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Sample records for allergenic extract manufacturers

  1. 21 CFR 680.2 - Manufacture of Allergenic Products.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 7 2014-04-01 2014-04-01 false Manufacture of Allergenic Products. 680.2 Section...) BIOLOGICS ADDITIONAL STANDARDS FOR MISCELLANEOUS PRODUCTS § 680.2 Manufacture of Allergenic Products. (a...) Cultures derived from microorganisms. Culture media into which organisms are inoculated for the...

  2. 21 CFR 680.2 - Manufacture of Allergenic Products.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 7 2011-04-01 2010-04-01 true Manufacture of Allergenic Products. 680.2 Section...) BIOLOGICS ADDITIONAL STANDARDS FOR MISCELLANEOUS PRODUCTS § 680.2 Manufacture of Allergenic Products. (a...) Cultures derived from microorganisms. Culture media into which organisms are inoculated for the...

  3. 21 CFR 680.2 - Manufacture of Allergenic Products.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 7 2013-04-01 2013-04-01 false Manufacture of Allergenic Products. 680.2 Section...) BIOLOGICS ADDITIONAL STANDARDS FOR MISCELLANEOUS PRODUCTS § 680.2 Manufacture of Allergenic Products. (a...) Cultures derived from microorganisms. Culture media into which organisms are inoculated for the...

  4. 21 CFR 680.2 - Manufacture of Allergenic Products.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 7 2012-04-01 2012-04-01 false Manufacture of Allergenic Products. 680.2 Section...) BIOLOGICS ADDITIONAL STANDARDS FOR MISCELLANEOUS PRODUCTS § 680.2 Manufacture of Allergenic Products. (a...) Cultures derived from microorganisms. Culture media into which organisms are inoculated for the...

  5. 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...

  6. Allergen extracts for immunotherapy: to mix or not to mix?

    PubMed

    Nony, Emmanuel; Martelet, Armelle; Jain, Karine; Moingeon, Philippe

    2016-03-01

    Allergen immunotherapy (AIT) is established as a curative treatment for allergic rhinitis, asthma, as well as insect venom allergy. AIT is based on the administration of natural allergen extracts via the subcutaneous or sublingual routes to reorient the immune system towards tolerogenic mechanisms. In this regard, since many patients are poly-allergic, mixtures of allergen extracts are often used with a potential risk to cause allergen degradation, thereby affecting treatment efficacy. Herein, we discuss the advantages and drawbacks of mixing homologous (i.e., related) or heterogeneous (i.e., unrelated) allergen extracts. We provide evidence for incompatibilities between mixes of grass pollen and house dust mite extracts containing bodies and feces, and summarize critical points to consider when mixing allergen extracts for AIT. PMID:26652799

  7. 21 CFR 680.2 - Manufacture of Allergenic Products.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 7 2010-04-01 2010-04-01 false Manufacture of Allergenic Products. 680.2 Section 680.2 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED...) Cultures derived from microorganisms. Culture media into which organisms are inoculated for the...

  8. Allergenic evaluation of Malassezia furfur crude extracts.

    PubMed

    Gandra, R F; Melo, T A; Matsumoto, F E; Pires, M F C; Croce, J; Gambale, W; Paula, C R

    2002-01-01

    Crude extracts of the lipophilic yeast Malassezia furfur were obtained from 2, 6, 10 and 28 day old cultures. The in vitro cultivation periods corresponded, respectively, to the lag phase, middle of the log phase, end of log phase and the decline phase of the growth curve, which was based on viable cell counts obtained with a fluorescent viability test. Biochemical analyses showed that the protein and carbohydrate contents were greater in day 10 extracts. Seventy patients with different allergic manifestations and 30 healthy volunteers were skin prick tested using the extracts. Of these, thirteen (18.57%) patients gave positive responses. SDS PAGE gradient electrophoretic profiles of the preparations indicated that the 28 day extracts contained the greatest number of protein bands with molecular weights ranging mostly between 30 and 94 kDa. Immunoblots incubated with individual patient sera showed that four IgE binding M. furfur allergens of approximately 88, 61, 52 and 39 kDa were present in the 28 day extracts. The components identified could be used for detecting IgE mediated responses to M. furfur among individuals affected with different allergic conditions. PMID:12650593

  9. [Evaluation of the total biological activity and allergenic composition of allergenic extracts].

