The crystal structure determination of Hyal/Fab 21E11 complex (2.6 A) enabled the identification of the Hyal-IgG interface which provides indirect information on the Hyal-IgE interaction (B-cell epitope). The epitope is composed of a linear array of nine residues (Arg138, His141-Arg148) located at the tip of a helix-turn-helix motive which protrudes away from the globular core and fits tightly into the deep surface pocket formed by the residues from the six complementarity determining regions (CDRs) of the Fab. The epitope is continuous and yet its conformation appears to be essential for Ab recognition, since the synthetic 15-mer peptide comprising the entire epitope (Arg138-Glu152) is neither recognized by mAb 21E11 nor by human IgEs. The structure of the complex provides the basis for the rational design of Hyal derivatives with reduced allergenic activity, which could be used in the development of safer allergen-specific immunotherapy. PMID:17374540

  16. Food allergens.

    PubMed

    Burks, W; Helm, R; Stanley, S; Bannon, G A

    2001-06-01

    A number of advances in the scientific knowledge concerning adverse food reactions have been made in the past few years. Understanding about the nature of the food allergen itself, the molecular characterization of the epitopes on these allergens, the pathophysiology of the clinical reaction, and the diagnostic methods have all been significantly enhanced.

  17. A hypoallergenic hybrid molecule with increased immunogenicity consisting of derivatives of the major grass pollen allergens, Phl p 2 and Phl p 6.

    PubMed

    Linhart, Birgit; Mothes-Luksch, Nadine; Vrtala, Susanne; Kneidinger, Michael; Valent, Peter; Valenta, Rudolf

    2008-07-01

    Allergen-specific immunotherapy is currently based on the administration of allergen extracts containing natural allergens. However, its broad application is limited by the poor quality of these extracts. Based on recombinant allergens, well-defined allergy vaccines for allergen-specific immunotherapy can be produced. Furthermore, they can be modified to reduce their allergenic activity and to avoid IgE-mediated side effects. Here, we demonstrate that the immunogenicity of two grass pollen-derived hypoallergenic allergen derivatives could be increased by engineering them as a single hybrid molecule. We used a hypoallergenic Phl p 2 mosaic, generated by fragmentation of the Phl p 2 sequence and reassembly of the resulting peptides in an altered order, and a truncated Phl p 6 allergen, to produce a hybrid protein. The hybrid retained the reduction of IgE reactivity and allergenic activity of its components as shown by ELISA and basophil activation assays. Immunization with the hybrid molecule demonstrated the increased immunogenicity of this molecule, leading to higher levels of allergen-specific IgG antibodies compared to the single components. These antibodies could inhibit patients' IgE binding to the wild-type allergens. Thus, the described strategy allows the development of safer and more efficacious vaccines for the treatment of grass pollen allergy.

  18. Mutants of the major ryegrass pollen allergen, Lol p 5, with reduced IgE-binding capacity: candidates for grass pollen-specific immunotherapy.

    PubMed

    Swoboda, Ines; De Weerd, Nicole; Bhalla, Prem L; Niederberger, Verena; Sperr, W R; Valent, Peter; Kahlert, Helga; Fiebig, Helmut; Verdino, Petra; Keller, Walter; Ebner, Christof; Spitzauer, Susanne; Valenta, Rudolf; Singh, Mohan B

    2002-01-01

    More than 400 million individuals are sensitized to grass pollen allergens. Group 5 allergens represent the most potent grass pollen allergens recognized by more than 80 % of grass pollen allergic patients. The aim of our study was to reduce the allergenic activity of group 5 allergens for specific immunotherapy of grass pollen allergy. Based on B- and T-cell epitope mapping studies and on sequence comparison of group 5 allergens from different grasses, point mutations were introduced by site-directed mutagenesis in highly conserved sequence domains of Lol p 5, the group 5 allergen from ryegrass. We obtained Lol p 5 mutants with low IgE-binding capacity and reduced allergenic activity as determined by basophil histamine release and by skin prick testing in allergic patients. Circular dichroism analysis showed that these mutants exhibited an overall structural fold similar to the recombinant Lol p 5 wild-type allergen. In addition, Lol p 5 mutants retained the ability to induce proliferation of group 5 allergen-specific T cell lines and clones. Our results demonstrate that a few point mutations in the Lol p 5 sequence yield mutants with reduced allergenic activity that represent potential vaccine candidates for immunotherapy of grass pollen allergy.

  19. MHC Class II-Restricted Presentation of the Major House Dust Mite Allergen Der p 1 Is GILT-Dependent: Implications for Allergic Asthma

    PubMed Central

    West, Laura Ciaccia; Grotzke, Jeff E.; Cresswell, Peter

    2013-01-01

    Gamma-interferon-inducible lysosomal thiol reductase (GILT) is known to reduce disulfide bonds present in proteins internalized by antigen presenting cells, facilitating optimal processing and presentation of peptides on Major Histocompatibility Complex class II molecules, as well as the subsequent activation of CD4-positive T lymphocytes. Here, we show that GILT is required for class II-restricted processing and presentation of immunodominant epitopes from the major house dust mite allergen Der p 1. In the absence of GILT, CD4-positive T cell responses to Der p 1 are significantly reduced, resulting in mitigated allergic airway inflammation in response to Der p 1 and house dust mite extracts in a murine model of asthma. PMID:23326313

  20. Cloning, expression, purification, crystallization and preliminary X-ray diffraction studies of a major group 7 allergen, Der f 7, from the dust mite Dermatophagoides farinae.

    PubMed

    Tan, Kang Wei; Kumar, Sundramurthy; Chew, Fook Tim; Mok, Yu Keung

    2011-12-01

    Der f 7 is a major group 7 allergen from the dust mite Dermatophagoides farinae that shows 86% sequence identity to the homologous allergen Der p 7 from D. pteronyssinus. Der f 7 was successfully overexpressed in an Escherichia coli expression system and purified to homogeneity using Ni-NTA affinity and size-exclusion column chromatography. SeMet-labelled Der f 7 was crystallized by the hanging-drop vapour-diffusion method using a reservoir solution consisting of 0.1 M bis-tris pH 7.4 and 28% polyethylene glycol monomethyl ether 2000 at 293 K. X-ray diffraction data were collected to 2.24 Å resolution using synchrotron radiation. The crystals belonged to the orthorhombic system, space group P2(1)2(1)2(1), with unit-cell parameters a = 50.19, b = 58.67, c = 123.81 Å. Based on the estimated Matthews coefficient (2.16 Å(3) Da(-1)), two molecules of Der f 7 could be present in the asymmetric unit of the crystal lattice.

  1. 26-Week oral safety study in macaques for transgenic rice containing major human T-cell epitope peptides from Japanese cedar pollen allergens.

    PubMed

    Domon, Eiji; Takagi, Hidenori; Hirose, Sakiko; Sugita, Koichi; Kasahara, Saori; Ebinuma, Hiroyasu; Takaiwa, Fumio

    2009-06-24

    A study of repeated oral administration of transgenic rice containing a hybrid peptide of major human T-cell epitopes (7Crp) from Japanese cedar pollen allergens was carried out in cynomolgus macaques over 26 weeks. The monkeys were divided into three groups, each comprising three males and three females, administered a high dose of transgenic rice, a low dose of transgenic rice, or a high dose of the parental rice strain. The transgenic rice 7crp#10 and the parental nontransgenic control were polished, steamed, mashed, and prepared in water at 40% (w/v). Monkeys were orally administered a high or low dose of transgenic rice or the nontransgenic control by gavage every day. No adverse effects on general behavior or body weight of animals were observed during the study. Analysis of blood from monkeys administered for 26 weeks showed that, with few exceptions, there were no significant differences in hematological or biochemical values between them. Additionally, neither pathological symptoms nor histopathological abnormalities were observed. Thus, it was concluded that oral administration of transgenic rice containing T-cell epitopes from Japanese cedar pollen allergens has no adverse effects.

  2. Mutational epitope analysis of Pru av 1 and Api g 1, the major allergens of cherry (Prunus avium) and celery (Apium graveolens): correlating IgE reactivity with three-dimensional structure.

    PubMed

    Neudecker, Philipp; Lehmann, Katrin; Nerkamp, Jörg; Haase, Tanja; Wangorsch, Andrea; Fötisch, Kay; Hoffmann, Silke; Rösch, Paul; Vieths, Stefan; Scheurer, Stephan

    2003-11-15

    Birch pollinosis is often accompanied by adverse reactions to food due to pollen-allergen specific IgE cross-reacting with homologous food allergens. The tertiary structure of Pru av 1, the major cherry (Prunus avium) allergen, for example, is nearly identical with Bet v 1, the major birch (Betula verrucosa) pollen allergen. In order to define cross-reactive IgE epitopes, we generated and analysed mutants of Pru av 1 and Api g 1.0101, the major celery (Apium graveolens) allergen, by immunoblotting, EAST (enzyme allergosorbent test), CD and NMR spectroscopy. The mutation of Glu45 to Trp45 in the P-loop region, a known IgE epitope of Bet v 1, significantly reduced IgE binding to Pru av 1 in a subgroup of cherry-allergic patients. The backbone conformation of Pru av 1 wild-type is conserved in the three-dimensional structure of Pru av 1 Trp45, demonstrating that the side chain of Glu45 is involved in a cross-reactive IgE epitope. Accordingly, for a subgroup of celery-allergic patients, IgE binding to the homologous celery allergen Api g 1.0101 was enhanced by the mutation of Lys44 to Glu. The almost complete loss of IgE reactivity to the Pru av 1 Pro112 mutant is due to disruption of its tertiary structure. Neither the mutation Ala112 nor deletion of the C-terminal residues 155-159 influenced IgE binding to Pru av 1. In conclusion, the structure of the P-loop partially explains the cross-reactivity pattern, and modulation of IgE-binding by site-directed mutagenesis is a promising approach to develop hypo-allergenic variants for patient-tailored specific immunotherapy.

  3. Fungal allergens.

    PubMed Central

    Horner, W E; Helbling, A; Salvaggio, J E; Lehrer, S B

    1995-01-01

    Airborne fungal spores occur widely and often in far greater concentrations than pollen grains. Immunoglobulin E-specific antigens (allergens) on airborne fungal spores induce type I hypersensitivity (allergic) respiratory reactions in sensitized atopic subjects, causing rhinitis and/or asthma. The prevalence of respiratory allergy to fungi is imprecisely known but is estimated at 20 to 30% of atopic (allergy-predisposed) individuals or up to 6% of the general population. Diagnosis and immunotherapy of allergy to fungi require well-characterized or standardized extracts that contain the relevant allergen(s) of the appropriate fungus. Production of standardized extracts is difficult since fungal extracts are complex mixtures and a variety of fungi are allergenic. Thus, the currently available extracts are largely nonstandardized, even uncharacterized, crude extracts. Recent significant progress in isolating and characterizing relevant fungal allergens is summarized in the present review. Particularly, some allergens from the genera Alternaria, Aspergillus, and Cladosporium are now thoroughly characterized, and allergens from several other genera, including some basidiomycetes, have also been purified. The availability of these extracts will facilitate definitive studies of fungal allergy prevalence and immunotherapy efficacy as well as enhance both the diagnosis and therapy of fungal allergy. PMID:7621398

  4. Establishment of recombinant major allergens Bet v 1 and Phl p 5a as Ph. Eur. reference standards and validation of ELISA methods for their measurement. Results from feasibility studies.

    PubMed

    Vieths, S; Barber, D; Chapman, M; Costanzo, A; Daas, A; Fiebig, H; Hanschmann, K M; Hrabina, M; Kaul, S; Ledesma, A; Moingeon, P; Reese, G; Schörner, C; van Ree, R; Weber, B; Buchheit, K H

    2012-04-01

    The potency of allergen extracts is determined as total allergenic activity without consideration of their composition and the units differ from one manufacturer to another, making it very difficult to compare the different products. Recently, purified major allergens have been obtained by recombinant DNA technology and produced under Good Manufacturing Practice (GMP) conditions. In principle, such recombinant allergens could be established as reference standards and could help for the standardisation of the major allergen content of allergen extracts. Two recombinant major allergens, one from birch pollen, rBet v 1, and one from Timothy grass pollen, Phl p 5a, have been selected at the end of the CREATE programme as a potential starting point for the establishment as European Pharmacopoeia (Ph. Eur.) Reference Standards through a project run by the Biological Standardisation Programme (BSP) of the European Directorate for the Quality of Medicines & HealthCare (EDQM). To this end, bulk candidate recombinant materials, produced under GMP conditions, were procured from two European manufacturers and subsequently formulated and lyophilised. Four ELISA systems from three different manufacturers were included in the project, two for Bet v 1 and two for Phl p 5a with the aim of establishing reference methods for determination of the respective major antigens both in natural allergen extracts as well as in recombinant allergen products. The project was run in 3 phases: a preparatory and preliminary testing phase (feasibility phase or Phase 1), an extended feasibility phase carried out in 3 laboratories (Phase 2) to confirm the transferability of the methods and an international collaborative study with a large number of participating laboratories (Phase 3). This article describes the work done in Phase 1 and Phase 2, i.e. the physico-chemical and biological characterisation of the recombinant candidate reference standards, the assessment of their suitability for the

  5. Outdoor allergens.

    PubMed Central

    Burge, H A; Rogers, C A

    2000-01-01

    Outdoor allergens are an important part of the exposures that lead to allergic disease. Understanding the role of outdoor allergens requires a knowledge of the nature of outdoor allergen-bearing particles, the distributions of their source, and the nature of the aerosols (particle types, sizes, dynamics of concentrations). Primary sources for outdoor allergens include vascular plants (pollen, fern spores, soy dust), and fungi (spores, hyphae). Nonvascular plants, algae, and arthropods contribute small numbers of allergen-bearing particles. Particles are released from sources into the air by wind, rain, mechanical disturbance, or active discharge mechanisms. Once airborne, they follow the physical laws that apply to all airborne particles. Although some outdoor allergens penetrate indoor spaces, exposure occurs mostly outdoors. Even short-term peak outdoor exposures can be important in eliciting acute symptoms. Monitoring of airborne biological particles is usually by particle impaction and microscopic examination. Centrally located monitoring stations give regional-scale measurements for aeroallergen levels. Evidence for the role of outdoor allergens in allergic rhinitis is strong and is rapidly increasing for a role in asthma. Pollen and fungal spore exposures have both been implicated in acute exacerbations of asthma, and sensitivity to some fungal spores predicts the existence of asthma. Synergism and/or antagonism probably occurs with other outdoor air particles and gases. Control involves avoidance of exposure (staying indoors, preventing entry of outdoor aerosols) as well as immunotherapy, which is effective for pollen but of limited effect for spores. Outdoor allergens have been the subject of only limited studies with respect to the epidemiology of asthma. Much remains to be studied with respect to prevalence patterns, exposure and disease relationships, and control. PMID:10931783

  6. Allergen databases and allergen semantics.

    PubMed

    Gendel, Steven M

    2009-08-01

    The efficacy of any specific bioinformatic analysis of the potential allergenicity of new food proteins depends directly on the nature and content of the databases that are used in the analysis. A number of different allergen-related databases have been developed, each designed to meet a different need. These databases differ in content, organization, and accessibility. These differences create barriers for users and prevent data sharing and integration. The development and application of appropriate semantic web technologies, (for example, a food allergen ontology) could help to overcome these barriers and promote the development of more advanced analytic capabilities.

  7. Allergenicity of processed food.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Food allergies have become a major public health issue in many countries. In the U.S. it is estimated that approximately 150 individuals die each year from accidental ingestion of an allergic food. As a result, the federal government recently passed the food allergen labeling law which went into ef...

  8. A combination vaccine for allergy and rhinovirus infections based on rhinovirus-derived surface protein VP1 and a nonallergenic peptide of the major timothy grass pollen allergen Phl p 1.

    PubMed

    Edlmayr, Johanna; Niespodziana, Katarzyna; Linhart, Birgit; Focke-Tejkl, Margarete; Westritschnig, Kerstin; Scheiblhofer, Sandra; Stoecklinger, Angelika; Kneidinger, Michael; Valent, Peter; Campana, Raffaela; Thalhamer, Josef; Popow-Kraupp, Theresia; Valenta, Rudolf

    2009-05-15

    Allergens and rhinovirus infections are among the most common elicitors of respiratory diseases. We report the construction of a recombinant combination vaccine for allergy and rhinovirus infections based on rhinovirus-derived VP1, the surface protein which is critically involved in infection of respiratory cells, and a nonallergenic peptide of the major grass pollen allergen Phl p 1. Recombinant hybrid molecules consisting of VP1 and a Phl p 1-derived peptide of 31 aa were expressed in Escherichia coli. The hybrid molecules did not react with IgE Abs from grass pollen allergic patients and lacked allergenic activity when exposed to basophils from allergic patients. Upon immunization of mice and rabbits, the hybrids did not sensitize against Phl p 1 but induced protective IgG Abs that cross-reacted with group 1 allergens from different grass species and blocked allergic patients' IgE reactivity to Phl p 1 as well as Phl p 1-induced basophil degranulation. Moreover, hybrid-induced IgG Abs inhibited rhinovirus infection of cultured human epithelial cells. The principle of fusing nonallergenic allergen-derived peptides onto viral carrier proteins may be used for the engineering of safe allergy vaccines which also protect against viral infections.

  9. Study of the epitope structure of purified Dac G I and Lol p I, the major allergens of Dactylis glomerata and Lolium perenne pollens, using monoclonal antibodies.

    PubMed

    Mourad, W; Mécheri, S; Peltre, G; David, B; Hébert, J

    1988-11-15

    The use of mAb allowed us to further analyze the cross-reactivity between purified Dac g I and Lol p I, the major allergens of Dactylis glomerata (cocksfoot) and Lolium perenne (Rye grass), respectively. It was first shown, using IEF, followed by immunoprinting, that serum IgE antibodies from most grass-sensitive patients recognize both Dac g I and Lol p I. Second, three different anti-Lol p I mAb, 290A-167, 348A-6, and 539A-6, and one anti-Dac g I mAb, P3B2 were all shown to react with Dac g I and Lol p I, indicating that the two molecules share common epitopes. Epitope specificity of the mAb was determined by competitive binding inhibition of a given labeled mAb to solid phase fixed Dac g I or Lol p I by the mAb. The results indicated that the four mAb are directed against four different and non-overlapping epitopes present on both allergens. Using double-binding RIA, our data strongly suggest that the common epitopes are not repetitive on both molecules. In addition to their similar physicochemical characteristics, such as isolectric points and m.w., Dac g I and Lol p I share four identical epitopes. Binding inhibition of human IgE to Lol p I and Dac g I by the mAb was also assessed. The results indicated that each mAb was able to inhibit such reactions to variable degree but no additive inhibition was observed when two mAb of different specificities were used in combination, suggesting that the human IgE binding site is partially shared by each epitope recognized by the four mAb.

  10. Stability of patch test allergens.

    PubMed

    Joy, Nicole Marie; Rice, Kristen R; Atwater, Amber Reck

    2013-01-01

    Patch testing is widely used in evaluating suspected contact dermatitis. One major component of a quality patch test result is a dependable, predictable allergen supply. The allergen needs to be present at a sufficient concentration to elicit a reaction in an allergic patient. To better understand the stability of patch-test allergens, we completed a systematic review of the literature. We found that there is variability in stability among patch-test allergens and that although a few have been shown to be stable, many degrade when in storage. In most cases, expiration dates should be honored. In addition, allergen panels should be prepared as close to the time of patch test application as is possible.

  11. Structural aspects of fungal allergens.

    PubMed

    Crameri, Reto

    2015-03-01

    Despite the increasing number of solved crystal structures of allergens, the key question why some proteins are allergenic and the vast majority is not remains unanswered. The situation is not different for fungal allergens which cover a wide variety of proteins with different chemical properties and biological functions. They cover enzymes, cell wall, secreted, and intracellular proteins which, except cross-reactive allergens, does not show any evidence for structural similarities at least at the three-dimensional level. However, from a diagnostic point of view, pure allergens biotechnologically produced by recombinant technology can provide us, in contrast to fungal extracts which are hardly producible as standardized reagents, with highly pure perfectly standardized diagnostic reagents.

  12. Stability of patch test allergens.

    PubMed

    Joy, Nicole Marie; Rice, Kristen R; Atwater, Amber Reck

    2013-01-01

    Patch testing is widely used in evaluating suspected contact dermatitis. One major component of a quality patch test result is a dependable, predictable allergen supply. The allergen needs to be present at a sufficient concentration to elicit a reaction in an allergic patient. To better understand the stability of patch-test allergens, we completed a systematic review of the literature. We found that there is variability in stability among patch-test allergens and that although a few have been shown to be stable, many degrade when in storage. In most cases, expiration dates should be honored. In addition, allergen panels should be prepared as close to the time of patch test application as is possible. PMID:24030367

  13. Fagopyrins and Protofagopyrins: Detection, Analysis, and Potential Phototoxicity in Buckwheat.

    PubMed

    Benković, Eva Tavčar; Kreft, Samo

    2015-06-24

    Buckwheat contains many healthy nutrients, and its consumption is therefore increasing. Buckwheat also contains fluorescent phototoxic fagopyrins. A systematic review of fagopyrins and the phototoxicity of buckwheat found that reliable quantitative data on fagopyrin toxicity are not yet available. Generally, buckwheat seeds, flour, and teas are safe in normal amounts. Diets extensively composed of buckwheat sprouts, herbs, and particularly flowers or of fagopyrin-rich buckwheat extracts may cause fagopyrism. A reference standard is needed, as it would enable the accurate evaluation of fagopyrin content in buckwheat products and would allow proper testing of their as yet unknown physical, chemical, and biological characteristics.

  14. Buckwheat phenolic metabolites in health and disease.

    PubMed

    Kreft, Marko

    2016-06-01

    Buckwheat (Fagopyrum esculentum Moench, F. tataricum Gaertner) groats and flour have been established globally as nutritional foods because of their high levels of proteins, polyphenols and minerals. In some regions, buckwheat herb is used as a functional food. In the present study, reports of in vitro studies, preclinical and clinical trials dealing with the effect of buckwheat and its metabolites were reviewed. There are numerous reports of potential health benefits of consuming buckwheat, which may be in the form of food, dietary supplements, home remedies or possibly pharmaceutical drugs; however, adverse effects, including those resulting from contamination, must be considered. There are reports of antioxidative activity of buckwheat, which contains high levels of rutin and quercetin. On the other hand, both cytotoxic and antigenotoxic effects have been shown. Reduction of hyperlipidaemia, reduction of blood pressure and improved weight regulation have been suggested. Consuming buckwheat may have a beneficial effect on diabetes, since lower postprandial blood glucose and insulin response have been reported. In addition, buckwheat metabolites, such as rutin, may have intrinsic protective effects in preserving insulin signalling. Rutin has also been suggested to have potential therapeutic applications for the treatment of Alzheimer's disease. The literature indicates that buckwheat is safe to consume and may have various beneficial effects on human health. PMID:27046048

  15. Allergy Work-Up Including Component-Resolved Diagnosis: How to Make Allergen-Specific Immunotherapy More Specific.

    PubMed

    Kleine-Tebbe, Jörg; Matricardi, Paolo M; Hamilton, Robert G

    2016-02-01

    Symptoms are recorded by obtaining a clinical history. Allergen sensitization is demonstrated by skin prick test or allergen-specific IgE serology. IgE sensitizations to allergen sources can be identified knowing the relationship between major aeroallergens and homologous allergen families. Some develop allergic sensitization to pan-allergens. Allergen extracts do not allow definitive separation of the sources. IgE antibody analysis of the major allergenic molecules facilitates differentiation of sensitizing allergen sources. IgE sensitizations to inhalant allergens are only relevant in the case of corresponding symptoms. In questionable cases, conjunctival or nasal provocation tests help induce confirmatory symptoms and identify relevant allergens for immunotherapy.

  16. A Cross-Reactive Human Single-Chain Antibody for Detection of Major Fish Allergens, Parvalbumins, and Identification of a Major IgE-Binding Epitope.

    PubMed

    Bublin, Merima; Kostadinova, Maria; Fuchs, Julian E; Ackerbauer, Daniela; Moraes, Adolfo H; Almeida, Fabio C L; Lengger, Nina; Hafner, Christine; Ebner, Christof; Radauer, Christian; Liedl, Klaus R; Valente, Ana Paula; Breiteneder, Heimo

    2015-01-01

    Fish allergy is associated with moderate to severe IgE-mediated reactions to the calcium binding parvalbumins present in fish muscle. Allergy to multiple fish species is caused by parvalbumin-specific cross-reactive IgE recognizing conserved epitopes. In this study, we aimed to produce cross-reactive single chain variable fragment (scFv) antibodies for the detection of parvalbumins in fish extracts and the identification of IgE epitopes. Parvalbumin-specific phage clones were isolated from the human ETH-2 phage display library by three rounds of biopanning either against cod parvalbumin or by sequential biopanning against cod (Gad m 1), carp (Cyp c 1) and rainbow trout (Onc m 1) parvalbumins. While biopanning against Gad m 1 resulted in the selection of clones specific exclusively for Gad m 1, the second approach resulted in the selection of clones cross-reacting with all three parvalbumins. Two clones, scFv-gco9 recognizing all three parvalbumins, and scFv-goo8 recognizing only Gad m 1 were expressed in the E. coli non-suppressor strain HB2151 and purified from the periplasm. scFv-gco9 showed highly selective binding to parvalbumins in processed fish products such as breaded cod sticks, fried carp and smoked trout in Western blots. In addition, the scFv-gco9-AP produced as alkaline phosphatase fusion protein, allowed a single-step detection of the parvalbumins. In competitive ELISA, scFv-gco9 was able to inhibit binding of IgE from fish allergic patients' sera to all three β-parvalbumins by up to 80%, whereas inhibition by scFv-goo8 was up to 20%. 1H/15N HSQC NMR analysis of the rGad m 1:scFv-gco9 complex showed participation of amino acid residues conserved among these three parvalbumins explaining their cross-reactivity on a molecular level. In this study, we have demonstrated an approach for the selection of cross-reactive parvalbumin-specific antibodies that can be used for allergen detection and for mapping of conserved epitopes. PMID:26579717

  17. A Cross-Reactive Human Single-Chain Antibody for Detection of Major Fish Allergens, Parvalbumins, and Identification of a Major IgE-Binding Epitope

    PubMed Central

    Fuchs, Julian E.; Ackerbauer, Daniela; Moraes, Adolfo H.; Almeida, Fabio C. L.; Lengger, Nina; Hafner, Christine; Ebner, Christof; Radauer, Christian; Liedl, Klaus R.; Valente, Ana Paula; Breiteneder, Heimo

    2015-01-01

    Fish allergy is associated with moderate to severe IgE-mediated reactions to the calcium binding parvalbumins present in fish muscle. Allergy to multiple fish species is caused by parvalbumin-specific cross-reactive IgE recognizing conserved epitopes. In this study, we aimed to produce cross-reactive single chain variable fragment (scFv) antibodies for the detection of parvalbumins in fish extracts and the identification of IgE epitopes. Parvalbumin-specific phage clones were isolated from the human ETH-2 phage display library by three rounds of biopanning either against cod parvalbumin or by sequential biopanning against cod (Gad m 1), carp (Cyp c 1) and rainbow trout (Onc m 1) parvalbumins. While biopanning against Gad m 1 resulted in the selection of clones specific exclusively for Gad m 1, the second approach resulted in the selection of clones cross-reacting with all three parvalbumins. Two clones, scFv-gco9 recognizing all three parvalbumins, and scFv-goo8 recognizing only Gad m 1 were expressed in the E. coli non-suppressor strain HB2151 and purified from the periplasm. scFv-gco9 showed highly selective binding to parvalbumins in processed fish products such as breaded cod sticks, fried carp and smoked trout in Western blots. In addition, the scFv-gco9-AP produced as alkaline phosphatase fusion protein, allowed a single-step detection of the parvalbumins. In competitive ELISA, scFv-gco9 was able to inhibit binding of IgE from fish allergic patients’ sera to all three β-parvalbumins by up to 80%, whereas inhibition by scFv-goo8 was up to 20%. 1H/15N HSQC NMR analysis of the rGad m 1:scFv-gco9 complex showed participation of amino acid residues conserved among these three parvalbumins explaining their cross-reactivity on a molecular level. In this study, we have demonstrated an approach for the selection of cross-reactive parvalbumin-specific antibodies that can be used for allergen detection and for mapping of conserved epitopes. PMID:26579717

  18. Allergens are not pathogens

    PubMed Central

    Weiss, Richard; Scheiblhofer, Sandra; Thalhamer, Josef

    2014-01-01

    Vaccination against infectious diseases has been one of the major breakthroughs in human medical history, saving the lives of millions of people each year. More recently, prophylactic vaccination against non-infectious diseases such as cancer, Alzheimer’s disease, diabetes, and type I allergy is being investigated. Particularly in case of IgE-driven allergic disorders, which afflict almost a quarter of the population in highly developed countries, preventative measures would represent a major improvement for patients’ health as well as an economic relief for public health services. As an alternative to allergen-specific immunotherapy, prophylactic vaccination against type I allergic diseases could slow down or even stop the progress of the allergy pandemic. Allergen-encoding gene-based vaccines, i.e., plasmid DNA and mRNA vaccines, provide the advantage of purity over crude allergen extracts, which involve the risk of de novo sensitizations. Furthermore, these formulations have been demonstrated to induce T helper 1 as well as T regulatory immune responses—a pre-requisite for prophylactic intervention against allergies. However, prophylactic vaccines against environmental allergens strikingly differ from conventional vaccines against infectious diseases or therapeutic approaches concerning the underlying immunological mechanisms. PMID:24280693

  19. Murine Model of Buckwheat Allergy by Intragastric Sensitization with Fresh Buckwheat Flour Extract

    PubMed Central

    Oh, Sejo; Lee, Kisun; Jang, Young-Ju; Sohn, Myung-Hyun; Lee, Kyoung-En; Kim, Kyu-Earn

    2005-01-01

    Food allergies affect about 4% of the Korean population, and buckwheat allergy is one of the most severe food allergies in Korea. The purpose of the present study was to develop a murine model of IgE-mediated buckwheat hypersensitivity induced by intragastric sensitization. Young female C3H/HeJ mice were sensitized and challenged intragastricly with fresh buckwheat flour (1, 5, 25 mg/dose of proteins) mixed in cholera toxin, followed by intragastric challenge. Anaphylactic reactions, antigen-specific antibodies, splenocytes proliferation assays and cytokine productions were evaluated. Oral buckwheat challenges of sensitized mice provoked anaphylactic reactions such as severe scratch, perioral/periorbital swellings, or decreased activity. Reactions were associated with elevated levels of buckwheat-specific IgE antibodies. Splenocytes from buckwheat allergic mice exhibited significantly greater proliferative responses to buckwheat than non-allergic mice. Buckwheat-stimulated IL-4, IL-5, and INF-γ productions were associated with elevated levels of buckwheat-specific IgE in sensitized mice. In this model, 1 mg and 5 mg dose of sensitization produced almost the same degree of Th2-directed immune response, however, a 25 mg dose showed blunted antibody responses. In conclusion, we developed IgE-mediated buckwheat allergy by intragastric sensitization and challenge, and this model could provide a good tool for future studies. PMID:16100445

  20. [Soybean allergens and hypoallergenic germplasm enhancement].

    PubMed

    Fang, Xu-Qian; Zhu, You-Lin; Qiu, Li-Juan

    2006-08-01

    Food allergy is a public sanitary problem which has received attention worldwide. It is becoming an increasingly interesting problem to decrease the concentration of allergens for improvement of the food security. Soybean allergens in seeds are composing of storage proteins, structure proteins, and disease-related proteins. Among them, Gly m Bd 28K, Gly m Bd 30K and Gly m Bd 60K are the major allergens located in 7S conglycinin fragments. By recognizing allergens' physicochemical property, hypersensitivity and gene structure, certain progresses had been made to reduce the concentration of allergens in soybean through food processing, traditional breeding and genetic engineering. The paper reviewed the sorts and characters of soybean allergens, the physicochemical property of the three immunodominant allergens and their gene structures. Progress in developing hypoallergenic cultivars was also discussed.

  1. Allergen Immunotherapy.

    PubMed

    Rael, Efren

    2016-09-01

    Allergies affect a large proportion of the population. Allergies can adversely affect productivity, sleep, and quality of life and can lead to life-threatening reactions. Allergies can spread to affect multiple organ systems. Allergen immunotherapy is the only therapy that can change the natural history of allergic disease. PMID:27545737

  2. Peanut allergens.

    PubMed

    Becker, Wolf-Meinhard; Jappe, Uta

    2014-01-01

    The earliest known evidence of peanut farming dates back 7,600 years. With a prevalence of roughly 1%, peanut allergy is a diagnostic and treatment challenge, but is also a very good model for studying all aspects of food allergy, including its molecular basis and pathomechanisms. Therefore, the very starting point for elucidating all these aspects is the identification of peanut allergens with subsequent clearing of their structure and their preparation as pure recombinant and/or natural allergens. This is the basis for in vitro diagnostic tests as well as the development of immunotherapeutic drugs. With regard to class I food allergy, peanut allergy affects by far the largest group of patients. In peanuts, 12 allergens have been identified and their molecular characteristics are described herein. Ara h 1, Ara h 3.01 and Ara h 3.02 (the former Ara h 4) belong to the cupin superfamily. The conglutins Ara h 2, Ara h 6 and Ara h 7, and the non-specific lipid transfer protein Ara h 9 belong to the prolamin superfamily. Ara h 5 (profilin) and Ara h 8 (Bet v 1-homologous protein) cause class II food allergies and are associated with inhalation allergy to pollen via the sequential and/or conformational similarity of molecules. Two peanut oleosins are listed as Ara h 10 and Ara h 11 and two defensins as Ara h 12 and Ara h 13 by the WHO/IUIS Allergen Nomenclature Subcommittee. The effect of the above-specified allergens has to be considered in the context of their matrix, which is influenced by processing factors and the individual's immune system. PMID:24925406

  3. IgG4 inhibits peanut-induced basophil and mast cell activation in peanut-tolerant children sensitized to peanut major allergens

    PubMed Central

    Santos, Alexandra F.; James, Louisa K.; Bahnson, Henry T.; Shamji, Mohammed H.; Couto-Francisco, Natália C.; Islam, Sabita; Houghton, Sally; Clark, Andrew T.; Stephens, Alick; Turcanu, Victor; Durham, Stephen R.; Gould, Hannah J.; Lack, Gideon

    2015-01-01

    explanation for the absence of clinical reactivity in PS patients sensitized to major peanut allergens. PMID:25670011

  4. Mapping and mutational analysis of the IgE-binding epitopes on Ara h 1, a legume vicilin protein and a major allergen in peanut hypersensitivity.

    PubMed

    Burks, A W; Shin, D; Cockrell, G; Stanley, J S; Helm, R M; Bannon, G A

    1997-04-15

    Peanut allergy is a significant health problem because of the prevelance and potential severity of the allergic reaction. Serum IgE from patients with documented peanut hypersensitivity reactions and overlapping peptides were used to identify the IgE-binding epitopes on the major peanut allergen, Ara h 1. At least twenty-three different linear IgE-binding epitopes, located throughout the length of the Ara h 1 protein, were identified. All of the epitopes were 6-10 amino acids in length, but there was no obvious sequence motif shared by all peptides. Four of the peptides appeared to be immunodominant IgE-binding epitopes in that they were recognized by serum from more than 80% of the patients tested and bound more IgE than any of the other Ara h 1 epitopes. Mutational analysis of the immunodominant epitopes revealed that single amino acid changes within these peptides had dramatic effects on IgE-binding characteristics. The identification and determination of the IgE-binding capabilities of core amino acids in epitopes on the Ara h 1 protein will make it possible to address the pathophysiologic and immunologic mechanisms regarding peanut hypersensitivity reactions specifically and food hypersensitivity in general.

  5. The Major Cow Milk Allergen Bos d 5 Manipulates T-Helper Cells Depending on Its Load with Siderophore-Bound Iron

    PubMed Central

    Roth-Walter, Franziska; Pacios, Luis F.; Gomez-Casado, Cristina; Hofstetter, Gerlinde; Roth, Georg A.; Singer, Josef; Diaz-Perales, Araceli; Jensen-Jarolim, Erika

    2014-01-01

    The mechanisms of allergic sensitization to milk are still elusive. The major allergen Bos d 5 belongs to the lipocalin-family and thus is able to transport numerous ligands. In this study we investigated its ability to bind to iron-siderophore complexes and tested the immune-modulatory properties of Bos d 5 in either forms. Structural and in silico docking analysis of Bos d 5 revealed that Bos d 5 is able to bind to iron via catechol-based flavonoids (quercetin, myricetin, luteolin) that act as siderophores as confirmed by spectral-analysis and iron staining. Calculated dissociation constants of docking analyses were below 1 µM by virtual addition of iron. When incubated with human peripheral blood mononuclear cells (PBMCs), only the apo-form of Bos d 5 led to an increase of CD4+positive cells and significantly elevated IL13 and IFNγ-levels. In contrast, holo-Bos d 5 decreased numbers of CD4 expressing cells and induced apoptosis. Taken together, our data give evidence that Bos d 5 is capable of binding iron via siderophores. Moreover, our data support for the first time the notion that the form of application (apo- or holo-form) is decisive for the subsequent immune response. The apo-form promotes Th2 cells and inflammation, whereas the holo-form appears to be immunosuppressive. PMID:25117976

  6. Detection of Ara h 1 (a major peanut allergen) in food using an electrochemical gold nanoparticle-coated screen-printed immunosensor.

    PubMed

    Alves, Rita C; Pimentel, Filipa B; Nouws, Henri P A; Marques, Raquel C B; González-García, María Begoña; Oliveira, M Beatriz P P; Delerue-Matos, Cristina

    2015-02-15

    A gold nanoparticle-coated screen-printed carbon electrode was used as the transducer in the development of an electrochemical immunosensor for Ara h 1 (a major peanut allergen) detection in food samples. Gold nanoparticles (average diameter = 32 nm) were electrochemically generated on the surface of screen-printed carbon electrodes. Two monoclonal antibodies were used in a sandwich-type immunoassay and the antibody-antigen interaction was electrochemically detected through stripping analysis of enzymatically (using alkaline phosphatase) deposited silver. The total time of the optimized immunoassay was 3h 50 min. The developed immunosensor allowed the quantification of Ara h 1 between 12.6 and 2000 ng/ml, with a limit of detection of 3.8 ng/ml, and provided precise (RSD <8.7%) and accurate (recovery >96.6%) results. The immunosensor was successfully applied to the analysis of complex food matrices (cookies and chocolate), being able to detect Ara h 1 in samples containing 0.1% of peanut.

  7. Impact of thermal processing on legume allergens.

    PubMed

    Verma, Alok Kumar; Kumar, Sandeep; Das, Mukul; Dwivedi, Premendra D

    2012-12-01

    Food induced allergic manifestations are reported from several parts of the world. Food proteins exert their allergenic potential by absorption through the gastrointestinal tract and can even induce life threatening anaphylaxis reactions. Among all food allergens, legume allergens play an important role in induction of allergy because legumes are a major source of protein for vegetarians. Most of the legumes are cooked either by boiling, roasting or frying before consumption, which can be considered a form of thermal treatment. Thermal processing may also include autoclaving, microwave heating, blanching, pasteurization, canning, or steaming. Thermal processing of legumes may reduce, eliminate or enhance the allergenic potential of a respective legume. In most of the cases, minimization of allergenic potential on thermal treatment has generally been reported. Thus, thermal processing can be considered an important tool by indirectly prevent allergenicity in susceptible individuals, thereby reducing treatment costs and reducing industry/office/school absence in case of working population/school going children. The present review attempts to explore various possibilities of reducing or eliminating allergenicity of leguminous food using different methods of thermal processing. Further, this review summarizes different methods of food processing, major legumes and their predominant allergenic proteins, thermal treatment and its relation with antigenicity, effect of thermal processing on legume allergens; also suggests a path that may be taken for future research to reduce the allergenicity using conventional/nonconventional methods.

  8. Seafood allergy and allergens: a review.

    PubMed

    Lehrer, S B; Ayuso, R; Reese, G

    2003-01-01

    Seafoods are composed of diverse sea organisms and humans are allergic to many of them. Tropomyosin is a major allergen in many shellfish, especially crustacea and mollusks. Interestingly, tropomyosin has also been identified as an important allergen in other invertebrates including dust mites and cockroaches, and it has been proposed by some to be an invertebrate pan allergen. Different regions of shrimp tropomyosin bind IgE; 5 major IgE-binding regions have been identified in shrimp tropomyosin containing 8 epitopes. Mutations of these shrimp allergenic epitopes can reduce seafood allergenicity; methods utilizing such mutations will provide safer vaccines for more effective treatment of seafood-allergic patients, and in the future less-allergenic seafood products for consumption.

  9. Structural basis for the specific recognition of the major antigenic peptide from the Japanese cedar pollen allergen Cry j 1 by HLA-DP5.

    PubMed

    Kusano, Seisuke; Kukimoto-Niino, Mutsuko; Satta, Yoko; Ohsawa, Noboru; Uchikubo-Kamo, Tomomi; Wakiyama, Motoaki; Ikeda, Mariko; Terada, Takaho; Yamamoto, Ken; Nishimura, Yasuharu; Shirouzu, Mikako; Sasazuki, Takehiko; Yokoyama, Shigeyuki

    2014-08-26

    The major allergen, Cry j 1, was isolated from Japanese cedar Cryptomeria japonica (Cry j) pollen and was shown to react with immunoglobulin E antibodies in the sera from pollinosis patients. We previously reported that the frequency of HLA-DP5 was significantly higher in pollinosis patients and the immunodominant peptides from Cry j 1 bound to HLA-DP5 to activate Th2 cells. In the present study, we determined the crystal structure of the HLA-DP5 heterodimer in complex with a Cry j 1-derived nine-residue peptide, at 2.4Å resolution. The peptide-binding groove recognizes the minimal peptide with 10 hydrogen bonds, including those between the negatively charged P1 pocket and the Lys side chain at the first position in the peptide sequence. We confirmed that HLA-DP5 exhibits the same Cry j 1-binding mode in solution, through pull-down experiments using structure-based mutations of Cry j 1. We also identified the characteristic residues of HLA-DP5 that are responsible for the distinct properties of the groove, by comparing the structure of HLA-DP5 and the previously reported structures of HLA-DP2 in complexes with pDRA of the self-antigen. The comparison revealed that the HLA-DP5·pCry j 1 complex forms several hydrogen bond/salt bridge networks between the receptor and the antigen that were not observed in the HLA-DP2·pDRA complex. Evolutionary considerations have led us to conclude that HLA-DP5 and HLA-DP2 represent two major groups of the HLA-DP family, in which the properties of the P1 and P4 pockets have evolved and acquired the present ranges of epitope peptide-binding specificities.

  10. [Allergen analysis].

    PubMed

    Röder, Martin; Weber, Wolfgang

    2016-07-01

    The fundamental requirement when testing for and ensuring compliance with legally required labelling regulations is the reliable analysis of food allergens. This can be carried out by means of either DNA (deoxyribonucleic acid) or protein detection. Protein detection has the advantage of directly detecting the allergenic component and can currently be carried out using immunological (enzyme-linked immunosorbent assay [ELISA])/lateral flow devices [LFD]) or mass spectrometry-based techniques. DNA detection is indirect, but allows the presence of food allergens to be validated through the use of another marker. Each method has its pros and cons, which have to be considered on a case-by-case basis. ELISA is quantitative, quick and easy to carry out and has high sensitivity. LFD testing is ideal for industrial applications, as the tests can be carried out on-site. Both antibody-based tests may have problems with processed foods and false positive results. Mass-spectrometric techniques show a lot of promise, but are currently still time-consuming and complex to carry out. They also run into problems with processed foods and their degree of sensitivity is matrix and parameter dependent. For these reasons, this technique is only occasionally used. Polymerase chain reaction (PCR) provides the highest specificity and, depending on the target sequence, a very good to good level of sensitivity. Despite the high stability of DNA, PCR is still subject to the influence of processing and matrix related factors. Due to natural variation and production-related changes in the structures relevant in the process of detection, all methods exhibit a relatively high level of uncertainty of measurement. At present, there is no method which provides the absolute correct quantification. However, by means of laboratory-based analyses it is possible to calibrate for the allergen in question and thus be able to make reliable measurements using methods that are already available. PMID

  11. Common food allergens and their IgE-binding epitopes.

    PubMed

    Matsuo, Hiroaki; Yokooji, Tomoharu; Taogoshi, Takanori

    2015-10-01

    Food allergy is an adverse immune response to certain kinds of food. Although any food can cause allergic reactions, chicken egg, cow's milk, wheat, shellfish, fruit, and buckwheat account for 75% of food allergies in Japan. Allergen-specific immunoglobulin E (IgE) antibodies play a pivotal role in the development of food allergy. Recent advances in molecular biological techniques have enabled the efficient analysis of food allergens. As a result, many food allergens have been identified, and their molecular structure and IgE-binding epitopes have also been identified. Studies of allergens have demonstrated that IgE antibodies specific to allergen components and/or the peptide epitopes are good indicators for the identification of patients with food allergy, prediction of clinical severity and development of tolerance. In this review, we summarize our current knowledge regarding the allergens and IgE epitopes in the well-researched allergies to chicken egg, cow's milk, wheat, shrimp, and peanut. PMID:26433529

  12. Mammalian-derived respiratory allergens - implications for diagnosis and therapy of individuals allergic to furry animals.

    PubMed

    Nilsson, Ola B; van Hage, Marianne; Grönlund, Hans

    2014-03-01

    Furry animals cause respiratory allergies in a significant proportion of the population. A majority of all mammalian allergens are spread as airborne particles, and several have been detected in environments where furry animals are not normally kept. The repertoire of allergens from each source belongs to a restricted number of allergen families. Classification of allergen families is particularly important for the characterization of allergenicity and cross-reactivity of allergens. In fact, major mammalian allergens are taken from only three protein families, i.e. the secretoglobin, lipocalin and kallikrein families. In particular, the lipocalin superfamily harbours major allergens in all important mammalian allergen sources, and cross-reactivity between lipocalin allergens may explain cross-species sensitization between mammals. The identification of single allergen components is of importance to improve diagnosis and therapy of allergic patients using component-resolved diagnostics and allergen-specific immunotherapy (ASIT) respectively. Major disadvantages with crude allergen extracts for these applications emphasize the benefits of careful characterization of individual allergens. Furthermore, detailed knowledge of the characteristics of an allergen is crucial to formulate attenuated allergy vaccines, e.g. hypoallergens. The diverse repertoires of individual allergens from different mammalian species influence the diagnostic potential and clinical efficacy of ASIT to furry animals. As such, detailed knowledge of individual allergens is essential for adequate clinical evaluation. This review compiles current knowledge of the allergen families of mammalian species, and discusses how this information may be used for improved diagnosis and therapy of individuals allergic to mammals. PMID:24041755

  13. Mammalian-derived respiratory allergens - implications for diagnosis and therapy of individuals allergic to furry animals.

    PubMed

    Nilsson, Ola B; van Hage, Marianne; Grönlund, Hans

    2014-03-01

    Furry animals cause respiratory allergies in a significant proportion of the population. A majority of all mammalian allergens are spread as airborne particles, and several have been detected in environments where furry animals are not normally kept. The repertoire of allergens from each source belongs to a restricted number of allergen families. Classification of allergen families is particularly important for the characterization of allergenicity and cross-reactivity of allergens. In fact, major mammalian allergens are taken from only three protein families, i.e. the secretoglobin, lipocalin and kallikrein families. In particular, the lipocalin superfamily harbours major allergens in all important mammalian allergen sources, and cross-reactivity between lipocalin allergens may explain cross-species sensitization between mammals. The identification of single allergen components is of importance to improve diagnosis and therapy of allergic patients using component-resolved diagnostics and allergen-specific immunotherapy (ASIT) respectively. Major disadvantages with crude allergen extracts for these applications emphasize the benefits of careful characterization of individual allergens. Furthermore, detailed knowledge of the characteristics of an allergen is crucial to formulate attenuated allergy vaccines, e.g. hypoallergens. The diverse repertoires of individual allergens from different mammalian species influence the diagnostic potential and clinical efficacy of ASIT to furry animals. As such, detailed knowledge of individual allergens is essential for adequate clinical evaluation. This review compiles current knowledge of the allergen families of mammalian species, and discusses how this information may be used for improved diagnosis and therapy of individuals allergic to mammals.

  14. Identification of a new IgE-binding epitope of peanut oleosin that cross-reacts with buckwheat.

    PubMed

    Kobayashi, Shoko; Katsuyama, Shinta; Wagatsuma, Tamae; Okada, Shinji; Tanabe, Soichi

    2012-01-01

    Peanut and buckwheat induce a severe allergic reaction, anaphylaxis, which is considered to be mediated by immunoglobulin E (IgE). We identified in this study a new IgE-binding epitope of the peanut allergen that cross-reacted with buckwheat. The phosphate-buffered saline-soluble fraction of buckwheat inhibited the binding between IgE and the peanut allergen. A cross-reactive peptide was isolated from the α-chymotrypsin hydrolysate of peanut. Based on the amino acid sequence and mass spectrometric analysis data, the peptide was identified as Ser-Asp-Gln-Thr-Arg-Thr-Gly-Tyr (SDQTRTGY); this sequence is identical to amino acids 2-9 in the N-terminal hydrophilic domain of oleosin 3 which is located on the surface of the lipid storage body. Synthetic SDQTRTGY was found to bind with IgE in the sera of all eight peanut-allergic patients tested. Since many foods of plant origin contain oleosin, the possibility of an anaphylactic cross-reaction in allergic patients should always be considered.

  15. Influence of ultrasonic treatment on the allergenic properties of Shrimp ( Penaeus vannamei) Allergen

    NASA Astrophysics Data System (ADS)

    Zhenxing, Li; Hong, Lin; Limin, Cao

    2006-04-01

    The present study was undertaken to determine whether high intensity ultrasound could reduce the allergic properties of shrimp allergens. Reducing the allergenic properties of these allergens will be beneficial to allergic individuals. Samples of shrimp protein extract and shrimp muscle were treated by high-intensity ultrasound with water bathing at 0°C or 50°C for different time periods. The treated and untreated samples were then analyzed by SDS-PAGE, Western blots and competitive inhibition ELISA (Ci-ELISA) to determine the shrimp allergenicity. The results show that high-intensity ultrasound has no effect on allergenicity when the extracts were treated at 0°C. However, a significant decrease was observed in the level of the major shrimp allergen, Pen a 1, when the samples were treated at 50°C. In the determination of allergenicity with Ci-ELISA, a reduction in IgE binding was also observed.

  16. Bacterial fermentation of recombinant major wasp allergen Antigen 5 using oxygen limiting growth conditions improves yield and quality of inclusion bodies.

    PubMed

    Kischnick, Stefanie; Weber, Bernhard; Verdino, Petra; Keller, Walter; Sanders, Ernst A; Anspach, F Birger; Fiebig, Helmut; Cromwell, Oliver; Suck, Roland

    2006-06-01

    A process for bacterial expression and purification of the recombinant major wasp allergen Antigen 5 (Ves v 5) was developed to produce protein for diagnostic and therapeutic applications for type 1 allergic diseases. Special attention was focused on medium selection, fermentation conditions, and efficient refolding procedures. A soy based medium was used for fermentation to avoid peptone from animal origin. Animal-derived peptone required the use of isopropyl-beta-D-thiogalactopyranoside (IPTG) for the induction of expression. In the case of soy peptone, a constitutive expression was observed, suggesting the presence of a component that mimics IPTG. Batch cultivation at reduced stirrer speed caused a reduced biomass due to oxygen limitation. However, subsequent purification and processing of inclusion bodies yielded significantly higher amount of product. Furthermore, the protein composition of the inclusion bodies differed. Inclusion bodies were denatured and subjected to diafiltration. Detailed monitoring of diafiltration enabled the determination of the transition point. Final purification was conducted using cation-exchange and size-exclusion chromatography. Purified recombinant Ves v 5 was analyzed by RP-HPLC, CD-spectroscopy, SDS-PAGE, and quantification ELISA. Up to 15 mg highly purified Ves v 5 per litre bioreactor volume were obtained, with endotoxin concentrations less than 20 EU mg(-1) protein and high comparability to the natural counterpart. Analytical results confirm the suitability of the recombinant protein for diagnostic and clinical applications. The results clearly demonstrate that not only biomass, but especially growth conditions play a key role in the production of recombinant Ves v 5. This has an influence on inclusion body formation, which in turn influences the renaturation rate and absolute product yield. This might also be true for other recombinant proteins that accumulate as inclusion bodies in Escherichia coli.

  17. A human monoclonal IgE antibody that binds to MGL_1304, a major allergen in human sweat, without activation of mast cells and basophils.

    PubMed

    Ishii, Kaori; Hiragun, Makiko; Hiragun, Takaaki; Kan, Takanobu; Kawaguchi, Tomoko; Yanase, Yuhki; Tanaka, Akio; Takahagi, Shunsuke; Hide, Michihiro

    MGL_1304, a major allergen in human sweat for patients with atopic dermatitis and/or cholinergic urticaria, is secreted from Malassezia globosa on human skin. The amounts of MGL_1304 and IgE against MGL_1304 are evaluated by the histamine release test using basophils or mast cells sensitized with serum containing IgE against MGL_1304, and enzyme linked sorbent assay (ELISA) using MGL_1304 and anti-MGL_1304 antibodies. Here, we identified a human monoclonal IgE (ABS-IgE) that binds to the high affinity IgE receptor (FcεRI) and MGL_1304 with high affinity (KD = 1.99 nM) but does not release histamine from basophils and mast cells. An ELISA using ABS-IgE as a standard IgE revealed that the amount of IgE against MGL_1304 (1000 U/ml) in the standard sera of patients with AD, employed in our previous report, is 32 ng/ml. A sandwich ELISA using ABS-IgE as a detection antibody showed approximately 10 times lower detection limit for MGL_1304 than ELISA in which MGL_1304 is directly bound to an ELISA plate. Moreover, ABS-IgE prevented histamine release from mast cells and basophils by neutralizing MGL_1304 not only in a free form in solution, but also on FcεRI expressed on the cell surface without cell activation. ABS-IgE may be used both to quantify the amount of MGL_1304 and anti-MGL_1304 IgE, and possibly for the treatment of diseases caused/aggravated by type I allergy to MGL_1304. PMID:26541454

  18. A human monoclonal IgE antibody that binds to MGL_1304, a major allergen in human sweat, without activation of mast cells and basophils.

    PubMed

    Ishii, Kaori; Hiragun, Makiko; Hiragun, Takaaki; Kan, Takanobu; Kawaguchi, Tomoko; Yanase, Yuhki; Tanaka, Akio; Takahagi, Shunsuke; Hide, Michihiro

    MGL_1304, a major allergen in human sweat for patients with atopic dermatitis and/or cholinergic urticaria, is secreted from Malassezia globosa on human skin. The amounts of MGL_1304 and IgE against MGL_1304 are evaluated by the histamine release test using basophils or mast cells sensitized with serum containing IgE against MGL_1304, and enzyme linked sorbent assay (ELISA) using MGL_1304 and anti-MGL_1304 antibodies. Here, we identified a human monoclonal IgE (ABS-IgE) that binds to the high affinity IgE receptor (FcεRI) and MGL_1304 with high affinity (KD = 1.99 nM) but does not release histamine from basophils and mast cells. An ELISA using ABS-IgE as a standard IgE revealed that the amount of IgE against MGL_1304 (1000 U/ml) in the standard sera of patients with AD, employed in our previous report, is 32 ng/ml. A sandwich ELISA using ABS-IgE as a detection antibody showed approximately 10 times lower detection limit for MGL_1304 than ELISA in which MGL_1304 is directly bound to an ELISA plate. Moreover, ABS-IgE prevented histamine release from mast cells and basophils by neutralizing MGL_1304 not only in a free form in solution, but also on FcεRI expressed on the cell surface without cell activation. ABS-IgE may be used both to quantify the amount of MGL_1304 and anti-MGL_1304 IgE, and possibly for the treatment of diseases caused/aggravated by type I allergy to MGL_1304.

  19. Quantitative methods for food allergens: a review.

    PubMed

    Kirsch, Stéphanie; Fourdrilis, Séverine; Dobson, Rowan; Scippo, Marie-Louise; Maghuin-Rogister, Guy; De Pauw, Edwin

    2009-09-01

    The quantitative detection of allergens in the food chain is a strategic health objective as the prevalence of allergy continues to rise. Food allergenicity is caused by proteins either in their native form or in forms resulting from food processing. Progress in mass spectrometry greatly opened up the field of proteomics. These advances are now available for the detection and the quantification of traces of allergenic proteins in complex mixtures, and complete the set of biological tests used until now, such as ELISA or PCR. We review methods classified according to their ability to simultaneously quantify and identify allergenic proteins and underline major advances in the mass-spectrometric methods.

  20. Biology of weed pollen allergens.

    PubMed

    Gadermaier, Gabriele; Dedic, Azra; Obermeyer, Gerhard; Frank, Susanne; Himly, Martin; Ferreira, Fatima

    2004-09-01

    Weeds represent a heterogeneous group of plants, usually defined by no commercial or aesthetic value. Important allergenic weeds belong to the plant families Asteraceae, Amaranthaceae, Urticaceae, Euphorbiaceae, and Plantaginaceae. Major allergens from ragweed, mugwort, feverfew, pellitory, goosefoot, Russian thistle, plantain, and Mercurialis pollen have been characterized to varying degrees. Four major families of proteins seem to be the major cause of allergic reactions to weed pollen: the ragweed Amb a 1 family of pectate lyases; the defensin-like Art v 1 family from mugwort, feverfew, and probably also from sunflower; the Ole e 1-like allergens Pla l 1 from plantain and Che a 1 from goosefoot; and the nonspecific lipid transfer proteins Par j 1 and Par j 2 from pellitory. As described for other pollens, weed pollen also contains the panallergens profilin and calcium-binding proteins, which are responsible for extensive cross-reactivity among pollen-sensitized patients.

  1. Identification, expression, and characterization of a major salivary allergen (Cul s 1) of the biting midge Culicoides sonorensis relevant for summer eczema in horses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salivary proteins of Culicoides biting midges are thought to play a key role in the induction of summer eczema (SE), a seasonal recurrent allergic dermatitis in horses. The present study describes the identification of a candidate allergen in artificially collected saliva of the North American speci...

  2. Extraction and analysis of coffee bean allergens.

    PubMed

    Lehrer, S B; Karr, R M; Salvaggio, J E

    1978-05-01

    Workers in the coffee industry can develop occupational allergic disease upon exposure to dust associated with coffee manufacturing. Since controversy exists as to the source or chemical nature of these allergens, the mouse model of reaginic antibody production was used to assess the potential sources of allergens in samples obtained from a local coffee manufacturing plant. Mice were immunized with extracts of coffee dust and beans and the resulting reaginic antibody response determined by the passive cutaneous anaphylaxis reaction. Cross-reacting allergens were detected in samples of coffee dust, cleaner can debris and green coffee beans, but not in chaff or roasted coffee beans. None of the allergens detected in coffee samples cross-reacted with extract of castor beans, although these extracts contained the potent castor bean allergen. Green coffee bean allergens partially purified by gel filtration were heterogeneous with respect to molecular size, although quite similar in their reactivity with reaginic antiserum. These results suggest that the green coffee bean is the major source of allergen in coffee manufacturing plants. This allergen is heterogeneous with respect to size and heat lability, and is immunochemically different from the castor bean allergen.

  3. Effects of buffer additives and thermal processing methods on the solubility of shrimp (Penaeus monodon) proteins and the immunoreactivity of its major allergen.

    PubMed

    Lasekan, Adeseye O; Nayak, Balunkeswar

    2016-06-01

    This study examines the potential of two buffer additives (Tween 20 and DTT) to improve the solubility of proteins from shrimp subjected to different heat treatments and the allergenicity of tropomyosin in the extracts. The concentration of soluble proteins extracted by all the buffers from processed shrimp was significantly reduced compared with untreated samples. The concentration of total soluble proteins from heat treated shrimp increased significantly when phosphate buffer containing both surfactant and reducing agent was used as the extraction buffer. However, the concentrations of heat-stable proteins in the buffers were mostly similar. The electrophoretic profile of extracted proteins showed that tropomyosin is very stable under the different heat treatment methods used in this study except for high pressure steaming where the intensity of tropomyosin band was reduced. Competitive inhibition ELISA showed that high pressure steaming reduced the allergenicity of tropomyosin compared with other heat treatments methods.

  4. Latex-allergic patients sensitized to the major allergen hevein and hevein-like domains of class I chitinases show no increased frequency of latex-associated plant food allergy

    PubMed Central

    Radauer, Christian; Adhami, Farzaneh; Fürtler, Irene; Wagner, Stefan; Allwardt, Dorothee; Scala, Enrico; Ebner, Christof; Hafner, Christine; Hemmer, Wolfgang; Mari, Adriano; Breiteneder, Heimo

    2011-01-01

    Allergies to certain fruits such as banana, avocado, chestnut and kiwi are described in 30–70% of latex-allergic patients. This association is attributed to the cross-reactivity between the major latex allergen hevein and hevein-like domains (HLDs) from fruit class I chitinases. We aimed to assess the extent of cross-reactivity between hevein and HLDs using sera from latex-allergic patients with and without plant food allergy. Hevein and HLDs of latex, banana, and avocado chitinases were expressed in Escherichia coli as fusion proteins with the maltose-binding protein and purified by affinity chromatography. IgE binding to these proteins was studied in sera from 59 latex-allergic patients and 20 banana-allergic patients without latex allergy by ELISA and ELISA inhibition. Additionally, 16,408 allergic patients’ sera were tested for IgE binding to hevein, latex chitinase, and wheat germ agglutinin using an allergen microarray. Hevein-specific IgE was detected in 34/59 (58%) latex-allergic patients’ sera. HLDs of latex, banana, and avocado chitinases were recognized by 21 (36%), 20 (34%), and 9 (15%) sera, respectively. In contrast, only one of 20 banana-allergic patients without latex allergy was sensitized to chitinase HLDs. In most tested latex-allergic patients’ sera, IgE binding to hevein was only partially reduced by preincubation with HLDs. Among hevein-sensitized, latex-allergic patients, the percentage of plant food allergy (15/34 = 44%) was equal to latex-allergic patients without hevein sensitization (11/25 = 44%). In the general allergic population, 230 of 16,408 sera (1.4%) reacted to hevein and/or a hevein-like allergen. Of these, 128 sera showed an isolated sensitization to hevein, whereas only 17 bound to latex chitinase or wheat germ agglutinin without hevein sensitization. In conclusion, the IgE response to HLDs is elicited by hevein as sensitizing allergen in most cases. Despite considerable cross-reactivity between these allergens, no

  5. Household Arthropod Allergens in Korea

    PubMed Central

    Jeong, Kyoung Yong

    2009-01-01

    Arthropods are important in human health, which can transmit pathogens to humans, parasitize, or produce important allergens. Allergy prevalence becomes higher in Korea recently as well as other developed countries in contrast to a decrease of infectious diseases. Allergic diseases caused by household arthropods have increased dramatically during the last few decades since human beings spend more their time for indoor activities in modernized life style. Household arthropods are one of the most common causes of allergic diseases. Biological characterization of household arthropods and researches on their allergens will provide better understanding of the pathogenesis of allergic diseases and suggest new therapeutic ways. Therefore, studies on arthropods of allergenic importance can be considered one of the major research areas in medical arthropodology and parasitology. Here, the biology of several household arthropods, including house dust mites and cockroaches, the 2 most well known arthropods living indoor together with humans worldwide, and characteristics of their allergens, especially the research activities on these allergens performed in Korea, are summarized. PMID:19885330

  6. Buckwheat honeys: screening of composition and properties.

    PubMed

    Pasini, Federica; Gardini, Silvia; Marcazzan, Gian Luigi; Caboni, Maria Fiorenza

    2013-12-01

    The quality of 10 buckwheat honeys, collected from Italian and est European beekeepers declaring to produce monofloral honey, were evaluated by means of their pollen, physicochemical, phenolic and volatile composition data. The results of the traditional analyses and in particular electrical conductivity, optical rotation, pH and sugar composition revealed some poorly pure samples that could not fit in the buckwheat tipology. Honey volatiles, analysed by solid phase microextraction (SPME) and gas chromatography-mass-spectrometry (GC/MS), showed more than 100 volatile compounds, most of them present in all honey samples but with quantitative variation. Besides many furfural derivates, 3-methylbutanoic acid was the main volatile compound found in most of honeys. Also the presence of 2- and 3-methylbutanal and pheynalcetaldehyde confirmed the typical buckwheat aroma of some studied samples, corroborating physicochemical data. The HPLC phenolic profile was similar across the samples and p-hydroxybenzoic and p-coumaric acids proved to be the main components. PMID:23871027

  7. Molecular and immunological characterization of shellfish allergens.

    PubMed

    Leung, P S; Chu, K H

    1998-03-15

    Shellfish (crustaceans and mollusks) have long been known as a common cause of allergic reactions to food. Like other food allergies, the allergic reactions to shellfish involve IgE-mediated Type I hypersensitivity. Biochemical and molecular studies have documented the major shrimp allergen is the muscle protein tropomyosin. Subsequent molecular cloning studies on lobsters and crabs have characterized this protein as the common allergen in crustaceans. There has also been strong immunological evidence that tropomyosin is a cross-reactive allergen among crustaceans and mollusks. This is further confirmed by recent studies on the identification of allergens in squid and abalone. The advances in the characterization of shellfish allergens will not only enhance our understanding on the physiological basis of shellfish allergy but also lay the groundwork for the development of diagnostic and therapeutic design in food allergies.

  8. Effect of oleic acid on the allergenic properties of peanut and cashew allergens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Oleic acid is the major fatty acid in peanuts and cashews. There is limited information about its effect on peanut and cashew allergens during heating. The objective was to determine if heat treatment with oleic acid changes the allergenic properties of these nut proteins. Peanut and cashew protein...

  9. Origin and Functional Prediction of Pollen Allergens in Plants.

    PubMed

    Chen, Miaolin; Xu, Jie; Devis, Deborah; Shi, Jianxin; Ren, Kang; Searle, Iain; Zhang, Dabing

    2016-09-01

    Pollen allergies have long been a major pandemic health problem for human. However, the evolutionary events and biological function of pollen allergens in plants remain largely unknown. Here, we report the genome-wide prediction of pollen allergens and their biological function in the dicotyledonous model plant Arabidopsis (Arabidopsis thaliana) and the monocotyledonous model plant rice (Oryza sativa). In total, 145 and 107 pollen allergens were predicted from rice and Arabidopsis, respectively. These pollen allergens are putatively involved in stress responses and metabolic processes such as cell wall metabolism during pollen development. Interestingly, these putative pollen allergen genes were derived from large gene families and became diversified during evolution. Sequence analysis across 25 plant species from green alga to angiosperms suggest that about 40% of putative pollen allergenic proteins existed in both lower and higher plants, while other allergens emerged during evolution. Although a high proportion of gene duplication has been observed among allergen-coding genes, our data show that these genes might have undergone purifying selection during evolution. We also observed that epitopes of an allergen might have a biological function, as revealed by comprehensive analysis of two known allergens, expansin and profilin. This implies a crucial role of conserved amino acid residues in both in planta biological function and allergenicity. Finally, a model explaining how pollen allergens were generated and maintained in plants is proposed. Prediction and systematic analysis of pollen allergens in model plants suggest that pollen allergens were evolved by gene duplication and then functional specification. This study provides insight into the phylogenetic and evolutionary scenario of pollen allergens that will be helpful to future characterization and epitope screening of pollen allergens. PMID:27436829

  10. Allergen structures and epitopes.

    PubMed

    Meno, K H

    2011-07-01

    Human type 1 hypersensitivity diseases such as allergic rhinoconjunctivitis are characterized by allergen-specific IgE antibodies produced in allergic individuals after allergen exposure. IgE antibodies bound to receptors on the surface of effector cells trigger an allergic response by interacting with three-dimensional (conformational) epitopes on the allergen surface. Crystal structures are available for complexes of antibody specifically bound to five allergens, from birch pollen, bee venom, cockroach, cow's milk and timothy grass pollen. The details of the antibody-allergen interaction extending all the way to atomic resolution are available from such complexes. In vitro investigations using recombinant monoclonal antibodies and human basophils show that binding affinity is a key to triggering the allergic response. Continued molecular characterization of antibody-allergen interactions is paving the way for the use of recombinant allergens in allergen-specific diagnosis and immunotherapy. PMID:21668845

  11. Variability of Offending Allergens of Allergic Rhinitis According to Age: Optimization of Skin Prick Test Allergens

    PubMed Central

    Lee, Ji-Eun; Ahn, Jae-Chul; Han, Doo Hee; Kim, Dong-Young; Kim, Jung-Whun; Cho, Sang-Heon; Park, Heung-Woo

    2014-01-01

    Purpose This study evaluates offending allergens in patients with allergic rhinitis (AR) according to age that establish a minimal panel for skin prick test (SPT) allergens required to identify if a patient is sensitized. Methods We retrospectively analyzed SPT results according to age to determine the minimum test battery panel necessary to screen at least 93%-95% of AR patients. Allergic skin tests (common airborne indoor and outdoor allergens) were performed on 7,182 patients from January 2007 to June 2011. All patients were classified into 9 groups according to age; subsequently, we investigated offending allergens by age group. Results A total of 5,032 (70.1%) patients were found sensitized to at least one of the 55 aeroallergen extracts tested. The annual ranking of offending allergens was not significantly different from each other over the past 5 years. House dust mites (HDM) were the most prevalent allergens ranked from first to third for all 5 years. The allergens in the minimum test panel differed slightly among all age groups; in addition, the types of sensitized allergen sources were more diverse in the older versus younger age group. HDM covered a larger proportion of the sensitized allergens in the younger age group versus the older age group. Testing with 5 allergens (Dermatophagoides farinae, Tetranychus urticae, oak, mugwort and cockroach) adequately identified over 90% of the sensitized patients. Conclusions A SPT with around 5-7 allergens adequately detected most of the sensitization in the majority of the age groups in Korea. However, this study suggests that physicians perform the SPT with appropriately selected allergens in each age category for the screening of AR. PMID:24404393

  12. Effects of phytic acid on peanut allergens and allergenic properties of extracts.

    PubMed

    Chung, Si-Yin; Champagne, Elaine T

    2007-10-31

    Phytic acid would form soluble and insoluble complexes with proteins. Our objective was to determine if phytic acid forms insoluble complexes with major peanut allergens, and if such reaction results in a peanut extract with a lower level of soluble allergens and allergenic property. Extracts from raw and roasted peanuts were treated with and without phytic acid at various pH values and then analyzed by SDS-PAGE and a competitive inhibition ELISA (ciELISA). The ciELISA measured IgE binding using a pooled serum from peanut-allergic individuals. Results showed that phytic acid formed complexes with the major peanut allergens (Ara h 1 and Ara h 2), which were insoluble in acidic and neutral conditions. Succinylation of the allergens inhibited complex formation, indicating that lysine residues were involved. A 6-fold reduction in IgE binding or allergenic potency of the extract was observed after treatment with phytic acid. It was concluded that phytic acid formed insoluble complexes with the major peanut allergens, and resulted in a peanut extract with reduced allergenic potency. Application of phytic acid to a peanut butter slurry presented a similar result, indicating that phytic acid may find use in the development of hypoallergenic peanut-based products.

  13. Quantitation of major allergens in dust samples from urban populations collected in different seasons in two climatic areas of the Basque region (Spain).

    PubMed

    Echechipía, S; Ventas, P; Audícana, M; Urrutia, I; Gastaminza, G; Polo, F; Fernández de Corres, L

    1995-06-01

    We present the results of allergen content evaluation in 80 dust samples from 31 homes of atopic patients from two climatic areas (humid and subhumid), collected in two seasons of the year (autumn and winter). Monoclonal antibody-based immunoassays were used to quantify Der p 1, Der f 1, Der 2, Lep d 1, and Fel d 1. The results were compared according to climate, season, and the type of sensitization (Pyroglyphidae mites, storage mites, or grass pollens). We underline the predominance of Dermatophagoides pteronyssinus (89% of samples) over D. farinae (16% of samples) in our environment. Der p 1 rates were higher in the humid area (Mann-Whitney P < 0.001), especially in the autumn (Wilcoxon P < 0.05). Lep d 1 was detected in 23% of samples and Lep d 1 levels were higher in the homes of patients sensitized to storage mites (Mann-Whitney P < 0.05), whereas this allergen was not detected in the homes of pollen-allergic patients. Fel d 1 was detected in nine of the 31 homes (16% of samples) although there was a cat in only one home. PMID:7573840

  14. Animal Allergens and Their Presence in the Environment

    PubMed Central

    Zahradnik, Eva; Raulf, Monika

    2014-01-01

    Exposure to animal allergens is a major risk factor for sensitization and allergic diseases. Besides mites and cockroaches, the most important animal allergens are derived from mammals. Cat and dog allergies affect the general population; whereas, allergies to rodents or cattle is an occupational problem. Exposure to animal allergens is not limited to direct contact to animals. Based on their aerodynamic properties, mammalian allergens easily become airborne, attach to clothing and hair, and can be spread from one environment to another. For example, the major cat allergen Fel d 1 was frequently found in homes without pets and in public buildings, including schools, day-care centers, and hospitals. Allergen concentrations in a particular environment showed high variability depending on numerous factors. Assessment of allergen exposure levels is a stepwise process that involves dust collection, allergen quantification, and data analysis. Whereas a number of different dust sampling strategies are used, ELISA assays have prevailed in the last years as the standard technique for quantification of allergen concentrations. This review focuses on allergens arising from domestic, farm, and laboratory animals and describes the ubiquity of mammalian allergens in the human environment. It includes an overview of exposure assessment studies carried out in different indoor settings (homes, schools, workplaces) using numerous sampling and analytical methods and summarizes significant factors influencing exposure levels. However, methodological differences among studies have contributed to the variability of the findings and make comparisons between studies difficult. Therefore, a general standardization of methods is needed and recommended. PMID:24624129

  15. Enrichment of gluten-free tarhana with buckwheat flour.

    PubMed

    Bilgiçli, Nermin

    2009-01-01

    Tarhana, a traditional fermented cereal food in Turkey, is mainly prepared with wheat flour and yoghurt. Buckwheat is a major ingredient in the daily diet of the celiac patients due to its gluten-free composition. In this research, gluten-free tarhana samples were prepared with buckwheat flour (BWF), rice flour and corn starch. Control tarhana was made of wheat flour. In gluten-free formulations, instead of wheat flour, 40% BWF, 30% rice flour and 30% corn starch in the first formulation and 60% BWF, 20% rice flour and 20% corn starch in the second formulation were handled. BWF substitution at a 60% level increased the ash and fat contents of tarhana samples, but affected the lightness value of the samples negatively. Potassium, magnesium and phosphorus contents of the gluten-free tarhana increased significantly (P <0.05) with increasing amount of BWF in the formulation. Sensory evaluation showed that BWF addition resulted in changes in consistency, taste, sourness and grittiness. Tarhana containing 40% BWF received the highest taste and overall acceptability scores.

  16. Peanut allergens: an overview.

    PubMed

    Sáiz, Jorge; Montealegre, Cristina; Marina, Maria Luisa; García-Ruiz, Carmen

    2013-01-01

    Peanut is recognized as a potent food allergen producing one of the most frequent food allergies. This fact has originated the publication of an elevated number of scientific reports dealing with peanut allergens and, especially, the prevalence of peanut allergy. For this reason, the information available on peanut allergens is increasing and the debate about peanut allergy is always renewed. This article reviews the information currently available on peanut allergens and on the techniques used for their chemical characterization. Moreover, a general overview on the current biotechnological approaches used to reduce or eliminate peanut allergens is also provided. PMID:23638932

  17. Chemical composition and health effects of Tartary buckwheat.

    PubMed

    Zhu, Fan

    2016-07-15

    Tartary buckwheat (Fagopyrum tataricum) contains a range of nutrients including bioactive carbohydrates and proteins, polyphenols, phytosterols, vitamins, carotenoids, and minerals. The unique composition of Tartary buckwheat contributes to their various health benefits such as anti-oxidative, anti-cancer, anti-hypertension, anti-diabetic, cholesterol-lowering, and cognition-improving. Compared with the more widely cultivated and utilised common buckwheat (F. esculentum), Tartary buckwheat tends to contain higher amounts of certain bioactive components such as rutin, therefore, showing higher efficiency in preventing/treating various disorders. This review summarises the current knowledge of the chemical composition of Tartary buckwheat, and their bio-functions as studied by both in vitro and in vivo models. Tartary buckwheat can be further developed as a sustainable crop for functional food production to improve human health.

  18. Chemical composition and health effects of Tartary buckwheat.

    PubMed

    Zhu, Fan

    2016-07-15

    Tartary buckwheat (Fagopyrum tataricum) contains a range of nutrients including bioactive carbohydrates and proteins, polyphenols, phytosterols, vitamins, carotenoids, and minerals. The unique composition of Tartary buckwheat contributes to their various health benefits such as anti-oxidative, anti-cancer, anti-hypertension, anti-diabetic, cholesterol-lowering, and cognition-improving. Compared with the more widely cultivated and utilised common buckwheat (F. esculentum), Tartary buckwheat tends to contain higher amounts of certain bioactive components such as rutin, therefore, showing higher efficiency in preventing/treating various disorders. This review summarises the current knowledge of the chemical composition of Tartary buckwheat, and their bio-functions as studied by both in vitro and in vivo models. Tartary buckwheat can be further developed as a sustainable crop for functional food production to improve human health. PMID:26948610

  19. Identification, expression and characterization of a major salivary allergen (Cul s 1) of the biting midge Culicoides sonorensis relevant for summer eczema in horses

    PubMed Central

    Langner, Kathrin F. A.; Jarvis, Donald L.; Nimtz, Manfred; Heselhaus, Julia E.; McHolland, Linda E.; Leibold, Wolfgang; Drolet, Barbara S.

    2009-01-01

    Salivary proteins of Culicoides biting midges are thought to play a key role in summer eczema (SE), a seasonal recurrent allergic dermatitis in horses. The present study describes the identification, expression and clinical relevance of a candidate allergen of the North American midge Culicoides sonorensis. Immunoblot analysis of midge saliva revealed a 66 kDa protein (Cul s 1) that was bound by IgE from several SE-affected (SE+) horses. Further characterization by fragmentation, mass spectrometry and bioinformatics identified Cul s 1 as maltase, an enzyme involved in sugar meal digestion. A cDNA encoding Cul s 1 was isolated and expressed as a polyhistidine-tagged fusion protein in a baculovirus/insect cell expression system. The clinical relevance of the affinity-purified recombinant Cul s 1 (rCul s 1) was investigated by immunoblotting, histamine release testing (HRT) and intradermal testing (IDT) in eight SE+ and eight control horses. Seven SE+ horses had rCul s 1-specific IgE, whereas only one control animal had IgE directed against this allergen. Furthermore, the HRT showed rCul s 1 induced basophil degranulation in samples from seven of eight SE+ horses but in none of the control animals. rCul s 1 also induced immediate (7/8), late-phase (8/8) and delayed (1/8) skin reactivity in IDT on all SE+ horses that had a positive test with the whole body extract (WBE) of C. sonorensis. None of the control horses showed immediate or delayed skin reactivity with rCul s 1, and only one control horse had a positive late-phase response, while several non-specific late-phase reactions were observed with the insect WBE. Thus, we believe rCul s 1 is the first specific salivary allergen of C. sonorensis to be described that promises to advance both in vitro and in vivo diagnosis and may contribute to the development of immunotherapy for SE in horses. PMID:18708061

  20. Taxonomy of Allergenic Fungi.

    PubMed

    Levetin, Estelle; Horner, W Elliott; Scott, James A

    2016-01-01

    The Kingdom Fungi contains diverse eukaryotic organisms including yeasts, molds, mushrooms, bracket fungi, plant rusts, smuts, and puffballs. Fungi have a complex metabolism that differs from animals and plants. They secrete enzymes into their surroundings and absorb the breakdown products of enzyme action. Some of these enzymes are well-known allergens. The phylogenetic relationships among fungi were unclear until recently because classification was based on the sexual state morphology. Fungi lacking an obvious sexual stage were assigned to the artificial, now-obsolete category, "Deuteromycetes" or "Fungi Imperfecti." During the last 20 years, DNA sequencing has resolved 8 fungal phyla, 3 of which contain most genera associated with important aeroallergens: Zygomycota, Ascomycota, and Basidiomycota. Advances in fungal classification have required name changes for some familiar taxa. Because of regulatory constraints, many fungal allergen extracts retain obsolete names. A major benefit from this reorganization is that specific immunoglobulin E (IgE) levels in individuals sensitized to fungi appear to closely match fungal phylogenetic relationships. This close relationship between molecular fungal systematics and IgE sensitization provides an opportunity to systematically look at cross-reactivity and permits representatives from each taxon to serve as a proxy for IgE to the group. PMID:26725152

  1. Taxonomy of Allergenic Fungi.

    PubMed

    Levetin, Estelle; Horner, W Elliott; Scott, James A

    2016-01-01

    The Kingdom Fungi contains diverse eukaryotic organisms including yeasts, molds, mushrooms, bracket fungi, plant rusts, smuts, and puffballs. Fungi have a complex metabolism that differs from animals and plants. They secrete enzymes into their surroundings and absorb the breakdown products of enzyme action. Some of these enzymes are well-known allergens. The phylogenetic relationships among fungi were unclear until recently because classification was based on the sexual state morphology. Fungi lacking an obvious sexual stage were assigned to the artificial, now-obsolete category, "Deuteromycetes" or "Fungi Imperfecti." During the last 20 years, DNA sequencing has resolved 8 fungal phyla, 3 of which contain most genera associated with important aeroallergens: Zygomycota, Ascomycota, and Basidiomycota. Advances in fungal classification have required name changes for some familiar taxa. Because of regulatory constraints, many fungal allergen extracts retain obsolete names. A major benefit from this reorganization is that specific immunoglobulin E (IgE) levels in individuals sensitized to fungi appear to closely match fungal phylogenetic relationships. This close relationship between molecular fungal systematics and IgE sensitization provides an opportunity to systematically look at cross-reactivity and permits representatives from each taxon to serve as a proxy for IgE to the group.

  2. Mycorrhizal status and diversity of fungal endophytes in roots of common buckwheat (Fagopyrum esculentum) and tartary buckwheat (F. tataricum).

    PubMed

    Likar, Matevz; Bukovnik, Urska; Kreft, Ivan; Chrungoo, Nikhil K; Regvar, Marjana

    2008-09-01

    To determine the mycorrhizal status and to identify the fungi colonising the roots of the plants, common buckwheat (Fagopyrum esculentum) and tartary buckwheat (F. tataricum) were inoculated with an indigenous fungal mixture from a buckwheat field. Root colonisation was characterised by the hyphae and distinct microsclerotia of dark septate endophytes, with occasional arbuscules and vesicles of arbuscular mycorrhizal fungi. Sequences of arbuscular mycorrhizal fungi colonising tartary buckwheat clustered close to the Glomus species group A. Sequences with similarity to the Ceratobasidium/Rhizoctonia complex, a putative dark septate endophyte fungus, were amplified from the roots of both common and tartary buckwheat. To the best of our knowledge, this is the first report of arbuscular mycorrhizal colonisation in tartary buckwheat and the first molecular characterisation of these fungi that can colonise both of these economically important plant species.

  3. Protocol for simultaneous isolation of three important banana allergens.

    PubMed

    Nikolic, Jasna; Mrkic, Ivan; Grozdanovic, Milica; Popovic, Milica; Petersen, Arnd; Jappe, Uta; Gavrovic-Jankulovic, Marija

    2014-07-01

    Banana fruit (Musa acuminata) has become an important food allergen source in recent years. So far, 5 IgE reactive banana proteins have been identified, and the major allergens are: Mus a 2 (a class I chitinase, 31kDa), Mus a 4 (thaumatin-like protein, 21kDa), and Mus a 5 (β-1,3-glucanase, 33kDa). Due to variations in allergen expression levels, diagnostic reagents for food allergy can be improved by using individual allergen components instead of banana allergen extracts. The purpose of this study was to optimize the purification protocol of the three major allergens present in banana fruit: Mus a 2, Mus a 4 and Mus a 5. By employing a three-step purification protocol (a combination of anion-exchange, cation-exchange and reversed-phase chromatography) three important banana allergens were obtained in sufficient yield and high purity. Characterization of the purified proteins was performed by both biochemical (2-D PAGE, mass fingerprint and N-terminal sequencing) and immunochemical (immunoblot) methods. IgE reactivity to the purified allergens was tested by employing sera of five allergic patients. The purified allergens displayed higher sensitivity in IgE detection than the routinely used extracts. The three purified allergens are good candidates for reagents in component-based diagnosis of banana allergy.

  4. Reduction of rutin loss in buckwheat noodles and their physicochemical characterization

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rutin in buckwheat flour is converted into quercetin by rutin-degrading enzymes when water is added. Thus, buckwheat was subjected to hydrothermal treatments for minimizing the rutin loss in buckwheat-based foods by water addition. When native buckwheat flour was mixed with water, the rutin conten...

  5. Hypolipidemic activity of common (Fagopyrum esculentum Moench) and tartary (Fagopyrum tataricum Gaertn.) buckwheat.

    PubMed

    Tomotake, Hiroyuki; Kayashita, Jun; Kato, Norihisa

    2015-08-15

    Buckwheat grain has well-balanced nutritional value, whereas its digestibility is relatively low. This review summarizes recent advances in studies on the hypolipidemic activity of buckwheat. The most remarkable function is a powerful hypocholesterolemic activity of buckwheat protein in rats, which is far stronger than that of soy protein. The cholesterol-lowering effect is mediated by mechanisms involving higher excretion of fecal sterols and lower digestibility of buckwheat protein. The insoluble fraction of buckwheat protein associates with cholesterol and reduces micelle cholesterol uptake in caco-2 cells. Furthermore, consumption of buckwheat protein suppresses cholesterol-induced gallstones and body fat in rodents. Buckwheat sprouts also have hypolipidemic activity in rats or type 2 diabetic mice. Tartary buckwheat bran extract reduced the serum level of total cholesterol and triglyceride in hyperlipidemic rats. The consumption of buckwheat seed reduced low-density lipoprotein cholesterol in the pastureland Mongolian population. Taken together, buckwheat may be beneficial for prevention of hyperlipidemia.

  6. Food processing and allergenicity.

    PubMed

    Verhoeckx, Kitty C M; Vissers, Yvonne M; Baumert, Joseph L; Faludi, Roland; Feys, Marcel; Flanagan, Simon; Herouet-Guicheney, Corinne; Holzhauser, Thomas; Shimojo, Ryo; van der Bolt, Nieke; Wichers, Harry; Kimber, Ian

    2015-06-01

    Food processing can have many beneficial effects. However, processing may also alter the allergenic properties of food proteins. A wide variety of processing methods is available and their use depends largely on the food to be processed. In this review the impact of processing (heat and non-heat treatment) on the allergenic potential of proteins, and on the antigenic (IgG-binding) and allergenic (IgE-binding) properties of proteins has been considered. A variety of allergenic foods (peanuts, tree nuts, cows' milk, hens' eggs, soy, wheat and mustard) have been reviewed. The overall conclusion drawn is that processing does not completely abolish the allergenic potential of allergens. Currently, only fermentation and hydrolysis may have potential to reduce allergenicity to such an extent that symptoms will not be elicited, while other methods might be promising but need more data. Literature on the effect of processing on allergenic potential and the ability to induce sensitisation is scarce. This is an important issue since processing may impact on the ability of proteins to cause the acquisition of allergic sensitisation, and the subject should be a focus of future research. Also, there remains a need to develop robust and integrated methods for the risk assessment of food allergenicity.

  7. Allergenicity attributes of different peanut market types.

    PubMed

    Koppelman, Stef J; Jayasena, Shyamali; Luykx, Dion; Schepens, Erik; Apostolovic, Danijela; de Jong, Govardus A H; Isleib, Thomas G; Nordlee, Julie; Baumert, Joe; Taylor, Steve L; Cheng, Hsiaopo; Maleki, Soheila

    2016-05-01

    Four different market classes of peanut (Runner, Virginia Spanish, and Valencia) are commonly consumed in Western countries, but for some consumers peanuts are a main cause of food-induced anaphylaxis. Limited information is available on the comparative allergenicity of these distinct market classes. The aim of this study was to compare allergenicity attributes of different peanut cultivars. The protein content and protein profiles were highly comparable for all tested cultivars. All cultivar samples contained the major allergens Ara h 1, Ara h 2, Ara h 3 and Ara h 6, as assessed by SDS-PAGE and RP-HPLC, although some minor differences in major allergen content were found between samples. All samples were reactive in commercial ELISAs for detection and quantification of peanut protein. IgE-binding potency differed between samples with a maximum factor of 2, indicating a highly comparable allergenicity. Based on our observations, we conclude that peanuts from the main market types consumed in Western countries are highly comparable in their allergenicity attributes, indicating that safety considerations with regard to peanut allergy are not dependent on the peanut cultivar in question.

  8. Removing peanut allergens by tannic acid.

    PubMed

    Chung, Si-Yin; Reed, Shawndrika

    2012-10-01

    Tannic acid (TA) forms insoluble complexes with proteins. The aims here were to remove major peanut allergens as insoluble TA complexes and determine if they would dissociate and release the allergens at pH 2 and 8 (gut pH). Release of the allergens in the gut could lead to absorption and consequently an allergic reaction. TA (0.25, 0.5, 1, and 2 mg/ml) was added to a peanut butter extract (5 mg/ml; pH 7.2), stirred, and centrifuged. The precipitates were then suspended in buffer at pH 2, centrifuged, re-suspended at pH 8, and centrifuged. Supernatants from each step were analysed by SDS-PAGE, ELISA, and Western blots. The effect of NaCl (1M) on complexes was also determined. Results showed that complexes formed at a TA concentration >0.5 mg/ml did not release major peanut allergens at pH 2 and 8, regardless of 1M NaCl being present or not. IgE binding of the extracts was reduced substantially, especially at a TA concentration of 1-2 mg/ml. Animal or clinical studies are still needed before TA can find an application in the development of low-allergen peanut products/beverages or the removal of peanut allergens due to accidental ingestion. PMID:25005968

  9. Allergenicity attributes of different peanut market types.

    PubMed

    Koppelman, Stef J; Jayasena, Shyamali; Luykx, Dion; Schepens, Erik; Apostolovic, Danijela; de Jong, Govardus A H; Isleib, Thomas G; Nordlee, Julie; Baumert, Joe; Taylor, Steve L; Cheng, Hsiaopo; Maleki, Soheila

    2016-05-01

    Four different market classes of peanut (Runner, Virginia Spanish, and Valencia) are commonly consumed in Western countries, but for some consumers peanuts are a main cause of food-induced anaphylaxis. Limited information is available on the comparative allergenicity of these distinct market classes. The aim of this study was to compare allergenicity attributes of different peanut cultivars. The protein content and protein profiles were highly comparable for all tested cultivars. All cultivar samples contained the major allergens Ara h 1, Ara h 2, Ara h 3 and Ara h 6, as assessed by SDS-PAGE and RP-HPLC, although some minor differences in major allergen content were found between samples. All samples were reactive in commercial ELISAs for detection and quantification of peanut protein. IgE-binding potency differed between samples with a maximum factor of 2, indicating a highly comparable allergenicity. Based on our observations, we conclude that peanuts from the main market types consumed in Western countries are highly comparable in their allergenicity attributes, indicating that safety considerations with regard to peanut allergy are not dependent on the peanut cultivar in question. PMID:26921497

  10. Postharvest biology, quality and shelf-life of buckwheat microgreens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Buckwheat microgreens are rich in antioxidants and provitamins/vitamins, including flavonoids, carotenoids, and a-tocopherol. However, short shelf life has limited their commercial use. The purpose of this study was to optimize storage conditions to extend the shelf life of buckwheat microgreens. St...

  11. Wind-pollination and the roles of pollen allergenic proteins.

    PubMed

    Songnuan, Wisuwat

    2013-12-01

    Over the past few decades, there has been an explosion of understanding of the molecular nature of major allergens contained within pollens from the most important allergenic plant species. Most major allergens belong to only a few protein families. Protein characteristics, cross-reactivity, structures, and IgE binding epitopes have been determined for several allergens. These efforts have led to significant improvements in specific immunotherapy, yet there has been little discussion about the physiological functions of these proteins. Even with large amounts of available information about allergenic proteins from pollens, the incidence of pollen allergy continuously increases worldwide. The reason for this increase is unclear and is most likely due to a combination of factors. One important culprit might be a change in the pollen itself. Knowledge about pollen biology and how pollen is changing as a result of more extreme environmental conditions might improve our understanding of the disease. This review focuses on the characteristics of plants producing allergenic pollens that are relevant to pollen allergy, including the phylogenetic relationships, pollen dispersal distances, amounts of pollen produced, amounts of protein in each type of pollen, and how allergenic proteins are released from pollens. In addition, the physiological roles of major allergenic protein families will be discussed to help us understand why some of these proteins become allergens and why GMO plants with hypoallergenic pollens may not be successful.

  12. Tree pollen allergens-an update from a molecular perspective.

    PubMed

    Asam, C; Hofer, H; Wolf, M; Aglas, L; Wallner, M

    2015-10-01

    It is estimated that pollen allergies affect approximately 40% of allergic individuals. In general, tree pollen allergies are mainly elicited by allergenic trees belonging to the orders Fagales, Lamiales, Proteales, and Pinales. Over 25 years ago, the gene encoding the major birch pollen allergen Bet v 1 was the first such gene to be cloned and its product characterized. Since that time, 53 tree pollen allergens have been identified and acknowledged by the WHO/IUIS allergen nomenclature subcommittee. Molecule-based profiling of allergic sensitization has helped to elucidate the immunological connections of allergen cross-reactivity, whereas advances in biochemistry have revealed structural and functional aspects of allergenic proteins. In this review, we provide a comprehensive overview of the present knowledge of the molecular aspects of tree pollen allergens. We analyze the geographic distribution of allergenic trees, discuss factors pivotal for allergic sensitization, and describe the role of tree pollen panallergens. Novel allergenic tree species as well as tree pollen allergens are continually being identified, making research in this field highly competitive and instrumental for clinical applications.

  13. Developments in allergen-specific immunotherapy: from allergen extracts to allergy vaccines bypassing allergen-specific immunoglobulin E and T cell reactivity.

    PubMed

    Focke, M; Swoboda, I; Marth, K; Valenta, R

    2010-03-01

    Allergen-specific immunotherapy (SIT) is the only specific and disease-modifying approach for the treatment of allergy but several disadvantages have limited its broad applicability. We argue that the majority of the possible disadvantages of SIT such as unwanted effects, poor efficacy and specificity as well as inconvenient application are related to the poor quality of natural allergen extracts, which are the active ingredients of all currently available allergy vaccines. Because of the progress made in the field of molecular allergen characterization, new allergy vaccines based on recombinant allergens, recombinant hypoallergenic allergen derivatives and allergen-derived T cell peptides have entered clinical testing and hold promise to reduce the side-effects and to increase the specificity as well as the efficacy of SIT. Here, we present a refined immunotherapy concept, which is based on the use of peptides derived from allergen surfaces that exhibit reduced, allergen-specific IgE as well as T cell reactivity. These peptides when fused to non-allergenic carriers give rise to allergen-specific protective IgG responses with T cell help from a non-allergenic carrier molecule. We summarize the experimental data demonstrating that such peptide vaccines can bypass allergen-specific IgE as well as T cell activation and may be administered at high doses without IgE- and T cell-mediated side-effects. Should these peptide vaccines prove efficacious and safe in clinical trials, it may become possible to develop convenient, safe and broadly applicable forms of SIT as true alternatives to symptomatic, drug-based allergy treatment.

  14. Mammalian airborne allergens.

    PubMed

    Aalberse, Rob C

    2014-01-01

    Historically, horse dandruff was a favorite allergen source material. Today, however, allergic symptoms due to airborne mammalian allergens are mostly a result of indoor exposure, be it at home, at work or even at school. The relevance of mammalian allergens in relation to the allergenic activity of house dust extract is briefly discussed in the historical context of two other proposed sources of house dust allergenic activity: mites and Maillard-type lysine-sugar conjugates. Mammalian proteins involved in allergic reactions to airborne dust are largely found in only 2 protein families: lipocalins and secretoglobins (Fel d 1-like proteins), with a relatively minor contribution of serum albumins, cystatins and latherins. Both the lipocalin and the secretoglobin family are very complex. In some instances this results in a blurred separation between important and less important allergenic family members. The past 50 years have provided us with much detailed information on the genomic organization and protein structure of many of these allergens. However, the complex family relations, combined with the wide range of post-translational enzymatic and non-enzymatic modifications, make a proper qualitative and quantitative description of the important mammalian indoor airborne allergens still a significant proteomic challenge. PMID:24925404

  15. Dimerization of lipocalin allergens

    PubMed Central

    Niemi, Merja H.; Rytkönen-Nissinen, Marja; Miettinen, Ilja; Jänis, Janne; Virtanen, Tuomas; Rouvinen, Juha

    2015-01-01

    Lipocalins are one of the most important groups of inhalant animal allergens. The analysis of structural features of these proteins is important to get insights into their allergenicity. We have determined two different dimeric crystal structures for bovine dander lipocalin Bos d 2, which was earlier described as a monomeric allergen. The crystal structure analysis of all other determined lipocalin allergens also revealed oligomeric structures which broadly utilize inherent structural features of the β-sheet in dimer formation. According to the moderate size of monomer-monomer interfaces, most of these dimers would be transient in solution. Native mass spectrometry was employed to characterize quantitatively transient dimerization of two lipocalin allergens, Bos d 2 and Bos d 5, in solution. PMID:26346541

  16. Ribonuclease activity of buckwheat plant (Fagopyrum esculentum) cultivars with different sensitivities to buckwheat burn virus.

    PubMed

    Sindarovska, Y R; Guzyk, O I; Yuzvenko, L V; Demchenko, O A; Didenko, L F; Grynevych, O I; Spivak, M Ya

    2014-01-01

    Ribonucleases (RNases) are present in base-level amounts in intact plants, but this level is able to increase greatly under stress conditions. The possible cause for such an increase is protection against plant RNA-virus attack. Buckwheat burn virus (BBV) is a highly virulent pathogen that belongs to Rhabdoviridae family. In our study, we have analyzed the correlation between RNase activity and resistance of different buckwheat cultivars to BBV infection. Two cultivars, Kara-Dag and Roksolana, with different sensitivities to BBV have been used. Kara-Dag is a cultivar with medium sensitivity to virus and Roksolana is a tolerant cultivar. It has been shown that the base level of RNase activity in Roksolana cultivar was in most cases higher than the corresponding parameter in Kara-Dag cultivar. Both infected and uninfected plants of Roksolana cultivar demonstrated high RNase activity during two weeks. Whereas infected plants of Kara-Dag cultivar demonstrated unstable levels of RNase activity. Significant decline in RNase activity was detected on the 7th day post infection with subsequent gradual increase in RNase activity. Decline of the RNase activity during the first week could promote the virus replication and therefore more successful infection of upper leaves of plants. Unstable levels of RNase activity in infected buckwheat plants may be explained by insufficiency of virus-resistant mechanisms that determines the medium sensitivity of the cultivar to BBV. Thus, plants of buckwheat cultivar having less sensitivity to virus, displayed in general higher RNase activity.