    PubMed

    Lombardero, M; González, R; Duffort, O; Juan, F; Ayuso, R; Ventas, P; Cortés, C; Carreira, J

    1986-01-01

    In the present study, a complete procedure is presented in order to standardize allergenic extracts, the meaning of which is the measurement of the total allergenic activity and the determination of the allergenic composition. The measurement of the biological activity comprises 2 steps: Preparation of Reference Extracts and determination of their "in vivo" activity. Evaluation of the total allergenic activity of extracts for clinical use. Reference extracts were prepared from the main allergens and their "in vivo" biological activity was determined by a quantitative skin prick test in a sample of at least 30 allergic patients. By definition, the protein concentration of Reference Extract that produces, in the allergic population, a geometric mean wheal of 75 mm.2 has an activity of 100 biological units (BUs). The determination of the biological activity of a problem extract is made by RAST inhibition. The sample is compared with the corresponding Reference Extract by this technique and, from this comparison, it is possible to quantify the activity of the problem extract in biologic units (BUs) with clinical significance. Likewise, different techniques have been used to determine the allergenic composition of extracts. These techniques comprise 2 steps: Separation of the components of the extract. Identification of the components that bind specific human IgE. The separation of the components of the extract has been carried out by isoelectric focusing (IEF) and electrophoresis in the presence of sodium dodecyl sulphate (SDS-PAGE). In order to identify the allergenic components, an immunoblotting technique has been employed. The separated components in the IEF gel or SDS-PAGE gel are transferred to a nitrocellulose sheet and later on, this membrane is overlaid with a serum pool from allergic patients and a mouse monoclonal anti-human IgE, labelled with 125I. Finally, the autoradiography of the nitrocellulose membrane is obtained. In this way it is possible to compare

  10. PARTIAL CHARACTERIZATION OF ALLERGENS IN EXTRACTS OF STACHYBOTRYS CHARTARUM

    EPA Science Inventory

    PARTIAL CHARACTERIZATION OF ALLERGENS IN EXTRACTS OF Stachybotrys chartarum. M E Viana1, MJ Selgrade2, and M D Ward2. 1NCSU, Raleigh, NC, USA. 2NHEERL, ORD, US EPA, RTP, NC, USA.

    Exposure to Stachybotrys chartarum has been associated with the development of serious health ...

  11. Use of pulsed ultraviolet light to reduce the allergenic potency of soybean extracts.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pulsed ultraviolet light (PUV), a non-thermal food processing technology, is reported to be able to inactivate enzymes and reduce allergen levels from peanut extracts. The objective of this study was to determine if PUV would reduce the allergen levels and allergenic potency of soy extracts. Soy ext...

  12. Allergenic extracts from Metarhizium canisopliae: obtainment and characterization.

    PubMed

    Barbieri, R T; Croce, J; Gandra, R F; Gagete, E; Paula, C R; Gambale, W

    2005-01-01

    Metarhizium anisopliae is used as a biopesticide for insects that damage agricultural plantations like sugar cane and forage plants. In a previous study the sensitization to this fungus of asthmatic patients coming from sugar cane areas was showed. The aims of this work were: to compare crude extracts obtained with Tris-HCl and Coca liquid from several growth phases of M. anisopliae concerning the total content of proteins and their electrophoretic analysis profile; to evaluate in vivo allergic sensitization in Balb/c mice and allergic patients from a sugar cane area, and to characterize the allergenic fractions in the sera of patients positive for the prick test by means of Western-blotting. The extract obtained with Coca liquid on the 16th day was the one that presented the greatest number of proteic fractions, including all those present in the other extracts. Twelve fractions were verified in this extract with approximate molecular weights from 94 to 14 kDa. The allergenicity of the extract obtained on the 16th day was proven by the production of IgE antibodies in Balb/c mice, with titres of 200. Prick tests carried out with the extract of the 16th day in 79 atopic individuals (from sugar cane area), 35 atopic individuals (from urban area) and 11 non- atopic individuals showed respective positivity of 29%, 9% and 0%. The allergenic characterization in vitro was performed by means of Western blotting, and the fractions that reacted with the positive individuals' sera were those of approximate molecular weights of 67 kDa (95%); 20 kDa (55%); 94 kDa (36%); 34 and 36 kDa (23%); 43 and 48 kDa (14%); 16 kDa (9%) and 54kDa (5%). It was concluded that the crude allergenic extract, obtained with Coca liquid from the 16th day growth of Metarhizium anisopliae, contains allergenic fractions and can be used in diagnostic screening tests. PMID:16047714

  13. Allergen nomenclature.

    PubMed Central

    Marsh, D. G.; Goodfriend, L.; King, T. P.; Lowenstein, H.; Platts-Mills, T. A.