  17. Enzymatic treatment of peanut kernels to reduce allergen levels

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study investigated the use of enzymatic treatment to reduce peanut allergens in peanut kernel by processing conditions, such as, pretreatment with heat and proteolysis at different enzyme concentrations and treatment times. Two major peanut allergens, Ara h 1 and Ara h 2, were used as indicator...

  18. Reducing peanut allergens by high pressure combined with polyphenol oxidase

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Polyphenol oxidase (PPO) has been shown to reduce major peanut allergens (Ara h 1 and Ara h 2). Because high pressure (HP) can increase enzyme activity, we postulated that further reduction of peanut allergens can be achieved through HP combined with PPO. Peanut extracts were treated with each of th...

  19. Allergens in School Settings: Results of Environmental Assessments in 3 City School Systems

    ERIC Educational Resources Information Center

    Abramson, Stuart L.; Turner-Henson, Anne; Anderson, Lise; Hemstreet, Mary P.; Bartholomew, L. Kay; Joseph, Christine L. M.; Tang, Shenghui; Tyrrell, Shellie; Clark, Noreen M.; Ownby, Dennis

    2006-01-01

    Environmental allergens are major triggers for pediatric asthma. While children's greatest exposure to indoor allergens is in the home, other public places where children spend a large amount of time, such as school and day care centers, may also be sources of significant allergen encounters. The purpose of this article is to describe schoolroom…

  20. Characterization of Three Glucosyltransferase Genes in Tartary Buckwheat and Their Expression after Cold Stress.

    PubMed

    Zhou, Jing; Li, Cheng-Lei; Gao, Fei; Luo, Xiao-Peng; Li, Qing-Qing; Zhao, Hai-Xia; Yao, Hui-Peng; Chen, Hui; Wang, An-Hu; Wu, Qi

    2016-09-21

    Anthocyanins confer the red color in the hypocotyl of tartary buckwheat sprouts. Uridine diphosphate (UDP)-glucose:flavonoid 3-O-glycosyltransferase (UFGT) stabilizes anthocyanin by attaching the glucosyl moiety from UDP-glucose to the C3 hydroxyl of anthocyanin. In this study, we characterized three UFGT-like genes, designated FtUFGT1, 2, and 3 from tartary buckwheat. The results revealed that FtUFGT1, FtUFGT2, and FtUFGT3 can convert cyanidin to cyanidin 3-O-glucoside, with specific activities of 20.01 × 10(-3), 8.93 × 10(-3), and 20.24 × 10(-3) IU/mg, respectively. The active-site residues of the C-terminal domains and the N-terminal domains are important for the donor and acceptor recognition of these proteins. The expression of the three FtUFGTs paralleled the tissue-specific anthocyanin accumulation. After cold treatment, the increased content of anthocyanin was accompanied by the up-regulated expression of the three FtUFGTs. Among these three UGFT gene members, FtUFGT3 showed the highest expression level and the highest specific activity, suggesting that FtUFGT3 might be the major gene involved in anthocyanin biosynthesis. These results suggested that the FtUFGT genes, FtUFGT3 in particular, might be important candidates for anthocyanin formation in tartary buckwheat sprouts. PMID:27571449

  1. Structural identification of anthocyanins and analysis of concentrations during growth and flowering in buckwheat (Fagopyrum esculentum Moench) petals.

    PubMed

    Suzuki, Tatsuro; Kim, Sun-Ju; Mohamed, Zaidul Islam Sarker; Mukasa, Yuji; Takigawa, Shigenobu; Matsuura-Endo, Chie; Yamauchi, Hiroaki; Hashimoto, Naoto; Noda, Takahiro; Saito, Tatsuya

    2007-11-14

    The anthocyanin profiles and variety/breeding-line differences of anthocyanin concentrations in petals of common buckwheat flowers have been studied. Four anthocyanins, cyanidin 3-O-glucoside, cyanidin 3-O-rutinoside, cyanidin 3-O-rhamnoside, and cyanidin 3-O-galactosyl-rhamnoside were isolated from the petals of common buckwheat (Fagopyrum esculentum Moench), separated using high performance liquid chromatography and identified using reversed-phase liquid chromatography-electrospray ionization-tandem mass spectrometry techniques. In every variety/breeding line tested, cyanidin 3-O-rutinoside was detected as the major anthocyanin and the next is cyanidin 3-O-glucoside whereas cyanidin 3-O-rhamnoside and cyanidin 3-O-galactosyl-rhamnoside were trace or not detectable in white and pink flowered buckwheat. Of all the varieties/breeding lines tested, Gan-Chao, a Chinese variety, contained the highest amount of anthocyanins. The largest part of cyanidin moiety was presented as a proanthocyanidin form (PAs-Cy). Anthocyanins and PAs-Cy in petals were increased along with increase of flower development stages. Therefore, fully developed petals of red flowered buckwheat, especially Gan-Chao, are promising as a new anthocyanin-rich material for food processing.

  2. Allergen-induced asthma

    PubMed Central

    Cockcroft, Donald W

    2014-01-01

    It was only in the late 19th century that specific allergens, pollen, animal antigens and, later, house dust mite, were identified to cause upper and lower airway disease. Early allergen challenge studies, crudely monitored before measurement of forced expiratory volume in 1 s became widespread in the 1950s, focused on the immediate effects but noted in passing prolonged and/or recurrent asthma symptoms. The late asthmatic response, recurrent bronchoconstriction after spontaneous resolution of the early responses occurring 3 h to 8 h or more postchallenge, has been identified and well characterized over the past 50 years. The associated allergen-induced airway hyper-responsiveness (1977) and allergen-induced airway inflammation (1985) indicate that these late sequelae are important in the mechanism of allergen-induced asthma. Allergens are now recognized to be the most important cause of asthma. A standardized allergen inhalation challenge model has been developed and is proving to be a valuable research tool in the investigation of asthma pathophysiology and of potential new pharmacological agents for the treatment of asthma. PMID:24791256

  3. Worldwide food allergy labeling and detection of allergens in processed foods.

    PubMed

    Taylor, Steve L; Baumert, Joseph L

    2015-01-01

    The labeling of allergenic foods is an important public health measure to assist food-allergic consumers in avoiding foods that can cause allergic reactions. The regulatory framework for such labeling depends upon the selection of priority allergenic foods, which vary among countries. Most countries include milk, eggs, fish, crustacean shellfish, peanuts, tree nuts, soybeans, and cereal sources of gluten on the priority allergenic foods list, as recommended by the Codex Alimentarius Commission. However, a variety of other foods appear on the priority lists of some countries but not on others. Sesame seeds, molluscan shellfish, buckwheat, and mustard are identified in two or more countries. In most countries, all ingredients derived from these priority allergen sources must also be declared on labels by source. However, exemptions exist for some ingredients in some countries but not in others. Detection methods are critical for the enforcement of allergen labeling regulations and for the investigation of allergic reactions in the community by public health officials. The development of detection methods has advanced considerably over the past several decades and will be briefly reviewed in this chapter. Because of the emphasis on labeling and the development of detection methods, the ingredient statement on packaged food labels now contains more information than ever before to assist food-allergic consumers. PMID:26022883

  4. Worldwide food allergy labeling and detection of allergens in processed foods.

    PubMed

    Taylor, Steve L; Baumert, Joseph L

    2015-01-01

    The labeling of allergenic foods is an important public health measure to assist food-allergic consumers in avoiding foods that can cause allergic reactions. The regulatory framework for such labeling depends upon the selection of priority allergenic foods, which vary among countries. Most countries include milk, eggs, fish, crustacean shellfish, peanuts, tree nuts, soybeans, and cereal sources of gluten on the priority allergenic foods list, as recommended by the Codex Alimentarius Commission. However, a variety of other foods appear on the priority lists of some countries but not on others. Sesame seeds, molluscan shellfish, buckwheat, and mustard are identified in two or more countries. In most countries, all ingredients derived from these priority allergen sources must also be declared on labels by source. However, exemptions exist for some ingredients in some countries but not in others. Detection methods are critical for the enforcement of allergen labeling regulations and for the investigation of allergic reactions in the community by public health officials. The development of detection methods has advanced considerably over the past several decades and will be briefly reviewed in this chapter. Because of the emphasis on labeling and the development of detection methods, the ingredient statement on packaged food labels now contains more information than ever before to assist food-allergic consumers.

  5. Allergens in the Lab.

    ERIC Educational Resources Information Center

    Fisher, Thomas M.

    1987-01-01

    Points out the health and legal implications related to laboratory substances that could cause allergic reactions. Presents a list of potential cosmetic allergens and irritants. Includes precautionary measures dealing with allergy situations. (ML)

  6. Allergens and thunderstorm asthma.

    PubMed

    Nasser, Shuaib M; Pulimood, Thomas B

    2009-09-01

    Thunderstorm-related asthma is increasingly recognized in many parts of the world. This review focuses on important advances in the understanding of the mechanism of the role of allergens, in particular fungal spores such as Alternaria, in asthma epidemics associated with thunderstorms. From our observations, we have proposed that the prerequisites for this phenomenon are as follows: 1) a sensitized, atopic, asthmatic individual; 2) prior airway hyperresponsiveness before a sudden, large allergen exposure; 3) a large-scale thunderstorm with cold outflow occurring at a time and location during an allergen season in which large numbers of asthmatics are outdoors; and 4) sudden release of large amounts of respirable allergenic fragments, particularly fungal spores such as Alternaria. PMID:19671382

  7. Allergens and thunderstorm asthma.

    PubMed

    Nasser, Shuaib M; Pulimood, Thomas B

    2009-09-01

    Thunderstorm-related asthma is increasingly recognized in many parts of the world. This review focuses on important advances in the understanding of the mechanism of the role of allergens, in particular fungal spores such as Alternaria, in asthma epidemics associated with thunderstorms. From our observations, we have proposed that the prerequisites for this phenomenon are as follows: 1) a sensitized, atopic, asthmatic individual; 2) prior airway hyperresponsiveness before a sudden, large allergen exposure; 3) a large-scale thunderstorm with cold outflow occurring at a time and location during an allergen season in which large numbers of asthmatics are outdoors; and 4) sudden release of large amounts of respirable allergenic fragments, particularly fungal spores such as Alternaria.

  8. Allergen-specific immunotherapy.

    PubMed

    Nelson, Harold S; Norman, Philip S

    2014-01-01

    Specific immunotherapy was introduced for the treatment of grass pollen-induced hay fever in 1911. The treatment was soon extended to other pollens as well as perennial allergens, and to the treatment of bronchial asthma. Definitive studies of its efficacy for both rhinitis and asthma came only many decades later. Understanding gradually emerged of the underlying immunologic mechanisms that include the generation of regulatory T lymphocytes, immune deviation from allergen-specific Th2 to Th1 responses, and a shift in allergen-specific antibody production from immunoglobulin (Ig) E to IgG4. Along with understanding of the immune basis came an appreciation that immunotherapy modifies allergic disease expression, producing protection against disease progression and symptomatic improvement that persists for years after the treatment is discontinued. Recent new directions for immunotherapy include sublingual administration of inhalant allergens and use of the oral route to treat food allergy.

  9. Identification and characterization of granule bound starch synthase I (GBSSI) gene of tartary buckwheat (Fagopyrum tataricum Gaertn.).

    PubMed

    Wang, Xun; Feng, Bo; Xu, Zhibin; Sestili, Francesco; Zhao, Guojun; Xiang, Chao; Lafiandra, Domenico; Wang, Tao

    2014-01-25

    Tartary buckwheat (Fagopyrum tataricum Gaertn.) is increasingly considered as an important functional food material because of its rich nutraceutical compounds. Reserve starch is the major component of tartary buckwheat seed. However, the gene sequences and the molecular mechanism of tartary buckwheat starch synthesis are unknown so far. In this study, the complete genomic sequence and full-size cDNA coding tartary buckwheat granule-bound starch synthase I (FtGBSSI), which is responsible for amylose synthesis, were isolated and analyzed. The genomic sequence of the FtGBSSI contained 3947 nucleotides and was composed of 14 exons and 13 introns. The cDNA coding sequence of FtGBSSI shared 63.3%-75.1% identities with those of dicots and 56.6%-57.5% identities with monocots (Poaceae). In deduced amino acid sequence of FtGBSSI, eight motifs conserved among plant starch synthases were identified. A cleavage at the site IVC↓G of FtGBSSI protein produces the chloroplast transit sequence of 78 amino acids and the mature protein of 527 amino acids. The FtGBSSI mature protein showed an identity of 73.4%-77.8% with dicot plants, and 67.6%-70.4% with monocot plants (Poaceae). The mature protein was composed of 20 α-helixes and 16 β-strands, and folds into two main domains, N- and C-terminal domains. The critical residues which are involved in ADP and sugar binding were predicted. These results will be useful to modulate starch composition of buckwheat kernels with the aim to produce novel improved varieties in future breeding programs.

  10. Lyral: a fragrance allergen.

    PubMed

    Militello, Giuseppe; James, William

    2005-03-01

    Fragrances are a common cause of contact dermatitis and account for a large percentage of reactions to cosmetic products. Novel fragrance compounds that may not be detected by the common fragrance screening agents (including balsam of Peru and fragrance mix) are continually being produced. Lyral is one of those allergens found in many cosmetic and household products. This review will discuss the recent literature and the significance of this allergen to allergic contact dermatitis.

  11. Monitoring for airborne allergens

    SciTech Connect

    Burge, H.A. )

    1992-07-01

    Monitoring for allergens can provide some information on the kinds and levels of exposure experienced by local patient populations, providing volumetric methods are used for sample collection and analysis is accurate and consistent. Such data can also be used to develop standards for the specific environment and to begin to develop predictive models. Comparing outdoor allergen aerosols between different monitoring sites requires identical collection and analysis methods and some kind of rational standard, whether arbitrary, or based on recognized health effects.32 references.

  12. [Allergen management in the food industry].

    PubMed

    Röder, Martin; Weber, Wolfgang

    2016-07-01

    Due to the lack of causative immunotherapies, individuals with food allergies have to rely on correct labelling for even minute amounts of allergenic constituents. It is not relevant to the allergic whether the source of the culprit food is an ingredient or an allergen that entered the food unintentionally, as is the case with so-called cross-contacts or hidden allergens.Efficient allergen management is the manufacturer's prerequisite for coping with allergenic foods in the food production environment and handling them in a way that avoids cross-contact. If it is technically not feasible to eliminate cross-contacts entirely, it must be ensured that these cross-contacts do not enter the final product without being detected.This article discusses measures that should be considered in allergen management. Examples include recording all relevant allergens in the production facility, staff sensitization and training, and taking into account all areas of production from incoming raw materials to outgoing goods.For the evaluation of unavoidable cross-contacts, it is possible today to draw on data from clinical trials for many of the substances that are subject to labelling. This data can be used to assess the risk of the final product.However, the data from threshold studies is not legally binding, so it is left to the manufacturer to assess the level up to which the food is safe for the allergic. In particular the non-harmonized approach of the EU member countries' food safety authorities currently represents a major obstacle, as this can lead to food recalls even though existing levels were evaluated as being safe according to the risk assessments performed. PMID:27299344

  13. Rutin, quercetin, and free amino acid analysis in buckwheat (Fagopyrum) seeds from different locations.

    PubMed

    Bai, C Z; Feng, M L; Hao, X L; Zhong, Q M; Tong, L G; Wang, Z H

    2015-12-29

    In this study, five common buckwheats and nine tartary buckwheats grown at different locations were analyzed for the contents of rutin, quercetin, and amino acids by high-performance liquid chromatography and spectrophotometry. The rutin content was higher than quercetin in buckwheat seeds. Rutin content was in the range from 0.05 (0.05 g per 100 g dry seeds) to 1.35% of buckwheat seeds. Quercetin content varied from 0.01 to 0.17% and in some common buckwheats it was even difficult to detect. Comparatively, tartary buckwheat seeds contained more rutin and quercetin than common buckwheat seeds. Meanwhile, the bran has higher rutin content than the farina in tartary buckwheat seeds, with a respective content of 0.45 to 1.19% and 0.14 to 0.67%. It was found that amino acid contents were around 1.79 to 12.65% (farina) and 5.74 to 7.89% (bran) in common buckwheats, and 1.73 to 5.63% (farina) and 2.64 to 16.78% (bran) in tartary buckwheat seeds. The highest total rutin content was found to be 1.35% in tartary buckwheat seeds from Sichuan, China. The highest total amounts of amino acid were detected to be 20.13% in tartary buckwheat seeds from Changzhi, Shanxi Province (China). Our results suggested that food products made of whole-buckwheat flour are healthier than those made of fine white flour.

  14. [Cross reactivity of food allergens and its clinical relevance].

    PubMed

    Moneret-Vautrin, Denise Anne

    2005-10-01

    Cross-reactions between food allergens and other allergens are a major focus of interest. They include cross-allergies between Betulaceae and Compositae pollen, and also between fruits and vegetables (Prunoideae and Apiaceae). Cross-allergies between animal allergens include mites, cockroaches and crustaceans, milk and meat, animal epithelia, meat and egg. Cross-reactivity results from homology between protein sequences, and is highly likely when this homology reaches about 70%. Phylogenetically similar proteins occur in all species and are known as pan allergens. Profilins, Bet v1 homologues, and lipid transfer proteins have varying degrees of clinical relevance. The involvement of cross-reactivity in the persistence of sensitization and in allergic disorders is unclear. The consequences of cross-reactivity during specific immunotherapy with total allergenic extracts are random. Interpretation of biological tests of IgE binding is also biased by cross-reactivity. The use of panels of major recombinant allergens should help to identify specific sensitization profiles as well as clinically relevant sensitization. Cross-reactivity between epitopes of inhalants and of food allergens may perpetuate and intensify allergic disorders. The consequences of cross-reactivity between allergens and autologous proteins are unknown. PMID:16669147

  15. 66. Buckwheat Launder, date unknown Historic Photograph, Photographer Unknown; Collection ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    66. Buckwheat Launder, date unknown Historic Photograph, Photographer Unknown; Collection of William Everett, Jr. (Wilkes-Barre, PA), photocopy by Joseph E.B. Elliot - Huber Coal Breaker, 101 South Main Street, Ashley, Luzerne County, PA

  16. Recombinant allergens for allergen-specific immunotherapy: 10 years anniversary of immunotherapy with recombinant allergens.

    PubMed

    Valenta, Rudolf; Linhart, B; Swoboda, I; Niederberger, V

    2011-06-01

    The broad applicability of allergen-specific immunotherapy for the treatment and eventually prevention of IgE-mediated allergy is limited by the poor quality and allergenic activity of natural allergen extracts that are used for the production of current allergy vaccines. Today, the genetic code of the most important allergens has been deciphered; recombinant allergens equalling their natural counterparts have been produced for diagnosis and immunotherapy, and a large panel of genetically modified allergens with reduced allergenic activity has been characterized to improve safety of immunotherapy and explore allergen-specific prevention strategies. Successful immunotherapy studies have been performed with recombinant allergens and hypoallergenic allergen derivatives and will lead to the registration of the first recombinant allergen-based vaccines in the near future. There is no doubt that recombinant allergen-based vaccination strategies will be generally applicable to most allergen sources, including respiratory, food and venom allergens and allow to produce safe allergy vaccines for the treatment of the most common forms of IgE-mediated allergies.

  17. Mechanisms of allergen-specific immunotherapy and immune tolerance to allergens.

    PubMed

    Akdis, Cezmi A; Akdis, Mübeccel

    2015-01-01

    Substantial progress in understanding mechanisms of immune regulation in allergy, asthma, autoimmune diseases, tumors, organ transplantation and chronic infections has led to a variety of targeted therapeutic approaches. Allergen-specific immunotherapy (AIT) has been used for 100 years as a desensitizing therapy for allergic diseases and represents the potentially curative and specific way of treatment. The mechanisms by which allergen-AIT has its mechanisms of action include the very early desensitization effects, modulation of T- and B-cell responses and related antibody isotypes as well as inhibition of migration of eosinophils, basophils and mast cells to tissues and release of their mediators. Regulatory T cells (Treg) have been identified as key regulators of immunological processes in peripheral tolerance to allergens. Skewing of allergen-specific effector T cells to a regulatory phenotype appears as a key event in the development of healthy immune response to allergens and successful outcome in AIT. Naturally occurring FoxP3(+) CD4(+)CD25(+) Treg cells and inducible type 1 Treg (Tr1) cells contribute to the control of allergen-specific immune responses in several major ways, which can be summarized as suppression of dendritic cells that support the generation of effector T cells; suppression of effector Th1, Th2 and Th17 cells; suppression of allergen-specific IgE, and induction of IgG4; suppression of mast cells, basophils and eosinophils and suppression of effector T cell migration to tissues. New strategies for immune intervention will likely include targeting of the molecular mechanisms of allergen tolerance and reciprocal regulation of effector and regulatory T cell subsets.

  18. Paprika rhinoconjunctivitis case reveals new occupational Capsicum allergens.

    PubMed

    Airaksinen, Liisa; Riekki, Riitta; Vuokko, Aki; Puustinen, Anne

    2015-07-01

    No allergens related to paprika or cayenne respiratory allergy have been identified thus far. We describe a previously healthy 28-year woman who developed work-related rhinoconjunctivitis after four years of kebab-restaurant work. The allergy was studied using skin prick tests, serum specific IgE and nasal provocation tests. Specific IgE protein reactions were studied by Western blot analysis. Paprika, cayenne and curry allergens were identified from the strongest immunoblot bands using tandem mass spectrometry. A positive skin prick test, high specific IgE and positive nasal provocation test confirmed occupational rhinoconjunctivitis from Capsicum spices. Defensin J1 and Vicilin were identified as major paprika and cayenne allergens in this case. Vicilin was detected also from the curry ingredients. Two new occupational respiratory allergens from the Capsicum species were identified. These differ from previously reported bell pepper allergens. We emphasize that substantial spice handling at work poses an allergy risk.

  19. Paprika rhinoconjunctivitis case reveals new occupational Capsicum allergens.

    PubMed

    Airaksinen, Liisa; Riekki, Riitta; Vuokko, Aki; Puustinen, Anne

    2015-07-01

    No allergens related to paprika or cayenne respiratory allergy have been identified thus far. We describe a previously healthy 28-year woman who developed work-related rhinoconjunctivitis after four years of kebab-restaurant work. The allergy was studied using skin prick tests, serum specific IgE and nasal provocation tests. Specific IgE protein reactions were studied by Western blot analysis. Paprika, cayenne and curry allergens were identified from the strongest immunoblot bands using tandem mass spectrometry. A positive skin prick test, high specific IgE and positive nasal provocation test confirmed occupational rhinoconjunctivitis from Capsicum spices. Defensin J1 and Vicilin were identified as major paprika and cayenne allergens in this case. Vicilin was detected also from the curry ingredients. Two new occupational respiratory allergens from the Capsicum species were identified. These differ from previously reported bell pepper allergens. We emphasize that substantial spice handling at work poses an allergy risk. PMID:25944018

  20. Analysis of U.S. Food and Drug Administration food allergen recalls after implementation of the food allergen labeling and consumer protection act.

    PubMed

    Gendel, Steven M; Zhu, Jianmei

    2013-11-01

    To avoid potentially life-threatening reactions, food allergic consumers rely on information on food labels to help them avoid exposure to a food or ingredient that could trigger a reaction. To help consumers in the United States obtain the information that they need, the Food Allergen Labeling and Consumer Protection Act of 2004 defined a major food allergen as being one of eight foods or food groups and any ingredient that contains protein from one of these foods or food groups. A food that contains an undeclared major food allergen is misbranded under the U.S. Food, Drug, and Cosmetic Act and is subject to recall. Food allergen labeling problems are the most common cause of recalls for U.S. Food and Drug Administration (FDA)-regulated food products. To help understand why food allergen recalls continue to occur at a high rate, information on each food allergen recall that occurred in fiscal years 2007 through 2012 was obtained from the FDA recall database. This information was analyzed to identify the food, allergen, root cause, and mode of discovery for each food allergen recall. Bakery products were the most frequently recalled food type, and milk was the most frequently undeclared major food allergen. Use of the wrong package or label was the most frequent problem leading to food allergen recalls. These data are the first reported that indicate the importance of label and package controls as public health measures.

  1. Origin and Functional Prediction of Pollen Allergens in Plants1[OPEN

    PubMed Central

    Chen, Miaolin; Xu, Jie; Ren, Kang; Searle, Iain

    2016-01-01

    Pollen allergies have long been a major pandemic health problem for human. However, the evolutionary events and biological function of pollen allergens in plants remain largely unknown. Here, we report the genome-wide prediction of pollen allergens and their biological function in the dicotyledonous model plant Arabidopsis (Arabidopsis thaliana) and the monocotyledonous model plant rice (Oryza sativa). In total, 145 and 107 pollen allergens were predicted from rice and Arabidopsis, respectively. These pollen allergens are putatively involved in stress responses and metabolic processes such as cell wall metabolism during pollen development. Interestingly, these putative pollen allergen genes were derived from large gene families and became diversified during evolution. Sequence analysis across 25 plant species from green alga to angiosperms suggest that about 40% of putative pollen allergenic proteins existed in both lower and higher plants, while other allergens emerged during evolution. Although a high proportion of gene duplication has been observed among allergen-coding genes, our data show that these genes might have undergone purifying selection during evolution. We also observed that epitopes of an allergen might have a biological function, as revealed by comprehensive analysis of two known allergens, expansin and profilin. This implies a crucial role of conserved amino acid residues in both in planta biological function and allergenicity. Finally, a model explaining how pollen allergens were generated and maintained in plants is proposed. Prediction and systematic analysis of pollen allergens in model plants suggest that pollen allergens were evolved by gene duplication and then functional specification. This study provides insight into the phylogenetic and evolutionary scenario of pollen allergens that will be helpful to future characterization and epitope screening of pollen allergens. PMID:27436829

  2. The evaluation of allergens and allergic diseases in children.

    PubMed

    Lee, C S; Tang, R B; Chung, R L

    2000-12-01

    Knowing the incidence of allergic diseases and their relationship with allergens is a crucial requirement for therapeutic judgment. We present our experience on the incidence, clinical features and allergens of the allergic diseases detected by multiple allergosorbent chemiluminescent assay (MAST-CLA) in children from 1997 to 1999 at the Taipei Veterans General Hospital. The incidence of bronchial asthma, allergic rhinitis and atopic dermatitis are significantly different when stratified by age groups. Among the enrolled 2008 patients, 980 (48.8%) patients have positive MAST-CLA results. Of these, 562 (57.3%) are male and 418 (42.7%) are female. A significant increase among patients with positive allergens is also found when stratified by age group. Inhalant allergen is the major allergen detected in our patients. House dust mites Dermatophagoides pteronyssinus (Dp) and Dermatophagoides farinae (Df), cockroaches, feathers, and dog dander show the highest incidence in the 7- to 12-year-old group. In the fungal group, Aspergillus and Penicillium also show a significant difference in the incidence among different age groups. Pollen allergens, on a whole, show significant difference in incidence among different age groups. The food allergen group shows variable significant difference in incidence. Crab, milk, and egg white show the highest significant incidence in the 2- to 6-year-old group. These results suggest that the incidence of allergens detected in allergic diseases varies among different age groups. PMID:11269366

  3. [Recombinant allergens. For routine use or still only science?].

    PubMed

    Schmid-Grendelmeier, P

    2010-11-01

    Component-resolved diagnosis of allergies allows disease-specific patterns of sensitization in some conditions such as allergic bronchopulmonary aspergillosis ABPA). By determination of IgE against important pollen allergens such as Bet v 1, Ole e 1 or Phl p1/Phl p 5, more precise guidance for allergen-specific immunotherapy may be achieved, as pollen extracts contain mostly these major allergens. Sensitizations against minor allergens such as profilins or polcalcins influence the outcome of IgE measurements against full allergen sources, but are often of limited clinical relevance. In food allergy, frequent cross reactivity between pollens such as birch pollen via Bet v 1/PR10 proteins can be identified. Sensitization against some storage proteins such as peanut (Ara h 2) or lipid transfer proteins of peach (Pru p 3) or hazelnut (Cor a 8) may indicate an increased risk of severe anaphylactic reactions. Exercise-induced anaphylaxis, unclear sensitizations against latex or double-positivity in insect allergy are other useful indications for component-resolved diagnosis. Microarray-based allergen chip diagnosis makes possible today the detection of IgE against more than 100 allergens in tiny amounts of serum and is very promising, but still needs evaluation and optimization in regard to allergen selection and sensitivity.

  4. Protein families: implications for allergen nomenclature, standardisation and specific immunotherapy.

    PubMed

    Breiteneder, Heimo

    2009-01-01

    Allergens are embedded into the protein universe as members of large families and superfamilies of related proteins which is a direct consequence of their shared evolution. The classification of allergens by protein families offers a valuable frame of reference that allows the design of experiments to study cross-reactivity and allergenic potency of proteins. Information on protein family membership also complements the current official IUIS allergen nomenclature. All presently known allergens belong to one of 140 (1.4%) of the 10,340 protein families currently described by version 23.0 of the Pfam database. This is indicative of a strong bias among allergens towards certain protein architectures that are able to induce an IgE response in an atopic immune system. However, even small variations in the structure of a protein alter its immunological characteristics. Various isoforms of the major birch pollen allergen Bet v 1 were shown to possess highly variant immunogenic and allergenic properties. Ber e 1 and SFA8, two 2S albumins, were revealed to display differential capacities to polarise an immune response. Such data will be exploited in the future for the design of allergy vaccines.

  5. Current Overview of Allergens of Plant Pathogenesis Related Protein Families

    PubMed Central

    Sinha, Mau; Singh, Rashmi Prabha; Kushwaha, Gajraj Singh; Iqbal, Naseer; Singh, Avinash; Kaushik, Sanket; Sharma, Sujata; Singh, Tej P.

    2014-01-01

    Pathogenesis related (PR) proteins are one of the major sources of plant derived allergens. These proteins are induced by the plants as a defense response system in stress conditions like microbial and insect infections, wounding, exposure to harsh chemicals, and atmospheric conditions. However, some plant tissues that are more exposed to environmental conditions like UV irradiation and insect or fungal attacks express these proteins constitutively. These proteins are mostly resistant to proteases and most of them show considerable stability at low pH. Many of these plant pathogenesis related proteins are found to act as food allergens, latex allergens, and pollen allergens. Proteins having similar amino acid sequences among the members of PR proteins may be responsible for cross-reactivity among allergens from diverse plants. This review analyzes the different pathogenesis related protein families that have been reported as allergens. Proteins of these families have been characterized in regard to their biological functions, amino acid sequence, and cross-reactivity. The three-dimensional structures of some of these allergens have also been evaluated to elucidate the antigenic determinants of these molecules and to explain the cross-reactivity among the various allergens. PMID:24696647

  6. Recombinant proteins in newly developed foods: identification of allergenic activity.

    PubMed

    Lehrer, S B; Reese, G

    1997-01-01

    A number of agricultural crops are being modified for various purposes using recombinant DNA technology. Since transferred genes may code for proteins that are ordinarily not present, there is concern about the potential allergenicity of these new varieties. The safety evaluation of transgenic foods is relatively easy when the allergenicity of the gene source is known. Recombinant allergens in genetically engineered or altered foods can be identified using traditional immunological assays such as RAST or ELISA inhibition or immunoblotting procedures. Our recent studies of two corn proteins (10 kD and HSZ) used to alter grain amino acid composition and of transgenic soybeans with an altered fatty acid profile are examples of this approach. Both 10 kD and HSZ did not bind IgE antibodies from sera of corn-reactive subjects by immunoblotting. Studies of wild-type and transgenic soybeans with high oleic acidic content by RAST inhibition and immunoblotting with pooled sera of soy-allergic individuals demonstrated no difference in the allergen content of both extracts. In contrast to these studies, a recent investigation by Nordlee et al. (1996) of transgenic soybeans which expressed a methionine/cysteine-rich protein from Brazil nuts identified this protein as a major Brazil nut allergen. These studies indicate that, when the gene source is from a known allergen or if the recipient contains allergens, it is possible to determine whether the allergen content of the transgenic line is altered relative to the nontransgenic varieties.

  7. Stability of food allergens to digestion in vitro.

    PubMed

    Astwood, J D; Leach, J N; Fuchs, R L

    1996-10-01

    An integral part of the safety assessment of genetically modified plants is consideration of possible human health effects, especially food allergy. Prospective testing for allergenicity of proteins obtained from sources with no prior history of causing allergy has been difficult because of the absence of valid methods and models. Food allergens may share physicochemical properties that distinguish them from nonallergens, properties that may be used as a tool to predict the inherent allergenicity of proteins newly introduced into the food supply by genetic engineering. One candidate property is stability to digestion. We have systematically evaluated the stability of food allergens that are active via the gastrointestinal tract in a simple model of gastric digestion, emphasizing the major allergens of plant-derived foods such as legumes (peanuts and soybean). Important food allergens were stable to digestion in the gastric model (simulated gastric fluid). For example, soybean beta-conglycinin was stable for 60 min. In contrast, nonallergenic food proteins, such as spinach ribulose bis-phosphate carboxylase/oxygenase, were digested in simulated gastric fluid within 15 sec. The data are consistent with the hypothesis that food allergens must exhibit sufficient gastric stability to reach the intestinal mucosa where absorption and sensitization (development of atopy) can occur. Thus, the stability to digestion is a significant and valid parameter that distinguishes food allergens from nonallergens.

  8. Positive reaction to allergen (image)

    MedlinePlus

    Allergic reaction is a sensitivity to a specific substance, called an allergen, that is contacted through the skin, inhaled into the lungs, swallowed or injected. The body's reaction to an allergen can be mild, such as ...

  9. [Current contact allergens].

    PubMed

    Geier, J; Uter, W; Lessmann, H; Schnuch, A

    2011-10-01

    Ever-changing exposure to contact allergens, partly due to statutory directives (e.g. nickel, chromate, methyldibromo glutaronitrile) or recommendations from industrial associations (e.g. hydroxyisohexyl 3-cyclohexene carboxaldehyde), requires on-going epidemiologic surveillance of contact allergy. In this paper, the current state with special focus in fragrances and preservatives is described on the basis of data of the Information Network of Departments of Dermatology (IVDK) of the year 2010. In 2010, 12,574 patients were patch tested in the dermatology departments belonging to the IVDK. Nickel is still the most frequent contact allergen. However the continuously improved EU nickel directive already has some beneficial effect; sensitization frequency in young women is dropping. In Germany, chromate-reduced cement has been in use now for several years, leading to a decline in chromate sensitization in brick-layers. Two fragrance mixes are part of the German baseline series; they are still relevant. The most important fragrances in these mixes still are oak moss absolute and hydroxyisohexyl 3-cyclohexene carboxaldehyde. However, in relation to these leading allergens, sensitization frequency to other fragrances contained in the mixes seems to be increasing. Among the preservatives, MCI/MI has not lost its importance as contact allergen, in contrast to MDBGN. Sources of MCI/MI sensitization obviously are increasingly found in occupational context. Methylisothiazolinone is a significant allergen in occupational settings, and less frequently in body care products.

  10. Multiple grass mixes as opposed to single grasses for allergen immunotherapy in allergic rhinitis.

    PubMed

    Gangl, K; Niederberger, V; Valenta, R

    2013-11-01

    Grass pollen allergy affects approximately 40% of allergic patients. Subcutaneous allergen immunotherapy (SCIT) is the only allergen-specific and disease-modifying treatment available. Currently available therapeutic vaccines for the treatment of grass pollen allergy are based on natural grass pollen extracts which are either made from pollen of one cross-reactive grass species or from several related grass species. Clinical studies have shown that SCIT performed with timothy grass pollen extract is effective for the treatment of grass pollen allergy. Moreover, it has been demonstrated that recombinant timothy grass pollen allergens contain the majority of relevant epitopes and can be used for SCIT in clinical trials. However, recent in vitro studies have suggested that mixes consisting of allergen extracts from several related grass species may have advantages for SCIT over single allergen extracts. Here, we review current knowledge regarding the disease-relevant allergens in grass pollen allergy, available clinical studies comparing SCIT with allergen extracts from timothy grass or from mixes of several related grass species of the Pooideae subfamily, in vitro cross-reactivity studies performed with natural allergen extracts and recombinant allergens and SCIT studies performed with recombinant timothy grass pollen allergens. In vitro and clinical studies performed with natural allergen extracts reveal no relevant advantages of using multiple grass mixes as opposed to single grass pollen extracts. Several studies analysing the molecular composition of natural allergen extracts and the molecular profile of patients' immune responses after SCIT with allergen extracts indicate that the major limitation for the production of a high quality grass pollen vaccine resides in intrinsic features of natural allergen extracts which can only be overcome with recombinant allergen-based technologies.

  11. [Allergenicity of lupin flour].

    PubMed

    Leduc, V; Moneret-Vautrin, D A; Guérin, L

    2002-06-01

    Lupin flour is used in human food for its high quality nutritional and functional qualities. The frequency of crossed allergy between lupin flour and peanuts, both members of the family of Leguminosae, is strong, since 68% of patients who are allergic to peanut have shown positive reactions to lupin flour when tested by TPO-DA. Cases of isolated allergy to lupin flour without pre-existence of peanut allergy as well as workplace asthma by inhalation are also rarely seen. The specific allergens of lupin and those that participate in crosses with peanut have been studied by SDS-PAGE and immunoblot. The diversity of allergens contained in different lupin flour has also been studied. Further, the detection of lupin flour in a "pizza" flour which induced a strong allergic reaction exposed its eventual implication as a masked allergen.

  12. Challenges in testing genetically modified crops for potential increases in endogenous allergen expression for safety.

    PubMed

    Panda, R; Ariyarathna, H; Amnuaycheewa, P; Tetteh, A; Pramod, S N; Taylor, S L; Ballmer-Weber, B K; Goodman, R E

    2013-02-01

    Premarket, genetically modified (GM) plants are assessed for potential risks of food allergy. The major risk would be transfer of a gene encoding an allergen or protein nearly identical to an allergen into a different food source, which can be assessed by specific serum testing. The potential that a newly expressed protein might become an allergen is evaluated based on resistance to digestion in pepsin and abundance in food fractions. If the modified plant is a common allergenic source (e.g. soybean), regulatory guidelines suggest testing for increases in the expression of endogenous allergens. Some regulators request evaluating endogenous allergens for rarely allergenic plants (e.g. maize and rice). Since allergic individuals must avoid foods containing their allergen (e.g. peanut, soybean, maize, or rice), the relevance of the tests is unclear. Furthermore, no acceptance criteria are established and little is known about the natural variation in allergen concentrations in these crops. Our results demonstrate a 15-fold difference in the major maize allergen, lipid transfer protein between nine varieties, and complex variation in IgE binding to various soybean varieties. We question the value of evaluating endogenous allergens in GM plants unless the intent of the modification was production of a hypoallergenic crop. PMID:23205714

  13. Challenges in testing genetically modified crops for potential increases in endogenous allergen expression for safety.

    PubMed

    Panda, R; Ariyarathna, H; Amnuaycheewa, P; Tetteh, A; Pramod, S N; Taylor, S L; Ballmer-Weber, B K; Goodman, R E

    2013-02-01

    Premarket, genetically modified (GM) plants are assessed for potential risks of food allergy. The major risk would be transfer of a gene encoding an allergen or protein nearly identical to an allergen into a different food source, which can be assessed by specific serum testing. The potential that a newly expressed protein might become an allergen is evaluated based on resistance to digestion in pepsin and abundance in food fractions. If the modified plant is a common allergenic source (e.g. soybean), regulatory guidelines suggest testing for increases in the expression of endogenous allergens. Some regulators request evaluating endogenous allergens for rarely allergenic plants (e.g. maize and rice). Since allergic individuals must avoid foods containing their allergen (e.g. peanut, soybean, maize, or rice), the relevance of the tests is unclear. Furthermore, no acceptance criteria are established and little is known about the natural variation in allergen concentrations in these crops. Our results demonstrate a 15-fold difference in the major maize allergen, lipid transfer protein between nine varieties, and complex variation in IgE binding to various soybean varieties. We question the value of evaluating endogenous allergens in GM plants unless the intent of the modification was production of a hypoallergenic crop.

  14. [Shrimp as an allergen source].

    PubMed

    Múnera, Marlon; Gómez, Luis; Puerta, Leonardo

    2013-01-01

    Allergy to shellfish is one of the most prevalent food allergies in several countries, especially the one induced by consuming or having contact with shrimp. Several shrimp species are known to induce allergy diseases. However, the whole spectrum of allergens they contain is unknown and few of them have been completely characterized. This study was done in order to know the recent advances in the characterization of shrimp allergens and its relationship with allergens from other arthropods of importance in allergic diseases. We emphasize the species Litopenaeus vannamei , the most consumed shrimp in Colombia. Well characterized shrimp allergens are named following an official classification; nevertheless, they are better known according to the biological function associated with them. Tropomiosin, the main and most studied allergen in different shrimp species, is involved in crossreactivity among shrimp and other arthropods like domestic mites. The other characterized allergens seem to have a minor participation in this cross-reactivity. The allergenic potential of L. vannamei is not well known and few of its allergens have been characterized, whilst others that were recently identified such as the hemocyanin and the fatty acid binding proteins are beginning to be studied. Preliminary results suggest that these allergens are involved in the cross-reactivity between shrimp and domestic mites, which deserves further evaluation. The molecular and immunological characterization of all allergens present in shrimp would help understanding its allergenic role.

  15. Bleach Neutralizes Mold Allergens

    ERIC Educational Resources Information Center

    Science Teacher, 2005

    2005-01-01

    Researchers at National Jewish Medical and Research Center have demonstrated that dilute bleach not only kills common household mold, but may also neutralize the mold allergens that cause most mold-related health complaints. The study, published in the Journal of Allergy and Clinical Immunology, is the first to test the effect on allergic…

  16. Tartary buckwheat (Fagopyrum tataricum Gaertn.) as a source of dietary rutin and quercitrin.

    PubMed

    Fabjan, Nina; Rode, Janko; Kosir, Iztok Joze; Wang, Zhuanhua; Zhang, Zheng; Kreft, Ivan

    2003-10-22

    Two samples of tartary buckwheat (Fagopyrum tataricum Gaertn.) from China and one from Luxembourg were studied by high-performance liquid chromatography (HPLC) to reveal the possibilities of growing tartary buckwheat herb as a possible source of rutin, quercetin, and quercitrin. The content of rutin was determined as up to 3% dry weight (DW) in tartary buckwheat herb. Quercitrin values were in the range of 0.01-0.05% DW. Only traces of quercetin were detected in just some of the samples. Tartary buckwheat seeds contained more rutin (about 0.8-1.7% DW) than common buckwheat seeds (0.01% DW). Rutin and quercetin content in seeds depends on variety and growing conditions. Tartary buckwheat seeds contained traces of quercitrin and quercetin, which were not found in common buckwheat seeds.

  17. MucoRice-cholera toxin B-subunit, a rice-based oral cholera vaccine, down-regulates the expression of α-amylase/trypsin inhibitor-like protein family as major rice allergens.

    PubMed

    Kurokawa, Shiho; Nakamura, Rika; Mejima, Mio; Kozuka-Hata, Hiroko; Kuroda, Masaharu; Takeyama, Natsumi; Oyama, Masaaki; Satoh, Shigeru; Kiyono, Hiroshi; Masumura, Takehiro; Teshima, Reiko; Yuki, Yoshikazu

    2013-07-01

    To develop a cold chain- and needle/syringe-free rice-based cholera vaccine (MucoRice-CTB) for human use, we previously advanced the MucoRice system by introducing antisense genes specific for endogenous rice storage proteins and produced a molecularly uniform, human-applicable, high-yield MucoRice-CTB devoid of plant-associated sugar. To maintain the cold chain-free property of this vaccine for clinical application, we wanted to use a polished rice powder preparation of MucoRice-CTB without further purification but wondered whether this might cause an unexpected increase in rice allergen protein expression levels in MucoRice-CTB and prompt safety concerns. Therefore, we used two-dimensional fluorescence difference gel electrophoresis and shotgun MS/MS proteomics to compare rice allergen protein expression levels in MucoRice-CTB and wild-type (WT) rice. Both proteomics analyses showed that the only notable change in the expression levels of rice allergen protein in MucoRice-CTB, compared with those in WT rice, was a decrease in the expression levels of α-amylase/trypsin inhibitor-like protein family such as the seed allergen protein RAG2. Real-time PCR analysis showed mRNA of RAG2 reduced in MucoRice-CTB seed. These results demonstrate that no known rice allergens appear to be up-reregulated by genetic modification of MucoRice-CTB, suggesting that MucoRice-CTB has potential as a safe oral cholera vaccine for clinical application.

  18. Peanut Allergy, Allergen Composition, and Methods of Reducing Allergenicity: A Review.

    PubMed

    Zhou, Yang; Wang, Jin-Shui; Yang, Xiao-Jia; Lin, Dan-Hua; Gao, Yun-Fang; Su, Yin-Jie; Yang, Sen; Zhang, Yan-Jie; Zheng, Jing-Jing

    2013-01-01

    Peanut allergy affects 1-2% of the world's population. It is dangerous, and usually lifelong, and it greatly decreases the life quality of peanut-allergic individuals and their families. In a word, peanut allergy has become a major health concern worldwide. Thirteen peanut allergens are identified, and they are briefly introduced in this paper. Although there is no feasible solution to peanut allergy at present, many methods have shown great promise. This paper reviews methods of reducing peanut allergenicity, including physical methods (heat and pressure, PUV), chemical methods (tannic acid and magnetic beads), and biological methods (conventional breeding, irradiation breeding, genetic engineering, enzymatic treatment, and fermentation). PMID:26904614

  19. Peanut Allergy, Allergen Composition, and Methods of Reducing Allergenicity: A Review

    PubMed Central

    Wang, Jin-shui; Yang, Xiao-jia; Lin, Dan-hua; Gao, Yun-fang; Su, Yin-jie; Yang, Sen; Zhang, Yan-jie; Zheng, Jing-jing

    2013-01-01

    Peanut allergy affects 1-2% of the world's population. It is dangerous, and usually lifelong, and it greatly decreases the life quality of peanut-allergic individuals and their families. In a word, peanut allergy has become a major health concern worldwide. Thirteen peanut allergens are identified, and they are briefly introduced in this paper. Although there is no feasible solution to peanut allergy at present, many methods have shown great promise. This paper reviews methods of reducing peanut allergenicity, including physical methods (heat and pressure, PUV), chemical methods (tannic acid and magnetic beads), and biological methods (conventional breeding, irradiation breeding, genetic engineering, enzymatic treatment, and fermentation). PMID:26904614

  20. Complete Chloroplast Genome Sequence of Tartary Buckwheat (Fagopyrum tataricum) and Comparative Analysis with Common Buckwheat (F. esculentum).

    PubMed

    Cho, Kwang-Soo; Yun, Bong-Kyoung; Yoon, Young-Ho; Hong, Su-Young; Mekapogu, Manjulatha; Kim, Kyung-Hee; Yang, Tae-Jin

    2015-01-01

    We report the chloroplast (cp) genome sequence of tartary buckwheat (Fagopyrum tataricum) obtained by next-generation sequencing technology and compared this with the previously reported common buckwheat (F. esculentum ssp. ancestrale) cp genome. The cp genome of F. tataricum has a total sequence length of 159,272 bp, which is 327 bp shorter than the common buckwheat cp genome. The cp gene content, order, and orientation are similar to those of common buckwheat, but with some structural variation at tandem and palindromic repeat frequencies and junction areas. A total of seven InDels (around 100 bp) were found within the intergenic sequences and the ycf1 gene. Copy number variation of the 21-bp tandem repeat varied in F. tataricum (four repeats) and F. esculentum (one repeat), and the InDel of the ycf1 gene was 63 bp long. Nucleotide and amino acid have highly conserved coding sequence with about 98% homology and four genes--rpoC2, ycf3, accD, and clpP--have high synonymous (Ks) value. PCR based InDel markers were applied to diverse genetic resources of F. tataricum and F. esculentum, and the amplicon size was identical to that expected in silico. Therefore, these InDel markers are informative biomarkers to practically distinguish raw or processed buckwheat products derived from F. tataricum and F. esculentum.

  1. Contact allergens for armpits--allergenic fragrances specified on deodorants.

    PubMed

    Klaschka, Ursula

    2012-11-01

    According to the so-called "26 allergens rule" 26 supposedly allergenic fragrances must be specified on the containers of cosmetic products if they are present above 0.001% in leave-on products and, 0.01% in rinse-off products. This declaration is meant to inform the consumers of potential risks of skin sensitizers in the products. As many consumers of deodorants suffer from allergic or irritant contact dermatitis in the axillae, the presence of allergens in deodorants deserves special attention. The objective of this study was to find answers to the following questions: Does compulsory labeling lead to omission of strong allergenic fragrances in deodorants? Is there a difference in the use patterns of strong and weak allergens? What is the quantitative exposure to fragrances by deodorants? Is the situation in Germany different from other European countries? Is there a difference between deodorants for men and for women? I tested the implementation of the "26 allergens rule" and compiled which allergenic fragrances are specified on the containers of deodorants. Three market studies were conducted in Germany in 2008, 2010 and 2011. The labels of a total number of 374 deodorants were analyzed as to whether any of the "26 allergens" were listed. The frequency of each allergen in the deodorants was compared with results from previous studies by other authors. It was found that up to 83% of the deodorants contain at least one of the "26 allergens" and that up to 30% of all products contain strong allergens above the threshold for labeling (0.001% in the product). The most frequently listed allergens are medium or weak allergens. In comparison with other authors, the frequency of the "26 allergens" in products is slightly smaller in these recent studies for the German market. There is no significant difference between deodorants for men and women, as far as the labeling of the "26 allergens" is concerned. The results show that the mandatory labeling procedure as designed

  2. Contact allergens for armpits--allergenic fragrances specified on deodorants.

    PubMed

    Klaschka, Ursula

    2012-11-01

    According to the so-called "26 allergens rule" 26 supposedly allergenic fragrances must be specified on the containers of cosmetic products if they are present above 0.001% in leave-on products and, 0.01% in rinse-off products. This declaration is meant to inform the consumers of potential risks of skin sensitizers in the products. As many consumers of deodorants suffer from allergic or irritant contact dermatitis in the axillae, the presence of allergens in deodorants deserves special attention. The objective of this study was to find answers to the following questions: Does compulsory labeling lead to omission of strong allergenic fragrances in deodorants? Is there a difference in the use patterns of strong and weak allergens? What is the quantitative exposure to fragrances by deodorants? Is the situation in Germany different from other European countries? Is there a difference between deodorants for men and for women? I tested the implementation of the "26 allergens rule" and compiled which allergenic fragrances are specified on the containers of deodorants. Three market studies were conducted in Germany in 2008, 2010 and 2011. The labels of a total number of 374 deodorants were analyzed as to whether any of the "26 allergens" were listed. The frequency of each allergen in the deodorants was compared with results from previous studies by other authors. It was found that up to 83% of the deodorants contain at least one of the "26 allergens" and that up to 30% of all products contain strong allergens above the threshold for labeling (0.001% in the product). The most frequently listed allergens are medium or weak allergens. In comparison with other authors, the frequency of the "26 allergens" in products is slightly smaller in these recent studies for the German market. There is no significant difference between deodorants for men and women, as far as the labeling of the "26 allergens" is concerned. The results show that the mandatory labeling procedure as designed

  3. Molecular and biochemical classification of plant-derived food allergens.

    PubMed

    Breiteneder, H; Ebner, C

    2000-07-01

    Molecular biology and biochemical techniques have significantly advanced the knowledge of allergens derived from plant foods. Surprisingly, many of the known plant food allergens are homologous to pathogenesis-related proteins (PRs), proteins that are induced by pathogens, wounding, or certain environmental stresses. PRs have been classified into 14 families. Examples of allergens homologous to PRs include chitinases (PR-3 family) from avocado, banana, and chestnut; antifungal proteins such as the thaumatin-like proteins (PR-5) from cherry and apple; proteins homologous to the major birch pollen allergen Bet v 1 (PR-10) from vegetables and fruits; and lipid transfer proteins (PR-14) from fruits and cereals. Allergens other than PR homologs can be allotted to other well-known protein families such as inhibitors of alpha-amylases and trypsin from cereal seeds, profilins from fruits and vegetables, seed storage proteins from nuts and mustard seeds, and proteases from fruits. As more clinical data and structural information on allergenic molecules becomes available, we may finally be able to answer what characteristics of a molecule are responsible for its allergenicity.

  4. New strategies for allergen immunotherapy.

    PubMed

    Carnés, Jerónimo; Robinson, Douglas S

    2008-06-01

    Specific allergen immunotherapy, consisting in the administration of increasing amounts of offending allergens into sensitive patients was first used nearly one hundred years ago and remains in use worldwide for treatment of allergic rhinitis and asthma. It has been recognised as the only effective treatment for type I allergic diseases when the appropriate quantities of allergens are used. The immunological mechanisms by which specific immunotherapy is effective include the modulation of T cells and the response of B-cells and is accompanied by significant decreases of specific IgE and increases in allergen specific IgG antibodies, mainly IgG4. While specific allergen injection immunotherapy is highly effective and the most common way of administration other routes such as oral or intranasal ways have been considered as and alternative to subcutaneous injections. During the last century, allergenic vaccines have been prepared using individual allergens adsorbed to different adjuvant substances. These vaccines have demonstrated efficacy and good results in different clinical trials. However, many novel approaches to allergen immunotherapy have been developed in the last years in order to increase the safety and efficacy of allergenic vaccines. In that way, different and modern vaccines have been prepared including more purified products such as depigmented allergen extracts; allergoids, consisting on big molecules of thousands of kDa, which contain all the individual allergens and show a significant decrease in severe adverse reactions; peptides or small aminoacid sequences; recombinant allergens; hypoallergenic vaccines where the IgE binding sites have been modified; or allergen-CpG fusion molecules. New presentations are under study and new treatments will be developed in the near future with the objective that the prevention of allergic disease may become a reality. The review article also discuss recent patent related to the field. PMID:19075996

  5. Reducing peanut allergens by high pressure combined with polyphenol oxidase

    NASA Astrophysics Data System (ADS)

    Chung, Si-Yin; Houska, Milan; Reed, Shawndrika

    2013-12-01

    Polyphenol oxidase (PPO) has been shown to reduce major peanut allergens. Since high pressure (HP) can increase enzyme activity, we postulated that further reduction of peanut allergens can be achieved through HP combined with PPO. Peanut extracts containing caffeic acid were treated with each of the following: (1) HP; (2) HP+PPO; (3) PPO; and (4) none. HP was conducted at 300 and 500 MPa, each for 3 and 10 min, 37 °C. After treatment, SDS-PAGE was performed and allergenic capacity (IgE binding) was determined colorimetrically in inhibition enzyme-linked immunosorbent assay and Western blots, using a pooled plasma from peanut-allergic patients. Data showed that HP alone had no effect on major peanut allergens. However, HP at 500 MPa combined with PPO (HP500/PPO) induced a higher (approximately twofold) reduction of major peanut allergens and IgE binding than PPO alone or HP300/PPO. There was no difference between treatment times. We concluded that HP500/PPO at 3-min enhanced a twofold reduction of the allergenic capacity of peanut extracts, as compared to PPO itself.

  6. Discovery and genetic analysis of non-bitter Tartary buckwheat (Fagopyrum tataricum Gaertn.) with trace-rutinosidase activity

    PubMed Central

    Suzuki, Tatsuro; Morishita, Toshikazu; Mukasa, Yuji; Takigawa, Shigenobu; Yokota, Satoshi; Ishiguro, Koji; Noda, Takahiro

    2014-01-01

    In a screening of about 500 lines of Tartary buckwheat, we identified lines that contained no detectable rutinosidase isozymes using an in-gel detection assay. We confirmed that seeds of these individuals had only a trace level of in-vitro rutinosidase activity. To investigate the heritability of the trace-rutinosidase characteristic, we analyzed the progeny of crosses between rutinosidase trace-lines, ‘f3g-162’, and the ‘Hokkai T8’. The F2 progeny clearly divided into two groups: those with rutinosidase activity under 1.5 nkat/g seed (trace-rutinosidase) and those with activity over 400 nkat/g seed (normal rutinosidase). The segregation pattern of this trait in F2 progeny exhibited 1 : 3 ratio (trace-rutinosidase : normal rutinosidase), suggesting that the trace-rutinosidase trait is conferred by a single recessive gene; rutinosidase-trace A (rutA). In addition, sensory panelists evaluated the bitterness of flour from trace-rutinosidase individuals and did not detect bitterness, whereas flour from normal rutinosidase individuals was found to have strong bitterness. Although at least three bitter compounds have been reported in Tartary buckwheat seeds, our present findings indicate that rutin hydrolysis is the major contributing factor to bitterness. In addition, the trace-rutinosidase line identified here, ‘f3g-162’, is a promising material for generating a non-bitter Tartary buckwheat variety. PMID:25914588

  7. Discovery and genetic analysis of non-bitter Tartary buckwheat (Fagopyrum tataricum Gaertn.) with trace-rutinosidase activity.

    PubMed

    Suzuki, Tatsuro; Morishita, Toshikazu; Mukasa, Yuji; Takigawa, Shigenobu; Yokota, Satoshi; Ishiguro, Koji; Noda, Takahiro

    2014-12-01

    In a screening of about 500 lines of Tartary buckwheat, we identified lines that contained no detectable rutinosidase isozymes using an in-gel detection assay. We confirmed that seeds of these individuals had only a trace level of in-vitro rutinosidase activity. To investigate the heritability of the trace-rutinosidase characteristic, we analyzed the progeny of crosses between rutinosidase trace-lines, 'f3g-162', and the 'Hokkai T8'. The F2 progeny clearly divided into two groups: those with rutinosidase activity under 1.5 nkat/g seed (trace-rutinosidase) and those with activity over 400 nkat/g seed (normal rutinosidase). The segregation pattern of this trait in F2 progeny exhibited 1 : 3 ratio (trace-rutinosidase : normal rutinosidase), suggesting that the trace-rutinosidase trait is conferred by a single recessive gene; rutinosidase-trace A (rutA). In addition, sensory panelists evaluated the bitterness of flour from trace-rutinosidase individuals and did not detect bitterness, whereas flour from normal rutinosidase individuals was found to have strong bitterness. Although at least three bitter compounds have been reported in Tartary buckwheat seeds, our present findings indicate that rutin hydrolysis is the major contributing factor to bitterness. In addition, the trace-rutinosidase line identified here, 'f3g-162', is a promising material for generating a non-bitter Tartary buckwheat variety.

  8. Designing a Multimer Allergen for Diagnosis and Immunotherapy of Dog Allergic Patients

    PubMed Central

    Nilsson, Ola B.; Neimert-Andersson, Theresa; Bronge, Mattias; Grundström, Jeanette; Sarma, Ranjana; Uchtenhagen, Hannes; Kikhney, Alexey; Sandalova, Tatyana; Holmgren, Erik; Svergun, Dmitri; Achour, Adnane; van Hage, Marianne; Grönlund, Hans

    2014-01-01

    Background Dog dander extract used for diagnosis and allergen-specific immunotherapy is often of variable and of poor quality. Objective To assemble four well-established dog allergen components into one recombinant folded protein for improved diagnosis and vaccination of allergy to dog. Methods A linked molecule, comprising the four dog lipocalin allergens Can f 1, Can f 2, Can f 4 and Can f 6 was constructed. The tetrameric protein was structurally characterized by small angle X-ray scattering, and compared with each single recombinant lipocalin allergen or an equimolar mix of the four allergens by analytical size exclusion chromatography, circular dichroism, allergen-specific IgE in serum by ELISA and allergen-dependent capacity to activate basophils. The immunogenicity of the fusion protein was evaluated in immunized mice by assessing splenocyte proliferation and antibody production. Results The linked tetrameric construct was produced as a soluble fusion protein, with the specific folds of the four individual allergens conserved. This multi-allergen molecule was significantly more efficient (p<0.001) than each single recombinant allergen in binding to dog-specific IgE, and the epitope spectrum was unaffected compared to an equimolar mix of the four allergens. Basophil degranulation revealed that the biologic activity of the linked molecule was retained. Immunization of mice with the linked construct induced comparable allergen-specific IgG responses with blocking capacity towards all included allergens and generated comparably low T-cell responses. Conclusion We provide the first evidence for a linked recombinant molecule covering the major dog allergens for potential use in diagnostics and allergy vaccination of dog allergic patients. PMID:25353166

  9. New treatments for allergen immunotherapy.

    PubMed

    Akdis, Mübeccel

    2014-01-01

    Allergen-specific immunotherapy (SIT) represents the only curative and specific way for the treatment of allergic diseases, which have reached a pandemic dimension in industrial countries affecting up to 20-30% of the population. Although applied for 100 years to cure allergy, SIT still faces several problems related to side effects and limited efficacy. Currently, allergen-SIT is performed with vaccines based on allergen extracts that can cause severe, often life threatening, anaphylactic reactions as well as new IgE sensitization to other allergens present in the extract. Low patient adherence and high costs due to long duration (3 to 5 years) of treatment have been commonly reported. Several strategies have been developed to tackle these issues and it became possible to produce recombinant allergen-SIT vaccines with reduced allergenic activity.

  10. New treatments for allergen immunotherapy

    PubMed Central

    2014-01-01

    Allergen-specific immunotherapy (SIT) represents the only curative and specific way for the treatment of allergic diseases, which have reached a pandemic dimension in industrial countries affecting up to 20-30% of the population. Although applied for 100 years to cure allergy, SIT still faces several problems related to side effects and limited efficacy. Currently, allergen-SIT is performed with vaccines based on allergen extracts that can cause severe, often life threatening, anaphylactic reactions as well as new IgE sensitization to other allergens present in the extract. Low patient adherence and high costs due to long duration (3 to 5 years) of treatment have been commonly reported. Several strategies have been developed to tackle these issues and it became possible to produce recombinant allergen-SIT vaccines with reduced allergenic activity. PMID:25258656

  11. Endogenous allergens in the regulatory assessment of genetically engineered crops.

    PubMed

    Graf, Lynda; Hayder, Hikmat; Mueller, Utz

    2014-11-01

    A scientific approach to the assessment of foods derived from genetically engineered (GE) crops is critical to maintaining objectivity and public confidence in regulatory decisions. Principles developed at the international level support regulators and enable robust and transparent safety assessments. A comparison of key constituents in the GE crop with a suitable comparator is an important element of an assessment. In Europe, endogenous allergens would be included in the comparative analysis, however this approach has been hindered by technical limitations on the ability to accurately measure identified allergenic proteins. Over recent years, improved proteomic methods have enabled researchers to focus on major allergenic proteins in conventional food crops, as information on natural variability is largely lacking. Emerging data for soybean indicate that variability in levels of major allergens already in the food supply is broad. This raises questions about the biological interpretation of differences between a GE plant and its conventional counterpart, in particular, whether any conclusions about altered allergenicity could be inferred. This paper discusses the scientific justification for requiring proteomic analysis of endogenous allergens as part of the evaluation. Ongoing scientific review and corresponding international discussion are integral to ensuring that data requirements address legitimate risk assessment questions.

  12. Relative immunogenicity of commonly allergenic foods versus rarely allergenic and nonallergenic foods in mice.

    PubMed

    Birmingham, Neil; Thanesvorakul, Sirinart; Gangur, Venu

    2002-12-01

    Food allergies affect 6 to 8% of children and 2% of adults in the United States. For reasons that are not clear, eight types of food account for a vast majority (approximately 90%) of food-induced hypersensitivity reactions. In this study, C57Bl/6 mice were used to test the hypothesis that commonly allergenic foods are intrinsically more immunogenic than rarely allergenic or nonallergenic foods in allergy-susceptible hosts. Groups of mice (n = 4 to 5) were injected intraperitoneally with the protein extracts (plus alum as an adjuvant) from chicken eggs, peanuts, almonds, filberts-hazelnuts, walnuts, soybeans, and wheat (commonly allergenic foods) and coffee, sweet potatoes, carrots, white potatoes, cherries, lettuce, and spinach (rarely allergenic and nonallergenic foods). Primary and secondary immune responses (as measured by specific IgG1 antibody serum levels) were measured by an enzyme-linked immunosorbent assay. Proteins from peanuts, almonds, filberts, sweet potatoes, cherries, and spinach elicited robust primary and/or secondary immune responses. Proteins from eggs, walnuts, and lettuce elicited poor primary responses but significant secondary responses. In contrast, wheat, soybeans, coffee, carrots, and white potatoes elicited barely detectable to poor primary and secondary immune responses. The order of the immunogenicity levels of these foods in mice is as follows: almonds = filberts > spinach (Rubisco) > peanuts > or = sweet potatoes > cherries > lettuce > walnuts > chicken eggs > carrots > or = white potatoes > wheat = coffee = soybeans. In summary, these data demonstrate for the first time that: (i) foods vary widely with regard to their relative immunogenicity in allergy-susceptible hosts and (ii) intrinsic immunogenicity in mice does not distinguish commonly allergenic foods from rarely allergenic or nonallergenic foods.

  13. Relationships between antioxidant compounds and antioxidant activities of tartary buckwheat during germination.

    PubMed

    Zhou, Xiaoli; Hao, Tingfeng; Zhou, Yiming; Tang, Wen; Xiao, Ying; Meng, Xiaoxiao; Fang, Xiang

    2015-04-01

    Relationships of changes between major non-enzymatic antioxidant compounds and antioxidant capacities of tartary buckwheat during germination were evaluated by means of correlation analysis and principal component analysis in this paper. The changes of antioxidant compounds, including vitamin C, vitamin E, flavonoids, carotenoids, and chlorophyll, and antioxidant activities were detected. A good accumulation in the content of vitamin C (0.71 mg/g), total flavonoids (19.53 mg rutin/g), and rutin (11.34 mg/g) was found after 7-day germination, but germination decreased the vitamin E activity. Germination improved the activities of buckwheat extracts to scavenge DPPH, ABTS, and superoxide free radicals by 107, 144, and 88 %, respectively. Furthermore, the correlation and principal component analysis showed that the vitamin C, total flavonoids, and rutin contents were closely related positively with free radicals scavenging properties, indicating that the compounds which play a key role in the elevated antioxidant activities during germination consisted of vitamin C, total flavonoids, and rutin, but not vitamin E and quercetin.

  14. Relationships between antioxidant compounds and antioxidant activities of tartary buckwheat during germination.

    PubMed

    Zhou, Xiaoli; Hao, Tingfeng; Zhou, Yiming; Tang, Wen; Xiao, Ying; Meng, Xiaoxiao; Fang, Xiang

    2015-04-01

    Relationships of changes between major non-enzymatic antioxidant compounds and antioxidant capacities of tartary buckwheat during germination were evaluated by means of correlation analysis and principal component analysis in this paper. The changes of antioxidant compounds, including vitamin C, vitamin E, flavonoids, carotenoids, and chlorophyll, and antioxidant activities were detected. A good accumulation in the content of vitamin C (0.71 mg/g), total flavonoids (19.53 mg rutin/g), and rutin (11.34 mg/g) was found after 7-day germination, but germination decreased the vitamin E activity. Germination improved the activities of buckwheat extracts to scavenge DPPH, ABTS, and superoxide free radicals by 107, 144, and 88 %, respectively. Furthermore, the correlation and principal component analysis showed that the vitamin C, total flavonoids, and rutin contents were closely related positively with free radicals scavenging properties, indicating that the compounds which play a key role in the elevated antioxidant activities during germination consisted of vitamin C, total flavonoids, and rutin, but not vitamin E and quercetin. PMID:25829633

  15. Mining Novel Allergens from Coconut Pollen Employing Manual De Novo Sequencing and Homology-Driven Proteomics.

    PubMed

    Saha, Bodhisattwa; Sircar, Gaurab; Pandey, Naren; Gupta Bhattacharya, Swati

    2015-11-01

    Coconut pollen, one of the major palm pollen grains is an important constituent among vectors of inhalant allergens in India and a major sensitizer for respiratory allergy in susceptible patients. To gain insight into its allergenic components, pollen proteins were analyzed by two-dimensional electrophoresis, immunoblotted with coconut pollen sensitive patient sera, followed by mass spectrometry of IgE reactive proteins. Coconut being largely unsequenced, a proteomic workflow has been devised that combines the conventional database-dependent analysis of tandem mass spectral data and manual de novo sequencing followed by a homology-based search for identifying the allergenic proteins. N-terminal acetylation helped to distinguish "b" ions from others, facilitating reliable sequencing. This led to the identification of 12 allergenic proteins. Cluster analysis with individual patient sera recognized vicilin-like protein as a major allergen, which was purified to assess its in vitro allergenicity and then partially sequenced. Other IgE-sensitive spots showed significant homology with well-known allergenic proteins such as 11S globulin, enolase, and isoflavone reductase along with a few which are reported as novel allergens. The allergens identified can be used as potential candidates to develop hypoallergenic vaccines, to design specific immunotherapy trials, and to enrich the repertoire of existing IgE reactive proteins.

  16. Mining Novel Allergens from Coconut Pollen Employing Manual De Novo Sequencing and Homology-Driven Proteomics.

    PubMed

    Saha, Bodhisattwa; Sircar, Gaurab; Pandey, Naren; Gupta Bhattacharya, Swati

    2015-11-01

    Coconut pollen, one of the major palm pollen grains is an important constituent among vectors of inhalant allergens in India and a major sensitizer for respiratory allergy in susceptible patients. To gain insight into its allergenic components, pollen proteins were analyzed by two-dimensional electrophoresis, immunoblotted with coconut pollen sensitive patient sera, followed by mass spectrometry of IgE reactive proteins. Coconut being largely unsequenced, a proteomic workflow has been devised that combines the conventional database-dependent analysis of tandem mass spectral data and manual de novo sequencing followed by a homology-based search for identifying the allergenic proteins. N-terminal acetylation helped to distinguish "b" ions from others, facilitating reliable sequencing. This led to the identification of 12 allergenic proteins. Cluster analysis with individual patient sera recognized vicilin-like protein as a major allergen, which was purified to assess its in vitro allergenicity and then partially sequenced. Other IgE-sensitive spots showed significant homology with well-known allergenic proteins such as 11S globulin, enolase, and isoflavone reductase along with a few which are reported as novel allergens. The allergens identified can be used as potential candidates to develop hypoallergenic vaccines, to design specific immunotherapy trials, and to enrich the repertoire of existing IgE reactive proteins. PMID:26426307

  17. Group 10 allergens (tropomyosins) from house-dust mites may cause covariation of sensitization to allergens from other invertebrates

    PubMed Central

    Inam, Muhammad; Ismail, Muhammad; Chaudhary, Farhana Riaz

    2012-01-01

    Group 10 allergens (tropomyosins) have been assumed to be a major cause of cross-reactivity between house-dust mites (HDMs) and other invertebrates. Despite all of the published data regarding the epidemiology, percent IgE binding and level of sensitization in the population, the role of tropomyosin as a cross-reactive allergen in patients with multiple allergy syndrome still remains to be elucidated. Homology between amino acid sequences reported in allergen databases of selected invertebrate tropomyosins was determined with Der f 10 as the reference allergen. The 66.9 and 54.4% identities were found with selected crustacean and insect species, respectively, whereas only 20.4% identity was seen with mollusks. A similar analysis was performed using reported B-cell IgE-binding epitopes from Met e1 (shrimp allergen) and Bla g7 (cockroach allergen) with other invertebrate tropomyosins. The percent identity in linear sequences was higher than 35% in mites, crustaceans, and cockroaches. The polar and hydrophobic regions in these groups were highly conserved. These findings suggest that tropomyosin may be a major cause of covariation of sensitization between HDMs, crustaceans, and some species of insects and mollusks. PMID:23342293

  18. Allergen-free probiotics.

    PubMed

    Mogna, Giovanni; Strozzi, Gian Paolo; Mogna, Luca

    2008-09-01

    Food sensitivities are constantly increasing in "westernized" countries and may pose serious health risks to sensitized individuals. Severe allergy episodes have also been reported after the intake of probiotic products containing milk protein residues, especially in children. The need for safe and effective probiotic strains and food supplements, which contain them, is now emerging clearly. The present work describes the way of achieving this aim by the avoidance of any kind of raw materials at risk, both in probiotic strain industrial manufacturing and finished product formulation. Allergen-free probiotics represent, without any doubt, an innovative and safe tool for human health.

  19. Facing Hymenoptera Venom Allergy: From Natural to Recombinant Allergens

    PubMed Central

    Perez-Riverol, Amilcar; Justo-Jacomini, Débora Lais; Zollner, Ricardo de Lima; Brochetto-Braga, Márcia Regina

    2015-01-01

    Along with food and drug allergic reactions, a Hymenoptera insect Sting (Apoidea, Vespidae, Formicidae) is one of the most common causes of anaphylaxis worldwide. Diagnoses of Hymenoptera venom allergy (HVA) and specific immunotherapy (SIT) have been based on the use of crude venom extracts. However, the incidence of cross-reactivity and low levels of sensibility during diagnosis, as well as the occurrence of nonspecific sensitization and undesired side effects during SIT, encourage the search for novel allergenic materials. Recombinant allergens are an interesting approach to improve allergy diagnosis and SIT because they circumvent major problems associated with the use of crude venom. Production of recombinant allergens depends on the profound molecular characterization of the natural counterpart by combining some “omics” approaches with high-throughput screening techniques and the selection of an appropriate system for heterologous expression. To date, several clinically relevant allergens and novel venom toxins have been identified, cloned and characterized, enabling a better understanding of the whole allergenic and envenoming processes. Here, we review recent findings on identification, molecular characterization and recombinant expression of Hymenoptera venom allergens and on the evaluation of these heterologous proteins as valuable tools for tackling remaining pitfalls on HVA diagnosis and immunotherapy. PMID:26184309

  20. Facing Hymenoptera Venom Allergy: From Natural to Recombinant Allergens.

    PubMed

    Perez-Riverol, Amilcar; Justo-Jacomini, Débora Lais; Zollner, Ricardo de Lima; Brochetto-Braga, Márcia Regina

    2015-07-09

    Along with food and drug allergic reactions, a Hymenoptera insect Sting (Apoidea, Vespidae, Formicidae) is one of the most common causes of anaphylaxis worldwide. Diagnoses of Hymenoptera venom allergy (HVA) and specific immunotherapy (SIT) have been based on the use of crude venom extracts. However, the incidence of cross-reactivity and low levels of sensibility during diagnosis, as well as the occurrence of nonspecific sensitization and undesired side effects during SIT, encourage the search for novel allergenic materials. Recombinant allergens are an interesting approach to improve allergy diagnosis and SIT because they circumvent major problems associated with the use of crude venom. Production of recombinant allergens depends on the profound molecular characterization of the natural counterpart by combining some "omics" approaches with high-throughput screening techniques and the selection of an appropriate system for heterologous expression. To date, several clinically relevant allergens and novel venom toxins have been identified, cloned and characterized, enabling a better understanding of the whole allergenic and envenoming processes. Here, we review recent findings on identification, molecular characterization and recombinant expression of Hymenoptera venom allergens and on the evaluation of these heterologous proteins as valuable tools for tackling remaining pitfalls on HVA diagnosis and immunotherapy.

  1. Comparison of phenolic compositions between common and tartary buckwheat (Fagopyrum) sprouts.

    PubMed

    Kim, Sun-Ju; Zaidul, I S M; Suzuki, Tatsuro; Mukasa, Yuji; Hashimoto, Naoto; Takigawa, Sigenobu; Noda, Takahiro; Matsuura-Endo, Chie; Yamauchi, Hiroaki

    2008-10-15

    The phenolic compositions of non-germinated/germinated seeds and seed sprouts (at 6-10 day-old) of common (Fagopyrum esculentum Möench) and tartary (Fagopyrum tataricum Gaertn.) buckwheats were investigated. Phenolic compounds, including chlorogenic acid, four C-glycosylflavones (orientin, isoorientin vitexin, isovitexin), rutin and quercetin, were determined in the seed sprouts by high-performance liquid chromatography (HPLC). In the edible parts of common buckwheat sprouts, individual phenolics significantly increased during sprout growth from 6 to 10 days after sowing (DAS), whereas in tartary buckwheat sprouts they did not. While the sum contents of phenolic compounds in the edible part (mean 24.4mg/g DW at 6-10 DAS) of tartary buckwheat sprouts were similar to those of common buckwheat sprouts, rutin contents in the non-germinated/germinated seeds (mean 14.7mg/g DW) and edible parts (mean 21.8mg/g DW) of tartary buckwheat were 49- and 5-fold, respectively, higher than those of common buckwheat. Extracts of the edible parts of both species showed very similar free radical-scavenging activities (mean 1.7μmol trolox eq/g DW), suggesting that the overall antioxidative activity might be affected by the combination of identified phenolics and unidentified (minor) components. Therefore, buckwheat seed sprouts are recommended for their high antioxidative activity, as well as being an excellent dietary source of phenolic compounds, particularly tartary buckwheat sprouts, being rich in rutin.

  2. Buckwheat Honey Attenuates Carbon Tetrachloride-Induced Liver and DNA Damage in Mice

    PubMed Central

    Cheng, Ni; Wu, Liming; Zheng, Jianbin; Cao, Wei

    2015-01-01

    Buckwheat honey, which is widely consumed in China, has a characteristic dark color. The objective of this study was to investigate the protective effects of buckwheat honey on liver and DNA damage induced by carbon tetrachloride in mice. The results revealed that buckwheat honey had high total phenolic content, and rutin, hesperetin, and p-coumaric acid were the main phenolic compounds present. Buckwheat honey possesses super DPPH radical scavenging activity and strong ferric reducing antioxidant power. Administration of buckwheat honey for 10 weeks significantly inhibited serum lipoprotein oxidation and increased serum oxygen radical absorbance capacity. Moreover, buckwheat honey significantly inhibited aspartate aminotransferase and alanine aminotransferase activities, which are enhanced by carbon tetrachloride. Hepatic malondialdehyde decreased and hepatic antioxidant enzymes (superoxide dismutase and glutathione peroxidase) increased in the presence of buckwheat honey. In a comet assay, lymphocyte DNA damage induced by carbon tetrachloride was significantly inhibited by buckwheat honey. Therefore, buckwheat honey has a hepatoprotective effect and inhibits DNA damage, activities that are primarily attributable to its high antioxidant capacity. PMID:26508989

  3. Purification and characterization of the main allergen of Plantago lanceolata pollen, Pla l 1.

    PubMed

    Calabozo, B; Barber, D; Polo, F

    2001-02-01

    English plantain (Plantago lanceolata) pollen is an important cause of pollinosis in the temperate regions of North America, Australia and Europe. However, very little is known about its allergen composition. The aim of this study was to identify plantain allergens, and to isolate and characterize a major allergen. Allergens were identified by immunoblotting with individual allergic patients' sera. Isolation of the major allergen was achieved by sequential reverse-phase and size-exclusion HPLC. Allergenic characterization was performed by ELISA and immunoblotting after SDS-PAGE with sera from plantain-allergic patients. N-terminal amino acid sequence was established by Edman degradation. Allergograms showed that 13 out of the 14 sera assayed had IgE to a group of proteins with a molecular weight in the range of 16-20 kd, that turned out to be different isoforms or variants of the major allergen Pla l l. Eighteen amino acid residues from the N-terminal end of one of the isoforms, and 10 of three others, were sequenced, and a partial sequence identity with Ole e 1 was found. Prevalence of specific IgE to purified Pla l 1 in plantain allergic patients was 86%, and represents about 80% of the total IgE-binding capacity of the plantain extract. The most relevant allergen from P.lanceolata pollen, Pla l 1, has been purified and characterized. This contributes to a greater knowledge of the allergen composition of this important weed, and clears the way for the standardization of plantain allergen products in terms of major allergen content.

  4. Absolute quantification of allergens from complex mixtures: a new sensitive tool for standardization of allergen extracts for specific immunotherapy.

    PubMed

    Seppälä, Ulla; Dauly, Claire; Robinson, Sarah; Hornshaw, Martin; Larsen, Jørgen Nedergaard; Ipsen, Henrik

    2011-04-01

    Products for specific diagnosis and immunotherapy of IgE-mediated allergies are currently based on natural extracts. Quantification of major allergen content is an important aspect of standardization as important allergens particularly impact vaccine potency. The aim of the study was to develop a mass spectrometry (MS) based assay for absolute quantification of Timothy (Phleum pratense) pollen allergens Phl p 1 and Phl p 5 in P. pratense extract. High-resolution and accurate mass (HRAM) MS was selected for its ability to detect peptides with high selectivity and mass accuracy (<3 ppm). Isotope labeled heavy peptides were used for absolute quantification of specific isoallergens of Phl p 1 and Phl p 5 at low femtomole level in P. pratense extract. Robustness and linearity of the method was demonstrated with intra day precision ≤ 5% (n = 3). Phl p 1b was shown to be 5 times less abundant than its variant Phl p 1a and Phl p 5b was shown to be 9 times more abundant than the Phl p 5a. The present study shows that allergen, and/or isoallergen specific, surrogate signature peptides analyzed with HRAM MS is a sensitive and accurate tool for identification and quantification of allergens from complex allergen sources.

  5. [Comparative immunological characteristics of Daphnia allergens].

    PubMed

    Berzhets, V M; Mochalov, A A; Sipitsyna, N E; Petrova, N S; Kanchurin, A Kh

    1986-08-01

    Materials on the study of Daphnia allergens are presented. Daphnia allergens have been shown to possess considerable sensitizing properties. The optimum method for the preparation of the allergen has been selected. The method of measuring the electrophoretic mobility of cells with a view to the evaluation of the specific activity of Daphnia allergen is proposed. PMID:2429484

  6. Allergen-induced airway responses.

    PubMed

    Gauvreau, Gail M; El-Gammal, Amani I; O'Byrne, Paul M

    2015-09-01

    Environmental allergens are an important cause of asthma and can contribute to loss of asthma control and exacerbations. Allergen inhalation challenge has been a useful clinical model to examine the mechanisms of allergen-induced airway responses and inflammation. Allergen bronchoconstrictor responses are the early response, which reaches a maximum within 30 min and resolves by 1-3 h, and late responses, when bronchoconstriction recurs after 3-4 h and reaches a maximum over 6-12 h. Late responses are followed by an increase in airway hyperresponsiveness. These responses occur when IgE on mast cells is cross-linked by an allergen, causing degranulation and the release of histamine, neutral proteases and chemotactic factors, and the production of newly formed mediators, such as cysteinyl leukotrienes and prostaglandin D2. Allergen-induced airway inflammation consists of an increase in airway eosinophils, basophils and, less consistently, neutrophils. These responses are mediated by the trafficking and activation of myeloid dendritic cells into the airways, probably as a result of the release of epithelial cell-derived thymic stromal lymphopoietin, and the release of pro-inflammatory cytokines from type 2 helper T-cells. Allergen inhalation challenge has also been a widely used model to study potential new therapies for asthma and has an excellent negative predictive value for this purpose. PMID:26206871

  7. Quantitative Proteomic Profiling of Peanut Allergens in Food Ingredients Used for Oral Food Challenges.

    PubMed

    Johnson, Philip E; Sayers, Rebekah L; Gethings, Lee A; Balasundaram, Anuradha; Marsh, Justin T; Langridge, James I; Mills, E N Clare

    2016-06-01

    Profiling allergens in complex food ingredients used in oral food challenges and immunotherapy is crucial for regulatory acceptance. Mass spectrometry based analysis employing data-independent acquisition coupled with ion mobility mass spectrometry-mass spectrometry (DIA-IM-MS) was used to investigate the allergen composition of raw peanuts and roasted peanut flour ingredients used in challenge meals. This comprehensive qualitative and quantitative analysis using label-free approaches identified and quantified 123 unique protein accessions. Semiquantitative analysis indicated that allergens Ara h 1 and Ara h 3 were the most abundant proteins and present in approximately equal amounts and were extracted in reduced amounts from roasted peanut flours. The clinically significant allergens Ara h 2 and 6 were less abundant, but relative quantification was unaffected by roasting. Ara h 5 was undetectable in any peanut sample, while the Bet v 1 homologue Ara h 8 and the lipid transfer protein allergen, Ara h 9, were detected in low abundance. The oleosin allergens, Ara h 10 and 11, were moderately abundant in the raw peanuts but were 100-fold less abundant in the defatted roasted peanut flour than the major allergens Ara h 1, 3, 2, and 6. Certain isoforms of the major allergens dominated the profile. The relative quantitation of the major peanut allergens showed little variation between different batches of roasted peanut flour. These data will support future development of targeted approaches for absolute quantification of peanut allergens which can be applied to both food ingredients used in clinical studies and extracts used for skin testing and to identify trace levels of allergens in foods. PMID:27064171

  8. Quantitative Proteomic Profiling of Peanut Allergens in Food Ingredients Used for Oral Food Challenges.

    PubMed

    Johnson, Philip E; Sayers, Rebekah L; Gethings, Lee A; Balasundaram, Anuradha; Marsh, Justin T; Langridge, James I; Mills, E N Clare

    2016-06-01

    Profiling allergens in complex food ingredients used in oral food challenges and immunotherapy is crucial for regulatory acceptance. Mass spectrometry based analysis employing data-independent acquisition coupled with ion mobility mass spectrometry-mass spectrometry (DIA-IM-MS) was used to investigate the allergen composition of raw peanuts and roasted peanut flour ingredients used in challenge meals. This comprehensive qualitative and quantitative analysis using label-free approaches identified and quantified 123 unique protein accessions. Semiquantitative analysis indicated that allergens Ara h 1 and Ara h 3 were the most abundant proteins and present in approximately equal amounts and were extracted in reduced amounts from roasted peanut flours. The clinically significant allergens Ara h 2 and 6 were less abundant, but relative quantification was unaffected by roasting. Ara h 5 was undetectable in any peanut sample, while the Bet v 1 homologue Ara h 8 and the lipid transfer protein allergen, Ara h 9, were detected in low abundance. The oleosin allergens, Ara h 10 and 11, were moderately abundant in the raw peanuts but were 100-fold less abundant in the defatted roasted peanut flour than the major allergens Ara h 1, 3, 2, and 6. Certain isoforms of the major allergens dominated the profile. The relative quantitation of the major peanut allergens showed little variation between different batches of roasted peanut flour. These data will support future development of targeted approaches for absolute quantification of peanut allergens which can be applied to both food ingredients used in clinical studies and extracts used for skin testing and to identify trace levels of allergens in foods.

  9. Air-conditioner filters enriching dust mites allergen

    PubMed Central

    Zhan, Xiaodong; Li, Chaopin; Xu, Haifeng; Xu, Pengfei; Zhu, Haibin; Diao, Jidong; Li, Na; Zhao, Beibei

    2015-01-01

    We detected the concentration of dust mites allergen (Der f1 & Der p1) in the air of different places before and after the starting of air-conditioners in Wuhu City, Anhui, China, and to discuss the relation between the dust mites allergen in air-conditioner filters and the asthma attack. The dust samples were collected from the air-conditioner filters in dining rooms, shopping malls, hotels and households respectively. Concentrations of dust mites major group allergen 1 (Der f 1, Der p1) were detected with enzyme linked immunosorbent assay (ELISA), and the dust mite immune activities were determined by dot-ELISA. The concentration of Der f1 in dining rooms, shopping malls, hotels and households was 1.52 μg/g, 1.24 μg/g, 1.31 μg/g and 1.46 μg/g respectively, and the concentration of Der p1 in above-mentioned places was 1.23 μg/g, 1.12 μg/g, 1.16 μg/g and 1.18 μg/g respectively. The concentration of Der f1 & Der p1 in air was higher after the air-conditioners starting one hours later, and the difference was significant (P<0.05, respectively). Additionally, dot-ELISA findings revealed that the allergen extracted from the dust was capable of reacting with IgE from the sera of asthma mice allergic to dust mites. The study concludes that air-conditioner filters can enrich dust mites major group allergen, and the allergens can induce asthma. The air-conditioner filters shall be cleaned or replaced regularly to prevent or reduce accumulation of the dust mites and its allergens. PMID:26064381

  10. Air-conditioner filters enriching dust mites allergen.

    PubMed

    Zhan, Xiaodong; Li, Chaopin; Xu, Haifeng; Xu, Pengfei; Zhu, Haibin; Diao, Jidong; Li, Na; Zhao, Beibei

    2015-01-01

    We detected the concentration of dust mites allergen (Der f1 & Der p1) in the air of different places before and after the starting of air-conditioners in Wuhu City, Anhui, China, and to discuss the relation between the dust mites allergen in air-conditioner filters and the asthma attack. The dust samples were collected from the air-conditioner filters in dining rooms, shopping malls, hotels and households respectively. Concentrations of dust mites major group allergen 1 (Der f 1, Der p1) were detected with enzyme linked immunosorbent assay (ELISA), and the dust mite immune activities were determined by dot-ELISA. The concentration of Der f1 in dining rooms, shopping malls, hotels and households was 1.52 μg/g, 1.24 μg/g, 1.31 μg/g and 1.46 μg/g respectively, and the concentration of Der p1 in above-mentioned places was 1.23 μg/g, 1.12 μg/g, 1.16 μg/g and 1.18 μg/g respectively. The concentration of Der f1 & Der p1 in air was higher after the air-conditioners starting one hours later, and the difference was significant (P<0.05, respectively). Additionally, dot-ELISA findings revealed that the allergen extracted from the dust was capable of reacting with IgE from the sera of asthma mice allergic to dust mites. The study concludes that air-conditioner filters can enrich dust mites major group allergen, and the allergens can induce asthma. The air-conditioner filters shall be cleaned or replaced regularly to prevent or reduce accumulation of the dust mites and its allergens. PMID:26064381

  11. Mammal-derived respiratory lipocalin allergens do not exhibit dendritic cell-activating capacity.

    PubMed

    Parviainen, S; Kinnunen, T; Rytkönen-Nissinen, M; Nieminen, A; Liukko, A; Virtanen, T

    2013-03-01

    Most mammal-derived respiratory allergens belong to the lipocalin family of proteins. Determinants of their allergenic capacity are still unknown. Innate immune cells, in particular dendritic cells, have been shown to be involved in the allergenicity of some proteins. As recognition by dendritic cells is one of the few plausible mechanisms for the allergenicity of proteins, we wanted to investigate their role in the allergenicity of lipocalin allergens. Therefore, we first incubated human monocyte-derived dendritic cells with immunologically functional recombinant allergens mouse Mus m 1, dog Can f 1 and 2, cow Bos d 2, horse Equ c 1 and natural Bos d 2. Then, the surface marker expression and cytokine production of dendritic cells and their capacity to promote T cell proliferation and Th2 immune deviation in naïve CD4(+) T cells were examined in vitro. We found that near to endotoxin-free lipocalin allergens had no effect on the activation, allostimulatory capacity or cytokine production of dendritic cells. The dendritic cells could not induce immune deviation in naïve CD4(+) T cells. In contrast, lipopolysaccharide activated the dendritic cells efficiently. However, lipocalin allergens were not able to modify the lipopolysaccharide-induced responses. We conclude that an important group of mammal-derived respiratory allergens, lipocalins, appear not to be able to activate dendritic cells, a major component involved in the allergenicity of some proteins. It is conceivable that this incapacity of lipocalin allergens to arouse innate immunity may be associated with their poor capacity to induce a strong T cell response, verified in several studies.

  12. Allergens in school settings: results of environmental assessments in 3 city school systems.

    PubMed

    Abramson, Stuart L; Turner-Henson, Anne; Anderson, Lise; Hemstreet, Mary P; Bartholomew, L Kay; Joseph, Christine L M; Tang, Shenghui; Tyrrell, Shellie; Clark, Noreen M; Ownby, Dennis

    2006-08-01

    Environmental allergens are major triggers for pediatric asthma. While children's greatest exposure to indoor allergens is in the home, other public places where children spend a large amount of time, such as school and day care centers, may also be sources of significant allergen encounters. The purpose of this article is to describe schoolroom allergen levels from 3 different geographic sites obtained from dust samples collected in the fall and in spring. Environmental dust samples were collected from elementary schools in Birmingham (AL), Detroit (MI), and Houston (TX), from 4 room locations, including the cafeteria, library, upper grades, and lower grades. Samples were assayed for dust mite (Dermatophagoides pteronyssinus and Dermatophagoides farinae), cat (Felis domesticus), and cockroach (Blatella germanica 2) allergen levels. Allergen levels varied by geographic location and type of schoolroom. Schoolroom settings differed by the type of flooring (hard and carpet), room characteristics and use (food service, library shelves with books, and general classroom with multiple types of materials [individual desks and different types of furniture]), and the average age of the schoolroom dwellers (younger vs. older children). Dust mite, cat, and cockroach allergens were present in all schoolrooms and all sites at varying levels by season and by type of room. Schools may be important sources of direct allergen exposure and reservoirs that could potentially contribute to allergic sensitization and disease exacerbation in children. Further studies are needed to carefully examine the environmental allergen load in schools and its effect on children.

  13. The allergens in cosmetics.

    PubMed

    de Groot, A C; Bruynzeel, D P; Bos, J D; van der Meeren, H L; van Joost, T; Jagtman, B A; Weyland, J W

    1988-10-01

    The ingredients responsible for allergy to cosmetics were determined in 119 patients suffering from cosmetic-related contact dermatitis. Most reactions (56.3%) were caused by skin care products, followed by nail cosmetics (13.4%), perfumes (8.4%), and hair cosmetics (5.9%). Preservatives were most frequently implicated (32.0%), followed by fragrances (26.5%) and emulsifiers (14.3%). By far the most important cosmetic allergen was Kathon CG, (a preservative system containing, as active ingredients, a mixture of methylisothiazolinone and methyl chloroisothiazolinone) reacting in 33 patients (27.7%). Other frequent causes of cosmetic-related contact allergic reactions were toluenesulfonamide/formaldehyde resin in nail hardener and/or nail lacquer (15 patients [12.6%]), and oleamidopropyl dimethylamine, an emulsifier in baby body lotion (13 patients [10.9%]).

  14. Grass Pollen Allergens

    PubMed Central

    Augustin, Rosa; Hayward, Barbara J.

    1962-01-01

    Cocksfoot and Timothy pollen extracts are each found to contain at least fifteen components antigenic in rabbits. Most of these can also be allergens for man, but only a few are regularly so. These `principal' allergens have now been isolated in highly purified form. Procedures are given for a simple method of preparing extracts for clinical purposes and for the partial separation, concentration and purification of the allergens by means of differential extractions of the pollens and by means of ultrafiltration, isoelectric precipitation and salt fractionations (at acid and neutral pH) of the extracts. Isoelectric precipitations gave highly pigmented acid complexes, two of which moved as single sharp peaks at pH 7.4 in free electrophoresis, but proved to be hardly active by skin tests. Acid NaCl fractionation of the remainder resulted for Cocksfoot and Timothy in the isolation of a nearly white powder (T21.111121112 = T21B) which was weight for weight 1000–10,000 times as active as the pollen from which it had been derived. The powders have retained their activity for 7 years. By gel diffusion tests, they were found to contain two antigens (one in each preparation) which were immunologically partially related, but the Timothy preparation contained in addition the `innermost' `twin' antigens specific for Timothy that we had discovered previously in the crude extracts by gel diffusion methods. Skin reactions could be elicited in hay-fever subjects by prick tests with concentrations of 10-9–10-8 g./ml., which is equivalent to intradermal injections of 10-11–10-10 mg. and represents a 300-fold purification with respect to the concentrates of crude pollen extracts prepared by ultrafiltration and dialysis. Fractionation on DEAE-cellulose of one of the highly purified Timothy preparations (T21.11112112 = T21A) and other, crude Timothy and Cocksfoot extracts resulted in considerable and reproducible separation of the various antigens, with no indication of the

  15. In silico allergenicity prediction of several lipid transfer proteins.

    PubMed

    Garino, Cristiano; Coïsson, Jean Daniel; Arlorio, Marco

    2016-02-01

    Non-specific lipid transfer proteins (nsLTPs) are common allergens and they are particularly widespread within the plant kingdom. They have a highly conserved three-dimensional structure that generate a strong cross-reactivity among the members of this family. In the last years several web tools for the prediction of allergenicity of new molecules based on their homology with known allergens have been released, and guidelines to assess potential allergenicity of proteins through bioinformatics have been established. Even if such tools are only partially reliable yet, they can provide important indications when other kinds of molecular characterization are lacking. The potential allergenicity of 28 amino acid sequences of LTPs homologs, either retrieved from the UniProt database or in silico deduced from the corresponding EST coding sequence, was predicted using 7 publicly available web tools. Moreover, their similarity degree to their closest known LTP allergens was calculated, in order to evaluate their potential cross-reactivity. Finally, all sequences were studied for their identity degree with the peach allergen Pru p 3, considering the regions involved in the formation of its known conformational IgE-binding epitope. Most of the analyzed sequences displayed a high probability to be allergenic according to all the software employed. The analyzed LTPs from bell pepper, cassava, mango, mungbean and soybean showed high homology (>70%) with some known allergenic LTPs, suggesting a potential risk of cross-reactivity for sensitized individuals. Other LTPs, like for example those from canola, cassava, mango, mungbean, papaya or persimmon, displayed a high degree of identity with Pru p 3 within the consensus sequence responsible for the formation, at three-dimensional level, of its major conformational epitope. Since recent studies highlighted how in patients mono-sensitized to peach LTP the levels of IgE seem directly proportional to the chance of developing cross

  16. Pollen Allergens for Molecular Diagnosis.

    PubMed

    Pablos, Isabel; Wildner, Sabrina; Asam, Claudia; Wallner, Michael; Gadermaier, Gabriele

    2016-04-01

    Pollen allergens are one of the main causes of type I allergies affecting up to 30% of the population in industrialized countries. Climatic changes affect the duration and intensity of pollen seasons and may together with pollution contribute to increased incidences of respiratory allergy and asthma. Allergenic grasses, trees, and weeds often present similar habitats and flowering periods compromising clinical anamnesis. Molecule-based approaches enable distinction between genuine sensitization and clinically mostly irrelevant IgE cross-reactivity due to, e. g., panallergens or carbohydrate determinants. In addition, sensitivity as well as specificity can be improved and lead to identification of the primary sensitizing source which is particularly beneficial regarding polysensitized patients. This review gives an overview on relevant pollen allergens and their usefulness in daily practice. Appropriate allergy diagnosis is directly influencing decisions for therapeutic interventions, and thus, reliable biomarkers are pivotal when considering allergen immunotherapy in the context of precision medicine.