    1986-01-01

    This article presents a nomenclature system for allergens which has been officially recommended by the International Union of Immunological Societies (IUIS). The nomenclature is based on proposals of the IUIS Sub-Committee for Allergen Nomenclature and is applicable to highly purified, well-characterized allergens and to non-purified or partially purified allergenic extracts. PMID:3492310

  14. Using phenolic compounds to reduce the allergenic properties of peanut extracts and peanut butter slurries.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Since phenolic compounds may form insoluble complexes with proteins, we determined that their interaction with peanut allergens leads to a reduction in the allergenic properties of peanut extracts and peanut butter slurries. Phenolics, such as, caffeic acid, chlorogenic acid, and ferulic acid were e...

  15. 12 Comprehensive Detection of Allergens in Grass Pollen Extracts by Mass Spectrometry

    PubMed Central

    Augustin, Steffen; Mitulski, Liane; Cromwell, Oliver; Reese, Gerald; Nandy, Andreas

    2012-01-01

    Background More than 40% of type 1-allergic individuals suffer from hypersensitivity to grass pollen. Patients are treated traditionally with specific immunotherapy using pollen extracts derived from one or several different Pooideae species. While for several species the most important allergens (group 1 and group 5) have been identified, other allergens have either not been identified or sequence data are still missing. We have used mass spectrometry (MS) together with genetic and immunological methods to identify allergens in various grass pollen extracts. Methods Pollen extracts of 6 different grass species (Phleum pratense, Holcus lanatus, Lolium perenne, Dactylus glomerata, Festuca pratensis, Poa pratensis) and a mixture thereof were analyzed. For identification of allergens by MS, extracts were subjected to enzymatic digestion. Resulting peptides were separated by liquid chromatography and analyzed by tandem mass spectrometry. Protein identification was performed by searching both the NCBIPlant release and an individually designed database. The presence of individual allergens was confirmed with allergen-specific monoclonal antibodies. Unknown sequences were determined following cDNA synthesis from pollen RNA and allergen sequence amplification by PCR. Results Fes p 1 and Fes p 5 were identified by the PCR approach. MS analysis of pollen extracts from the 6 individual species resulted in detection of all known allergens including the newly identified Fes p 1 and Fes p 5. Based on the homology of allergens from different grass species, previously unknown sequences of representatives of groups 2, 3, 4, 7, 11, 12 and 13 were detected by MS in investigated extracts with high sequence coverage. Group 6 allergens could not be identified in some of the analyzed extracts. These findings are supported by immunological analyses and thus demonstrate the specificity of the applied method. Members of all allergen groups were identified in an extract mix prepared from

  16. Comparison of different extraction solutions for the analysis of allergens in hen's egg.

    PubMed

    Hildebrandt, S; Steinhart, H; Paschke, A

    2008-06-01

    An important requirement for the correct procedure of allergen analysis in hen's egg is to obtain complete and unaltered protein extracts. Besides the aim of a quantitative extraction of the allergens from the matrix, it is equally important not to alter their allergenic potential during the extraction process. This paper describes and compares six extraction solutions for the analysis of whole-egg proteins and allergens. These requirements were examined via protein determination according to Bradford [Bradford, M. M. (1976). Rapid and sensitive method for quantitation of microgram quantities of protein utilizing principle of protein-dye binding. Analytical Biochemistry, 72, 248-254] and Kjeldahl [Meyer, A. H. (2006). Lebensmittelrecht, Verlag C.H. Beck München, Stand: 1. February 2006, § 64, Lebensmittel- und Futtermittelgesetzbuch, Amtliche Sammlung von Untersuchungsmethoden, Nr. L 06.00-7] as well as the EAST-inhibition method. It could be demonstrated that the extraction with a urea solution (8M) led to significant interferences during the protein determination, and substantially reduced the allergenic potential of egg proteins. With all other extraction solutions adequate protein contents could be extracted. The highest protein content was achieved by the extraction with phosphate buffered saline followed by a Tween 20 solution, physiological saline, water, and acetate buffer. The results show that none of these extracts - except for the urea solution (8M) - was altered in its' allergenic potential. PMID:26065775

  17. 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.