  17. Are specific allergen sensitivities inherited?

    PubMed

    Misiak, Rana Tawil; Wegienka, Ganesa; Zoratti, Edward

    2010-09-01

    A family history of an allergic condition is a well-accepted risk factor for the development of an allergic condition in an individual, particularly for allergic disorders such as asthma, eczema, and allergic rhinitis. However, the question of whether specific allergen sensitization is inherited requires a complicated answer, as environmental exposure plays an important role in the development of allergen-specific IgE. This article summarizes the findings of recent studies in the literature regarding what is known about the inheritance of specific allergens. Overall, properly collected and analyzed data appear to both support and refute the hypothesis that specific allergen sensitization is inherited, even when attempting to account for the complexities of varying study methodologies and the evaluation of diverse populations and communities. PMID:20574668

  18. [Sensitisation to 'poffertjes' as a result of sleeping on a pillow containing buckwheat].

    PubMed

    van Ginkel, C J W

    2002-03-30

    A 19-year-old man suffered an anaphylactic reaction after eating 'poffertjes' (small Dutch pancakes). This reaction appeared to be the result of an IgE-mediated allergy to buckwheat, a principal ingredient of 'poffertjes'. It is highly likely that the patient was sensitised by sleeping on a pillow stuffed with buckwheat husk. PMID:11957384

  19. Characterization of cookies made from wheat flour blended with buckwheat flour and effect on antioxidant properties.

    PubMed

    Jan, Ulfat; Gani, Adil; Ahmad, Mudasir; Shah, Umar; Baba, Waqas N; Masoodi, F A; Maqsood, Sajid; Gani, Asir; Wani, Idress Ahmed; Wani, S M

    2015-10-01

    Buckwheat flour was incorporated into wheat flour at different levels (0, 20, 40, 60, 80, and 100 %) and the physicochemical, functional and antioxidant properties of the blended flour were studied. This study also investigated the effect of buckwheat on the retention of antioxidant properties of cookies during baking. The results showed significant variation in physicochemical and functional properties of the blended flour. The addition of buckwheat flour into wheat flour also increased the antioxidant properties of blended flour proportionally, but metal chelating properties decreased. The incorporation of buckwheat in wheat flour helped in better retention of antioxidant potential of cookies during baking process as buckwheat cookies (100 % buckwheat) showed greater percentage increase in antioxidant properties than control (100 % wheat). Quality characteristics of cookies such as hardness and spread ratio decreased, while as non-enzymatic browning (NEB) increased significantly with increase in the proportion of buckwheat flour in wheat flour. The Overall acceptability of cookies by sensory analysis was highest at 40 % level of blending. This study concluded that addition of buckwheat in wheat flour, may not only improve the physico-chemical and functional properties of the blended flour but may also enhance the nutraceutical potential of the product prepared from it.

  20. Characterization of cookies made from wheat flour blended with buckwheat flour and effect on antioxidant properties.

    PubMed

    Jan, Ulfat; Gani, Adil; Ahmad, Mudasir; Shah, Umar; Baba, Waqas N; Masoodi, F A; Maqsood, Sajid; Gani, Asir; Wani, Idress Ahmed; Wani, S M

    2015-10-01

    Buckwheat flour was incorporated into wheat flour at different levels (0, 20, 40, 60, 80, and 100 %) and the physicochemical, functional and antioxidant properties of the blended flour were studied. This study also investigated the effect of buckwheat on the retention of antioxidant properties of cookies during baking. The results showed significant variation in physicochemical and functional properties of the blended flour. The addition of buckwheat flour into wheat flour also increased the antioxidant properties of blended flour proportionally, but metal chelating properties decreased. The incorporation of buckwheat in wheat flour helped in better retention of antioxidant potential of cookies during baking process as buckwheat cookies (100 % buckwheat) showed greater percentage increase in antioxidant properties than control (100 % wheat). Quality characteristics of cookies such as hardness and spread ratio decreased, while as non-enzymatic browning (NEB) increased significantly with increase in the proportion of buckwheat flour in wheat flour. The Overall acceptability of cookies by sensory analysis was highest at 40 % level of blending. This study concluded that addition of buckwheat in wheat flour, may not only improve the physico-chemical and functional properties of the blended flour but may also enhance the nutraceutical potential of the product prepared from it. PMID:26396378

  1. Sensitization to fungal allergens: Resolved and unresolved issues.

    PubMed

    Fukutomi, Yuma; Taniguchi, Masami

    2015-10-01

    Exposure and sensitization to fungal allergens can promote the development and worsening of allergic diseases. Although numerous species of fungi have been associated with allergic diseases in the literature, the significance of fungi from the genera Alternaria, Cladosporium, Penicillium, Aspergillus, and Malassezia has been well documented. However, it should be emphasized that the contribution of different fungal allergens to allergic diseases is not identical, but species-specific. Alternaria and Cladosporium species are considered to be important outdoor allergens, and sensitization and exposure to species of these genera is related to the development of asthma and rhinitis, as well as epidemics of asthma exacerbation, including life-threatening asthma exacerbation. In contrast, xerophilic species of Penicillium and Aspergillus, excluding Aspergillus fumigatus, are implicated in allergic diseases as indoor allergens. A. fumigatus has a high capacity to colonize the bronchial tract of asthmatic patients, causing severe persistent asthma and low lung function, and sometimes leading to allergic bronchopulmonary aspergillosis. Malassezia are common commensals of healthy skin, although they are also associated with atopic dermatitis, especially on the head and neck, but not with respiratory allergies. Despite its importance in the management of allergic diseases, precise recognition of species-specific IgE sensitization to fungal allergens is often challenging because the majority of fungal extracts exhibit broad cross-reactivity with taxonomically unrelated fungi. Recent progress in gene technology has contributed to the identification of specific and cross-reactive allergen components from different fungal sources. However, data demonstrating the clinical relevance of IgE reactivity to these allergen components are still insufficient. PMID:26433528

  2. Mechanisms and characteristics of airway sensitization to indoor allergens.

    PubMed

    Liccardi, G; Cazzola, M; Russo, M; Gilder, J A; D'Amato, M; D'Amato, G

    2001-02-01

    The increasing prevalence of allergic respiratory diseases, and particularly of bronchial asthma, has been linked to changes induced by human activities in outdoor and indoor environments. People living in industrialized countries spend most of their time indoors: in private homes, offices and means of transport. Indoor environments are not a refuge from outdoor air pollution. Modern systems for energy saving such as insulated windows and doors reduce the indoor natural ventilation and consequently increase the rate of indoor humidity. These conditions may determine an increase in the level of indoor pollutants (tobacco smoke, gases produced by cooling processes etc.) and of allergens derived from mites, domestic animals and cockroaches. Upholstered furniture, wall-to-wall carpets, central heating systems and/or humidifiers may also contribute to the growth of mite populations. The increasing levels of exposure to pollutants and allergens in indoor environments represents a risk factor for the development of airway sensitization, especially if these materials are inhaled early in life. The major cat allergen Fel d 1 is considered an ubiquitous allergen, since it has been found in many indoor environments where a cat has never been kept. The clothing of cat owners seems to help spread Fel d 1 in cat-free environments. Sensitization to cockroach allergens is very common in patients living in urban areas where unhygenic conditions may favour the growth of cockroach populations. Monitoring of the levels of allergens and strategies of allergen and pollutant avoidance in indoor environments are the main ways to reduce the prevalence of respiratory allergies induced by these materials.

  3. Biochemical and immunological characterization of recombinant allergen Lol p 1.

    PubMed

    Tamborini, E; Faccini, S; Lidholm, J; Svensson, M; Brandazza, A; Longhi, R; Groenlund, H; Sidoli, A; Arosio, P

    1997-11-01

    Pollen from perennial rye grass (Lolium perenne), a major cause of type-I allergy worldwide, contains a complex mixture of allergenic proteins among which Lol p 1 is one of the most important. We describe the expression, purification and characterization of a recombinant Lol p 1 overproduced in Escherichia coli. The recombinant allergen, expressed in high yields and purified in milligram amounts, bound to specific IgE antibodies from human sera, induced histamine release from sensitized human basophils, and elicited rabbit antisera that recognize specifically recombinant Lol p 1 and natural Lol p 1 of pollen extract. Recombinant Lol p 1 was used to develop ImmunoCAP assays for analysis of 150 sera that were Radioallergosorbent test positive to L. perenne pollen. In 130 of them (87%) the assay detected a significant level of IgE antibodies to Lol p 1, reaching on average 37% of the level obtained with a test for IgE to the whole grass pollen extract. To map epitopes on Lol p 1, we produced three deletion mutants [des-(116-240)-Lol p 1, des-(1-88)-Lol p 1 and des-(133-189)-Lol p 1], which were efficiently expressed in bacteria. These all showed a strong reactivity with the specific rabbit IgG antibodies, but lacked most or all the allergenic properties of recombinant Lol p 1. A study of the antigenic structure of Lol p 1 was performed using the three deletion mutants and a set of 17-18-residue overlapping synthetic peptides covering the whole allergen sequence. The results indicate that human IgE and rabbit IgG antibodies bind to distinct regions of Lol p 1, and that at least some important IgE epitopes are mainly conformational. The findings suggest that recombinant allergens constitute useful reagents for further development of serological diagnosis of allergy, and that it should be possible to produce immunogenic fragments of allergenic proteins without allergenic properties.

  4. Current codex guidelines for assessment of potential protein allergenicity.

    PubMed

    Ladics, G S

    2008-10-01

    A rigorous safety assessment process exists for GM crops. It includes evaluation of the introduced protein as well as the crop containing such protein with the goal of demonstrating the GM crop is "as-safe-as" non-transgenic crops in the food supply. One of the major issues for GM crops is the assessment of the expressed protein for allergenic potential. Currently, no single factor is recognized as an identifier for protein allergenicity. Therefore, a weight-of-evidence approach, which takes into account a variety of factors and approaches for an overall assessment of allergenic potential, is conducted [Codex Alimentarious Commission, 2003. Alinorm 03/34: Joint FAO/WHO Food Standard Programme, Codex Alimentarious Commission, Twenty-Fifth Session, Rome, Italy, 30 June-5 July, 2003. Appendix III, Guideline for the conduct of food safety assessment of foods derived from recombinant-DNA plants, and Appendix IV, Annex on the assessment of possible allergenicity, pp. 47-60]. This assessment is based on what is known about allergens, including the history of exposure and safety of the gene(s) source; protein structure (e.g., amino acid sequence identity to human allergens); stability to pepsin digestion in vitro [Thomas, K. et al., 2004. A multi-laboratory evaluation of a common in vitro pepsin digestion assay protocol used in assessing the safety of novel proteins. Regul. Toxicol. Pharmacol. 39, 87-98]; an estimate of exposure of the novel protein(s) to the gastrointestinal tract where absorption occurs (e.g., protein abundance in the crop, processing effects); and when appropriate, specific IgE binding studies or skin prick testing. Additional approaches may be considered (e.g., animal models; targeted sera screening) as the science evolves; however, such approaches have not been thoroughly evaluated or validated for predicting protein allergenicity.

  5. Fungi: the neglected allergenic sources.

    PubMed

    Crameri, R; Garbani, M; Rhyner, C; Huitema, C

    2014-02-01

    Allergic diseases are considered the epidemics of the twentieth century estimated to affect more than 30% of the population in industrialized countries with a still increasing incidence. During the past two decades, the application of molecular biology allowed cloning, production and characterization of hundreds of recombinant allergens. In turn, knowledge about molecular, chemical and biologically relevant allergens contributed to increase our understanding of the mechanisms underlying IgE-mediated type I hypersensitivity reactions. It has been largely demonstrated that fungi are potent sources of allergenic molecules covering a vast variety of molecular structures including enzymes, toxins, cell wall components and phylogenetically highly conserved cross-reactive proteins. Despite the large knowledge accumulated and the compelling evidence for an involvement of fungal allergens in the pathophysiology of allergic diseases, fungi as a prominent source of allergens are still largely neglected in basic research as well as in clinical practice. This review aims to highlight the impact of fungal allergens with focus on asthma and atopic dermatitis.

  6. The Contribution of Buckwheat Genetic Resources to Health and Dietary Diversity.

    PubMed

    Sytar, Oksana; Brestic, Marian; Zivcak, Marek; Tran, Lam-Son Phan

    2016-06-01

    Despite several reports on the beneficial effects of buckwheat in prevention of human diseases, little attention has been devoted to the variability of biochemical and physiological traits in different buckwheat genetic resources. This review describes the biochemical evaluation of buckwheat genetic resources and the identification of elite genotypes for plant breeding and exploitation. The various types of bioactive compounds present in different varieties provide basic background information needed for the efficient production of buckwheat foods with added value. In this review, we will provide an integrated view of the biochemistry of bioactive compounds of buckwheat plants of different origin, especially of fagopyrin, proteins and amino acids, as well as of other phenolic compounds including rutin and chlorogenic acid. In addition to the genetic background, the effect of different growth conditions is discussed. The health effects of fagopyrin, phenolic acids, specific proteins and rutin are also presented. PMID:27252586

  7. Trace element water improves the antioxidant activity of buckwheat (Fagopyrum esculentum Moench) sprouts.

    PubMed

    Liu, Chia-Ling; Chen, Yih-Shyuan; Yang, Joan-Hwa; Chiang, Been-Huang; Hsu, Cheng-Kuang

    2007-10-31

    Buckwheat (Fagopyrum esculentum Moench) was grown in trace element water (TEW) (100, 200, 300, 400, and 500 ppm) and deionized water (DIW) to evaluate whether the beneficial effects of trace elements on the antioxidant activity could be accomplished with the supplement of TEW. At 300 ppm, TEW significantly increased the Cu, Zn, Mn, and Fe contents in buckwheat sprout but not the Se content. However, the levels of rutin, isoorientin, vitexin, and isovitexin did not differ between buckwheat sprouts grown in TEW and DIW. The ethanolic extract from buckwheat sprout grown in 300 ppm of TEW showed higher ferrous ion chelating activity and inhibitory activity toward lipid peroxidation than that grown in DIW. The extract in the TEW group also enhanced intracellular superoxide dismutase activity and lowered reactive oxygen species and superoxide anion in the human Hep G2 cell. It was concluded that TEW could increase the antioxidant activities of buckwheat sprouts.

  8. [Different uses of Fagopyrum esculentum Moench (buckwheat) in Japan and China: what ancient medical documents reveal].

    PubMed

    Tatsumi, Nami; Marui, Eiji

    2012-03-01

    The purpose of this thesis is to demonstrate that buckwheat has been recognized, both in Japan and China, as a crop that is useful in many ways: as an agricultural crop, and for the healing powers and properties that, according to traditional Chinese medicine, it has. A comparative study of ancient documents pertaining to medicine in these countries has made it clear that this is the case. Buckwheat, however, has been used quite differently in each country. As is shown in some ancient Chinese documents pertaining to medicine, China has treated buckwheat primarily as a medicine for clinical use rather than as an edible crop. Nowadays, buckwheat is eaten only in some regions of China. Although it came to Japan from China as a medicine, in Japan buckwheat gradually became a popular food crop. It has become an important component of traditional Japanese cuisine thanks in part to government support and the strong demand that developed in Japanese society.

  9. Impact of Wild Loci on the Allergenic Potential of Cultivated Tomato Fruits.

    PubMed

    Ghiani, Alessandra; D'Agostino, Nunzio; Citterio, Sandra; Raiola, Assunta; Asero, Riccardo; Barone, Amalia; Rigano, Maria Manuela

    2016-01-01

    Tomato (Solanum lycopersicum) is one of the most extensively consumed vegetables but, unfortunately, it is also able to induce allergic reactions. In the past, it has been shown that the choice of tomato cultivar significantly influenced the allergic reaction of tomato allergic subjects. In this study we investigated the allergenic potential of the cultivated tomato line M82 and of two selected lines carrying small chromosome regions from the wild species Solanum pennellii (i.e. IL7-3 and IL12-4). We evaluated the positive interactions of IgEs of allergic subjects in order to investigate the different allergenic potential of the lines under investigation. We used proteomic analyses in order to identify putative tomato allergens. In addition, bioinformatic and transcriptomic approaches were applied in order to analyse the structure and the expression profiles of the identified allergen-encoding genes. These analyses demonstrated that fruits harvested from the two selected introgression lines harbour a different allergenic potential as those from the cultivated genotype M82. The different allergenicity found within the three lines was mostly due to differences in the IgE recognition of a polygalacturonase enzyme (46 kDa), one of the major tomato allergens, and of a pectin methylesterase (34 kDa); both the proteins were more immunoreactive in IL7-3 compared to IL12-4 and M82. The observed differences in the allergenic potential were mostly due to line-dependent translational control or post-translational modifications of the allergens. We demonstrated, for the first time, that the introgression from a wild species (S. pennellii) in the genomic background of a cultivated tomato line influences the allergenic properties of the fruits. Our findings could support the isolation of favorable wild loci promoting low allergenic potential in tomato. PMID:27182705

  10. Impact of Wild Loci on the Allergenic Potential of Cultivated Tomato Fruits

    PubMed Central

    Ghiani, Alessandra; D’Agostino, Nunzio; Citterio, Sandra; Raiola, Assunta; Asero, Riccardo; Barone, Amalia; Rigano, Maria Manuela

    2016-01-01

    Tomato (Solanum lycopersicum) is one of the most extensively consumed vegetables but, unfortunately, it is also able to induce allergic reactions. In the past, it has been shown that the choice of tomato cultivar significantly influenced the allergic reaction of tomato allergic subjects. In this study we investigated the allergenic potential of the cultivated tomato line M82 and of two selected lines carrying small chromosome regions from the wild species Solanum pennellii (i.e. IL7-3 and IL12-4). We evaluated the positive interactions of IgEs of allergic subjects in order to investigate the different allergenic potential of the lines under investigation. We used proteomic analyses in order to identify putative tomato allergens. In addition, bioinformatic and transcriptomic approaches were applied in order to analyse the structure and the expression profiles of the identified allergen-encoding genes. These analyses demonstrated that fruits harvested from the two selected introgression lines harbour a different allergenic potential as those from the cultivated genotype M82. The different allergenicity found within the three lines was mostly due to differences in the IgE recognition of a polygalacturonase enzyme (46 kDa), one of the major tomato allergens, and of a pectin methylesterase (34 kDa); both the proteins were more immunoreactive in IL7-3 compared to IL12-4 and M82. The observed differences in the allergenic potential were mostly due to line-dependent translational control or post-translational modifications of the allergens. We demonstrated, for the first time, that the introgression from a wild species (S. pennellii) in the genomic background of a cultivated tomato line influences the allergenic properties of the fruits. Our findings could support the isolation of favorable wild loci promoting low allergenic potential in tomato. PMID:27182705

  11. Allergenic airborne pollen and spores in Anchorage, Alaska

    SciTech Connect

    Anderson, J.H.

    1985-05-01

    Major aeroallergens in Anchorage are birch, alder, poplar, spruce, grass pollen, Cladosporium, and unspecified fungus spores. Lesser pollens are sorrel, willow, pine, juniper, sedge, lamb's-quarters, wormwood, plantain, and others. The aero-flora is discussed in terms of the frequency of allergenically significant events and within-season and year-to-year dynamics.

  12. Allergenic airborne pollen and spores in Anchorage, Alaska.

    PubMed

    Anderson, J H

    1985-05-01

    Major aeroallergens in Anchorage are birch, alder, poplar, spruce, grass pollen, Cladosporium, and unspecified fungus spores. Lesser pollens are sorrel, willow, pine, juniper, sedge, lamb's-quarters, wormwood, plantain, and others. The aero-flora is discussed in terms of the frequency of allergenically significant events and within-season and year-to-year dynamics.

  13. Crystal structure of peanut (Arachis hypogaea) allergen Ara h 5

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Profilins from numerous species are known to be allergens, including food allergens, such as peanut (Arachis hypogaea) allergen Ara h 5, and pollen allergens, such as birch allergen Bet v 2. Patients with pollen allergy can also cross-react to peanut. Structural characterization of allergens will al...

  14. Allergenicity of orange juice and orange seeds: a clinical study.

    PubMed

    Zhu, S L; Ye, S T; Yu, Y

    1989-06-01

    Oranges are considered to be common allergenic fruits in China. They may induce severe food allergy in sensitive individuals. Allergic histories were analyzed in 26 orange-sensitive patients. Intradermal tests with extracts of orange juice and seeds were performed in 16 out of the 26 patients. P-K test was performed in one patient. The allergic history analysis suggested that clinical symptoms of some orange-allergic subjects were different from other fruit allergies but similar to nut and other oil plant seed allergies. The skin test and P-K test showed that the major allergenic components of orange reside in orange seeds instead of orange juice. Systemic reactions developed in 5 patients after intradermal tests with 1:20-200 (w/v) orange seed extracts. We considered that orange seed contains high potent allergens which may induce orange sensitivity due to careless chewing of orange seeds. PMID:2751771

  15. Characterization of a new oriental-mustard (Brassica juncea) allergen, Bra j IE: detection of an allergenic epitope.

    PubMed Central

    Monsalve, R I; Gonzalez de la Peña, M A; Menendez-Arias, L; Lopez-Otin, C; Villalba, M; Rodriguez, R

    1993-01-01

    Bra j IE, a major allergen from oriental-mustard (Brassica juncea) seeds, has been isolated and characterized. Its primary structure has been elucidated. This protein is composed of two chains (37 and 92 amino acids) linked by disulphide bridges. The amino acid sequence obtained is closely related to that previously determined for Sin a I, an allergen isolated from yellow mustard (Sinapis alba). A common epitope has been detected in the large chain of both Bra j IE and Sin a I by means of electroblotting and immunodetection with 2B3, which is a monoclonal antibody raised against the yellow-mustard allergen. A histidine residue of the large chain of both mustard allergens has been found to be essential for the recognition by 2B3 antibody. A synthetic multiantigenic peptide containing this His was recognized by 2B3 as well as by sera of mustard-hypersensitive individuals. Therefore this antigenic determinant must be involved in the allergenicity of these proteins. Images Figure 3 Figure 7 Figure 8 PMID:7688955

  16. Beneficial Influence of Short-Term Germination on Decreasing Allergenicity of Peanut Proteins.

    PubMed

    Li, Yingchao; Sun, Xiulan; Ma, Zhezhe; Cui, Yan; Du, Chao; Xia, Xiuhua; Qian, He

    2016-01-01

    Most allergenic storage proteins in peanuts are degraded during seed germination. By altering this natural physiological process, it might be possible to reduce peanut protein allergenicity. However, little is known about the change in allergenic proteins and their corresponding immunocreactivity, and the effects of major environmental conditions on their allergenicity during germination. In this study, the influence of different germination conditions (temperature and light) on the degradation of Ara h1 and allergenicity changes of peanut seeds was evaluated by ELISA and Western blotting. The results showed that the 40- and 65-kDa proteins in peanut seeds degraded rapidly during the time course, beginning at 60 (at 25 °C) and 108 h (at 20 °C), and the corresponding immunocreactivity of Ara h1 decreased approximately one-third after 5 to 7 d of germination. Compared with the cotyledons, the embryonic axes had a higher proportion of Ara h1, which was then degraded relatively faster during germination, resulting in a significant reduction in its allergenicity. Although a higher temperature improved the seed germination rate, it affected sprout quality (as did light); therefore, 25 °C and dark surroundings were suitable conditions under which peanut sprouts were processed; neither factor significantly affected the allergenicity of Ara h1. These results provided a theoretical basis for studies using biological methods to reduce peanut allergenicity.

  17. Beneficial Influence of Short-Term Germination on Decreasing Allergenicity of Peanut Proteins.

    PubMed

    Li, Yingchao; Sun, Xiulan; Ma, Zhezhe; Cui, Yan; Du, Chao; Xia, Xiuhua; Qian, He

    2016-01-01

    Most allergenic storage proteins in peanuts are degraded during seed germination. By altering this natural physiological process, it might be possible to reduce peanut protein allergenicity. However, little is known about the change in allergenic proteins and their corresponding immunocreactivity, and the effects of major environmental conditions on their allergenicity during germination. In this study, the influence of different germination conditions (temperature and light) on the degradation of Ara h1 and allergenicity changes of peanut seeds was evaluated by ELISA and Western blotting. The results showed that the 40- and 65-kDa proteins in peanut seeds degraded rapidly during the time course, beginning at 60 (at 25 °C) and 108 h (at 20 °C), and the corresponding immunocreactivity of Ara h1 decreased approximately one-third after 5 to 7 d of germination. Compared with the cotyledons, the embryonic axes had a higher proportion of Ara h1, which was then degraded relatively faster during germination, resulting in a significant reduction in its allergenicity. Although a higher temperature improved the seed germination rate, it affected sprout quality (as did light); therefore, 25 °C and dark surroundings were suitable conditions under which peanut sprouts were processed; neither factor significantly affected the allergenicity of Ara h1. These results provided a theoretical basis for studies using biological methods to reduce peanut allergenicity. PMID:26618608

  18. Analysis of green coffee bean and castor bean allergens using RAST inhibition.

    PubMed

    Lehrer, S B; Karr, R M; Salvaggio, J E

    1981-07-01

    Coffee workers with occupational allergic symptoms and positive skin tests to green coffee bean and factor dust antigens have elevated serum IgE antibodies (by radioallergosorbent test--RAST) to green coffee and castor bean allergens. These antibodies were used in a RAST inhibition assay to analyse coffee and castor allergens. Bean allergens were extracted by homogenization in PBS, centrifugation and concentration of supernates by ultrafiltration. Green coffee bean allergens, fractionated by gel filtration and Pevikon block electrophoresis, were shown to be very heterogeneous with a molecular weight range of 50 000 to 500 000 daltons. Castor allergens were more homogeneous with a molecular weight of 14 000 daltons and were partially purified by Pevikon block electrophoresis, gel filtration and isoelectrofocusing. Chemical analysis showed that protein was the major component in both allergen extracts. However, proteolytic enzymes could only partially destroy allergenic activity. Such isolation and characterization of these allergens should result in better methods of diagnosis and treatment of coffee workers with occupational allergic disease.

  19. Determination of storage conditions for shrimp extracts: analysis of specific IgE-allergen profiles.

    PubMed

    Piboonpocanun, Surapon; Boonchoo, Siribangon; Pariyaprasert, Wipada; Visitsunthorn, Nualanong; Jirapongsananuruk, Orathai

    2010-03-01

    The consumption of shrimp is a common cause of food hypersensitivity reactions. Shrimp allergy is diagnosed using a skin prick test (SPT) as well as by food challenges. Due to the lack of a wide variety of commercial shrimp extracts for SPTs, we selected various shrimp species for the preparation of local shrimp extracts. However, optimal storage conditions for the shrimp extracts which also maintains allergenic potency has not yet been identified. The objective of the present study was to determine the potency of the shrimp extracts under different storage conditions and durations. Specific IgE-allergen profiles of eight shrimp-allergic patients were investigated by using sera incubated with extracts prepared from lyophilized raw or boiled shrimp, which were stored at 4 degress C or -20 degress C for up to 4 weeks. When stored at -20 degress C, most allergens were preserved after 4 weeks. However, storage at 4 degress C results in few allergens remaining after 2 weeks. Boiled-shrimp extracts stored at 4 degree C and -20 degress C contained higher amounts of IgE-allergen complexes than raw-shrimp extracts. Moreover, in both raw and boiled shrimp extracts, the IgE bound 36-40 kDa allergens constituted the major proteins since they were observed in all IgE-allergen profiles. In conclusion, we recommend that shrimp extracts are stored at -20 degress C for 4 weeks to prevent the loss of allergens.

  20. Recombinant allergens: what does the future hold?

    PubMed

    Valenta, Rudolf; Niespodziana, Katarzyna; Focke-Tejkl, Margit; Marth, Katharina; Huber, Hans; Neubauer, Angela; Niederberger, Verena

    2011-04-01

    This year we are celebrating not only the centenary of allergen-specific immunotherapy but also the 10-year anniversary of the first administration of recombinant allergen-based vaccines to allergic patients. By using recombinant DNA technology, defined and safe allergy vaccines can be produced that allow us to overcome many, if not all, of the problems associated with the use of natural allergen extracts, such as insufficient quality, allergenic activity, and poor immunogenicity. Here we provide an update of clinical studies with recombinant allergen-based vaccines, showing that some of these vaccines have undergone successful clinical evaluation up to phase III studies. Furthermore, we introduce a strategy for allergen-specific immunotherapy based on recombinant fusion proteins consisting of viral carrier proteins and allergen-derived peptides without allergenic activity, which holds the promise of being free of side effects and eventually being useful for prophylactic vaccination.

  1. Changes in Atmospheric CO2 Influence the Allergenicity of Aspergillus fumigatus fungal spore

    NASA Astrophysics Data System (ADS)

    Lang-Yona, N.; Levin, Y.; Dannemoller, K. C.; Yarden, O.; Peccia, J.; Rudich, Y.

    2013-12-01

    Increased allergic susceptibility has been documented without a comprehensive understanding for its causes. Therefore understanding trends and mechanisms of allergy inducing agents is essential. In this study we investigated whether elevated atmospheric CO2 levels can affect the allergenicity of Aspergillus fumigatus, a common allergenic fungal species. Both direct exposure to changing CO2 levels during fungal growth, and indirect exposure through changes in the C:N ratios in the growth media were inspected. We determined the allergenicity of the spores through two types of immunoassays, accompanied with genes expression analysis, and proteins relative quantification. We show that fungi grown under present day CO2 levels (392 ppm) exhibit 8.5 and 3.5 fold higher allergenicity compared to fungi grown at preindustrial (280 ppm) and double (560 ppm) CO2 levels, respectively. A corresponding trend is observed in the expression of genes encoding for known allergenic proteins and in the major allergen Asp f1 concentrations, possibly due to physiological changes such as respiration rates and the nitrogen content of the fungus, influenced by the CO2 concentrations. Increased carbon and nitrogen levels in the growth medium also lead to a significant increase in the allergenicity, for which we propose two different biological mechanisms. We suggest that climatic changes such as increasing atmospheric CO2 levels and changes in the fungal growth medium may impact the ability of allergenic fungi such as Aspergillus fumigatus to induce allergies. The effect of changing CO2 concentrations on the total allergenicity per 10^7 spores of A. fumigatus (A), the major allergen Asp f1 concentration in ng per 10^7 spores (B), and the gene expression by RT-PCR (C). The error bars represent the standard error of the mean.

  2. Optimal recovery of high-purity rutin crystals from the whole plant of Fagopyrum esculentum Moench (buckwheat) by extraction, fractionation, and recrystallization.

    PubMed

    Kim, Kyoung Heon; Lee, Ki Won; Kim, Dong Young; Park, Hyung Hwan; Kwon, Ik Boo; Lee, Hyong Joo

    2005-10-01

    Rutin, one of the flavonoids derived from plants, is increasingly in demand in the food, pharmaceutical, and cosmetic industries due to its various biological and physiological activities including antioxidation, anti-inflammation, and anti-hypertension. The whole plant of buckwheat (Fagopyrum esculentum Moench) is a major source of natural rutin. This study developed a low-cost process encompassing the efficient extraction, fractionation, and recrystallization to obtain high-purity rutin from buckwheat, and it could improve the economic utilization of this abundant low-value agricultural product. The sequential separation and purification procedures established in this study involved extraction with 50% (v/v) aqueous ethanol at 80 degrees C for 1 h followed by elution with water and aqueous ethanols at 20% and 30% (v/v) on a styrene-based resin column, and recrystallization at 4 degrees C for 12 h. These conditions resulted in the recovery of 92% of total rutin with over 95% purity. In the present study, high-purity rutin was obtained from whole buckwheat through low-cost processes, the separation and purification strategy established in this study could provide valuable information to the relevant industries.

  3. Identification of tartary buckwheat tea aroma compounds with gas chromatography-mass spectrometry.

    PubMed

    Qin, Peiyou; Ma, Tingjun; Wu, Li; Shan, Fang; Ren, Guixing

    2011-08-01

    Tartary buckwheat tea, which is an important and healthy product, has a distinct malty aroma. However, its characteristic aroma compounds have not been elucidated. The aims of present study were identification and quantification of its aroma compounds. The analyses were performed by gas chromatography-mass spectrometry (GC-MS) after 3 different isolation techniques. Seventy-seven compounds were identified. Among these compounds, 35 were quantified by available standards. The compounds with a high probability of contribution to the tartary buckwheat tea aroma (OAV ≥ 10) were as follows: 2,5-dimethyl-4-hydroxy-3(2H)-furanone, nonanal, 2,3-diethyl-5-methylpyrazine, benzeneacetaldehyde, maltol, 2,5-dimethylpyrazine, 2-ethyl-5-methylpyrazine, trimethylpyrazine. Some nutritional and bioactive compounds were also identified in this study, such as linoleic acid, niacin, vanillic acid, 7-hydroxycoumarin, butylated hydroxytoluene. Practical Application: Tartary buckwheat, one type of buckwheat, has gained much attention from nutritionists and medical doctors in recent years. It is rich in rutin, quercetin, and other nutrients that are good for health. Tartary buckwheat-based product such as tartary buckwheat tea is an important and popular healthy product in China, Japan,South Korea, European countries as well as in American countries. It has a distinct malty aroma. The present study first identified and quantified of its aroma compounds. The results will draw attention to other researchers in food flavor and buckwheat filed. PMID:22417522

  4. Physiological characterization of aluminum tolerance and accumulation in tartary and wild buckwheat.

    PubMed

    Wang, Hua; Chen, Rong Fu; Iwashita, Takashi; Shen, Ren Fang; Ma, Jian Feng

    2015-01-01

    Ionic aluminum (Al) is toxic for plant growth, but some plant species are able to accumulate Al at high concentrations without showing toxicity symptoms. In order to determine whether other species in the genus Fagopyrum are able to accumulate Al like common buckwheat (Fagopyrum esculentum), we investigated the external and internal detoxification mechanisms of Al in two self-compatible species: tartary (Fagopyrum tataricum) and wild buckwheat (Fagopyrum homotropicum). Both tartary and wild buckwheat showed high Al tolerance comparable to common buckwheat. Furthermore, these two species also secreted oxalate rapidly from the roots in response to Al in a time-dependent manner. Both tartary and wild buckwheat accumulated > 1 mg g(-1) Al in the leaves after short-term exposure to Al. Analysis with (27) Al-nuclear magnetic resonance (NMR) revealed that Al was present in the form of Al-oxalate (1 : 3 ratio) in the roots and leaves, but in the form of Al-citrate (1 : 1 ratio) in the xylem sap in both species. These results indicate that similar to common buckwheat, both tartary and wild buckwheat detoxify Al externally and internally, respectively, by secreting oxalate from the roots and by forming the Al-oxalate complex, which is a nonphytotoxic form. These features of Al response and accumulation may be conserved in genus Fagopyrum.

  5. Plantlet Regeneration of Tartary Buckwheat (Fagopyrum tataricum Gaertn.) in Vitro Tissue Cultures.

    PubMed

    Wang, Cheng-Long; Dong, Xue-Ni; Ding, Meng-Qi; Tang, Yi-Xiong; Zhu, Xue-Mei; Wu, Yan-Min; Zhou, Mei-Liang; Shao, Ji-Rong

    2016-01-01

    Tartary buckwheat is an ancient annual dicotyledonous herb, which is widely distributed around the world, specifically in the high altitude area of southwestern China and in the hill region of Himalayan. The plantlet regeneration of tartary buckwheat via somatic embryogenesis or multiple shoot induction was investigated in two different tartary buckwheats, Yuanzi and Xichang. The regeneration ability of Yuanzi was better than Xichang tartary buckwheat, and the hypocotyls were better than cotyledons as tartary buckwheat plantlet regeneration explants via somatic embryogenesis. The most suitable medium for callus induction was Murashige and Skoog basal medium added 2 mg/L 2, 4- dichlorophenoxyacetic acid and 1 mg/L Kinetin, which could reach up to 98.96% callus induction percentage. The plantlet regeneration percentage from callus of tartary buckwheat could reach up to 55.77%, which induced on 2.0 mg/L Benzyladenine and 1.0 mg/L KT in MS basal medium. In addition, maximum of multiple shoot induction percentage was 69.05%, which was observed in case of Yuanzi tartary buckwheat in MS basal medium with added 3.0 mg/L 6-BA and 1.0 mg/L Thidiazuron. Roots induction of regenerated plants were achieved on 1/2 MS basal medium with added 1mg/L Indole-3-Butytric acid, which has 75% survival after transferred regenerated plants to soil under field conditions.

  6. Development of a sandwich enzyme-linked immunosorbent assay (ELISA) for detection of buckwheat residues in food.

    PubMed

    Panda, Rakhi; Taylor, Steve L; Goodman, Richard E

    2010-08-01

    Buckwheat is a pseudocereal (an eudicot with seed qualities and uses similar to those of monocot cereals, family Poaceae) that is consumed in some Asian countries as a staple, and in some western countries as a health food. Allergic reactions to buckwheat are common in some countries. The objective was to develop a specific and sensitive sandwich enzyme-linked immunosorbent assay (ELISA) to detect traces of buckwheat that might inadvertently contaminate other foods in order to assure accurate labeling and consumer protection. Buckwheat-specific antibodies produced in 3 species of animals were tested for specificity and titer by direct ELISA and immunoblot. A sandwich ELISA was developed utilizing pooled rabbit antibuckwheat sera to capture buckwheat proteins and pooled goat antibuckwheat sera, followed by enzyme-labeled rabbit antigoat immunoglobulin G (IgG), to detect bound buckwheat proteins. The lower limit of quantification (LOQ) of the sandwich ELISA was 2 parts per million (ppm) of buckwheat in the presence of complex food matrices. The ELISA is highly specific with no cross-reactivity to any of 80 food ingredients and matrices tested. Validation studies conducted with buckwheat processed into noodles and muffins showed greater than 90% and 60% recovery, respectively. The percent recovery of buckwheat from noodles was similar to that achieved with a commercial buckwheat ELISA kit (ELISA Systems Pty. Ltd., Windsor, Queensland, Australia) at high buckwheat concentrations. However, the sensitivity of this ELISA was greater than the commercial ELISA. This newly developed ELISA is sufficiently specific and sensitive to detect buckwheat residues in processed foods to protect buckwheat-allergic subjects from potential harm. Practical Application: Buckwheat is becoming a common food ingredient in a number of processed foods due to potentially beneficial nutritional properties, without the celiac disease inducing glutenin proteins of wheat and related cereals. However

  7. Current challenges facing the assessment of the allergenic capacity of food allergens in animal models.

    PubMed

    Bøgh, Katrine Lindholm; van Bilsen, Jolanda; Głogowski, Robert; López-Expósito, Iván; Bouchaud, Grégory; Blanchard, Carine; Bodinier, Marie; Smit, Joost; Pieters, Raymond; Bastiaan-Net, Shanna; de Wit, Nicole; Untersmayr, Eva; Adel-Patient, Karine; Knippels, Leon; Epstein, Michelle M; Noti, Mario; Nygaard, Unni Cecilie; Kimber, Ian; Verhoeckx, Kitty; O'Mahony, Liam

    2016-01-01

    Food allergy is a major health problem of increasing concern. The insufficiency of protein sources for human nutrition in a world with a growing population is also a significant problem. The introduction of new protein sources into the diet, such as newly developed innovative foods or foods produced using new technologies and production processes, insects, algae, duckweed, or agricultural products from third countries, creates the opportunity for development of new food allergies, and this in turn has driven the need to develop test methods capable of characterizing the allergenic potential of novel food proteins. There is no doubt that robust and reliable animal models for the identification and characterization of food allergens would be valuable tools for safety assessment. However, although various animal models have been proposed for this purpose, to date, none have been formally validated as predictive and none are currently suitable to test the allergenic potential of new foods. Here, the design of various animal models are reviewed, including among others considerations of species and strain, diet, route of administration, dose and formulation of the test protein, relevant controls and endpoints measured. PMID:27313841

  8. Removing peanut allergens by tannic acid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tannic acid (TA) is known to bind and form insoluble complexes with proteins, including peanut allergens; however, whether such complexes would dissociate and release the allergens at pH 2 and 8 (i.e., gastric and intestinal pH) is not clear. Release of the allergens in the gut could lead to absorpt...

  9. Grass pollen allergens globally: the contribution of subtropical grasses to burden of allergic respiratory diseases.

    PubMed

    Davies, J M

    2014-06-01

    Grass pollens of the temperate (Pooideae) subfamily and subtropical subfamilies of grasses are major aeroallergen sources worldwide. The subtropical Chloridoideae (e.g. Cynodon dactylon; Bermuda grass) and Panicoideae (e.g. Paspalum notatum; Bahia grass) species are abundant in parts of Africa, India, Asia, Australia and the Americas, where a large and increasing proportion of the world's population abide. These grasses are phylogenetically and ecologically distinct from temperate grasses. With the advent of global warming, it is conceivable that the geographic distribution of subtropical grasses and the contribution of their pollen to the burden of allergic rhinitis and asthma will increase. This review aims to provide a comprehensive synthesis of the current global knowledge of (i) regional variation in allergic sensitivity to subtropical grass pollens, (ii) molecular allergenic components of subtropical grass pollens and (iii) allergic responses to subtropical grass pollen allergens in relevant populations. Patients from subtropical regions of the world show higher allergic sensitivity to grass pollens of Chloridoideae and Panicoideae grasses, than to temperate grass pollens. The group 1 allergens are amongst the allergen components of subtropical grass pollens, but the group 5 allergens, by which temperate grass pollen extracts are standardized for allergen content, appear to be absent from both subfamilies of subtropical grasses. Whilst there are shared allergenic components and antigenic determinants, there are additional clinically relevant subfamily-specific differences, at T- and B-cell levels, between pollen allergens of subtropical and temperate grasses. Differential immune recognition of subtropical grass pollens is likely to impact upon the efficacy of allergen immunotherapy of patients who are primarily sensitized to subtropical grass pollens. The literature reviewed herein highlights the clinical need to standardize allergen preparations for both

  10. Importance of indoor allergens in the induction of allergy and elicitation of allergic disease.

    PubMed

    Custovic, A; Simpson, A; Woodcock, A

    1998-01-01

    During the last few decades, many countries have experienced an increase in the prevalence and severity of asthma. Over the same period, the population in the developed world has retreated indoors, and homes have become better insulated and more energy efficient, resulting in a warm and humid environment with low ventilation rates, ideally suited to house-dust-mite population growth throughout the year. Increasing exposure and increasing sensitivity to indoor allergens represent a progressively higher risk factor for the development of asthma. The development of sensitivity to indoor allergens and the symptoms and severity of asthma in later childhood are directly related to the exposure to allergens in infancy. It was relatively straightforward to demonstrate a quantitative linear dose-response relationship between exposure to house-dust-mite allergens and subsequent sensitization. However, showing the same for exposure and asthma severity has been more difficult, as the relationship between exposure and asthma symptoms in already sensitized individuals is much more complex than in the case of exposure and sensitization. Nevertheless, sensitized individuals are likely to have more severe asthma if exposed to high allergen levels than if their level of exposure is low. Sensitization to house-dust mites is a major independent risk factor for asthma in all areas where climate is conducive to mite population growth. The relevance of allergens other that mite is not consistent between different areas, and depends on the climate, habits, and socio-economic features of the local community. It would appear that presence of mite allergens in homes "overshadows" other allergens (e.g., cat, dog, or cockroach) as a risk factor for sensitization and subsequent development of allergic disease. It is possible that this is the consequence of the difference in inherent potency between allergen sources, and the question of why mite allergens are so potent in inducing sensitization

  11. Pectate Lyase Pollen Allergens: Sensitization Profiles and Cross-Reactivity Pattern

    PubMed Central

    Bernardi, Maria Livia; Gadermaier, Gabriele; Weiss, Richard; Ebner, Christof; Yokoi, Hidenori; Takai, Toshiro; Didierlaurent, Alain; Rafaiani, Chiara; Briza, Peter; Mari, Adriano; Behrendt, Heidrun; Wallner, Michael; Ferreira, Fátima

    2015-01-01

    Background Pollen released by allergenic members of the botanically unrelated families of Asteraceae and Cupressaceae represent potent elicitors of respiratory allergies in regions where these plants are present. As main allergen sources the Asteraceae species ragweed and mugwort, as well as the Cupressaceae species, cypress, mountain cedar, and Japanese cedar have been identified. The major allergens of all species belong to the pectate lyase enzyme family. Thus, we thought to investigate cross-reactivity pattern as well as sensitization capacities of pectate lyase pollen allergens in cohorts from distinct geographic regions. Methods The clinically relevant pectate lyase pollen allergens Amb a 1, Art v 6, Cup a 1, Jun a 1, and Cry j 1 were purified from aqueous pollen extracts, and patients´ sensitization pattern of cohorts from Austria, Canada, Italy, and Japan were determined by IgE ELISA and cross-inhibition experiments. Moreover, we performed microarray experiments and established a mouse model of sensitization. Results In ELISA and ELISA inhibition experiments specific sensitization pattern were discovered for each geographic region, which reflected the natural allergen exposure of the patients. We found significant cross-reactivity within Asteraceae and Cupressaceae pectate lyase pollen allergens, which was however limited between the orders. Animal experiments showed that immunization with Asteraceae allergens mainly induced antibodies reactive within the order, the same was observed for the Cupressaceae allergens. Cross-reactivity between orders was minimal. Moreover, Amb a 1, Art v 6, and Cry j 1 showed in general higher immunogenicity. Conclusion We could cluster pectate lyase allergens in four categories, Amb a 1, Art v 6, Cup a 1/Jun a 1, and Cry j 1, respectively, at which each category has the potential to sensitize predisposed individuals. The sensitization pattern of different cohorts correlated with pollen exposure, which should be considered for

  12. Emergent and unusual allergens in cosmetics.

    PubMed

    Pascoe, David; Moreau, Linda; Sasseville, Denis

    2010-01-01

    Allergic contact dermatitis from cosmetics is a common problem that is occasionally caused by new or rare allergens. When a patient has a positive patch test to a cosmetic product but to none of the common or commercially available allergens, it is important to further patch-test this patient to the ingredients of the product. Thorough testing with the breakdown of ingredients, usually obtained through cooperation with the manufacturer, often allows identification of the culprit allergen in the cosmetic product. In this article, we discuss emerging or rare allergens discovered by this method, including nail lacquer and lipstick allergens, copolymers, shellac, alkyl glucosides, glycols, protein derivatives, idebenone, and octocrylene. PMID:20487655

  13. 18 Proteomic and Immunological Characterization of Ragweed Allergens

    PubMed Central

    Augustin, Steffen; Stock, Marion; Cromwell, Oliver; Nandy, Andreas; Reese, Gerald

    2012-01-01

    Background The prevalence of sensitization to ragweed has risen in North America and across Europe. Although the pectate lyase Amb a 1, the major allergen of ragweed, was identified as long ago as the 1960s, little is known about the allergenicity of the 5 Amb a 1 isoallergens and other allergens present in ragweed pollen. Ragweed extracts and purified Amb a 1 isoallergens have now been characterized for their allergenic potential to determine whether a single Amb a 1 isoallergen, several isoallergens or a combination with other allergens should be included in a recombinant SIT vaccine. Methods Extracts from North American short ragweed (Ambrosia artemisiifolia) pollen were investigated by mass spectrometry (MS), 2D-PAGE and immunoblotting. Furthermore, Amb a 1 isoallergens were purified and IgE reactivity determined by immunoblotting and IgE inhibition. Results 2D-PAGE and MS of ragweed extract proved the presence of all 5 known Amb a 1 isoallergens, of which Amb a 1.01 represents the dominant form. Additionally all other ragweed allergens known by sequence (Amb a 3, Amb a 4, Amb a 5, Amb a 6, Amb a 8, Amb a 9, Amb a 10) were identified. The highest IgE reactivity by immunoblotting was observed for Amb a 1.01 followed by Amb a 1.03; other Amb a 1 isoallergens as well as other detected ragweed allergens showed only weak IgE reactivity. All isoallergens with the exception of Amb a 1.04, which is only of low abundance in ragweed extract, were purified. Similar to the immunoblot analysis with crude extract, the purified isoallergens Amb a 1.02 and Amb a 1.05 showed weak IgE binding, whereas Amb a 1.01 and Amb a 1.03 had high IgE reactivity. First IgE inhibition experiments suggest that Amb a 1.01 contains all relevant IgE epitopes. Conclusions Amb a 1.01 is the most abundant Amb a 1 isoallergen, and presumably the most important ragweed allergen. However, a larger panel of ragweed-allergic subjects has to be analyzed with regard to IgE and T cell reactivities, to be

  14. Cationic inhibitors of serine proteinases from buckwheat seeds.

    PubMed

    Tsybina, T A; Dunaevsky, Y E; Musolyamov, A K; Egorov, T A; Belozersky, M A

    2001-09-01

    Preparations of low molecular weight protein inhibitors of serine proteinases have been obtained from buckwheat (Fagopyrum esculentum) seeds by chromatography of seed extract on trypsin-Sepharose 4B, Mono-Q, and Mono-S ion exchangers (FPLC regime). Their molecular masses, determined by mass spectrometry, were 5203 (BWI-1c), 5347 (BWI-2c), 7760 (BWI-3c), and 6031 daltons (BWI-4c). All of the inhibitors possess high pH- and thermal stability in the pH range 2-12. In addition to trypsin, BWI-3c and BWI-4c inhibited chymotrypsin and subtilisin-like bacterial proteases. The N-terminal sequences of all of the inhibitors were determined: BWI-1c (23 residues), BWI-2c (33 residues), BWI-3c (18 residues), and BWI-4c (20 residues). In their physicochemical properties and N-terminal amino acid sequences, the buckwheat seed trypsin inhibitors BWI-3c and BWI-4c appear to belong to potato proteinase inhibitor I family. PMID:11703172

  15. Mite, cat, and cockroach exposure, allergen sensitisation, and asthma in children: a case-control study of three schools

    PubMed Central

    Sporik, R.; Squillace, S.; Ingram, J. M.; Rakes, G.; Honsinger, R.; Platts-Mills, T.

    1999-01-01

    BACKGROUND—The amount of allergen necessary to sensitise genetically "at risk" children is unclear. The relation between allergen exposure and asthma is also uncertain.
METHODS—To ensure a wide range of allergen exposures the data from case-control studies of asthma in children aged 12-14 years attending three schools in Los Alamos, New Mexico and Central Virginia were combined. Skin prick tests to indoor and outdoor allergens and bronchial hyperreactivity to histamine were assessed in children with and without symptoms of asthma. The concentration of mite, cat, and cockroach allergens in dust from the children's homes was used as a marker of exposure.
RESULTS—Three hundred and thirty two children (157 with asthmatic symptoms and 175 controls) were investigated. One hundred and eighty three were classified as atopic on the basis of allergen skin prick tests and 68 as asthmatic (symptoms plus bronchial responsiveness). The prevalence and degree of sensitisation to mite and cockroach, but not cat, was strongly associated in atopic children with increasing domestic concentrations of these allergens. Asthma was strongly associated with sensitisation to indoor allergens (p<10-6) and weakly to outdoor allergens (p = 0.026). There was an association between current asthma and the concentration of mite allergen amongst atopic children (p = 0.008) but not amongst those who were specifically mite sensitised (p = 0.16).
CONCLUSIONS—The domestic reservoir concentration of mite and cockroach, but not cat, allergen was closely related to the prevalence of sensitisation in atopic children. However, the prevalence of current asthma had a limited relationship to these allergen measurements, suggesting that other factors play a major part in determining which allergic individuals develop asthma.

 PMID:10413718

  16. Identification of Aspergillus (A. flavus and A. niger) Allergens and Heterogeneity of Allergic Patients' IgE Response.

    PubMed

    Vermani, Maansi; Vijayan, Vannan Kandi; Agarwal, Mahendra Kumar

    2015-08-01

    Aspergillus species (A. flavus and A. niger) are important sources of inhalant allergens. Current diagnostic modalities employ crude Aspergillus extracts which only indicate the source to which the patient has been sensitized, without identifying the number and type of allergens in crude extracts. We report a study on the identification of major and minor allergens of the two common airborne Aspergillus species and heterogeneity of patients' IgE response to them. Skin prick tests were performed on 300 patients of bronchial asthma and/or allergic rhinitis and 20 healthy volunteers. Allergen specific IgE in patients' sera was estimated by enzyme allergosorbent test (EAST). Immunoblots were performed to identify major/minor allergens of Aspergillus extracts and to study heterogeneity of patients'IgE response to them. Positive cutaneous responses were observed in 17% and 14.7% of patients with A. flavus and A. niger extracts, respectively. Corresponding EAST positivity was 69.2% and 68.7%. In immunoblots, 5 allergenic proteins were identified in A. niger extract, major allergens being 49, 55.4 and 81.5 kDa. Twelve proteins bound patients' IgE in A. flavus extract, three being major allergens (13.3, 34 and 37 kDa). The position and slopes of EAST binding and inhibition curves obtained with individual sera varied from patient to patient. The number and molecular weight of IgE-binding proteins in both the Aspergillus extracts varied among patients. These results gave evidence of heterogeneity of patients' IgE response to major/minor Aspergillus allergens. This approach will be helpful to identify disease eliciting molecules in the individual patients (component resolved diagnosis) and may improve allergen-specific immunotherapy.

  17. Responsiveness of basophil granulocytes of horses suffering from chronic obstructive pulmonary disease to various allergens.

    PubMed

    Dirscherl, P; Grabner, A; Buschmann, H

    1993-10-01

    As basophils are the major effector cells of allergic reactions, confirmation of the allergic etiology of chronic obstructive pulmonary disease (COPD) was sought by the demonstration of a specific in vitro response of equine basophilic blood cells to some potential allergens (Aspergillus, Cladosporidium, Mucor, Penicillium, extracts of dust particles of hay and straw). The allergen induced degranulation of basophils and the histamine and protease release from basophils during incubation with the allergens were tested. By evaluating the results obtained from 14 COPD horses and eight controls it could be shown that the sensitivity of the basophils of affected horses was increased, particularly against the allergen extract of Mucor mucedo and Mucor spinosus. Further a greater percentage of COPD horses reacted positively with the Mucor allergen extract. The mitogenic stimulation of lymphocytes by PHA and by the allergen extracts used gave comparable results in affected and control horses. Thus the in vitro stimulation of basophils may be an easily to perform testing device for the identification of potential allergens involved in the pathogenesis of equine COPD.

  18. Is ragweed pollen allergenicity governed by environmental conditions during plant growth and flowering?

    PubMed Central

    Ghiani, Alessandra; Ciappetta, Silvia; Gentili, Rodolfo; Asero, Riccardo; Citterio, Sandra

    2016-01-01

    Pollen allergenicity is one of the main factors influencing the prevalence and/or severity of allergic diseases. However, how genotype and environment contribute to ragweed pollen allergenicity has still to be established. To throw some light on the factors governing allergenicity, in this work 180 ragweed plants from three Regions (Canada, France, Italy) were grown in both controlled (constant) and standard environmental conditions (seasonal changes in temperature, relative humidity and light). Pollen from single plants was characterized for its allergenic potency and for the underlying regulation mechanisms by studying the qualitative and quantitative variations of the main isoforms of the major ragweed allergen Amb a 1. Results showed a statistically higher variability in allergenicity of pollen from standard conditions than from controlled conditions growing plants. This variability was due to differences among single plants, regardless of their origin, and was not ascribed to differences in the expression and IgE reactivity of individual Amb a 1 isoforms but rather to quantitative differences involving all the studied isoforms. It suggests that the allergenic potency of ragweed pollen and thus the severity of ragweed pollinosis mainly depends on environmental conditions during plant growth and flowering, which regulate the total Amb a 1 content. PMID:27457754

  19. Effects of an acaricide on mite allergen levels in the homes of asthmatic children.

    PubMed

    Chew, F T; Goh, D Y; Lee, B W

    1996-10-01

    Previous study by the current authors has shown that treating homes with D'Allergen, an acaricidal agent, can reduce bronchial hyper-reactivity in asthmatic children with house dust mite allergy. In the present study, the effects of a single D'Allergen treatment on the levels of major dust mite allergens, Der p I and Der f I was evaluated, and the duration of its effectiveness in the environment determined. Twenty randomly selected homes were treated with the acaricide and ten remained untreated. Dust samples were collected from mattresses, upholstered sofas and carpets of these homes before and 1, 2 and 4 months after treatment. The samples were then assayed for Der p I and Der f I allergens using a sandwich enzyme immunoassay. The results showed that D'Allergen was effective in reducing dust mite allergen levels in all three niches by 1.5-22.3 times below baseline values. This effect, however, was only present for 2 months, and the dust mite allergen levels increased to those of the baseline by the fourth month after treatment. These results indicated that repeated applications of the acaricide were required at 2-3 monthly intervals to obtain optimal effectiveness.

  20. Safety evaluation of standardized allergen extract of Japanese cedar pollen for sublingual immunotherapy.

    PubMed

    Mitobe, Yuko; Yokomoto, Yasuki; Ohashi-Doi, Katsuyo

    2015-04-01

    Japanese cedar (JC) pollinosis is caused by Japanese cedar pollen (JCP) and most common seasonal allergic disease in Japan. Subcutaneous immunotherapy (SCIT) with allergen extract of JCP (JCP-allergen extract) is well established for JC pollinosis treatment with improvement of symptoms. However, major drawbacks for SCIT are repeated painful injections, frequent hospital visits and anaphylactic risk. Currently, sublingual immunotherapy (SLIT) has received much attention as an advanced alternative application with lower incidence of systemic reactions because the liquid or tablet form of allergen is placed under the tongue. The aim of this study was safety evaluation of standardized JCP-allergen extract currently developed for SLIT in JC pollinosis. JCP-allergen extract showed no potential genotoxicity. No systemic effects were observed in rats administered JCP-allergen extract orally for 26 weeks followed by 4-week recovery period. Mild local reactions such as hyperplasia and increased globule leukocytes resulting from vehicle (glycerin)-induced irritation were observed in stomach. No-observed-adverse-effect level was greater than 10,000 JAU/kg/day for systemic toxicity, equivalent to 300-fold the human dose. No local irritation was found in rabbits oral mucosae by 7-day sublingual administration. These results demonstrate the safe profile of standardized JCP-allergen extract, suggesting it is suitable for SLIT in JC pollinosis.

  1. Is ragweed pollen allergenicity governed by environmental conditions during plant growth and flowering?

    NASA Astrophysics Data System (ADS)

    Ghiani, Alessandra; Ciappetta, Silvia; Gentili, Rodolfo; Asero, Riccardo; Citterio, Sandra

    2016-07-01

    Pollen allergenicity is one of the main factors influencing the prevalence and/or severity of allergic diseases. However, how genotype and environment contribute to ragweed pollen allergenicity has still to be established. To throw some light on the factors governing allergenicity, in this work 180 ragweed plants from three Regions (Canada, France, Italy) were grown in both controlled (constant) and standard environmental conditions (seasonal changes in temperature, relative humidity and light). Pollen from single plants was characterized for its allergenic potency and for the underlying regulation mechanisms by studying the qualitative and quantitative variations of the main isoforms of the major ragweed allergen Amb a 1. Results showed a statistically higher variability in allergenicity of pollen from standard conditions than from controlled conditions growing plants. This variability was due to differences among single plants, regardless of their origin, and was not ascribed to differences in the expression and IgE reactivity of individual Amb a 1 isoforms but rather to quantitative differences involving all the studied isoforms. It suggests that the allergenic potency of ragweed pollen and thus the severity of ragweed pollinosis mainly depends on environmental conditions during plant growth and flowering, which regulate the total Amb a 1 content.

  2. Flavonoids, Antioxidant Activity and Aroma Compounds Analysis from Different Kinds of Tartary Buckwheat Tea

    PubMed Central

    Peng, L. X.; Zou, L.; Wang, J. B.; Zhao, J. L.; Xiang, D. B.; Zhao, G.

    2015-01-01

    The rutin, quercetin concentrations, antioxidant activity, and aroma compounds in different commercial tartary buckwheat tea were analyzed in our study. Results revealed that the materials and the processing protocol affected the chemical composition and activity of tartary buckwheat tea. Rutin and quercetin concentrations, antioxidant activity were significantly different in various kinds of tartary buckwheat tea, where the whole bran tea and the whole plant tea had the lower rutin, but higher quercetin concentrations and higher antioxidant activity. The whole embryo tea had the converse results. There was strong correlation between quercetin concentration and antioxidant activity (r>0.98, P<0.05). Meanwhile, Twenty eight different aroma compounds in tartary buckwheat tea were identified by gas chromatography-mass spectrometry. Those compounds were mainly composed of pyrazine, aldehydes, fatty acids and ketones. The main type of aroma compounds in different tartary buckwheat tea were similar, but their relative contents were different. The implications to the quality control of buckwheat tea were extensively discussed. PMID:26997692

  3. Hypolipidemic and antioxidant effects of buckwheat leaf and flower mixture in hyperlipidemic rats.

    PubMed

    Ðurendić-Brenesel, Maja; Popović, Tamara; Pilija, Vladimir; Arsić, Aleksandra; Milić, Miljan; Kojić, Danijela; Jojić, Nikola; Milić, Nataša

    2013-05-01

    As a source of biologically active compounds, buckwheat has beneficial effects in nutrition due to its high content of flavonoids, particularly rutin. Aim of our study was to examine effects of buckwheat on plasma lipid status and phospholipids fatty acids composition, histological and parameters of oxidative stress in Wistar rats fed a high-fat diet. This study showed that buckwheat leaf and flower (BLF) mixture supplementation significantly reduce weight gain, plasma lipid concentrations and atherogenic index in rats fed a high-fat diet. Treatment of the high-fat group of animals with buckwheat significantly increased percentage of n-6 fatty acids as well as eicosapentaenoic acid (EPA) and decreased percentage of saturated fatty acids (SFA) and oleic acid. Buckwheat antioxidant effects diminished negative influence of high-fat diet in hyperlipidemic rats, while pathohistological analysis of liver confirmed changes after high-fat consumption. Our results showed hypolipidemic, antiatherogenic and antioxidative features of buckwheat leaf and flower mixture, and these parts of the plant with the highest rutin content could be beneficial in prevention and curing of hyperlipidemia.

  4. Allergenicity assessment strategy for novel food proteins and protein sources.

    PubMed

    Verhoeckx, Kitty; Broekman, Henrike; Knulst, André; Houben, Geert

    2016-08-01

    To solve the future food insecurity problem, alternative and sustainable protein sources (e.g. insects, rapeseed, fava bean and algae) are now being explored for the production of food and feed. To approve these novel protein sources for future food a comprehensive risk assessment is needed according to the European food legislation. Allergenicity risk assessment might pose some major difficulties, since detailed guidance on how to assess the allergenic potential of novel foods is not available. At present, the approach relies mostly on the guidance of allergenicity assessment for genetically modified (GM) plant foods. The most recent one was proposed by EFSA (2010 and 2011); "weight-of-evidence approach". However this guidance is difficult to interpret, not completely applicable or validated for novel foods and therefore needs some adjustments. In this paper we propose a conceptual strategy which is based on the "weight-of-evidence approach" for food derived from GM plants and other strategies that were previously published in the literature. This strategy will give more guidance on how to assess the allergenicity of novel food proteins and protein sources.

  5. Allergenicity assessment strategy for novel food proteins and protein sources.

    PubMed

    Verhoeckx, Kitty; Broekman, Henrike; Knulst, André; Houben, Geert

    2016-08-01

    To solve the future food insecurity problem, alternative and sustainable protein sources (e.g. insects, rapeseed, fava bean and algae) are now being explored for the production of food and feed. To approve these novel protein sources for future food a comprehensive risk assessment is needed according to the European food legislation. Allergenicity risk assessment might pose some major difficulties, since detailed guidance on how to assess the allergenic potential of novel foods is not available. At present, the approach relies mostly on the guidance of allergenicity assessment for genetically modified (GM) plant foods. The most recent one was proposed by EFSA (2010 and 2011); "weight-of-evidence approach". However this guidance is difficult to interpret, not completely applicable or validated for novel foods and therefore needs some adjustments. In this paper we propose a conceptual strategy which is based on the "weight-of-evidence approach" for food derived from GM plants and other strategies that were previously published in the literature. This strategy will give more guidance on how to assess the allergenicity of novel food proteins and protein sources. PMID:27012375

  6. Review of statutory and voluntary labelling of food allergens.

    PubMed

    Boden, Mark; Dadswell, Ruth; Hattersley, Sue

    2005-11-01

    Food allergy represents an increasingly important health problem, with prevalence in Western Europe continuing to rise. While some reactions are mild, others can include life-threatening anaphylactic shock. It is estimated that food allergies affect 1-2% of the adult population and < or =8% of children. Relatively few foods are to blame for a large majority of allergic reactions to food in the UK, with most reactions being to milk, eggs, peanuts (Arachis hypogea), nuts, fish, shellfish, soyabean, sesame (Sesamum indicum L.) and wheat. There is currently no cure for food allergy and the few available treatments are focused on relieving the specific symptoms. Consumers with food allergies and food intolerances rely on food labelling to enable them to make informed choices about the foods they eat. Whilst there have recently been important advances in the labelling of food allergens, these advances relate only to requirements for the labelling of the deliberate use of specified food allergens in foods sold pre-packed. In other areas the development of guidance for food manufacturers and retailers on how to assess the risks of possible allergen cross-contamination during food production and manufacture, and then to determine appropriate advisory labelling, is well advanced. Work to address the issue of how to provide appropriate allergen information for foods sold loose, or in catering establishments, is also in progress.

  7. Dermatophagoides farinae allergen levels from two different sources within the same home: evaluation of two different collection techniques.

    PubMed

    Twiggs, J T; Gray, R L; DeJongh, S; Marx, J J

    1991-05-01

    The allergens produced by the dust mites of the Dermatophagoides genus are now recognized as one of the major allergens in house dust. Quantification of those allergens helps to evaluate the effectiveness of avoidance measures and specific treatments aimed at the reduction of the mites and their allergens. The present study was undertaken to compare the level of D. farinae allergen collected by a standard vacuum cleaner with a disposable bag and the level of allergen collected by a hand-held vacuum equipped with a special filter. Samples were obtained from the main living area of 18 homes of patients allergic to D. farinae. Using a hand-held vacuum especially equipped with a removable micropore filter, a 2-m2 area of carpet was vacuumed for two minutes and an identical sample was collected from the major upholstered piece of furniture in the room. A third sample was obtained with an upright vacuum fitted with a clean disposable bag. The dust collected was extracted in phosphate buffered saline and resultant extracts were assayed for D. farinae allergen content by microtiter plate RAST inhibition assay. Results from these assays showed a large interhome variability regardless of sampling technique, but intrahome sampling showed remarkable consistency. A linear regression analysis demonstrated all of the comparisons correlated at a probability level of P less than .05. We conclude that dust mite allergen levels collected in a disposable bag vacuum cleaner are reliable indicators of the level of house dust mites present in the homes of patients.

  8. House Dust Mite Allergy in Korea: The Most Important Inhalant Allergen in Current and Future

    PubMed Central

    Jeong, Kyoung Yong; Park, Jung-Won

    2012-01-01

    The house-dust mite (HDM), commonly found in human dwellings, is an important source of inhalant and contact allergens. In this report, the importance of HDM allergy in Korea and the characteristics of allergens from dust mite are reviewed with an emphasis on investigations performed in Korea. In Korea, Dermatophagoides farinae is the dominant species of HDM, followed by D. pteronyssinus. Tyrophagus putrescentiae is also found in Korea, but its role in respiratory allergic disease in Korea is controversial. The relatively low densities of mite populations and concentrations of mite major allergens in dust samples from Korean homes, compared to westernized countries, are thought to reflect not only different climatic conditions, but also cultural differences, such as the use of 'ondol' under-floor heating systems in Korean houses. HDM are found in more than 90% of Korean houses, and the level of exposure to HDM is clinically significant. About 40%-60% of Korean patients suffering from respiratory allergies, and more than 40% of patients suffering from atopic dermatitis, are sensitized to HDM. Mite allergens can be summarized according to their inherent auto-adjuvant activities and/or their binding affinities to the adjuvant-like substances: proteolytic enzymes, lipid binding proteins, chitin binding proteins, and allergens not associated with adjuvant-like activity. In general, allergens with a strong adjuvant-like activity or adjuvant-binding activity elicit potent IgE reactivity. In Korea, Der f 2 is the most potent allergen, followed by Der f 1. Immune responses are modulated by the properties of the allergen itself and by the adjuvant-like substances that are concomitantly administered with the antigens. Characterization of allergenic molecules and elucidation of mechanisms by which adjuvant-like molecules modulate allergic reactions, not only in Korea but also worldwide, will provide valuable information on allergic diseases, and are necessary for the

  9. Allergen Microarray Indicates Pooideae Sensitization in Brazilian Grass Pollen Allergic Patients

    PubMed Central

    Moreira, Priscila Ferreira de Sousa; Gangl, Katharina; Vieira, Francisco de Assis Machado; Ynoue, Leandro Hideki; Linhart, Birgit; Flicker, Sabine; Fiebig, Helmut; Swoboda, Ines; Focke-Tejkl, Margarete; Taketomi, Ernesto Akio; Valenta, Rudolf; Niederberger, Verena

    2015-01-01

    Background Grass pollen, in particular from Lolium multiflorum is a major allergen source in temperate climate zones of Southern Brazil. The IgE sensitization profile of Brazilian grass pollen allergic patients to individual allergen molecules has not been analyzed yet. Objective To analyze the IgE sensitization profile of a Brazilian grass pollen allergic population using individual allergen molecules. Methods We analyzed sera from 78 grass pollen allergic patients for the presence of IgE antibodies specific for 103 purified micro-arrayed natural and recombinant allergens by chip technology. IgE-ELISA inhibition experiments with Lolium multiflorum, Phleum pratense extracts and a recombinant fusion protein consisting of Phl p 1, Phl p 2, Phl p 5 and Phl p 6 were performed to investigate cross-reactivities. Results Within the Brazilian grass pollen allergic patients, the most frequently recognized allergens were Phl p 1 (95%), Phl p 5 (82%), Phl p 2 (76%) followed by Phl p 4 (64%), Phl p 6 (45%), Phl p 11 (18%) and Phl p 12 (18%). Most patients were sensitized only to grass pollen allergens but not to allergens from other sources. A high degree of IgE cross-reactivity between Phleum pratense, Lolium multiflorum and the recombinant timothy grass fusion protein was found. Conclusions Component-resolved analysis of sera from Brazilian grass pollen allergic patients reveals an IgE recognition profile compatible with a typical Pooideae sensitization. The high degree of cross-reactivity between Phleum pratense and Lolium multiflorum allergens suggests that diagnosis and immunotherapy can be achieved with timothy grass pollen allergens in the studied population. PMID:26067084

  10. A microfluidic biosensor using graphene oxide and aptamer-functionalized quantum dots for peanut allergen detection.

    PubMed

    Weng, Xuan; Neethirajan, Suresh

    2016-11-15

    The increasing prevalence of food allergies and the intake of packing foods in the past two decades urge the need for more rapid, accurate, and sensitive assays to detect potential allergens in food in order to control the allergen content. Most of the commercial analytical tools for allergen detection rely on immunoassays such as ELISA. As far as disadvantages, ELISA can be time-consuming and expensive. Biosensors appear as a suitable alternative for the detection of allergens because they are rapid, highly sensitive, selective, less expensive, environmentally friendly, and easy to handle. In this study, we developed a microfluidic system integrated with a quantum dots (Qdots) aptamer functionalized graphene oxide (GO) nano-biosensor for simple, rapid, and sensitive food allergen detection. The biosensor utilized Qdots-aptamer-GO complexes as probes to undergo conformational change upon interaction with the food allergens, resulting in fluorescence changes due to the fluorescence quenching and recovering properties of GO by adsorption and desorption of aptamer-conjugated Qdots. This one-step 'turn on' homogenous assay in a ready-to-use microfluidic chip took ~10min to achieve a quantitative detection of Ara h 1, one of the major allergens appearing in peanuts. The results suggested this system had remarkable sensitivity and selectivity. The integration of a microfluidics platform in a homemade miniaturized optical analyzer provides a promising way for the rapid, cost-effective, and accurate on-site determination of food allergens. This biosensor can also be extended to the detection of other food allergens with a selection of corresponding aptamers. PMID:27240012

  11. Recombinant allergen profiles and health-related quality of life in seasonal allergic rhinitis.

    PubMed

    Canis, Martin; Gröger, Moritz; Becker, Sven; Klemens, Christine; Kramer, Matthias F

    2010-01-01

    We evaluated birch- and timothy-allergic patients for allergen-specific IgE profiles and health-related quality of life (HRQL). We examined 395 patients with seasonal allergic rhinitis against birch or timothy pollen. Sera were analyzed for IgE reactivity to recombinant allergens (Bet v 1, Bet v 2, Bet v 4; Phl p 1/p 5b, Phl p 7, and Phl p 12) and to native pollen extracts (t3 and g6). Subgroup-specific analyses were performed. Patients with cosensitization against structurally unrelated allergens were termed polysensitized. Patients allergic solely to birch or timothy were labeled monosensitized. HRQL was evaluated using an established questionnaire. In patients polysensitized against native birch pollen (n = 233) the prevalence of allergens was 86% for Bet v 1, 15% for Bet v 2, and 5% for Bet v 4. Similar for timothy (n = 256), the prevalence of allergens was 87% rPhl p 1/p 5b, 5% for rPhl p 7, and 14% for rPhl p 12. Values for birch-monosensitized patients (n = 42) were Bet v 1, 100%, and Bet v 2 and Bet v 4, 0%. Values for timothy-monosensitized patients (n = 35) were Phl p 1/p 5b, 100%; rPhl p 7, 0%; and rPhl p 12, 3%. No difference in HRQL existed between patients sensitized solely against major versus minor allergens in birch-allergic patients. Polysensitized cohorts showed sensitization profiles comparable with published data. Monosensitized patients showed IgE against major allergens in 100% of cases. Patients sensitized solely against major or minor allergens showed no differences in HRQL.

  12. Food allergens: molecular and immunological aspects, allergen databases and cross-reactivity.

    PubMed

    Lorenz, Anne-Regine; Scheurer, Stephan; Vieths, Stefan

    2015-01-01

    The currently known food allergens are assigned to a relatively small number of protein families. Food allergens grouped into protein families share common functional and structural features that can be attributed to the allergenic potency and potential cross-reactivity of certain proteins. Molecular data, in terms of structural information, biochemical characteristics and clinical relevance for each known allergen, including isoforms and variants, are mainly compiled into four open-access databases. Allergens are designated according to defined criteria by the World Health Organization and the International Union of Immunological Societies Allergen Nomenclature Sub-committee. Food allergies are caused by primary sensitisation to the disease-eliciting food allergens (class I food allergen), or they can be elicited as a consequence of a primary sensitisation to inhalant allergens and subsequent IgE cross-reaction to homologous proteins in food (class II food allergens). Class I and class II allergens display different clinical significance in children and adults and are characterised by different molecular features. In line with this, high stability when exposed to gastrointestinal digestion and heat treatment is attributed to many class I food allergens that frequently induce severe reactions. The stability of a food allergen is determined by its molecular characteristics and can be influenced by structural (chemical) modifications due to thermal processing. Moreover, the immunogenicity and allergenicity of food allergens further depends on specific T cell and B cell epitopes. Although the T cell epitope pattern can be highly diverse for individual patients, several immuno-prominent T cell epitopes have been identified. Such conserved T cell epitopes and IgE cross-reactive B cell epitopes contribute to cross-reactivity between food allergens of the same family and to clinical cross-reactivity, similar to the birch pollen-food syndrome. PMID:26022861

  13. Detection of castor allergens in castor wax.

    PubMed

    Lehrer, S B; Karr, R M; Müller, D J; Salvaggio, J E

    1980-01-01

    The presence of castor bean allergens in castor wax products was determined by in vivo and in vitro analysis of castor wax extracts. Allergens were detected in one extract of castor wax by the PCA reaction in mice, the RAST inhibition reaction, and skin prick test in castor bean sensitive individuals. However, these allergens in the wax were of much lower potency than those in the bean, and were not detectable in a deodorant product utilizing castor wax.

  14. The direct peptide reactivity assay: selectivity of chemical respiratory allergens.

    PubMed

    Lalko, Jon F; Kimber, Ian; Gerberick, G Frank; Foertsch, Leslie M; Api, Anne Marie; Dearman, Rebecca J

    2012-10-01

    It is well known that some chemicals are capable of causing allergic diseases of the skin and respiratory tract. Commonly, though not exclusively, chemical allergens are associated with the selective development of skin or respiratory sensitization. The reason for this divergence is unclear, although it is hypothesized that the nature of interactions between the chemical hapten and proteins is influential. The direct peptide reactivity assay (DPRA) has been developed as a screen for the identification of skin-sensitizing chemicals, and here we describe the use of this method to explore whether differences exist between skin and respiratory allergens with respect to their peptide-binding properties. Known skin and respiratory sensitizers were reacted with synthetic peptides containing either lysine (Lys) or cysteine (Cys) for 24 h. The samples were analyzed by HPLC/UV, and the loss of peptide from the reaction mixture was expressed as the percent depletion compared with the control. The potential for preferential reactivity was evaluated by comparing the ratio of Lys to Cys depletion (Lys:Cys ratio). The results demonstrate that the majority of respiratory allergens are reactive in the DPRA, and that in contrast to most skin-sensitizing chemicals, preferentially react with the Lys peptide. These data suggest that skin and respiratory chemical allergens can result in different protein conjugates, which may in turn influence the quality of induced immune responses. Overall, these investigations reveal that the DPRA has considerable potential to be incorporated into tiered testing approaches for the identification and characterization of chemical respiratory allergens. PMID:22713598

  15. Recombinant expression systems for allergen vaccines.

    PubMed

    Singh, Mohan B; Bhalla, Prem L

    2006-01-01

    Allergen immunotherapy of future is likely to be based on allergy vaccines that contain engineered allergens modified to abolish or substantially reduce their IgE-binding activity in order to remove the risk of unwanted anaphylactic responses. The development of efficient systems for the production of recombinant allergens in sufficient quantities is requirement for establishing use of engineered allergens as components of allergy vaccines. This review outlines relative advantages and disadvantages of various heterologous systems for production of recombinant allergens. Microbial systems are most convenient and cost effective platforms for the production of recombinant allergens. However, lack of post-translational processing implies that some allergens have to be expressed in eukaryotic systems for proper folding and post-translational modifications such as glycosylation. Yeast systems can yield high levels of recombinant allergens but often are associated with hyper- glycosylation problems. Mammalian cell culture systems offer suitable post -translational modifications but are nearly hundred fold more expensive than microbial systems. The use of plants as bio-factories for production of recombinant allergens is emerging as a very attractive option as plants-based production system offer several advantages over other expression systems such as post translational processing of proteins, low production costs, scale up ability and enhanced safety due to absence of animal or human pathogens.

  16. Protein digestibility and relevance to allergenicity.

    PubMed

    Bannon, Gary; Fu, Tong-Jen; Kimber, Ian; Hinton, Dennis M

    2003-06-01

    In January 2001 a Joint Food and Agriculture Organization of the United Nations/World Health Organization Expert Consultation Committee on Allergenicity of Foods Derived from Biotechnology published a report outlining in detail an approach for assessing the allergenic potential of novel proteins. One component of this decision tree is a determination of whether the protein of interest is resistant to proteolytic digestion. Although these (Italic)in vitro(/Italic) methodologies have been useful, the correlation between resistance to proteolysis and allergenic activity is not absolute. Two views and highlights of supporting research regarding the relationship of resistance to digestion and allergenicity are presented in this article.

  17. Mesoscale atmospheric transport of ragweed pollen allergens from infected to uninfected areas

    NASA Astrophysics Data System (ADS)

    Grewling, Ł.; Bogawski, P.; Jenerowicz, D.; Czarnecka-Operacz, M.; Šikoparija, B.; Skjøth, C. A.; Smith, M.

    2016-10-01

    Allergenic ragweed ( Ambrosia spp.) pollen grains, after being released from anthers, can be dispersed by air masses far from their source. However, the action of air temperature, humidity and solar radiation on pollen grains in the atmosphere could impact on the ability of long distance transported (LDT) pollen to maintain allergenic potency. Here, we report that the major allergen of Ambrosia artemisiifolia pollen (Amb a 1) collected in ambient air during episodes of LDT still have immunoreactive properties. The amount of Amb a 1 found in LDT ragweed pollen grains was not constant and varied between episodes. In addition to allergens in pollen sized particles, we detected reactive Amb a 1 in subpollen sized respirable particles. These findings suggest that ragweed pollen grains have the potential to cause allergic reactions, not only in the heavily infested areas but, due to LDT episodes, also in the regions unaffected by ragweed populations.

  18. Mesoscale atmospheric transport of ragweed pollen allergens from infected to uninfected areas

    NASA Astrophysics Data System (ADS)

    Grewling, Ł.; Bogawski, P.; Jenerowicz, D.; Czarnecka-Operacz, M.; Šikoparija, B.; Skjøth, C. A.; Smith, M.

    2016-02-01

    Allergenic ragweed (Ambrosia spp.) pollen grains, after being released from anthers, can be dispersed by air masses far from their source. However, the action of air temperature, humidity and solar radiation on pollen grains in the atmosphere could impact on the ability of long distance transported (LDT) pollen to maintain allergenic potency. Here, we report that the major allergen of Ambrosia artemisiifolia pollen (Amb a 1) collected in ambient air during episodes of LDT still have immunoreactive properties. The amount of Amb a 1 found in LDT ragweed pollen grains was not constant and varied between episodes. In addition to allergens in pollen sized particles, we detected reactive Amb a 1 in subpollen sized respirable particles. These findings suggest that ragweed pollen grains have the potential to cause allergic reactions, not only in the heavily infested areas but, due to LDT episodes, also in the regions unaffected by ragweed populations.

  19. Can we explain the allergenicity of peanuts on the basis of the three-dimensional structure of its allergens and use the information to devise means of eliminating peanut allergy?

    PubMed

    Mikita, Cecilia P; Padlan, Eduardo A

    2012-11-01

    Helical bundles are found in all the known structures of peanut allergens. The peptide fragments, which survive gastrointestinal digestion of the allergens and are absorbed intact, are hypothesized to reassociate and form stable helical bundles in the circulation, which could elicit a specific IgE response resulting in peanut allergy. The hypothesis is supported by the finding of a diminished allergenicity of an isoform of the peanut allergen, Ara h 3, which has a major deletion. A very probable consequence of this deletion is the reduced tendency to form a stable helix bundle. The discovery of structurally disrupted isoforms of the other peanut allergens and the breeding of plants that contain only those isoforms could lead to the elimination of peanut allergy. PMID:22910770

  20. Buckwheat as a Functional Food and Its Effects on Health.

    PubMed

    Giménez-Bastida, Juan Antonio; Zieliński, Henryk

    2015-09-16

    Buckwheat (BW) is a gluten-free pseudocereal that belongs to the Polygonaceae family. BW grain is a highly nutritional food component that has been shown to provide a wide range of beneficial effects. Health benefits attributed to BW include plasma cholesterol level reduction, neuroprotection, anticancer, anti-inflammatory, antidiabetic effects, and improvement of hypertension conditions. In addition, BW has been reported to possess prebiotic and antioxidant activities. In vitro and animal studies suggest that BW's bioactive compounds, such as D-chiro-inositol (DCI), BW proteins (BWP), and BW flavonoids (mainly rutin and quercetin) may be partially responsible for the observed effects. The purpose of this paper is to review the recent research regarding the health benefits of BW, in vitro and in vivo, focusing on the specific role of its bioactive compounds and on the mechanisms by which these effects are exerted.

  1. Molecular and Immunological Characterization of the First Allergenic Lipocalin in Hamster

    PubMed Central

    Torres, José Alberto; de las Heras, Manuel; Maroto, Aroa Sanz; Vivanco, Fernando; Sastre, Joaquín; Pastor-Vargas, Carlos

    2014-01-01

    The most frequent pet allergy is to cat and dog, but in recent years, it has become increasingly popular to have other pets, and the risk of exposure to new allergens is more prevalent. The list of new pets includes hamsters, and one of the most popular hamsters is the Siberian hamster (Phodopus sungorus). The aim of this study was the characterization and cloning of the major allergen from this hamster. The study of its allergenicity and cross-reactivity could improve the specific diagnosis and treatment for hamster-allergic patients. Thirteen Siberian hamster-allergic patients were recruited at the outpatient clinic. Protein extracts were prepared from the hair, urine, and salivary glands of four hamster species (European, golden, Siberian, and Roborovski). IgE-binding proteins were detected by immunoblotting and identified by mass spectrometry. The recombinant protein was produced in Escherichia coli and then purified by metal chelate affinity chromatography. The allergenic properties of the recombinant protein were tested by ELISA and immunoblotting, and biological activity was tested according to capacity for basophil activation. Three IgE-binding proteins were identified in extracts obtained from Siberian hamster hair, urine, and salivary glands. All proteins corresponded to the same protein, which was identified as a lipocalin. This lipocalin had no cross-reactivity with common and golden hamsters. The recombinant allergen was cloned and purified, showing similar IgE reactivity in vitro to Siberian hamster protein extracts. Also, the recombinant allergen was capable of producing biological activation in vivo. The major Siberian hamster allergen was cloned, and allergenic properties were characterized, providing a new tool for specific diagnosis of allergy to Siberian hamster. PMID:24993820

  2. Tartary buckwheat (Fagopyrum tataricum Gaertn.) starch, a side product in functional food production, as a potential source of retrograded starch.

    PubMed

    Gao, Jinfeng; Kreft, Ivan; Chao, Guimei; Wang, Ying; Liu, Xiaojin; Wang, Li; Wang, Pengke; Gao, Xiaoli; Feng, Baili

    2016-01-01

    A starch rich fraction is a side product in Tartary buckwheat processing. This study investigated the fractions that are of technological and nutritional interest. Tartary buckwheat starch granules had a diameter of 3-14 μm, and presented a typical type "A" X-ray diffraction pattern. They contained nearly 39.0% amylose. The solubility of Tartary buckwheat starch was much lower at 70-90 °C (ranging within 9.9-10.4% at 90 °C) than that in maize (up to 49.3%) and potato (up to 85.0%) starch. The starch of one variety of Tartary buckwheat had significantly lower solubility at 70 °C and 80 °C than that of common buckwheat. The starch peak viscosity and breakdown were higher and pasting time was shorter in Tartary buckwheat than in that of the starch of common buckwheat. Tartary buckwheat starch had unique pasting and physicochemical properties, and is thereby capable of being exploited as a suitable raw material of retrograded starch in food processing.

  3. Functional characterization of steam jet-cooked buckwheat flour as a fat replacer in cake-baking

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fancy buckwheat flour was thermo-mechanically modified by steam jet-cooking and the resulting product was evaluated as a fat replacer for the use in cakes with reduced-fat content. Steam jet-cooking caused the integrity of buckwheat flour components to be disrupted, significantly changing the physi...

  4. Effect of environmental intervention on mouse allergen levels in homes of inner-city Boston children with asthma

    PubMed Central

    Cronin, Beth; Wood, Robert A.; Eggleston, Peyton A.; Shih, Mei-Chiung; Song, Leslie; Tachdjian, Raffi; Oettgen, Hans C.

    2005-01-01

    Background: Recent studies have suggested that mouse allergen exposure and sensitization are common in urban children with asthma. The effectiveness of environmental intervention in reducing mouse allergen exposure has not been established. Objective: To evaluate whether environmental intervention of mouse extermination and cleaning results in a reduction in mouse allergen levels. Methods: Eighteen homes of children with positive mouse allergen skin test results and at least mild persistent asthma in urban Boston, MA, with evidence of mouse infestation or exposure were randomized in a 2:1 ratio (12 intervention and 6 control homes). The intervention homes received an integrated pest management intervention, which consisted of filling holes with copper mesh, vacuuming and cleaning, and using low-toxicity pesticides and traps. Dust samples were collected and analyzed for major mouse allergen (Mus m 1) and cockroach allergen (Bla g 1) at baseline and 1, 3, and 5 months after the intervention was started and compared with control homes. Results: Mouse allergen levels were significantly decreased compared with control homes by the end of the intervention period at month 5 in the kitchen and bedroom (kitchen intervention, 78.8% reduction; control, 319% increase; P = .02; bedroom intervention, 77.3% reduction; control, 358% increase; P < .01; and living room intervention, 67.6% reduction; control, 32% reduction; P = .07). Conclusions: Mouse allergen levels were significantly reduced during a 5-month period using an integrated pest management intervention. PMID:15104193

  5. Guilt by intimate association: what makes an allergen an allergen?

    PubMed

    Karp, Christopher L

    2010-05-01

    Why specific, ubiquitous, otherwise innocuous environmental proteins tend to provoke maladaptive, T(H)2-polarized immune responses in susceptible hosts is a fundamental mechanistic question for those interested in the pathogenesis, therapy, and prevention of allergic disease. The current renaissance in the study of innate immunity has provided important insights into this question. The theme emerging from recent studies is that direct (dys)functional interactions with pathways of innate immune activation that evolved to signal the presence of microbial infection are central to the molecular basis for allergenicity. This article reviews these data.

  6. Advances in allergen immunotherapy: aiming for complete tolerance to allergens.

    PubMed

    Akdis, Cezmi A; Akdis, Mübeccel

    2015-03-25

    Allergen-specific immunotherapy (AIT) has been used for more than 100 years as a tolerance-inducing therapy for allergic diseases and represents a potentially curative method of treatment. AIT functions through multiple mechanisms, including regulating T and B cell responses, changing antibody isotypes, and decreasing mediator release and migration of eosinophils, basophils, and mast cells to affected tissues. Despite the relative success of AIT, attempts are being made to improve this therapy in order to overcome problems in standardization, efficacy, safety, long duration of treatment, and costs. These have led to the development of biotechnological products with successful clinical results.

  7. Development of buckwheat and teff sourdoughs with the use of commercial starters.

    PubMed

    Moroni, Alice V; Arendt, Elke K; Morrissey, John P; Dal Bello, Fabio

    2010-08-15

    In this study we investigated the suitability of commercial starters for the production of gluten free sourdoughs. For this purpose, four different laboratory scale sourdoughs were developed from the flours buckwheat or teff. Two different starters (SA, SB) were used to start the fermentations, which were carried out under two technological conditions. Sourdoughs were propagated by back-slopping until the stability was reached. The composition of the stable sourdoughs was investigated by culture dependent techniques and the development of the dominant biota was monitored by PCR-DGGE. Unique and complex LAB and yeasts communities were detected in each sourdough, comprising strains which originated from the flours. The competitiveness of the starter LAB varied according to the substrate and the fermentation conditions applied. Among the LAB present in both SA and SB, L. helveticus and L. paracasei strains did not persist in buckwheat or teff sourdoughs. Lc. argentinum was competitive only in buckwheat sourdoughs, whereas L. reuteri persisted only in teff sourdough. L. fermentum and L. helveticus present in both starters dominated only the sourdoughs fermented at the higher temperature. Remarkably, the starter yeasts were outcompeted by spontaneous yeast strains, i.e. Kazachstania barnetti and Saccharomyces cerevisiae in teff sourdoughs, whereas no yeasts were isolated from buckwheat sourdoughs. The isolation of autochthonous LAB and yeasts from the stable teff and buckwheat sourdoughs indicates that both flours represent an important reservoir for the isolation of novel and competitive starters for the production of gluten free sourdough bread.

  8. Effects of germination on the nutritional properties, phenolic profiles, and antioxidant activities of buckwheat.

    PubMed

    Zhang, Ge; Xu, Zhicun; Gao, Yuanyuan; Huang, Xianxiao; Zou, Yanping; Yang, Tiankui

    2015-05-01

    Germination is considered to be an effective process for improving the nutritional quality and functionality of cereals. In this study, changes of nutritional ingredients, antinutritional components, chemical composition, and antioxidant activities of buckwheat seeds over 72 h of germination were investigated, and the reasons for these changes are discussed. With the prolonged germination time, the contents of crude protein, reducing sugar, total phenolics, total flavonoids, and condensed tannins increased significantly, while the levels of crude fat, phytic acid, and the activity of trypsin inhibitor decreased. Phenolic compounds, such as rutin, vitexin, isovitexin, orientin, isoorientin, chlorogenic acid, trans-3-hydroxycinnamic acid, and p-hydroxybenzoic acid increased significantly during the germination process, which may be due to the activation of phenylalanine ammonialyase. The improvement of flavonoids led to significant enhancement of the antioxidant activities of germinated buckwheat. Germinated buckwheat had better nutritional value and antioxidant activities than ungerminated buckwheat, and it represented an excellent natural source of flavonoids and phenolic compounds, especially rutin and C-glycosylflavones. Therefore, germinated buckwheat could be used as a promising functional food for health promotion.

  9. Assembly of the draft genome of buckwheat and its applications in identifying agronomically useful genes

    PubMed Central

    Yasui, Yasuo; Hirakawa, Hideki; Ueno, Mariko; Matsui, Katsuhiro; Katsube-Tanaka, Tomoyuki; Yang, Soo Jung; Aii, Jotaro; Sato, Shingo; Mori, Masashi

    2016-01-01

    Buckwheat (Fagopyrum esculentum Moench; 2n = 2x = 16) is a nutritionally dense annual crop widely grown in temperate zones. To accelerate molecular breeding programmes of this important crop, we generated a draft assembly of the buckwheat genome using short reads obtained by next-generation sequencing (NGS), and constructed the Buckwheat Genome DataBase. After assembling short reads, we determined 387,594 scaffolds as the draft genome sequence (FES_r1.0). The total length of FES_r1.0 was 1,177,687,305 bp, and the N50 of the scaffolds was 25,109 bp. Gene prediction analysis revealed 286,768 coding sequences (CDSs; FES_r1.0_cds) including those related to transposable elements. The total length of FES_r1.0_cds was 212,917,911 bp, and the N50 was 1,101 bp. Of these, the functions of 35,816 CDSs excluding those for transposable elements were annotated by BLAST analysis. To demonstrate the utility of the database, we conducted several test analyses using BLAST and keyword searches. Furthermore, we used the draft genome as a reference sequence for NGS-based markers, and successfully identified novel candidate genes controlling heteromorphic self-incompatibility of buckwheat. The database and draft genome sequence provide a valuable resource that can be used in efforts to develop buckwheat cultivars with superior agronomic traits. PMID:27037832

  10. Evaluating variability of allergens in commodity crops.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Crops with significant food allergen issues, include legumes, peanut and soybean, cereal grains, such as wheat and maize, and tree nuts (walnut, Brazil nut, among other phylogenetically diverse species) (Teuber et al. 2006). Officially recognized allergenic proteins may include one or multiple prot...

  11. Recombinant allergen-based provocation testing☆

    PubMed Central

    Niederberger, Verena; Eckl-Dorna, Julia; Pauli, Gabrielle

    2014-01-01

    Over the last 25 years, recombinant allergens from all important allergen sources have been cloned and are now available as recombinant proteins. These molecules can be produced in practically unlimited amounts without biological or batch-to-batch variability. It has been shown in provocation tests that recombinant allergens have similar clinical effects as their natural counterparts. With the help of these tools it is possible to reveal the precise reactivity profiles of patients and to uncover and differentiate cross-reactivity from genuine sensitization to an allergen source. Although it has been shown some time ago that it would be possible to replace crude allergen extracts with recombinant allergens for skin prick testing, and even though the use of allergen components can improve routine diagnosis, these tools are still not available for clinical routine applications. The use of provocation tests is a crucial step in the development of new, hypoallergenic vaccines for therapy of allergic disease. Here we describe important provocation methods (skin prick test, intradermal test, atopy patch test, nasal provocation, colonoscopic provocation test) and give an overview of the clinical provocation studies which have been performed with recombinant allergens so far. PMID:23920475

  12. Reducing food allergenicity at the molecular level.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Food allergens are a significant worldwide public health issue. Estimates for the prevalence of food allergies are around 1-2 % of the total population and up to 8 % of children; although, the prevalence may vary between populations and age groups. Peanuts are one of the most allergenic foods. The...

  13. 2S Albumin Storage Proteins: What Makes them Food Allergens?

    PubMed Central

    Moreno, F. Javier; Clemente, Alfonso

    2008-01-01

    2S albumin storage proteins are becoming of increasing interest in nutritional and clinical studies as they have been reported as major food allergens in seeds of many mono- and di-cotyledonous plants. This review describes the main biochemical, structural and functional properties of these proteins thought to play a role in determining their potential allergenicity. 2S albumins are considered to sensitize directly via the gastrointestinal tract (GIT). The high stability of their intrinsic protein structure, dominated by a well-conserved skeleton of cysteine residues, to the harsh conditions present in the GIT suggests that these proteins are able to cross the gut mucosal barrier to sensitize the mucosal immune system and/or elicit an allergic response. The flexible and solvent-exposed hypervariable region of these proteins is immunodominant and has the ability to bind IgE from allergic patients´ sera. Several linear IgE-binding epitopes of 2S albumins spanning this region have been described to play a major role in allergenicity; the role of conformational epitopes of these proteins in food allergy is far from being understood and need to be investigated. Finally, the interaction of these proteins with other components of the food matrix might influence the absorption rates of immunologically reactive 2S albumins but also in their immune response. PMID:18949071

  14. Airborne seafood allergens as a cause of occupational allergy and asthma.

    PubMed

    Lopata, Andreas L; Jeebhay, Mohamed F

    2013-06-01

    Occupational allergy and asthma is a serious adverse health outcome affecting seafood-processing workers. Allergic reactions are directed to two major seafood groups: fish and shellfish, with the latter group comprising crustaceans and molluscs. Several allergenic proteins have been identified in these different groups, but few have been characterised on a molecular level. Parvalbumin appears to be the major fish allergen, while tropomyosin the major crustacean allergen. Other IgE-binding proteins have also been identified in molluscs and other seafood-associated agents (e.g. Anisakis sp), although their molecular nature has not been characterised. Aerosolised allergens can be identified and quantified using immunological and chemical approaches, detecting levels as low as 10 ng/m(3). This contemporary review discusses interesting and recent findings in the area of occupational seafood allergy including high-risk occupations, environmental risk factors for airborne exposures, major and minor allergens implicated and innovative approaches in diagnosing and managing occupational allergy and asthma associated with seafood processing.

  15. Comparison of international food allergen labeling regulations.

    PubMed

    Gendel, Steven M

    2012-07-01

    Food allergy is a significant public health issue worldwide. Regulatory risk management strategies for allergic consumers have focused on providing information about the presence of food allergens through label declarations. A number of countries and regulatory bodies have recognized the importance of providing this information by enacting laws, regulations or standards for food allergen labeling of "priority allergens". However, different governments and organizations have taken different approaches to identifying these "priority allergens" and to designing labeling declaration regulatory frameworks. The increasing volume of the international food trade suggests that there would be value in supporting sensitive consumers by harmonizing (to the extent possible) these regulatory frameworks. As a first step toward this goal, an inventory of allergen labeling regulations was assembled and analyzed to identify commonalities, differences, and future needs.