  18. Removing Peanut Allergen Ara h 1 from Peanut Extracts Using p-Aminobenzamidine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rationale: Ara h 1 is one of 3 major allergens in peanut. Removing Ara h 1 from a peanut extract may produce a hypoallergenic peanut extract for immunotherapy and other purposes. Methods: Peanut extracts were treated overnight with and without 10 mM p-aminobenzamidine (pABA, a protease inhibitor) i...

  19. Effects of pulsed UV-light on peanut allergens in extracts and liquid peanut butter.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pulsed ultraviolet (PUV)-light, a non-thermal technology, was used to treat both peanut extracts and liquid peanut butter. The objective was to determine if such treatment would lead to a reduction in the allergenic potency of the peanut extract and butter. Peanut samples were PUV treated, using a X...

  20. Quantification of Sesquiterpene Lactones in Asteraceae Plant Extracts: Evaluation of their Allergenic Potential

    PubMed Central

    Salapovic, Helena; Geier, Johannes; Reznicek, Gottfried

    2013-01-01

    Sesquiterpene lactones (SLs), mainly those with an activated exocyclic methylene group, are important allergens in Asteraceae (Compositae) plants. As a screening tool, the Compositae mix, consisting of five Asteraceae plant extracts with allergenic potential (feverfew, tansy, arnica, yarrow, and German chamomile) is part of several national patch test baseline series. However, the SL content of the Compositae mix may vary due to the source material. Therefore, a simple spectrophotometric method for the quantitative measurement of SLs with the α-methylene-γ-butyrolactone moiety was developed, giving the percentage of allergenic compounds in plant extracts. The method has been validated and five Asteraceae extracts, namely feverfew (Tanacetum parthenium L.), tansy (Tanacetum vulgare L.), arnica (Arnica montana L.), yarrow (Achillea millefolium L.), and German chamomile (Chamomilla recutita L. Rauschert) that have been used in routine patch test screening were evaluated. A good correlation could be found between the results obtained using the proposed spectrophotometric method and the corresponding clinical results. Thus, the introduced method is a valuable tool for evaluating the allergenic potential and for the simple and efficient quality control of plant extracts with allergenic potential. PMID:24106675

  1. Multiplex Assay for Protein Profiling and Potency Measurement of German Cockroach Allergen Extracts

    PubMed Central

    Khurana, Taruna; Dobrovolskaia, Ekaterina; Shartouny, Jessica R.; Slater, Jay E.

    2015-01-01

    Background German cockroach (GCr) allergens induce IgE responses and may cause asthma. Commercial GCr allergen extracts are variable and existing assays may not be appropriate for determining extract composition and potency. Objective Our aim was to develop a multiplex antibody/bead-based assay for assessment of GCr allergen extracts. Methods Single chain fragment variable (scFv) antibodies against GCr were obtained by screening libraries derived from naïve human lymphocytes and hyperimmunized chicken splenocytes and bone marrow. Selected clones were sequenced and characterized by immunoblotting. Eighteen scFv antibodies (17 chicken, 1 human) coupled to polystyrene beads were used in this suspension assay; binding of targeted GCr allergens to antibody-coated beads was detected using rabbit antisera against GCr, and against specific allergens rBla g 1, rBla g 2, and rBla g 4. The assay was tested for specificity, accuracy, and precision. Extracts were also compared by IgE competition ELISA. Results Chicken scFv’s generated eight different binding patterns to GCr proteins from 14 to 150 kDa molecular weight. Human scFv’s recognized a 100 kDa GCr protein. The multiplex assay was found to be specific and reproducible with intra-assay coefficient of variation (CV) of 2.64% and inter-assay CV of 10.0%. Overall potencies of various GCr extracts were calculated using mean logEC50s for eight selected scFvs. Overall potency measures were also analyzed by assessing the contributions to potency of each target. Conclusions An scFv antibody-based multiplex assay has been developed capable of simultaneously measuring different proteins in a complex mixture, and to determine the potencies and compositions of allergen extracts. PMID:26444288