  16. From allergen genes to allergy vaccines.

    PubMed

    Valenta, Rudolf; Ferreira, Fatima; Focke-Tejkl, Margarete; Linhart, Birgit; Niederberger, Verena; Swoboda, Ines; Vrtala, Susanne

    2010-01-01

    IgE-mediated allergy is a hypersensitivity disease affecting more than 25% of the population. The structures of the most common allergens have been revealed through molecular cloning technology in the past two decades. On the basis of this knowledge of the sequences and three-dimensional structures of culprit allergens, investigators can now analyze the immune recognition of allergens and the mechanisms of allergic inflammation in allergic patients. Allergy vaccines have been constructed that are able to selectively target the aberrant immune responses in allergic patients via different pathways of the immune system. Here we review various types of allergy vaccines that have been developed based on allergen structures, results from their clinical application in allergic patients, and future strategies for allergen-specific immunotherapy and allergy prophylaxis.

  17. Requirements for acquiring a high-quality house dust mite extract for allergen immunotherapy

    PubMed Central

    Frati, Franco; Incorvaia, Cristoforo; David, Marie; Scurati, Silvia; Seta, Simona; Padua, Guglielmo; Cattaneo, Eleonora; Cavaliere, Carlo; Di Rienzo, Alessia; Dell’Albani, Ilaria; Puccinelli, Paola

    2012-01-01

    The house dust mite is a major cause of respiratory allergy worldwide. The management of mite allergy is based on avoidance measures, drug treatment, and allergen immunotherapy, but only allergen immunotherapy is able to modify the natural history of the disease. Injectable subcutaneous immunotherapy was introduced a century ago, while sublingual immunotherapy was proposed in the 1980s and emerged in the ensuing years as an effective and safe option to subcutaneous immunotherapy. However, the quality of the extracts to be used in allergen immunotherapy is crucial for the success of treatment. The mite extract for sublingual immunotherapy known as Staloral 300 was developed to offer optimal characteristics concerning the mite culture medium, standardization, and allergen dose. Double-blind, placebo-controlled trials with Staloral 300 have provided a substantial part of the clinical evidence analyzed in a meta-analysis of the efficacy of allergen immunotherapy in mite-induced rhinitis and asthma. Safety and tolerability are very good, mild local reactions in the mouth being the most common side effect. This makes it feasible to carry out sublingual immunotherapy for the 3–5-year duration needed to achieve long-lasting tolerance to the specific allergen. The performance of Staloral 300 may provide optimal conditions for an effective and safe sublingual immunotherapy in patients with mite-induced respiratory allergy. PMID:22654506

  18. Requirements for acquiring a high-quality house dust mite extract for allergen immunotherapy.

    PubMed

    Frati, Franco; Incorvaia, Cristoforo; David, Marie; Scurati, Silvia; Seta, Simona; Padua, Guglielmo; Cattaneo, Eleonora; Cavaliere, Carlo; Di Rienzo, Alessia; Dell'Albani, Ilaria; Puccinelli, Paola

    2012-01-01

    The house dust mite is a major cause of respiratory allergy worldwide. The management of mite allergy is based on avoidance measures, drug treatment, and allergen immunotherapy, but only allergen immunotherapy is able to modify the natural history of the disease. Injectable subcutaneous immunotherapy was introduced a century ago, while sublingual immunotherapy was proposed in the 1980s and emerged in the ensuing years as an effective and safe option to subcutaneous immunotherapy. However, the quality of the extracts to be used in allergen immunotherapy is crucial for the success of treatment. The mite extract for sublingual immunotherapy known as Staloral 300 was developed to offer optimal characteristics concerning the mite culture medium, standardization, and allergen dose. Double-blind, placebo-controlled trials with Staloral 300 have provided a substantial part of the clinical evidence analyzed in a meta-analysis of the efficacy of allergen immunotherapy in mite-induced rhinitis and asthma. Safety and tolerability are very good, mild local reactions in the mouth being the most common side effect. This makes it feasible to carry out sublingual immunotherapy for the 3-5-year duration needed to achieve long-lasting tolerance to the specific allergen. The performance of Staloral 300 may provide optimal conditions for an effective and safe sublingual immunotherapy in patients with mite-induced respiratory allergy.

  19. Characterization of a new subtype of allergen in dermatophagoides farinae—Der f 28

    PubMed Central

    Lin, Jian-Li; Wang, Yuan-Yuan; Xiao, Xiao-Jun; Wu, Yu-Lan; Sun, Bao-Qing; Gao, An-Jian; Liu, Zhi-Gang

    2015-01-01

    Background House dust mites (HDMs) are the major sources of indoor allergens which induce asthma, dermatitis, rhinitis, and some other allergic diseases. Close to 30 sub-allergens have been identified. Methods Through analyzing the full genome sequence of dust mite, a new allergen whose primary structure belongs to the heat shock protein family was identified. The sequence of this allergen was determined by cDNA cloning. The allergenicity was assayed by skin prick test, Western-blot and enzyme-linked immunosorbent assay (ELISA). Results r-Der f 28 bound to serum IgE from mite allergic patients. Positive responses to r-Der f 28 were shown in 11.5% by skin prick testing from 26 DM-allergic patients. Airway hyperresponsiveness, serum specific IgE and IL-4 were significantly increased in allergic asthma mouse model sensitized to r-Der f 28. Conclusions Der f 28 is a new subtype of allergen in dermatophagoides farinae. PMID:26623108

  20. Mapping of Lol p I allergenic epitopes by using murine monoclonal antibodies.

    PubMed

    Mourad, W; Bernier, D; Jobin, M; Hébert, J

    1989-11-01

    Murine monoclonal antibodies (MAbs) against three non-overlapping epitopes of Lol p I allergen were previously produced and subsequently used for purification of the allergen. In the present study, these MAbs were further characterized, and the biological activity of the purified allergen assessed. The three MAbs were of the IgG isotype and carried a kappa light chain. Their affinity constants were in the range of 7.4-15.1 x 10(-9) mol/l. Purified Lol p I kept its biological activity, as shown by its ability to induce histamine release by basophils of Lol p I-sensitive patients. The profiles of histamine release induced by either Lol p I or crude Lolium perenne extracts were comparable. This observation suggests that human IgE bound to basophils are polyspecific which has been confirmed by immunoblot and inhibition assay. Our data indicated also that Lol p I possesses a major allergenic epitope recognized by all human serum IgE tested. This epitope seems to be partially shared by those recognized by the three MAbs. Finally, preincubation of Lol p I with either one of the Mabs did not affect significantly the basophil-histamine release induced by the purified allergen. This suggests that Lol p I possesses allergenic sites other than the one shared by MAbs and IgE Abs.

  1. Using magnetic beads to reduce reanut allergens from peanut extracts.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ferric irons (Fe3+) and phenolic compounds have been shown to bind to peanut allergens. An easy way to isolate peanut allergens is by use of magnetic beads attached with or without phenolics to capture peanut allergens or allergen-Fe3+ complexes, thus, achieving the goal of producing peanut extracts...

  2. Effects of NO2 and Ozone on Pollen Allergenicity

    PubMed Central

    Frank, Ulrike; Ernst, Dieter

    2016-01-01

    This mini-review summarizes the available data of the air pollutants NO2 and ozone on allergenic pollen from different plant species, focusing on potentially allergenic components of the pollen, such as allergen content, protein release, IgE-binding, or protein modification. Various in vivo and in vitro studies on allergenic pollen are shown and discussed. PMID:26870080

  3. Functional properties of gluten-free pasta produced from amaranth, quinoa and buckwheat.

    PubMed

    Schoenlechner, Regine; Drausinger, Julian; Ottenschlaeger, Veronika; Jurackova, Katerina; Berghofer, Emmerich

    2010-12-01

    The use of amaranth, quinoa and buckwheat for the production of gluten-free pasta was investigated in the present study. The aim of the work was to produce pasta of good textural quality, in particular, low cooking loss, optimal cooking weight and texture firmness. The results demonstrated that pasta produced from amaranth had decreased texture firmness and cooking time, while pasta from quinoa mainly showed increased cooking loss. In buckwheat pasta the least negative effects were observed. By combination of all three raw materials to one flour blend in the ratio of 60% buckwheat, 20% amaranth and 20% quinoa, dough matrix was improved. After decreasing dough moisture to 30%, addition of an increased amount of egg white powder of 6% and addition of 1.2% emulsifier (distilled monoglycerides) texture firmness as well as cooking quality of gluten-free pasta produced from such a flour blend reached acceptable values comparable to wheat pasta.

  4. Modeling bulk canopy resistance from climatic variables for predicting hourly evapotranspiration of maize and buckwheat

    NASA Astrophysics Data System (ADS)

    Yan, Haofang; Shi, Haibin; Hiroki, Oue; Zhang, Chuan; Xue, Zhu; Cai, Bin; Wang, Guoqing

    2015-06-01

    This study presents models for predicting hourly canopy resistance ( r c) and evapotranspiration (ETc) based on Penman-Monteith approach. The micrometeorological data and ET c were observed during maize and buckwheat growing seasons in 2006 and 2009 in China and Japan, respectively. The proposed models of r c were developed by a climatic resistance ( r *) that depends on climatic variables. Non-linear relationships between r c and r * were applied. The measured ETc using Bowen ratio energy balance method was applied for model validation. The statistical analysis showed that there were no significant differences between predicted ETc by proposed models and measured ETc for both maize and buckwheat crops. The model for predicting ETc at maize field showed better performance than predicting ETc at buckwheat field, the coefficients of determination were 0.92 and 0.84, respectively. The study provided an easy way for the application of Penman-Monteith equation with only general available meteorological database.

  5. Functional properties of gluten-free pasta produced from amaranth, quinoa and buckwheat.

    PubMed

    Schoenlechner, Regine; Drausinger, Julian; Ottenschlaeger, Veronika; Jurackova, Katerina; Berghofer, Emmerich

    2010-12-01

    The use of amaranth, quinoa and buckwheat for the production of gluten-free pasta was investigated in the present study. The aim of the work was to produce pasta of good textural quality, in particular, low cooking loss, optimal cooking weight and texture firmness. The results demonstrated that pasta produced from amaranth had decreased texture firmness and cooking time, while pasta from quinoa mainly showed increased cooking loss. In buckwheat pasta the least negative effects were observed. By combination of all three raw materials to one flour blend in the ratio of 60% buckwheat, 20% amaranth and 20% quinoa, dough matrix was improved. After decreasing dough moisture to 30%, addition of an increased amount of egg white powder of 6% and addition of 1.2% emulsifier (distilled monoglycerides) texture firmness as well as cooking quality of gluten-free pasta produced from such a flour blend reached acceptable values comparable to wheat pasta. PMID:20972627

  6. Extraction of rutin from buckwheat (Fagopyrum esculentumMoench) seeds and determination by capillary electrophoresis.

    PubMed

    Kreft, S; Knapp, M; Kreft, I

    1999-11-01

    The content of the flavonoid rutin was determined in different milling fractions of buckwheat seeds and in buckwheat stems, leaves, and flowers. The extraction was performed by using a solvent containing 60% of ethanol and 5% of ammonia in water. The extracts were analyzed by capillary electrophoresis (running buffer of 50 mM borate (pH 9.3), 100 mM sodium dodecyl sulfate; determination at 380 nm). In bran fractions the concentration of rutin was 131-476 ppm, and in flour fractions 19-168 ppm. On average, about 300, 1000, and 46000 ppm of rutin were found in leaves, stems, and flowers, respectively. The results indicate that buckwheat could be an important nutritional source of flavonoids, especially in countries with a low mean daily flavonoid intake.

  7. Attempt to remove peanut allergens from peanut extracts, using IgE-attached magnetic beads.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Immunoglobulin E (IgE) antibodies from sera of peanut-allergic individuals are known to bind specifically to major peanut allergens, Ara h 1 and Ara h 2. The objective of this study was to determine the efficiency of magnetic beads (Dynabeads) attached with IgE antibodies in the removal of major pea...

  8. New routes for allergen immunotherapy

    PubMed Central

    Johansen, Pål; von Moos, Seraina; Mohanan, Deepa; Kündig, Thomas M.; Senti, Gabriela

    2012-01-01

    IgE-mediated allergy is a highly prevalent disease in the industrialized world. Allergen-specific immunotherapy (SIT) should be the preferred treatment, as it has long lasting protective effects and can stop the progression of the disease. However, few allergic patients choose to undergo SIT, due to the long treatment time and potential allergic adverse events. Since the beneficial effects of SIT are mediated by antigen presenting cells inducing Th1, Treg and antibody responses, whereas the adverse events are caused by mast cells and basophils, the therapeutic window of SIT may be widened by targeting tissues rich in antigen presenting cells. Lymph nodes and the epidermis contain high density of dendritic cells and low numbers of mast cells and basophils. The epidermis has the added benefit of not being vascularised thereby reducing the chances of anaphylactic shock due to leakage of allergen. Hence, both these tissues represent highly promising routes for SIT and are the focus of discussion in this review. PMID:23095873

  9. Characterization of the soluble allergenic proteins of cashew nut (Anacardium occidentale L.).

    PubMed

    Teuber, Suzanne S; Sathe, Shridhar K; Peterson, W Rich; Roux, Kenneth H

    2002-10-23

    The allergens associated with cashew food allergy have not been well-characterized. We sought to identify the major allergens in cashew nut by performing IgE immunoblots to dissociated and reduced or nonreduced cashew protein extracts, followed by sequencing of the peptides of interest. Sera from 15 subjects with life-threatening reactions to cashews and 8 subjects who tolerate cashews but have life-threatening reactions to other tree nuts were compared. An aqueous cashew protein extract containing albumin/globulin was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and subjected to IgE immunoblotting using patient sera. Selected IgE reactive bands were subjected to N-terminal amino acid sequencing. Each of the 15 sera from cashew-allergic subjects showed IgE binding to the cashew protein extract. The dominant IgE-binding antigens in the reduced preparations included peptides in the 31-35 kD range, consistent with the large subunits of the major storage 13S globulin (legumin-like protein). Low-molecular-weight polypeptides of the 2S albumin family, with similarity to the major walnut allergen Jug r 1, also bound IgE. The sera from eight patients who tolerate cashew but displayed allergies to other tree nuts showed only minimal or no IgE binding to cashew. Cashew food allergy is associated with the presence of IgE directed against the major seed storage proteins in cashew, including the 13S globulin (legumin group) and 2S albumins, both of which represent major allergen classes in several plant seeds. Thus, the legumin-group proteins and 2S albumins are again identified as major food allergens, which will help further research into seed protein allergenicity.

  10. Pharmacokinetic study of eplerenone in rats after long-term coadministration with buckwheat tea.

    PubMed

    Zou, Liang; Jia, Kun; Li, Rui; Wang, Ping; Lin, Jun-Zhi; Chen, Hui-Juan; Zhao, Gang; Peng, Lian-Xin

    2016-04-01

    The aim of this study was to investigate the effect of long-term intake of Tartary buckwheat tea on the pharmacokinetics (PK) of eplerenone in rats. A validated high-performance liquid chromatography-mass spectrometry (HPLC-MS) method was established to determine the eplerenone in plasma, and the portal vein absorption model was applied to conduct the pharmacokinetic study. Two groups of animals-buckwheat tea group and control group-were involved in this study. Plasma samples were obtained at different time points after administration, and were separated on Shimadzu HPLC-MS 2020 instruments. The method showed good linearity (r=0.9988) over a wide dynamic range (0.20-50 μg/mL). Within- and between-batch precisions ranged from 2.13% to 7.90%. The extraction recovery rates ranged from 91.96% to 94.96%. The data showed that in the Tartarian buckwheat group the area under the curve and maximum concentration of eplerenone were reduced compared with those of the blank group (p<0.01), but the time to reach peak concentrations of eplerenone (p<0.01) was prolonged. The results suggested that long-term consumption of Tartary buckwheat tea might induce the activities of the hepatic drug metabolizing enzyme, which can accelerate the metabolism of eplerenone. According to the results, the dosage of eplerenone should be adjusted in hypertension treatment trials when administered with Tartary buckwheat or Tartary buckwheat-containing dietary supplements to avoid potential drug interactions.

  11. Effects of methyl jasmonate on accumulation of flavonoids in seedlings of common buckwheat (Fagopyrum esculentum Moench).

    PubMed

    Horbowicz, M; Wiczkowski, W; Koczkodaj, Danuta; Saniewski, M

    2011-09-01

    The jasmonates, which include jasmonic acid and its methyl ester (MJ), play a central role in regulating the biosynthesis of many secondary metabolites, including flavonoids, and also are signaling molecules in environmental stresses. Synthesis of anthocyanins pigments is a final part of flavonoids pathway route. Accumulation of the pigments in young seedlings is stimulated by various environmental stresses, such as high-intensity light, wounding, pathogen attack, drought, sugar and nutrient deficiency. The anthocyanins take part in defense system against excess of light and UV-B light, and therefore it is probably main reason why young plant tissues accumulate enlarged levels of the pigments. The effects of exogenously applied MJ on level of anthocyanins, glycosides of apigenin, luteolin, quercetin and proanthocyanidins in seedlings of common buckwheat (Fagopyrum esculentum Moench) were studied. MJ decreased contents of all the found cyanidin glycosides and its aglycone in hypocotyls of buckwheat seedlings. However contents of particular anthocyanins in cotyledons of buckwheat seedlings treated with the plant hormone were not significantly different from the control. Applied doses of MJ did not affect levels of quercetin, apigenin and luteolin glycosides in the analyzed parts of buckwheat seedlings: cotyledons and hypocotyls. On the other hand, treatment of buckwheat seedlings with MJ clearly stimulated of proanthocyanidins biosynthesis in hypocotyls. We suggest that methyl jasmonate induces in hypocotyls of buckwheat seedlings the leucocyanidin reductase or anthocyanidin reductase, possible enzymes in proanthocyanidins synthesis, and/or inhibits anthocyanidin synthase, which transforms leucocyanidin into cyanidin. According to our knowledge this is the first report regarding the effect of methyl jasmonate on enhancing the accumulation of proanthocyanidins in cultivated plants.

  12. Will genetically modified foods be allergenic?

    PubMed

    Taylor, S L; Hefle, S L

    2001-05-01

    Foods produced through agricultural biotechnology, including such staples as corn, soybeans, canola, and potatoes, are already reaching the consumer marketplace. Agricultural biotechnology offers the promise to produce crops with improved agronomic characteristics (eg, insect resistance, herbicide tolerance, disease resistance, and climatic tolerance) and enhanced consumer benefits (eg, better taste and texture, longer shelf life, and more nutritious). Certainly, the products of agricultural biotechnology should be subjected to a careful and complete safety assessment before commercialization. Because the genetic modification ultimately results in the introduction of new proteins into the food plant, the safety, including the potential allergenicity, of the newly introduced proteins must be assessed. Although most allergens are proteins, only a few of the many proteins found in foods are allergenic under the typical circumstances of exposure. The potential allergenicity of the introduced proteins can be evaluated by focusing on the source of the gene, the sequence homology of the newly introduced protein to known allergens, the expression level of the novel protein in the modified crop, the functional classification of the novel protein, the reactivity of the novel protein with IgE from the serum of individuals with known allergies to the source of the transferred genetic material, and various physicochemical properties of the newly introduced protein, such as heat stability and digestive stability. Few products of agricultural biotechnology (and none of the current products) will involve the transfer of genes from known allergenic sources. Applying such criteria provides reasonable assurance that the newly introduced protein has limited capability to become an allergen.

  13. Allergenic tropomyosins and their cross-reactivities.

    PubMed

    Jeong, Kyoung Yong; Hong, Chein-Soo; Yong, Tai-Soon

    2006-01-01

    The ingestion or inhalation of some proteins may lead to adverse immune reactions. Allergens may trigger allergic reactions in genetically predisposed individuals when they are absorbed through the skin or make contact with mucous membranes. An allergic disease often deteriorates the quality of life and may sometimes be life-threatening due to anaphylactic shock. A number of allergens have been characterized from various multicellular organisms to date. It is thought to be reasonable to pay a special attention to the substance which is highly cross-reactive and which causes adverse responses in the molecules that are not sensitized but similar to the sensitized allergen. Tropomyosin has been described as an important food allergen in shrimp, lobster, crab, oysters, squid, and other invertebrates. Allergic reactions to shellfish and mollusks are often cross-reactive, which may be explained by the highly conserved amino acid sequences of tropomyosins among invertebrates, but vertebrate tropomyosins are not known to be allergenic. Several tropomyosins from domestic arthropods have been reported to be allergenic. Recently, it was suggested that an infection of helminthic parasites might lead to sensitization to tropomyosin and elicit allergic reactions to other invertebrates. Much effort has been made to characterize these allergenic tropomyosins from various sources. We will discuss the physicochemical characteristics and the potential application of tropomyosin for the diagnosis and therapeutics of allergic disorders.

  14. Germination of white radish, buckwheat and qing-geng-cai under low pressure in closed environment.

    PubMed

    Hinokuchi, Tsutomu; Oshima, Satoshi; Hashimoto, Hirofumi

    2004-11-01

    In order to cultivate plants under low pressure in closed environment, the germination rate of seeds of white radish was investigated under low pressure, low oxygen partial pressure and condition of pure oxygen. The result of these experiments showed that the germination rate was affected by the oxygen partial pressure. From this fact, it is possible to lower the total pressure by using only the pure oxygen in germination. Furthermore, the germination rates of seeds of buckwheat and qing-geng-cai were also investigated in pure oxygen for the comparison. Consequently, though tendency in germination rate of white radish was similar to qing-geng-cai, it was different from buckwheat.

  15. Silages containing buckwheat and chicory: quality, digestibility and nitrogen utilisation by lactating cows.

    PubMed

    Kälber, Tasja; Kreuzer, Michael; Leiber, Florian

    2012-02-01

    The suitability of silages containing buckwheat (Fagopyrum esculentum) and chicory (Cichorium intybus) for the nutrition of dairy cows was determined. Buckwheat and chicory were sown in mixture with ryegrass (Lolium multilorum), and a pure ryegrass culture served as a control forage. Swards were harvested 55 d after sowing and were ensiled after wilting, without additives in small round bales. Finally, buckwheat and chicory made up the dietary dry matter (DM) proportions of 0.46 and 0.34, respectively. Concentrates were restricted to 2 kg/d. Diets were fed to 3 x 6 late-lactating cows for 15 d at ad libitum access. During the collection period (days 10-15) amounts of feed intake and faeces, urine and milk were recorded and samples were taken. Ensilability was good for buckwheat and ryegrass swards, but was so less for the chicory sward, which was rich in total ash. The buckwheat silage was rich in acid detergent fibre (445 g/kg DM) and lignin (75.7 g/kg DM) and contained less crude protein (135 g/kg DM) and ether extract (15.8 g/kg DM) than the other silages. Consistent with that, the apparent digestibility of the organic matter and fibre were lowest when feeding this silage. The potassium concentrations in the chicory and ryegrass silages were high (61 g/ kg) and lower in buckwheat (47 g/kg). No significant treatment effects on intake, body weight, milk yield or milk composition as well as plasma beta-hydroxybutyrate and non-esterified fatty acids occurred. Being lowest in nitrogen (N) content, the buckwheat silage resulted in the lowest urine N losses and the most efficient N utilisation for milk protein synthesis, but this at cost of body N retention. The results show that silages containing buckwheat and chicory may be used as components of the forage part of dairy cows' diets even though they were found to have a lower feeding value than ryegrass silage.

  16. Effect of processing on the flavonoid content in buckwheat (Fagopyrum esculentum Möench) grain.

    PubMed

    Dietrych-Szostak, D; Oleszek, W

    1999-10-01

    Six flavonoids have been isolated and identified in buckwheat grain. These are rutin, orientin, vitexin, quercetin, isovitexin, and isoorientin. Rutin and isovitexin are the only flavonoid components of buckwheat seeds while hulls contain all six identified compounds. The total flavonoid concentration in the seeds was 18.8 and in the hulls 74 mg/100 g of dry matter. Dehulling the grain by using different temperature regimes resulted in drastic reductions of the total flavonoid concentration in the grain (by 75% of the control) and smaller but significant (15-20%) reduction in the hulls.

  17. Allergen Component Testing in the Diagnosis of Food Allergy.

    PubMed

    Schussler, Edith; Kattan, Jacob

    2015-09-01

    IgE-mediated food allergies are an important public health problem, affecting 5 % of adults and 8 % of children, with numerous studies indicating that the prevalence is increasing. Food allergic reactions can range in severity from mild to severe and life threatening. Accurate diagnosis of food allergy is necessary not only to provide appropriate and potentially life-saving preventive measures but also to prevent unwarranted dietary restrictions. The diagnosis of food allergy has traditionally been based on clinical history and food specific IgE (sIgE) testing, including skin prick testing (SPT), serum tests, or both. These tests tend to be extremely sensitive, but positive test results to foods that are tolerated are common. Studies of allergen component-resolved diagnostics (CRD) show that adjuvant use of this modality may provide a more accurate assessment in the diagnosis of food allergy, though the reported benefits are questionable for a number of major allergens. Furthermore, diagnostic cutoff values have been difficult to determine for allergens where component testing has been demonstrated to be beneficial.

  18. The antigenicity and allergenicity of microparticulated proteins: Simplesse.

    PubMed

    Sampson, H A; Cooke, S

    1992-10-01

    New technologies are allowing the food industry to develop products from standard foods which may not be recognized in its modified form by food allergic patients. One such product, Simplesse, has been formulated by microparticulation of egg white and/or cows' milk proteins and is used as a fat substitute in many fat-laden foods. The purpose of this study was to determine whether the process of microparticulation altered the allergenicity/antigenicity of egg white and cows' milk proteins compared to the starting materials. Soluble protein fractions of Simplesse and its respective starting materials were compared to egg white, cows' milk protein, an ultra-filtered egg white/condensed milk mixture, and/or a whey concentrate by SDS-polyacrylamide gel electrophoresis. In addition, sera from 16 patients with documented egg and/or cows' milk hypersensitivity and two controls who were not allergic to egg or milk were used to assess potential allergenicity/antigenicity of these products by immunoblot (Western blot) analysis. There were heterogeneous IgE and IgG binding patterns to the food fractions among these food allergic patients suggesting differing sensitivity patterns among the individuals tested. However, utilizing both SDS-PAGE and immunoblot analyses, the major allergens in the microparticulated products were the same as those found in the starting materials, egg and cows' milk. In addition, there was no evidence of 'novel' protein fractions in the Simplesse test materials compared to the starting materials. PMID:1464052

  19. IgE reactivity to hen egg white allergens in dogs with cutaneous adverse food reactions.

    PubMed

    Shimakura, Hidekatsu; Uchiyama, Jumpei; Saito, Taku; Miyaji, Kazuki; Fujimura, Masato; Masuda, Kenichi; Okamoto, Noriaki; DeBoer, Douglas J; Sakaguchi, Masahiro

    2016-09-01

    Dogs with cutaneous adverse food reactions (CAFR) often have specific IgE to food allergens. Egg white, which is majorly composed of ovomucoid, ovalbumin, ovotransferrin, and lysozyme, is a food allergen in dogs. Information of the IgE reactivity to purified egg white allergens supports accurate diagnosis and efficiency treatment in humans. However, to the best of our knowledge, there have been no studies on the IgE reactivity to purified egg white allergens in dogs. Here, we investigated the IgE reactivity to crude and purified allergens of hen egg white in dogs with CAFR. First, when we examined serum samples from 82 dogs with CAFR for specific IgE to crude egg white by ELISA, 9.8% (8/82) of the dogs with CAFR showed the IgE reactivity to crude egg white. We then used sera from the eight dogs with positive IgE reactivity to crude egg white to examine the IgE reactivity to four purified allergens, ovomucoid, ovalbumin, ovotransferrin, and lysozyme, by ELISA. We found that 75% (6/8) of the dogs showed IgE reactivity to both ovomucoid and ovalbumin, and that 37.5% (3/8) of the dogs showed IgE reactivity to ovotransferrin. None (0/8) showed IgE reactivity to lysozyme. Moreover, validating these results, the immunoblot analyses were performed using the sera of the three dogs showing the highest IgE reactivity to crude egg white. Both anti-ovomucoid and anti-ovalbumin IgE were detected in the sera of these dogs, while anti-ovotransferrin IgE was not detected. Considering these, ovomucoid and ovalbumin appears to be the major egg white allergens in dogs with CAFR. PMID:27436445

  20. Display of wasp venom allergens on the cell surface of Saccharomyces cerevisiae

    PubMed Central

    2010-01-01

    Background Yeast surface display is a technique, where the proteins of interest are expressed as fusions with yeast surface proteins and thus remain attached to the yeast cell wall after expression. Our purpose was to study whether allergens expressed on the cell surface of baker's yeast Saccharomyces cerevisiae preserve their native allergenic properties and whether the yeast native surface glycoproteins interfere with IgE binding. We chose to use the major allergens from the common wasp Vespula vulgaris venom: phospholipase A1, hyaluronidase and antigen 5 as the model. Results The proteins were expressed on the surface as fusions with a-agglutinin complex protein AGA2. The expression was confirmed by fluorescent cytometry (FACS) after staining the cells with antibody against a C-tag attached to the C-terminal end of the allergens. Phospholipase A1 and hyaluronidase retained their enzymatic activities. Phospholipase A1 severely inhibited the growth of the yeast cells. Antigen 5 - expressing yeast cells bound IgE antibodies from wasp venom allergic patient sera but not from control sera as demonstrated by FACS. Moreover, antigen 5 - expressing yeast cells were capable of mediating allergen-specific histamine release from human basophils. Conclusions All the three major wasp venom allergens were expressed on the yeast surface. A high-level expression, which was observed only for antigen 5, was needed for detection of IgE binding by FACS and for induction of histamine release. The non-modified S. cerevisiae cells did not cause any unspecific reaction in FACS or histamine release assay despite the expression of high-mannose oligosaccharides. In perspective the yeast surface display may be used for allergen discovery from cDNA libraries and possibly for sublingual immunotherapy as the cells can serve as good adjuvant and can be produced in large amounts at a low price. PMID:20868475

  1. Anaphylaxis to buckwheat in an atopic child: a risk factor for severe allergy to nuts and seeds?

    PubMed

    Varga, Eva-Maria; Kollmann, Dagmar; Zach, Maximilian; Bohle, Barbara

    2011-01-01

    Common buckwheat (Fagopyrum esculentum) is known to cause severe anaphylactic reactions in adult individuals. However, type I allergy to buckwheat is rarely seen in children. We report on a 7-year-old boy who developed a grade III anaphylactic reaction after consumption of a cake containing buckwheat flour. Prior to this incident, the boy had developed severe allergic reactions to hazelnuts and suffered from an oral allergy syndrome to poppy seed. Analysis of the patient's IgE reactivity by immunoblotting experiments revealed that he was sensitized to members of the 2S albumin and 11S globulin protein families in buckwheat. Additionally, cross-reactivity was found between the 11S globulins in buckwheat, poppy and hazelnut. IgE inhibition experiments indicated that the 11S globulin in buckwheat was the initial sensitizing protein. We conclude that 11S globulins in buckwheat have the potential to induce IgE antibodies cross-reactive with 11S globulins in other, botanically unrelated foods and may induce anaphylactic reactions.

  2. Antigenic Determinants of the Bilobal Cockroach Allergen Bla g 2.

    PubMed

    Woodfolk, Judith A; Glesner, Jill; Wright, Paul W; Kepley, Christopher L; Li, Mi; Himly, Martin; Muehling, Lyndsey M; Gustchina, Alla; Wlodawer, Alexander; Chapman, Martin D; Pomés, Anna

    2016-01-29

    Bla g 2 is a major indoor cockroach allergen associated with the development of asthma. Antigenic determinants on Bla g 2 were analyzed by mutagenesis based on the structure of the allergen alone and in complex with monoclonal antibodies that interfere with IgE antibody binding. The structural analysis revealed mechanisms of allergen-antibody recognition through cation-π interactions. Single and multiple Bla g 2 mutants were expressed in Pichia pastoris and purified. The triple mutant K132A/K251A/F162Y showed an ∼100-fold reduced capacity to bind IgE, while preserving the native molecular fold, as proven by x-ray crystallography. This mutant was still able to induce mast cell release. T-cell responses were assessed by analyzing Th1/Th2 cytokine production and the CD4(+) T-cell phenotype in peripheral blood mononuclear cell cultures. Although T-cell activating capacity was similar for the KKF mutant and Bla g 2 based on CD25 expression, the KKF mutant was a weaker inducer of the Th2 cytokine IL-13. Furthermore, this mutant induced IL-10 from a non-T-cell source at higher levels that those induced by Bla g 2. Our findings demonstrate that a rational design of site-directed mutagenesis was effective in producing a mutant with only 3 amino acid substitutions that maintained the same fold as wild type Bla g 2. These residues, which were involved in IgE antibody binding, endowed Bla g 2 with a T-cell modulatory capacity. The antigenic analysis of Bla g 2 will be useful for the subsequent development of recombinant allergen vaccines.

  3. House dust mite and cat allergen in different indoor environments.

    PubMed

    Custovic, A; Taggart, S C; Woodcock, A

    1994-12-01

    Allergy to house dust mites (HDM) and domestic pets is a major cause of asthma. People in developed countries spend more than 90% of their time indoors. We have measured levels of HDM allergen Der pI and cat allergen Fel dI in public buildings and public transport. Dust samples were collected by vacuuming a 1 m2 area for 2 min from five schools, six hotels, four cinemas, six pubs, three buses, two trains and 12 domestic households without a cat. Der pI and Fel dI were assayed with monoclonal antibodies in a two-site immunometric ELISA. Der pI concentration was significantly higher in the private homes than in comparable sites in public places except for cinema seats (where high values were found) compared with domestic sofas. Der pI > 2000 ng/g of fine dust was found in 30% of the upholstered seats, 9% having a concentration > 10,000 ng/g. Fel dI levels were significantly higher in the dust from upholstered seats (geometric mean 14.88 micrograms/g) than in carpeted floors (geometric mean 0.73 micrograms/g), and in public places than in private homes. Fel dI > 8 micrograms/g was found in 79% of the upholstered seats or furniture sampled in public buildings or public transport. In conclusion, upholstered seats from public buildings and public transport constitute an allergen reservoir for continuous contamination of the indoor environment which could compromise the effects of allergen avoidance employed at home.

  4. The indoor air and asthma: the role of cat allergens

    PubMed Central

    Kelly, Libby A.; Erwin, Elizabeth A.; Platts-Mills, Thomas A. E.

    2012-01-01

    Purpose of review The objective is to discuss recent progress in our understanding of the role of the indoor environment in asthma, focusing on the special role of cat allergens. Recent findings Sensitization to Fel d 1 is the dominant event in inhalant responses to cat; however, there are also IgE responses to the lipocalin (Fel d 4), to cat albumin (Fel d 2), and to the oligosaccharide galactose-alpha-1,3-galactose (alpha-gal) on cat IgA (Fel d 5w) and other molecules. The dose response and routes of sensitization for these allergens are now thought to be diverse. It is important to remember that exposure outside a house with a cat is sufficient to cause sensitization. Furthermore, the only solid evidence about a role in asthma relates to Fel d 1. Recently, it has been shown that tolerance associated with early exposure to cats can persist to age 18 and that IgE to alpha-gal (on cat IgA) is not related to asthma. In addition, a recent study of anti-IgE reinforces the evidence that IgE antibodies to indoor allergens make a major contribution to asthma severity. Summary Exposure to Fel d 1 in a home with a cat is far higher than the levels necessary to induce an allergic (IgE antibody) response. In keeping with that, children may develop tolerance, which can be long-lived. In addition, there is increasing evidence that IgE antibodies to an inhalant allergen, such as Fel d 1, dust mite, or cockroach, are causally related to lung inflammation and asthma. PMID:22081090

  5. Antigenic Determinants of the Bilobal Cockroach Allergen Bla g 2.

    PubMed

    Woodfolk, Judith A; Glesner, Jill; Wright, Paul W; Kepley, Christopher L; Li, Mi; Himly, Martin; Muehling, Lyndsey M; Gustchina, Alla; Wlodawer, Alexander; Chapman, Martin D; Pomés, Anna

    2016-01-29

    Bla g 2 is a major indoor cockroach allergen associated with the development of asthma. Antigenic determinants on Bla g 2 were analyzed by mutagenesis based on the structure of the allergen alone and in complex with monoclonal antibodies that interfere with IgE antibody binding. The structural analysis revealed mechanisms of allergen-antibody recognition through cation-π interactions. Single and multiple Bla g 2 mutants were expressed in Pichia pastoris and purified. The triple mutant K132A/K251A/F162Y showed an ∼100-fold reduced capacity to bind IgE, while preserving the native molecular fold, as proven by x-ray crystallography. This mutant was still able to induce mast cell release. T-cell responses were assessed by analyzing Th1/Th2 cytokine production and the CD4(+) T-cell phenotype in peripheral blood mononuclear cell cultures. Although T-cell activating capacity was similar for the KKF mutant and Bla g 2 based on CD25 expression, the KKF mutant was a weaker inducer of the Th2 cytokine IL-13. Furthermore, this mutant induced IL-10 from a non-T-cell source at higher levels that those induced by Bla g 2. Our findings demonstrate that a rational design of site-directed mutagenesis was effective in producing a mutant with only 3 amino acid substitutions that maintained the same fold as wild type Bla g 2. These residues, which were involved in IgE antibody binding, endowed Bla g 2 with a T-cell modulatory capacity. The antigenic analysis of Bla g 2 will be useful for the subsequent development of recombinant allergen vaccines. PMID:26644466

  6. Local reactions from subcutaneous allergen immunotherapy.

    PubMed

    Coop, Christopher A

    2013-12-01

    Local reactions from subcutaneous allergen immunotherapy are very common during the course of immunotherapy. These local reactions are not bothersome to patients. Local reactions from immunotherapy also do not predict future local or systemic reactions. This review discusses the studies that show that local reactions are not predictive of future reactions and that dose adjustments for local reactions from allergen immunotherapy are unnecessary. The article also focuses on factors that lead to patient noncompliance with immunotherapy and evaluates methods to prevent local reactions from subcutaneous allergen immunotherapy. PMID:24283844

  7. Contact Dermatitis, Patch Testing, and Allergen Avoidance.

    PubMed

    Burkemper, Nicole M

    2015-01-01

    In patients presenting with a complaint of rash, contact dermatitis is often the underlying diagnosis making it an entity with which health care providers should be familiar. Contact dermatitis can be divided into irritant contact dermatitis and allergic contact dermatitis. In a patient suspected of having allergic contact dermatitis, patch testing can be done to identify specific allergens. Education focused on allergen avoidance and safe products is an integral part of treatment for the contact dermatitis patient. Knowledge of the most common allergens is helpful for clinicians to be able to provide this education.

  8. Contact Dermatitis, Patch Testing, and Allergen Avoidance.

    PubMed

    Burkemper, Nicole M

    2015-01-01

    In patients presenting with a complaint of rash, contact dermatitis is often the underlying diagnosis making it an entity with which health care providers should be familiar. Contact dermatitis can be divided into irritant contact dermatitis and allergic contact dermatitis. In a patient suspected of having allergic contact dermatitis, patch testing can be done to identify specific allergens. Education focused on allergen avoidance and safe products is an integral part of treatment for the contact dermatitis patient. Knowledge of the most common allergens is helpful for clinicians to be able to provide this education. PMID:26455061

  9. Local reactions from subcutaneous allergen immunotherapy.

    PubMed

    Coop, Christopher A

    2013-12-01

    Local reactions from subcutaneous allergen immunotherapy are very common during the course of immunotherapy. These local reactions are not bothersome to patients. Local reactions from immunotherapy also do not predict future local or systemic reactions. This review discusses the studies that show that local reactions are not predictive of future reactions and that dose adjustments for local reactions from allergen immunotherapy are unnecessary. The article also focuses on factors that lead to patient noncompliance with immunotherapy and evaluates methods to prevent local reactions from subcutaneous allergen immunotherapy.

  10. Cloning and expression of biologically active Plantago lanceolata pollen allergen Pla l 1 in the yeast Pichia pastoris.

    PubMed Central

    Calabozo, Belén; Díaz-Perales, Araceli; Salcedo, Gabriel; Barber, Domingo; Polo, Florentino

    2003-01-01

    The glycoprotein Pla l 1 is the major allergen from English plantain (Plantago lanceolata) pollen, which is a common cause of pollinosis in temperate areas. Three complete cDNAs for Pla l 1 isoforms were isolated by PCR using specific 3' and 5' primers. All three Pla l 1 cDNAs code for a 25-residue leader peptide and a 131-residue mature protein that contains two polymorphic positions, an N-glycosylation site at position 107 and six cysteine residues involved in three disulphide bridges. The allergen variant Pla l 1.0101 was produced in Pichia pastoris at a yield of 20 mg per litre of culture as a mixture of non-glycosylated (17 kDa), glycosylated (23 kDa) and dimeric (32-39 kDa) forms. Recombinant Pla l 1 (rPla l 1) was purified by affinity chromatography with an anti-natural Pla l 1 (anti-nPla l 1) monoclonal antibody, and its molecular and immunological properties were compared with the natural allergen by CD spectroscopic analysis, enzymic deglycosylation, lectin-binding assay, immunodetection and ELISA-inhibition assays using sera from plantain-allergic patients. The recombinant allergen is properly folded, as deduced from CD spectra, and the immunodominant allergenic epitopes of the natural allergen are preserved in rPla l 1. These results allow us to conclude that P. pastoris is a convenient system for the efficient production of biologically active rPla l 1, which could have a potential use for clinical purposes. Furthermore, a sequence similarity of Pla l 1 to the major allergen from the olive tree pollen, Ole e 1, is revealed in this work, and the allergenic cross-reactivity between both allergens has been studied. PMID:12646046

  11. Cloning and expression of biologically active Plantago lanceolata pollen allergen Pla l 1 in the yeast Pichia pastoris.

    PubMed

    Calabozo, Belén; Díaz-Perales, Araceli; Salcedo, Gabriel; Barber, Domingo; Polo, Florentino

    2003-06-15

    The glycoprotein Pla l 1 is the major allergen from English plantain (Plantago lanceolata) pollen, which is a common cause of pollinosis in temperate areas. Three complete cDNAs for Pla l 1 isoforms were isolated by PCR using specific 3' and 5' primers. All three Pla l 1 cDNAs code for a 25-residue leader peptide and a 131-residue mature protein that contains two polymorphic positions, an N-glycosylation site at position 107 and six cysteine residues involved in three disulphide bridges. The allergen variant Pla l 1.0101 was produced in Pichia pastoris at a yield of 20 mg per litre of culture as a mixture of non-glycosylated (17 kDa), glycosylated (23 kDa) and dimeric (32-39 kDa) forms. Recombinant Pla l 1 (rPla l 1) was purified by affinity chromatography with an anti-natural Pla l 1 (anti-nPla l 1) monoclonal antibody, and its molecular and immunological properties were compared with the natural allergen by CD spectroscopic analysis, enzymic deglycosylation, lectin-binding assay, immunodetection and ELISA-inhibition assays using sera from plantain-allergic patients. The recombinant allergen is properly folded, as deduced from CD spectra, and the immunodominant allergenic epitopes of the natural allergen are preserved in rPla l 1. These results allow us to conclude that P. pastoris is a convenient system for the efficient production of biologically active rPla l 1, which could have a potential use for clinical purposes. Furthermore, a sequence similarity of Pla l 1 to the major allergen from the olive tree pollen, Ole e 1, is revealed in this work, and the allergenic cross-reactivity between both allergens has been studied.

  12. Characterization of the allergenic potential of proteins: an assessment of the kiwifruit allergen actinidin.

    PubMed

    Dearman, Rebecca J; Beresford, Lorna; Foster, Emily S; McClain, Scott; Kimber, Ian

    2014-05-01

    Assessment of the potential allergenicity (IgE-inducing properties) of novel proteins is an important challenge in the overall safety assessment of foods. Resistance to digestion with pepsin is commonly measured to characterize allergenicity, although the association is not absolute. We have previously shown that specific IgE antibody production induced by systemic [intraperitoneal (i.p.)] exposure of BALB/c strain mice to a range of proteins correlates with allergenic potential for known allergens. The purpose of the present study was to explore further the utility of these approaches using the food allergen, actinidin. Recently, kiwifruit has become an important allergenic foodstuff, coincident with its increased consumption, particularly as a weaning food. The ability of the kiwifruit allergen actinidin to stimulate antibody responses has been compared with the reference allergen ovalbumin, and with the non-allergen bovine haemoglobin. Haemoglobin was rapidly digested by pepsin whereas actinidin was resistant unless subjected to prior chemical reduction (reflecting intracellular digestion conditions). Haemoglobin stimulated detectable IgG antibody production at relatively high doses (10%), but failed to provoke detectable IgE. In contrast, actinidin was both immunogenic and allergenic at relatively low doses (0.25% to 1%). Vigorous IgG and IgG1 antibody and high titre IgE antibody responses were recorded, similar to those provoked by ovalbumin. Thus, actinidin displays a marked ability to provoke IgE, consistent with allergenic potential. These data provide further encouragement that in tandem with analysis of pepsin stability, the induction of IgE after systemic exposure of BALB/c strain mice provides a useful approach for the prospective identification of protein allergens.

  13. Allergenic fragments of ryegrass (Lolium perenne) pollen allergen Lol p IV.

    PubMed

    Jaggi, K S; Ekramoddoullah, A K; Kisil, F T

    1989-01-01

    To facilitate studies on establishing the nature of structure/function relationships of allergens, ryegrass pollen allergen, Lol p IV, was cleaved into smaller fragments by cyanogen bromide (CNBr) and the resulting peptides were further digested with trypsin. The resulting peptides were then fractionated by high performance liquid chromatography (HPLC) on a C-18 reverse phase column. The allergenic activity of the HPLC fractions was evaluated in terms of their ability to inhibit the binding of 125I-Lol p IV to serum IgE antibodies of a grass-allergic patient. Many of these fractions inhibited the binding between the native allergen and IgE antibodies in a dose-dependent manner. The inhibitions were specific, i.e., the fractions did not inhibit the binding between 125I-Lol p I (a group-I ryegrass pollen allergen) and the IgE antibodies present in the allergic human serum. The possibility that the allergenic peptide fractions were contaminated by the native undegraded allergen, which might have accounted for the observed inhibition, was ruled out by the fact that the native allergen could not be detected by SDS-PAGE and the elution profiles of allergenically active peptides did not coincide with that of native allergen. One of the allergenic sites recognized by monoclonal antibody (Mab) 90, i.e., site A, was located in HPLC fractions 90-100 while another allergenic site B (recognized by Mab 12) appeared to be lost following the sequential digestion of Lol p IV with CNBr and trypsin.

  14. Evolution of nutrient ingredients in tartary buckwheat seeds during germination.

    PubMed

    Yiming, Zhou; Hong, Wang; Linlin, Cui; Xiaoli, Zhou; Wen, Tang; Xinli, Song

    2015-11-01

    Evolution of nutrient components and the antioxidative activity of seed sprouts of tartary buckwheat (Fagopyrum tataricum L. Gaertn) were investigated in the course of germination. Results showed that the contents of total flavonoids increased with germination time and leveled off after the third germination day with the changing trend of rutin and quercetin opposite to each other. The decrease of total protein and total sugar contents in the germinated seeds was accompanied respectively by an increase of amino acid and reducing sugar contents. The contents of vitamin C (Vc) and B1(V(B1)) exhibited a minimum with no appreciable changes found for vitamin B(2) (V(B2)) and B(6) (V(B6)). The contents of total chlorophyll, chlorophyll A and B all exhibited a maximum on the fifth germination day. The contents of fatty acids had no regular changing trend with germination time. The free radical-scavenging activities of the seeds increased with germination time and were caused by an increase in their antioxidative activity.

  15. Genomic Characterization of Phenylalanine Ammonia Lyase Gene in Buckwheat.

    PubMed

    Thiyagarajan, Karthikeyan; Vitali, Fabio; Tolaini, Valentina; Galeffi, Patrizia; Cantale, Cristina; Vikram, Prashant; Singh, Sukhwinder; De Rossi, Patrizia; Nobili, Chiara; Procacci, Silvia; Del Fiore, Antonella; Antonini, Alessandro; Presenti, Ombretta; Brunori, Andrea

    2016-01-01

    Phenylalanine Ammonia Lyase (PAL) gene which plays a key role in bio-synthesis of medicinally important compounds, Rutin/quercetin was sequence characterized for its efficient genomics application. These compounds possessing anti-diabetic and anti-cancer properties and are predominantly produced by Fagopyrum spp. In the present study, PAL gene was sequenced from three Fagopyrum spp. (F. tataricum, F. esculentum and F. dibotrys) and showed the presence of three SNPs and four insertion/deletions at intra and inter specific level. Among them, the potential SNP (position 949th bp G>C) with Parsimony Informative Site was selected and successfully utilised to individuate the zygosity/allelic variation of 16 F. tataricum varieties. Insertion mutations were identified in coding region, which resulted the change of a stretch of 39 amino acids on the putative protein. Our Study revealed that autogamous species (F. tataricum) has lower frequency of observed SNPs as compared to allogamous species (F. dibotrys and F. esculentum). The identified SNPs in F. tataricum didn't result to amino acid change, while in other two species it caused both conservative and non-conservative variations. Consistent pattern of SNPs across the species revealed their phylogenetic importance. We found two groups of F. tataricum and one of them was closely related with F. dibotrys. Sequence characterization information of PAL gene reported in present investigation can be utilized in genetic improvement of buckwheat in reference to its medicinal value. PMID:26990297

  16. Genomic Characterization of Phenylalanine Ammonia Lyase Gene in Buckwheat

    PubMed Central

    Thiyagarajan, Karthikeyan; Vitali, Fabio; Tolaini, Valentina; Galeffi, Patrizia; Cantale, Cristina; Vikram, Prashant; Singh, Sukhwinder; De Rossi, Patrizia; Nobili, Chiara; Procacci, Silvia; Del Fiore, Antonella; Antonini, Alessandro; Presenti, Ombretta; Brunori, Andrea

    2016-01-01

    Phenylalanine Ammonia Lyase (PAL) gene which plays a key role in bio-synthesis of medicinally important compounds, Rutin/quercetin was sequence characterized for its efficient genomics application. These compounds possessing anti-diabetic and anti-cancer properties and are predominantly produced by Fagopyrum spp. In the present study, PAL gene was sequenced from three Fagopyrum spp. (F. tataricum, F. esculentum and F. dibotrys) and showed the presence of three SNPs and four insertion/deletions at intra and inter specific level. Among them, the potential SNP (position 949th bp G>C) with Parsimony Informative Site was selected and successfully utilised to individuate the zygosity/allelic variation of 16 F. tataricum varieties. Insertion mutations were identified in coding region, which resulted the change of a stretch of 39 amino acids on the putative protein. Our Study revealed that autogamous species (F. tataricum) has lower frequency of observed SNPs as compared to allogamous species (F. dibotrys and F. esculentum). The identified SNPs in F. tataricum didn’t result to amino acid change, while in other two species it caused both conservative and non-conservative variations. Consistent pattern of SNPs across the species revealed their phylogenetic importance. We found two groups of F. tataricum and one of them was closely related with F. dibotrys. Sequence characterization information of PAL gene reported in present investigation can be utilized in genetic improvement of buckwheat in reference to its medicinal value. PMID:26990297

  17. Evolution of nutrient ingredients in tartary buckwheat seeds during germination.

    PubMed

    Yiming, Zhou; Hong, Wang; Linlin, Cui; Xiaoli, Zhou; Wen, Tang; Xinli, Song

    2015-11-01

    Evolution of nutrient components and the antioxidative activity of seed sprouts of tartary buckwheat (Fagopyrum tataricum L. Gaertn) were investigated in the course of germination. Results showed that the contents of total flavonoids increased with germination time and leveled off after the third germination day with the changing trend of rutin and quercetin opposite to each other. The decrease of total protein and total sugar contents in the germinated seeds was accompanied respectively by an increase of amino acid and reducing sugar contents. The contents of vitamin C (Vc) and B1(V(B1)) exhibited a minimum with no appreciable changes found for vitamin B(2) (V(B2)) and B(6) (V(B6)). The contents of total chlorophyll, chlorophyll A and B all exhibited a maximum on the fifth germination day. The contents of fatty acids had no regular changing trend with germination time. The free radical-scavenging activities of the seeds increased with germination time and were caused by an increase in their antioxidative activity. PMID:25976817

  18. Genomic Characterization of Phenylalanine Ammonia Lyase Gene in Buckwheat.

    PubMed

    Thiyagarajan, Karthikeyan; Vitali, Fabio; Tolaini, Valentina; Galeffi, Patrizia; Cantale, Cristina; Vikram, Prashant; Singh, Sukhwinder; De Rossi, Patrizia; Nobili, Chiara; Procacci, Silvia; Del Fiore, Antonella; Antonini, Alessandro; Presenti, Ombretta; Brunori, Andrea

    2016-01-01

    Phenylalanine Ammonia Lyase (PAL) gene which plays a key role in bio-synthesis of medicinally important compounds, Rutin/quercetin was sequence characterized for its efficient genomics application. These compounds possessing anti-diabetic and anti-cancer properties and are predominantly produced by Fagopyrum spp. In the present study, PAL gene was sequenced from three Fagopyrum spp. (F. tataricum, F. esculentum and F. dibotrys) and showed the presence of three SNPs and four insertion/deletions at intra and inter specific level. Among them, the potential SNP (position 949th bp G>C) with Parsimony Informative Site was selected and successfully utilised to individuate the zygosity/allelic variation of 16 F. tataricum varieties. Insertion mutations were identified in coding region, which resulted the change of a stretch of 39 amino acids on the putative protein. Our Study revealed that autogamous species (F. tataricum) has lower frequency of observed SNPs as compared to allogamous species (F. dibotrys and F. esculentum). The identified SNPs in F. tataricum didn't result to amino acid change, while in other two species it caused both conservative and non-conservative variations. Consistent pattern of SNPs across the species revealed their phylogenetic importance. We found two groups of F. tataricum and one of them was closely related with F. dibotrys. Sequence characterization information of PAL gene reported in present investigation can be utilized in genetic improvement of buckwheat in reference to its medicinal value.

  19. Mechanisms of immune tolerance to allergens: role of IL-10 and Tregs

    PubMed Central

    Akdis, Cezmi A.; Akdis, Mübeccel

    2014-01-01

    During the past 20 years, major advances have been made in understanding the molecular and cellular mechanisms of allergen tolerance in humans. The demonstration of T cell tolerance, particularly that mediated by the immune-suppressive functions of IL-10, led to a major conceptual change in this area. Currently, the known essential components of allergen tolerance include the induction of allergen-specific regulatory subsets of T and B cells, the immune-suppressive function of secreted factors, such as IL-10 and TGF-β, the production of IgG4 isotype allergen–specific blocking antibodies, and decreased allergic inflammatory responses by mast cells, basophils, and eosinophils in inflamed tissues. PMID:25365074

  20. The distribution of dust mite allergen in the houses of patients with asthma

    SciTech Connect

    Tovey, E.R.; Chapman, M.D.; Wells, C.W.; Platts-Mills, T.A.

    1981-11-01

    Using an inhibition radioimmunoassay for the major allergen from Dermatophagoides pteronyssinus (antigen P1), we studied the distribution of this dust allergen in the houses of patients with asthma. Both bed and floor dust samples contained a wide range of antigen P1, 100 to 100,000 ng/g of fine dust, and this concentration correlated well with the number of mite bodies (r . 0.81, p less than 0.001). We were unable to detect antigen P1 in the air of undisturbed rooms. However, during domestic activity, between 1 and 30 ng were collected on a filter than sampled air for 45 min at 17 L/min. Using a cascade impactor it was shown that greater than 80% of the airborne antigen P1 was associated with particles greater than 10 mu in diameter. Some of the particles containing allergen could be identified because they formed precipitin rings when impacted onto agarose containing rabbit antimite antiserum. These particles had the physical appearance of mite feces, which are the major source of antigen P1 in mite cultures. The results suggested that natural exposure to this dust allergen allows occasional fecal particles to enter the lungs and that these particles contain very concentrated allergen.

  1. Common ragweed (Ambrosia artemisiifolia L.): allergenicity and molecular characterization of pollen after plant exposure to elevated NO2.

    PubMed

    Zhao, Feng; Elkelish, Amr; Durner, Jörg; Lindermayr, Christian; Winkler, J Barbro; Ruёff, Franziska; Behrendt, Heidrun; Traidl-Hoffmann, Claudia; Holzinger, Andreas; Kofler, Werner; Braun, Paula; von Toerne, Christine; Hauck, Stefanie M; Ernst, Dieter; Frank, Ulrike

    2016-01-01

    Ragweed pollen is the main cause of allergenic diseases in Northern America, and the weed has become a spreading neophyte in Europe. Climate change and air pollution are speculated to affect the allergenic potential of pollen. The objective of this study was to investigate the effects of NO2 , a major air pollutant, under controlled conditions, on the allergenicity of ragweed pollen. Ragweed was exposed to different levels of NO2 throughout the entire growing season, and its pollen further analysed. Spectroscopic analysis showed increased outer cell wall polymers and decreased amounts of pectin. Proteome studies using two-dimensional difference gel electrophoresis and liquid chromatography-tandem mass spectrometry indicated increased amounts of several Amb a 1 isoforms and of another allergen with great homology to enolase Hev b 9 from rubber tree. Analysis of protein S-nitrosylation identified nitrosylated proteins in pollen from both conditions, including Amb a 1 isoforms. However, elevated NO2 significantly enhanced the overall nitrosylation. Finally, we demonstrated increased overall pollen allergenicity by immunoblotting using ragweed antisera, showing a significantly higher allergenicity for Amb a 1. The data highlight a direct influence of elevated NO2 on the increased allergenicity of ragweed pollen and a direct correlation with an increased risk for human health.

  2. Peach ( Prunus persica L. Batsch) allergen-encoding genes are developmentally regulated and affected by fruit load and light radiation.

    PubMed

    Botton, Alessandro; Andreotti, Carlo; Costa, Guglielmo; Ramina, Angelo

    2009-01-28

    The fruits of Rosaceae species may frequently induce allergic reactions in both adults and children, especially in the Mediterranean area. In peach, true allergens and cross-reactive proteins may cause hypersensitive reactions involving a wide diversity of symptoms. Three known classes of allergenic proteins, namely, Pru p 1, Pru p 3, and Pru p 4, have been reported to be mostly involved, but an exhaustive survey of the proteins determining the overall allergenic potential, their biological functions, and the factors affecting the expression of the related genes is still missing. In the present study, the expression profiles of some selected genes encoding peach allergen isoforms were studied during fruit growth and development and upon different fruit load and light radiation regimens. The results indicate that the majority of allergen-encoding genes are expressed at their maximum during the ripening stage, therefore representing a potential risk for peach consumers. Nevertheless, enhancing the light radiation and decreasing the fruit load achieved a reduction of the transcription rate of most genes and a possible decrease of the overall allergenic potential at harvest. According to these data, new growing practices could be set up to obtain hypoallergenic peach fruits and eventually combined with the cultivation of hypoallergenic genotypes to obtain a significant reduction of the allergenic potential.

  3. Common ragweed (Ambrosia artemisiifolia L.): allergenicity and molecular characterization of pollen after plant exposure to elevated NO2.

    PubMed

    Zhao, Feng; Elkelish, Amr; Durner, Jörg; Lindermayr, Christian; Winkler, J Barbro; Ruёff, Franziska; Behrendt, Heidrun; Traidl-Hoffmann, Claudia; Holzinger, Andreas; Kofler, Werner; Braun, Paula; von Toerne, Christine; Hauck, Stefanie M; Ernst, Dieter; Frank, Ulrike

    2016-01-01

    Ragweed pollen is the main cause of allergenic diseases in Northern America, and the weed has become a spreading neophyte in Europe. Climate change and air pollution are speculated to affect the allergenic potential of pollen. The objective of this study was to investigate the effects of NO2 , a major air pollutant, under controlled conditions, on the allergenicity of ragweed pollen. Ragweed was exposed to different levels of NO2 throughout the entire growing season, and its pollen further analysed. Spectroscopic analysis showed increased outer cell wall polymers and decreased amounts of pectin. Proteome studies using two-dimensional difference gel electrophoresis and liquid chromatography-tandem mass spectrometry indicated increased amounts of several Amb a 1 isoforms and of another allergen with great homology to enolase Hev b 9 from rubber tree. Analysis of protein S-nitrosylation identified nitrosylated proteins in pollen from both conditions, including Amb a 1 isoforms. However, elevated NO2 significantly enhanced the overall nitrosylation. Finally, we demonstrated increased overall pollen allergenicity by immunoblotting using ragweed antisera, showing a significantly higher allergenicity for Amb a 1. The data highlight a direct influence of elevated NO2 on the increased allergenicity of ragweed pollen and a direct correlation with an increased risk for human health. PMID:26177592

  4. Evaluation of the potential allergenicity of the enzyme microbial transglutaminase using the 2001 FAO/WHO Decision Tree.

    PubMed

    Pedersen, Mona H; Hansen, Tine K; Sten, Eva; Seguro, Katsuya; Ohtsuka, Tomoko; Morita, Akiko; Bindslev-Jensen, Carsten; Poulsen, Lars K

    2004-11-01

    All novel proteins must be assessed for their potential allergenicity before they are introduced into the food market. One method to achieve this is the 2001 FAO/WHO Decision Tree recommended for evaluation of proteins from genetically modified organisms (GMOs). It was the aim of this study to investigate the allergenicity of microbial transglutaminase (m-TG) from Streptoverticillium mobaraense. Amino acid sequence similarity to known allergens, pepsin resistance, and detection of protein binding to specific serum immunoglobulin E (IgE) (RAST) have been evaluated as recommended by the decision tree. Allergenicity in the source material was thought unlikely, since no IgE-mediated allergy to any bacteria has been reported. m-TG is fully degraded after 5 min of pepsin treatment. A database search showed that the enzyme has no homology with known allergens, down to a match of six contiguous amino acids, which meets the requirements of the decision tree. However, there is a match at the five contiguous amino acid level to the major codfish allergen Gad c1. The potential cross reactivity between m-TG and Gad c1 was investigated in RAST using sera from 25 documented cod-allergic patients and an extract of raw codfish. No binding between patient IgE and m-TG was observed. It can be concluded that no safety concerns with regard to the allergenic potential of m-TG were identified.

  5. Inactivation of allergens and toxins.

    PubMed

    Morandini, Piero

    2010-11-30

    Plants are replete with thousands of proteins and small molecules, many of which are species-specific, poisonous or dangerous. Over time humans have learned to avoid dangerous plants or inactivate many toxic components in food plants, but there is still room for ameliorating food crops (and plants in general) in terms of their allergens and toxins content, especially in their edible parts. Inactivation at the genetic rather than physical or chemical level has many advantages and classical genetic approaches have resulted in significant reduction of toxin content. The capacity, offered by genetic engineering, of turning off (inactivating) specific genes has opened up the possibility of altering the plant content in a far more precise manner than previously available. Different levels of intervention (genes coding for toxins/allergens or for enzymes, transporters or regulators involved in their metabolism) are possible and there are several tools for inactivating genes, both direct (using chemical and physical mutagens, insertion of transposons and other genetic elements) and indirect (antisense RNA, RNA interference, microRNA, eventually leading to gene silencing). Each level/strategy has specific advantages and disadvantages (speed, costs, selectivity, stability, reversibility, frequency of desired genotype and regulatory regime). Paradigmatic examples from classical and transgenic approaches are discussed to emphasize the need to revise the present regulatory process. Reducing the content of natural toxins is a trade-off process: the lesser the content of natural toxins, the higher the susceptibility of a plant to pests and therefore the stronger the need to protect plants. As a consequence, more specific pesticides like Bt are needed to substitute for general pesticides.

  6. [Governmental batch sample testing of allergen products].

    PubMed

    Bartel, D; Führer, F; Vieths, S

    2012-03-01

    Allergen products for specific immunotherapy of type I allergies were first authorized for the German market in the 1970s. In addition to finished products manufactured in advance and in batches, so-called named patient products have recently been defined as Medicinal Products by the German Medicinal Products Act ("Arzneimittelgesetz", AMG 14th Revision 2005). Some allergen products previously marketed as named patient products are now required to obtain marketing authorization according to the German ordinance for therapy allergens. Products have to be batch released by the competent German Federal Agency, the Paul-Ehrlich-Institut (PEI). Samples of product batches are delivered to the PEI in order to perform experimental quality controls. With regard to named patient products, PEI tests batch samples of the bulk extract preparations used for manufacturing of the respective, named patient products. The institute releases approximately 2,800 allergen product batches annually.

  7. 21 CFR 680.1 - Allergenic Products.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... use of the mold as a source material for Allergenic Products, in accordance with 21 CFR 601.2, the... capable of determining the good health of the animals. (iii) Immunization against tetanus. Animals of...

  8. 21 CFR 680.1 - Allergenic Products.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... use of the mold as a source material for Allergenic Products, in accordance with 21 CFR 601.2, the... capable of determining the good health of the animals. (iii) Immunization against tetanus. Animals of...

  9. 21 CFR 680.1 - Allergenic Products.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... use of the mold as a source material for Allergenic Products, in accordance with 21 CFR 601.2, the... capable of determining the good health of the animals. (iii) Immunization against tetanus. Animals of...

  10. 21 CFR 680.1 - Allergenic Products.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... use of the mold as a source material for Allergenic Products, in accordance with 21 CFR 601.2, the... capable of determining the good health of the animals. (iii) Immunization against tetanus. Animals of...

  11. 21 CFR 680.1 - Allergenic Products.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... use of the mold as a source material for Allergenic Products, in accordance with 21 CFR 601.2, the... capable of determining the good health of the animals. (iii) Immunization against tetanus. Animals of...

  12. Identification of crustacea allergens by crossed radioimmunoelectrophoresis.

    PubMed

    Lehrer, S B; McCants, M L; Salvaggio, J E

    1985-01-01

    Crossed immunoelectrophoresis (CIE) detected 18 precipitating antigens in extracts of shrimp. Of these antigens, crossed-line immunoelectrophoresis (CLIE) of shrimp extract demonstrated that 5 cross-reacted with crayfish, 3 with lobster and 1 with crab extract. Allergens present in the shrimp CIE plates were identified by crossed radioimmunoelectrophoresis (CRIE) using sera from 6 study subjects who were skin-test and RAST positive to shrimp extract. Of the 7 allergens detected, 3 (precipitins 1, 3 and 6) reacted with most of the 6 sera tested from shrimp-sensitive subjects. Precipitins 1 and 6 appear to be common crustacea allergens (present in shrimp, crayfish, lobster and crab) whereas precipitin 3 may be a specific allergen since it is present only in shrimp.

  13. Purification and molecular structure of digalactosyl myo-inositol (DGMI), trigalactosyl myo-inositol (TGMI), and fagopyritol B3 from common buckwheat seeds by NMR.

    PubMed

    Gui, Wei; Lemley, Bethan A; Keresztes, Ivan; Condo, Anthony M; Steadman, Kathryn J; Obendorf, Ralph L

    2013-10-18

    Three galactosyl cyclitols, digalactosyl myo-inositol (assigned the trivial name DGMI), trigalactosyl myo-inositol (assigned the trivial name TGMI), and trigalactosyl d-chiro-inositol (fagopyritol B3), were isolated from common buckwheat (Fagopyrum esculentum Moench) seeds. Structures of the three compounds were determined by 2D NMR spectroscopy. DGMI is α-d-galactopyranosyl-(1→6)-α-d-galactopyranosyl-(1→1)-1l-myo-inositol, TGMI is α-d-galactopyranosyl-(1→6)-α-d-galactopyranosyl-(1→6)-α-d-galactopyranosyl-(1→1)-1l-myo-inositol, and fagopyritol B3 is α-d-galactopyranosyl-(1→6)-α-d-galactopyranosyl-(1→6)-α-d-galactopyranosyl-(1→2)-1d-chiro-inositol. DGMI and TGMI are higher oligomers of galactinol, a major galactosyl donor in plant seeds. Fagopyritol B3 is a higher oligomer of the fagopyritol B series and one of six fagopyritols found in buckwheat seeds and in the bran milling fraction. Samples of TGMI and fagopyritol B3 also contained the compounds N-(β-glucopyranosyl)-nicotinic acid and β-d-apiofuranosyl-(1→6)-β-d-(1-O-methyl)-glucopyranoside.

  14. Allergen-specific immunotherapy in atopic eczema.

    PubMed

    Darsow, Ulf; Forer, Ingeborg; Ring, Johannes

    2011-08-01

    Aeroallergens are relevant eliciting factors of allergic rhinoconjunctivitis and bronchial asthma but also of atopic eczema. The use of allergen-specific immunotherapy as in respiratory atopic diseases is controversial in patients with atopic eczema, but refined diagnostic methods to characterize subgroups of patients with relevant allergies and the results of smaller controlled studies give rise to new approaches in this field. This article reviews the theoretical problems and practical results associated with allergen-specific immunotherapy in atopic eczema. PMID:21461718

  15. Molecular and immunological characterization of allergens from the entomopathogenic fungus Beauveria bassiana

    PubMed Central

    Westwood, Greg S; Huang, Shih-Wen; Keyhani, Nemat O

    2006-01-01

    Background Entomopathogenic fungi such as Beauveria bassiana are considered promising biological control agents for a variety of arthropod pests. Beauveria species, however, have the potential to elicit allergenic reactions in humans, although no specific allergens have been characterized to date. Methods Four putative allergens were identified within B. bassiana expressed sequence tag (EST) datasets. IgE-reactivity studies were performed using sera from patients displaying mold allergies against recombinant B. bassiana proteins expressed in E. coli. Results Full length cDNA and genomic nucleotide sequences of four potential B. bassiana allergens were isolated. BLASTX search results led to their putative designation as follows; Bb-Eno1, with similarity to fungal enolases; Bb-f2, similar to the Aspergillus fumigatus major allergen, Asp f2 and to a fibrinogen binding mannoprotein; Bb-Ald, similar to aldehyde dehydrogenases; and Bb-Hex, similar to N-acetyl-hexosaminadases. All four genes were cloned into E. coli expression systems and recombinant proteins were produced. Immunoblots of E. coli extracts probed with pooled as well as individual human sera from patients displaying mould allergies demonstrated IgE reactivity versus recombinant Bb-Eno1 and Bb-Ald. Conclusion Four putative Beauveria bassiana allergens were identified. Recombinant proteins corresponding to two of the four, Bb-Eno1 and Bb-Ald were bound by sera IgEs derived from patients with fungal allergies. These data confirm the potential allergenicity of B. bassiana by identification of specific human IgE reactive epitopes. PMID:16995945

  16. Chemical changes in rubber allergens during vulcanization.

    PubMed

    Bergendorff, Ola; Persson, Christina; Lüdtke, Anna; Hansson, Christer

    2007-09-01

    Allergic contact dermatitis to rubber is caused by residues of chemicals used in manufacturing a rubber product. Several different additives are used to achieve a final product of the desired characteristics. Accelerators such as thiurams, dithiocarbamates, and mercaptobenzothiazoles are often among the additives responsible for allergic reactions recognized by dermatologists. The chemistry of the vulcanization process is complicated; as it occurs at an elevated temperature with a mixture of reactive chemicals, the compositions of the initial and final products differ. This paper investigates the changes in composition of common allergens during vulcanization, doing so by chemically analysing various rubber formulations at different stages of the process. Major changes were found in which added chemicals were consumed and new ones produced. An important observation is that thiuram disulfides rarely appear in the final rubber although they may have been used as additives. Instead, thiurams are often converted to dithiocarbamates or to products formed by addition to mercaptobenzothiazole structures, if these have been used together with thiurams as accelerators. PMID:17680862

  17. Chemical changes in rubber allergens during vulcanization.

    PubMed

    Bergendorff, Ola; Persson, Christina; Lüdtke, Anna; Hansson, Christer

    2007-09-01

    Allergic contact dermatitis to rubber is caused by residues of chemicals used in manufacturing a rubber product. Several different additives are used to achieve a final product of the desired characteristics. Accelerators such as thiurams, dithiocarbamates, and mercaptobenzothiazoles are often among the additives responsible for allergic reactions recognized by dermatologists. The chemistry of the vulcanization process is complicated; as it occurs at an elevated temperature with a mixture of reactive chemicals, the compositions of the initial and final products differ. This paper investigates the changes in composition of common allergens during vulcanization, doing so by chemically analysing various rubber formulations at different stages of the process. Major changes were found in which added chemicals were consumed and new ones produced. An important observation is that thiuram disulfides rarely appear in the final rubber although they may have been used as additives. Instead, thiurams are often converted to dithiocarbamates or to products formed by addition to mercaptobenzothiazole structures, if these have been used together with thiurams as accelerators.

  18. Teff, buckwheat, quinoa and amaranth: Ancient whole grain gluten-free egg-free pasta

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This report demonstrates innovative ancient whole grains, gluten-free, egg-free pasta (no chemicals added) made using a kitchen counter-top appliance. Whole grain, fusilli pasta was prepared with teff, buckwheat, quinoa and amaranth flours. These ancient grains are called “Super Foods” due to thei...

  19. Metabolism of secondary metabolites isolated from Tartary buckwheat and its extract.

    PubMed

    Ren, Qiang; Li, Yingfei; Wu, Caisheng; Wang, Caihong; Jin, Ying; Zhang, Jinlan

    2014-07-01

    The metabolism of Tartary buckwheat was investigated using a strategy from single bioactive compounds to complex Tartary buckwheat extract. Firstly, the metabolites of different structural compounds were investigated by an in situ liver-intestinal perfusion model and the metabolic pathways were proposed. Furthermore, Tartary buckwheat extract in rats was elucidated on the basis of the metabolism information of single compounds. High-performance liquid chromatography coupled with high resolution mass and multiple-stage mass spectrometry (HPLC-HRMS/MS(n)) was performed to characterise and identify 19 metabolites in perfusate and intestinal content after administration of single compounds to an in situ liver/intestinal perfusion model and 16 metabolites and 6 components in rat faeces, urine, bile, and plasma after oral administration of Tartary buckwheat to rats. Five new metabolites were identified as the glucuronidation and sulfation products of N-trans-feruloyltyramine and the methylation product of quercetin-3-O-[β-d-xyloxyl-(1→2)-α-l-rhamnoside]. The metabolic pathways of phenylpropanoid glycosides and N-trans-feruloyltyramine were proposed for the first time.

  20. Antioxidant activity of commercial buckwheat flours and their free and bound phenolic compositions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Buckwheat flours (Whole, Farinetta, Supreme, and Fancy) were investigated for their compositions, free and bound phenolic contents, antioxidant activities, and flavonoid contents using spectrophotometer and LC-ESI-IT- MS (LC-MS). Farinetta flour contained the highest oil, protein, and free and boun...

  1. Ancient whole grain Gluten-free egg-free Teff, Buckwheat, Quinoa and Amaranth pasta (abstract)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This report demonstrates innovative ancient whole grain, gluten-free, egg-free pasta (no chemicals added) made using a kitchen counter-top appliance. Whole grain, fusilli pasta was prepared with teff, buckwheat, quinoa and amaranth flours. These ancient grains are called “Super Foods” due to thei...

  2. Distribution of Vitamin E, squalene, epicatechin, and rutin in common buckwheat plants (Fagopyrum esculentum Moench).

    PubMed

    Kalinova, Jana; Triska, Jan; Vrchotova, Nadezda

    2006-07-26

    Buckwheat leaves and young parts of the plant are consumed in some countries as a vegetable. Green flour, obtained by milling of the dried plants, is used as a natural food colorant. The distribution of vitamin E, squalene, epicatechin, and rutin (as the most important antioxidants) within buckwheat plants, as well as changes of their content within leaves during the growing season, were determined by GC-MS and HPLC analyses. alpha-Tocopherol was found as the main component of vitamin E in all parts of the plant; epicatechin and squalene were also detected. For the use of buckwheat as an antioxidant source in the human diet, the most suitable part of the plants seems to be the leaves and the flowers at the stage of full flowering due to the considerable amounts of rutin and epicatechin. alpha-Tocopherol content correlates positively with temperature, drought, and duration of solar radiation. Certain differences appear among varieties of buckwheat, especially in their squalene and rutin contents.

  3. Rheological and pasting properties of buckwheat (Fagopyrum esculentum Moench) flours with and without jet-cooking

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pasting, rheological and water-holding properties of buckwheat (Fagopyrum esculentum) flour obtained from whole achenes separated into three particle sizes, and three commercial flours (Fancy, Supreme and Farinetta) were measured with or without jet-cooking. Fancy had instantaneous paste viscosity ...

  4. Ecology of Kearney’s Buckwheat (Eriogonum nummulare) for Restoration Efforts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Eriogonum are a highly complex group of plants with a long history of changes in scientific names. These distinctive plants often have unique associations with endemic animals. One such case that has received much interest involves Kearney’s Buckwheat (Eriogonum nummulare) and an endemic Sand Mo...

  5. Characterization of cDNA encoding resveratrol synthase and accumulation of resveratrol in tartary buckwheat.

    PubMed

    Kim, Yeon Bok; Thwe, Aye Aye; Kim, YeJi; Yeo, Sun Kyung; Lee, Chanhui; Park, Sang Un

    2013-11-01

    Resveratrol synthase (RS) is the key enzyme for biosynthesis of resveratrol which come from coumaroyl-coenzyme A (CoA) and malonyl-CoA. Here, we report the cloning and characterization of a RS gene and accumulation of resveratrol in tartary buckwheat (Fagopyrum tataricum). FtRS was composed of 1173 bp open reading frame and 390 amino acid residues and had a theoretical molecular weight and isoelectric point value of 43.70 kDa and 6.24, respectively. The FtRS expression levels were examined in sprouts and different organs of two tartary buckwheat cultivars, Hokkai T8 (T8) and Hokkai T10 (T10). FtRS transcript levels and resveratrol contents were higher under the dark condition compared with light condition. The expression levels of different organs of T10 was not observed significant variations compared to different organs of T8. Interestingly, resveratrol was detected in the sprouts developmental stages, but no resveratrol could not detect in any other organs of both T8 and T10. Therefore, we suggest that the resveratrol content in tartary buckwheat sprouts may be attributed mainly to the dark condition. The characterization of FtRS will be helpful for better understanding of the resveratrol biosynthesis in tartary buckwheat. PMID:24427944

  6. [Occurrence of allergens on hospital premises].

    PubMed

    Andersen, U M; Osterballe, O

    2000-01-24

    The occurrence of allergens from the house-dust mites Der p 1, Der f 1 and Der m, and from dogs (Can f 1) and cats (Fel d 1) was assessed in Viborg Hospital. Three hundred samples collected in a standardized manner were analysed for allergens by ELISA technique. In only one dust sample was the total occurrence of mite allergens marginally above the sensitization threshold level of 2,000 ng mite allergens/g dust. For Fel d 1 a threshold level for sensitization or symptoms of 8,000 ng Fel d 1/g dust has been proposed; none of the dust samples contained this concentration. A low occurrence of Can f 1 was found. One dust sample contained 8,902 ng Can f 1, while the remainder exhibited lower concentrations. Efficient cleaning and adequate ventilation can reduce allergens in public buildings, but it is impossible to remove all allergens from upholstered furniture. Avoidance of such furniture in wards and outpatient departments which receive allergic patients might be considered.

  7. Cross-reactivity of peanut allergens.

    PubMed

    Bublin, Merima; Breiteneder, Heimo

    2014-04-01

    Peanut seeds are currently widely used as source of human food ingredients in the United States of America and in European countries due to their high quality protein and oil content. This article describes the classification and molecular biology of peanut seed allergens with particular reference to their cross-reactivities. Currently, the IUIS allergen nomenclature subcommittee accepts 12 peanut allergens. Two allergens belong to the cupin and four to the prolamin superfamily, and six are distributed among profilins, Bet v 1-like proteins, oleosins, and defensins. Clinical observations frequently report an association of peanut allergy with allergies to legumes, tree nuts, seeds, fruits and pollen. Molecular cross-reactivity has been described between members of the Bet v 1-like proteins, the non-specific lipid transfer proteins, and the profilins. This review also addresses the less well-studied cross-reactivity between cupin and prolamin allergens of peanuts and of other plant food sources and the recently discovered cross-reactivity between peanut allergens of unrelated protein families. PMID:24554241

  8. High pressure effects on allergen food proteins.

    PubMed

    Somkuti, Judit; Smeller, László

    2013-12-15

    There are several proteins, which can cause allergic reaction if they are inhaled or ingested. Our everyday food can also contain such proteins. Food allergy is an IgE-mediated immune disorder, a growing health problem of great public concern. High pressure is known to affect the structure of proteins; typically few hundred MPa pressure can lead to denaturation. That is why several trials have been performed to alter the structure of the allergen proteins by high pressure, in order to reduce its allergenicity. Studies have been performed both on simple protein solutions and on complex food systems. Here we review those allergens which have been investigated under or after high pressure treatment by methods capable of detecting changes in the secondary and tertiary structure of the proteins. We focus on those allergenic proteins, whose structural changes were investigated by spectroscopic methods under pressure in correlation with the observed allergenicity (IgE binding) changes. According to this criterion we selected the following allergen proteins: Mal d 1 and Mal d 3 (apple), Bos d 5 (milk), Dau c 1 (carrot), Gal d 2 (egg), Ara h 2 and Ara h 6 (peanut), and Gad m 1 (cod).

  9. Assessment of allergenicity to fungal allergens of Rohtak city, Haryana, India

    PubMed Central

    Kochar, Sanjeeta; Ahlawat, Manisha; Chaudhary, Dhruva

    2014-01-01

    Fungal spores are known as one of the important bioparticles causing allergic manifestation in human beings. Hence, knowledge of season and prevalence of the airborne allergens to which the patients are exposed is a prerequisite for proper diagnosis and treatment of allergic disorders in hypersensitive individuals. Keeping this in view, aerial survey was performed in the atmosphere of Rohtak city for 2 consecutive years (March 2008–February 2010), using a volumetric petri plate sampler. A total of 45 fungal spore types were recorded during the survey period. In the present study, February–April and July–November were identified as the peak seasons for Rohtak city. Cladosporium was the main contributor to the total fungal load with 25.14% followed by Alternaria (18.05%), Aspergillus niger (7.66%), Curvularia (5.31%), and Epicoccum (5.29%). Fifteen dominant viable fungal spore types were represented in the form of a fungal calendar. An attempt has also been made to assess the allergenicity of some of the fungal types recorded from the atmosphere of Rohtak city. The magnitude of variations observed in markedly positive skin reactions (2+ and above) varied from 17.3 to 2.3%. Penicillium oxalicum showed a markedly positive reaction in maximum number of patients (26; 17.3%) followed by Rhizopus nigricans (23; 15.3%). ELISA was performed with the sera of patients showing markedly positive skin reactions and the sera were classified into four groups based on percent binding. The majority of the sera showed 0–15% binding to different antigenic extracts, while sera showing >60% binding were least in number. Greater than 30% binding was observed against antigens of Rhizopus nigricans, Epicoccum purpurascens, Penicillium oxalicum, Curvularia lunata, Aspergillus flavus, Candida albicans and Neurospora sitophila. The concordance between positive skin reaction and serum-specific IgE antibodies ranged from 16.7 to 69.2%. PMID:24988378

  10. [Animal models for assessment of GMO allergenicity: advantages and limitations].

    PubMed

    Adel-Patient, K; Wal, J M

    2004-03-01

    Incidence of IgE-mediated allergic reactions to foods is increasing as well as the severity of associated symptoms and numerous foods are now incriminated, probably in relation with modifications of dietary habits and increased exposure to new or modified food ingredients. Therefore, the introduction on the market of food composed of or derived from genetically modified organisms (GMOs) raised the question of their potential allergenicity. Particularly with regards to the allergenicity of a newly expressed protein, it is necessary to obtain, from several steps in the risk assessment process, a cumulative body of evidence which minimises any uncertainty. This may include the use of animal model despite no fully reliable validated model is available yet. Such animal models should allow to address 3 major issues: Is the novel protein a sensitizer, i.e. does it possess intrinsic properties that allow to sensitize a predisposed individual? Is the protein an elicitor i.e. is it able to elicit an allergic reaction in a sensitised individual? And is the protein an adjuvant, i.e. can it facilitate or enhance the sensitisation to an other protein? Animal models under investigation currently include mice, rats and guinea pigs but models such as dogs and swine also appeared a few years ago. The aim is to mimic the mechanism and characteristics of the sensitisation phase and/or the elicitation phase of the allergic reaction as it occurs in atopic humans. They are necessary because sensitisation studies can obviously not be done in human and because in vitro tests cannot reproduce the complexity of the immune system. We propose a mouse model which mimics both phases of the allergic reaction. It has permitted to evidence that biochemical and clinical manifestations occuring during the active phases of the allergic reaction differ according to the structure of the allergen used for the challenge. This may allow to compare the allergenic potential of a genetically modified protein

  11. Effect of roasting time of buckwheat groats on the formation of Maillard reaction products and antioxidant capacity.

    PubMed

    Małgorzata, Wronkowska; Konrad, Piskuła Mariusz; Zieliński, Henryk

    2016-04-01

    Changes in the formation of Maillard reaction products and antioxidant capacity of buckwheat, induced by roasting at 160 °C for 30, 40 and 50 min, were evaluated in the study. Furozine, was detected after roasting, in all buckwheat samples. Increase of FIC, the presence of significant amounts of CML and enhanced browning were observed, along with increasing times of roasting. The formation of acrylamide in the obtained buckwheat products was also significantly connected with the time of roasting. A significant degradation was observed in natural antioxidants, as affected by heat treatment time. The colour parameter changed significantly with the increasing of roasting time. Overall, 30min of roasting was beneficial from a nutritional point of view for the obtained buckwheat product.

  12. Biodiversity of lactic acid bacteria and yeasts in spontaneously-fermented buckwheat and teff sourdoughs.

    PubMed

    Moroni, Alice V; Arendt, Elke K; Dal Bello, Fabio

    2011-05-01

    In this study, four different laboratory scale gluten-free (GF) sourdoughs were developed from buckwheat or teff flours. The fermentations were initiated by the spontaneous biota of the flours and developed under two technological conditions (A and B). Sourdoughs were propagated by continuous back-slopping until the stability was reached. The composition of the stable biota occurring in each sourdough was assessed using both culture-dependent and -independent techniques. Overall, a broad spectrum of lactic acid bacteria (LAB) and yeasts species, belonging mainly to the genera Lactobacillus, Pediococcus, Leuconostoc, Kazachstania and Candida, were identified in the stable sourdoughs. Buckwheat and teff sourdoughs were dominated mainly by obligate or facultative heterofermentative LAB, which are commonly associated with traditional wheat or rye sourdoughs. However, the spontaneous fermentation of the GF flours resulted also in the selection of species which are not consider endemic to traditional sourdoughs, i.e. Pediococcus pentosaceus, Leuconostoc holzapfelii, Lactobacillus gallinarum, Lactobacillus vaginalis, Lactobacillus sakei, Lactobacillus graminis and Weissella cibaria. In general, the composition of the stable biota was strongly affected by the fermentation conditions, whilst Lactobacillus plantarum dominated in all buckwheat sourdoughs. Lactobacillus pontis is described for the first time as dominant species in teff sourdough. Among yeasts, Saccharomyces cerevisiae and Candida glabrata dominated teff sourdoughs, whereas the solely Kazachstania barnetti was isolated in buckwheat sourdough developed under condition A. This study allowed the identification and isolation of LAB and yeasts species which are highly competitive during fermentation of buckwheat or teff flours. Representatives of these species can be selected as starters for the production of sourdough destined to GF bread production.

  13. Buckwheat achenes antioxidant profile modulates Aspergillus flavus growth and aflatoxin production.

    PubMed

    Chitarrini, G; Nobili, C; Pinzari, F; Antonini, A; De Rossi, P; Del Fiore, A; Procacci, S; Tolaini, V; Scala, V; Scarpari, M; Reverberi, M

    2014-10-17

    Buckwheat (Fagopyrum spp.) is a "pseudo-cereal" of great interest in the production of healthy foods since its flour, derived from achenes, is enriched with bioactive compounds and, due to the absence of gluten, may be used in composition of celiac diets. Amongst buckwheat species, F. tataricum achenes possess a larger amount of the antioxidant flavenol rutin than the common buckwheat F. esculentum. Ongoing climate change may favor plant susceptibility to the attack by pathogenic, often mycotoxigenic, fungi with consequent increase of mycotoxins in previously unexploited feeds and foodstuffs. In particular, Aspergillus flavus, under suitable environmental conditions such as those currently occurring in Italy, may produce aflatoxin B1 (AFB1), the most carcinogenic compound of fungal origin which is classified by IARC as Category 1. In this study, the viable achenes of two buckwheat species, F. tataricum (var. Golden) and F. esculentum (var. Aelita) were inoculated with an AFB1-producing A. flavus NRRL 3357 to analyze their relative performances against fungal invasion and toxin contamination. Notably, we sought the existence of a correlation between the amount of tocols/flavonols in the achenes of buckwheat, infected and non-infected with A. flavus, and to analyze the ability of the pathogen to grow and produce toxin during achene infection. Results suggest that achenes of F. tataricum, the best producer of antioxidant compounds in this study, are less susceptible to A. flavus infection and consequently, but not proportionally, to mycotoxin contamination compared with F. esculentum. Moreover, rutin-derived quercetin appears to be more efficient in inhibiting aflatoxin biosynthesis than the parent compound.

  14. Alt a 15 is a new cross-reactive minor allergen of Alternaria alternata.

    PubMed

    Gabriel, M F; Postigo, I; Gutiérrez-Rodríguez, A; Suñén, E; Guisantes, J A; Fernández, J; Tomaz, C T; Martínez, J

    2016-02-01

    Alternaria alternata is one of the most common saprophytes worldwide that is clinically and epidemiologically associated with severe asthma. Therefore, the identification and characterization of all A. alternata allergens are of major clinical importance. This study describes a new cross-reactive A. alternata allergen that was officially named Alt a 15 by the official Allergen Nomenclature Subcommittee. The complete coding region for Alt a 15 was amplified using 5' and 3' rapid amplification of cDNA ends and PCR. The recombinant protein was produced in Escherichia coli as a 65-kDa fusion protein, and the protein sequence exhibits high homology with several important fungal allergens. Immunoblotting analyses revealed that IgE antibodies from A. alternata-sensitized patients (n=59) bound to rAlt a 15 with a prevalence of 10.2%. All patients who presented sIgE to rAlt a 15 were apparently poly-sensitized to A. alternata and C. lunata. The extensive cross-reactivity between A. alternata and C. lunata serine proteases was confirmed using immunoblotting inhibition assays. Overall, Alt a 15 is an important new cross-reactive allergen of A. alternata that explains some allergies to A. alternata without Alt a 1 sensitization and initial diagnostic errors for allergies to Alternaria. This molecule may improve the accuracy of the diagnosis, the understanding, and the management of IgE-mediated fungal diseases.

  15. Identification and Characterization of a New Pecan [Carya illinoinensis (Wangenh.) K. Koch] Allergen, Car i 2.

    PubMed

    Zhang, Yuzhu; Lee, BoRam; Du, Wen-Xian; Lyu, Shu-Chen; Nadeau, Kari C; Grauke, Larry J; Zhang, Yan; Wang, Shuo; Fan, Yuting; Yi, Jiang; McHugh, Tara H

    2016-05-25

    The 7S vicilin and 11S legumin seed storage globulins belong to the cupin protein superfamily and are major food allergens in many foods from the "big eight" food allergen groups. Here, for the first time, pecan vicilin was found to be a food allergen. Western blot experiments revealed that 30% of 27 sera used in this study and 24% of the sera from 25 patients with double-blind, placebo controlled clinical pecan allergy contained IgE antibodies specific to pecan vicilin. This allergen consists of a low-complexity region at its N-terminal and a structured domain at the C-terminal that contains two cupin motifs and forms homotrimers. The crystal structure of recombinant pecan vicilin was determined. The refined structure gave R/Rfree values of 0.218/0.262 for all data to 2.65 Å. There were two trimeric biological units in the crystallographic asymmetric unit. Pecan vicilin is also a copper protein. These data may facilitate the understanding of the nutritional value and the allergenicity relevance of the copper binding property of seed storage proteins in tree nuts. PMID:27128197

  16. Comparative study on the allergenicity of different Litopenaeus vannamei extract solutions

    NASA Astrophysics Data System (ADS)

    Wu, Lisha; Lin, Haixin; Wang, Guoying; Lu, Zongchao; Chen, Guanzhi; Lin, Hong; Li, Zhenxing

    2013-11-01

    Allergen extracts are widely used for allergy diagnosis and treatment. The application of shrimp extract is hampered due to the low protein concentration and the inconsistent allergenicity. Extracting solutions are considered to be the primary limiting factor of protein extraction from crustaceans. This study aimed to select an optimal solution for shrimp protein extraction by comparing the allergenicity of different shrimp extracts. The effect of 7 existing or modified extracting solutions were evaluated, including the glycerol-NaCl solution, the glycerol Cocaine's solution, the buffered saline solution, the Cocaine's solution, the Glucose leaching solution, 1 mol L-1 KCl solution, and 0.01 mol L-1 phosphate buffered saline solution with and without dithiothreitolor (DTT). The quantitative (protein concentration) and qualitative parameters (SDS-PAGE protein patterns and immuno-reactivity) were determined using the sodium dodecyl sulfate polyacrylamide gel electrophoresis, enzyme linked immunosorbent assay and immunoblotting assay. Results showed that the 1 mol L-1 KCl solution with DTT was optimal for shrimp protein extraction, which yielded high concentration and allergenicity in the protein extract, including major and minor allergens. The 1 mol L-1 KCl solution with DDT is proposed for preparation of shrimp extract and associated allergy diagnosis, as well as potential applications for other crustaceans.

  17. Food allergens profiling with an imaging surface plasmon resonance-based biosensor.

    PubMed

    Rebe Raz, Sabina; Liu, Hong; Norde, Willem; Bremer, Maria G E G

    2010-10-15

    Food allergy is a growing health concern, which currently affects approximately 4% of adults and 8% of infants. For consumer protection purposes, food producers are required by law to disclose on the product label whether a major allergen is used during the production process. The commonly employed monitoring methods are highly laborious, time-consuming, and often expensive when screening for multiple allergens. Here, we utilize imaging surface plasmon resonance (iSPR) in combination with antibody array for rapid, quantitative, and multianalyte food allergens detection. We demonstrate how the use of this technology provides a complete allergen profile within short measurement time and with adequate sensitivity. The successful applicability of this approach is demonstrated by analyzing cookies and dark chocolate products from different manufacturers. Hazelnut content of the tested food products is also determined by enzyme linked immunosorbent assay and is found to correlate well with the hazelnut content determined by iSPR. This newly developed method opens the door to automated and high-throughput allergen analysis, ultimately aiming at providing the consumer with safer food.

  18. Identification and Characterization of a New Pecan [Carya illinoinensis (Wangenh.) K. Koch] Allergen, Car i 2.

    PubMed

    Zhang, Yuzhu; Lee, BoRam; Du, Wen-Xian; Lyu, Shu-Chen; Nadeau, Kari C; Grauke, Larry J; Zhang, Yan; Wang, Shuo; Fan, Yuting; Yi, Jiang; McHugh, Tara H

    2016-05-25

    The 7S vicilin and 11S legumin seed storage globulins belong to the cupin protein superfamily and are major food allergens in many foods from the "big eight" food allergen groups. Here, for the first time, pecan vicilin was found to be a food allergen. Western blot experiments revealed that 30% of 27 sera used in this study and 24% of the sera from 25 patients with double-blind, placebo controlled clinical pecan allergy contained IgE antibodies specific to pecan vicilin. This allergen consists of a low-complexity region at its N-terminal and a structured domain at the C-terminal that contains two cupin motifs and forms homotrimers. The crystal structure of recombinant pecan vicilin was determined. The refined structure gave R/Rfree values of 0.218/0.262 for all data to 2.65 Å. There were two trimeric biological units in the crystallographic asymmetric unit. Pecan vicilin is also a copper protein. These data may facilitate the understanding of the nutritional value and the allergenicity relevance of the copper binding property of seed storage proteins in tree nuts.

  19. Allergic and nonallergic interactions between house dust mite allergens and airway mucosa.

    PubMed

    Roche, N; Chinet, T C; Huchon, G J

    1997-03-01

    Asthma and allergy are extremely frequent diseases, affecting 5-10% and 30% of the population, respectively. The prevalence of asthma has increased in many developed countries, which may be due to several factors, including increased exposure to house dust mite (HDM) allergens. HDM to which humans are most frequently sensitized are Dermatophagoides pteronyssinus, Dermatophagoides farinae, and Euroglyphus maynei. These mites multiply in carpets, bedding and upholstered furniture in a hot and humid atmosphere. The allergens are digestive enzymes of the mites. Several epidemiological studies have shown that an increase in exposure to HDMs is associated with an increase in the prevalence of sensitization and asthma, whereas mite avoidance leads to a decrease in respiratory symptoms of sensitized asthmatic subjects. Sensitized subjects have specific immunoglobulin G and E (IgG and IgE) humoral responses, as well as proliferative T-cell responses to HDM allergens. Experimental exposure to HDM allergens induces bronchoalveolar inflammatory responses, that are characterized by the recruitment and activation of eosinophils, mastocytes, neutrophils, monocytes and lymphocytes. The cysteine protease activity of Der p 1 (a major allergen of D. pteronyssinus) has been shown to increase airway mucosal permeability, and may thereby contribute to the pathogenesis of airway inflammation and hyperresponsiveness by nonimmunological mechanisms. These epidemiological and experimental data support the recommendations for mite avoidance, especially in persons at high risk of developing asthma.

  20. Expression, purification and characterization of Der f 27, a new allergen from dermatophagoides farinae.

    PubMed

    Lin, Jianli; Li, Meng; Liu, Yulin; Jiang, Congli; Wu, Yulan; Wang, Yuanyuan; Gao, Anjian; Liu, Zhigang; Yang, Pingchang; Liu, Xiaoyu

    2015-01-01

    The house dust mite (HDM), Dermatophagoidesfarinae (D. farina), is one of the most important indoor allergen sources and a major elicitor of allergic asthma; itscharacterization is important in the diagnosis and immunotherapy of mite allergen-relevant diseases. This study aims to characterize a novel allergen, the D. farinae-derived serpin (Der f 27). In this study, the total RNA of D. farinae was extracted, and the Der f 27 gene was cloned and expressed. The allergenicity of recombinant Der f 27 protein was determined by enzyme-linked immunosorbent assay, and Western-blotting with the sera of asthma patients, and skin prick test (SPT) in allergic human subjects. A r-Der f 27 allergic asthma mouse model was established. The cloned Der f 27 gene has been presented at the Gene Bank with an accession number of KM009995. The IgE levels of r-Der f 27 in the serum from r-Der f 27 SPT positive allergic patients were 3 folds more than healthy subjects. The Der f 27 SPT positive ratewas 42.1% in 19 DM-SPT positive patients. Airway hyperresponsiveness, serum specific IgE, and levels of interleukin-4 in the spleen cell culture supernatant were significantly increased in allergic asthma mice sensitized to r-Der f 27. In conclusion, Der f 27 is a new subtype of house mite allergen.