  2. Comparison of different immobilized systems in the removal of peanut allergens from peanut extracts.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to determine which of the magnetic-bead systems (Ca2+, Fe3+, caffeic acid, hydrophobic) would bind and separate peanut allergens from other proteins in a peanut extract more efficiently. Commercial Ca2+ and hydrophobic magnetic beads, and caffeic-beads (prepared by at...

  3. ASSESSMENT OF A CRUDE FUNGAL (METARHIZIUM ANISOPLIAE) EXTRACT AND IT'S COMPONENTS FOR ALLERGENICITY

    EPA Science Inventory

    ASSESSMENT OF A CRUDE FUNGAL (METARHIZIUM ANISOPLIAE) EXTRACT AND IT'S COMPONENTS FOR ALLERGENICITY. M D W Ward1, M E Viana2, L B Copeland1, and MJ K Selgrade1. 1US EPA, ORD, NHEERL, RTP, NC, USA. 2NCSU, College of Veterinary Medicine, Raleigh, NC, USA.
    Metarhizium anisopli...

  4. Tracing tree nut allergens in chocolate: A comparison of DNA extraction protocols.

    PubMed

    Costa, Joana; Melo, Vítor S; Santos, Cristina G; Oliveira, M Beatriz P P; Mafra, Isabel

    2015-11-15

    The present work aimed at comparing different DNA extraction methods, from chocolate matrices, for the effective application in molecular techniques to detect tree nut allergens. For this study, DNA from almond or hazelnut model chocolates was extracted using seven selected protocols: the in-house methods of CTAB-PVP (cetyltrimethylammonium bromide-polyvinylpyrrolidone), Wizard with and without RNase, Wizard-PVP with and without RNase, and the Wizard Magnetic and Nucleospin kits. The extracts were assessed for their suitability for amplification by qualitative PCR and real-time PCR. From the evaluated protocols, Nucleospin presented the best results for almond and hazelnut amplification, achieving a limit of detection of 0.005% (w/w) with high PCR efficiency, linearity and range of amplification. These results highlight the importance of the DNA extraction protocol in the case of food allergens from complex matrices, such as chocolate, in which sensitivity is a key parameter. PMID:25977052

  5. 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

  6. Inhalation of stable dust extract prevents allergen induced airway inflammation and hyperresponsiveness

    PubMed Central

    Peters, M; Kauth, M; Schwarze, J; Körner‐Rettberg, C; Riedler, J; Nowak, D; Braun‐Fahrländer, C; von Mutius, E; Bufe, A; Holst, O

    2006-01-01

    Background Recent epidemiological studies have shown that growing up on a traditional farm provides protection from the development of allergic disorders such as hay fever and allergic asthma. We present experimental evidence that substances providing protection from the development of allergic diseases can be extracted from dust collected in stables of animal farms. Methods Stable dust was collected from 30 randomly selected farms located in rural regions of the Alps (Austria, Germany and Switzerland). The dust was homogenised with glass beads and extracted with physiological sodium chloride solution. This extract was used to modulate immune response in a well established mouse model of allergic asthma. Results Treatment of mice by inhalation of stable dust extract during sensitisation to ovalbumin inhibited the development of airway hyperresponsiveness and airway eosinophilia upon challenge, as well as the production of interleukin 5 by splenocytes and of antigen specific IgG1 and IgE. Dust extract also suppressed the generation of human dendritic cells in vitro. The biological activity of the dust extract was not exclusively mediated by lipopolysaccharide. Conclusions Stable dust from animal farms contains strong immune modulating substances. These substances can interfere with the development of both cellular and humoral immunity against allergens, thus suppressing allergen sensitisation, airway inflammation, and airway hyperresponsiveness in a murine model of allergic asthma. PMID:16244088

  7. COMPARISON OF SUBCUTANEOUS AND ORAL ROUTES OF EXPOSURE FOR EVALUATING ALLERGENICITY OF FOOD EXTRACTS

    EPA Science Inventory

    Evaluation of the potential for food allergenicity of any given protein is limited by the lack of an appropriate animal model. In this study we examined the intrinsic allergenicity of foods known to be allergenic (peanut, egg) or non-allergenic (spinach) by exposing mice either s...

  8. Food allergen analysis for processed food using a novel extraction method to eliminate harmful reagents for both ELISA and lateral-flow tests.

    PubMed

    Ito, Kaori; Yamamoto, Takayuki; Oyama, Yuriko; Tsuruma, Rieko; Saito, Eriko; Saito, Yoshikazu; Ozu, Takeshi; Honjoh, Tsutomu; Adachi, Reiko; Sakai, Shinobu; Akiyama, Hiroshi; Shoji, Masahiro

    2016-09-01

    Enzyme-linked immunosorbent assay (ELISA) is commonly used to determine food allergens in food products. However, a significant number of ELISAs give an erroneous result, especially when applied to highly processed food. Accordingly, an improved ELISA, which utilizes an extraction solution comprising the surfactant sodium lauryl sulfate (SDS) and reductant 2-mercaptoethanol (2-ME), has been specially developed to analyze food allergens in highly processed food by enhancing analyte protein extraction. Recently, however, the use of 2-ME has become undesirable. In the present study, a new extraction solution containing a human- and eco-friendly reductant, which is convenient to use at the food manufacturing site, has been established. Among three chemicals with different reducing properties, sodium sulfite, tris(3-hydroxypropyl)phosphine, and mercaptoethylamine sodium sulfite was selected as a 2-ME substitute. The protein extraction ability of SDS/0.1 M sodium sulfite solution was comparable to that of SDS/2-ME solution. Next, the ELISA performance for egg, milk, wheat, peanut, and buckwheat was evaluated by using model-processed foods and commercially available food products. The data showed that the SDS/0.1 M sulfite ELISA significantly correlated with the SDS/2-ME ELISA for all food allergens examined (p < 0.01), thereby establishing the validity of the SDS/0.1 M sulfite ELISA performance. Furthermore, the new SDS/0.1 M sulfite solution was investigated for its applicability to the lateral-flow (LF) test. The result demonstrated the successful analysis of food allergens in processed food, showing consistency with the SDS/0.1 M sulfite ELISA results. Accordingly, a harmonized analysis system for processed food comprising a screening LF test and a quantitative ELISA with identical extraction solution has been established. The ELISA based on the SDS/0.1 M sulfite extraction solution has now been authorized as the revised official method for food allergen

  9. Impact of the vulcanization process on the structural characteristics and IgE recognition of two allergens, Hev b 2 and Hev b 6.02, extracted from latex surgical gloves.

    PubMed

    Galicia, Christian; Mendoza-Hernández, Guillermo; Rodríguez-Romero, Adela

    2015-06-01

    Latex allergy is a health problem that mainly affects medical environments, causing anaphylactic shocks in extreme cases. Sensitization and reactions to this material is closely linked to the use of latex gloves. The objective of this study was to purify two of the major allergens from latex surgical gloves to study the biochemical and structural changes that could be generated during the product manufacture and to compare their IgE recognition with the non-processed allergens. Glycosylated allergen Hev b 2 (β-1,3-glucanase) and Hev b 6.02 (hevein) were purified from glove extracts using affinity (Concanavalin A) and reversed-phase chromatographies, respectively. ELISA experiments were performed with both proteins and sera from allergic patients to assess the IgE recognition, which was heterogeneous. Crystallographic methods were used to obtain the 3D structure of Hev b 6.02 from surgical gloves, which did not show evident modification when compared with the protein from the natural non-processed form. Despite having the same crystallographic structure, the IgE from some patients showed different recognition when the glove and the natural allergen were used in ELISA. Furthermore, using electrophoretic techniques, we identified three forms of Hev b 2: one corresponding to the complete polypeptide chain with posttranslational modifications, and two glycosylated fragments. The mixture of these three forms showed stronger recognition by IgE from latex-allergic patients than the pure non-processed allergen. In conclusion, IgE from subjects sensitized to latex products showed different recognition between the allergens obtained from a natural source and the processed material, even when the structure was maintained. This demonstrates the importance of using processed allergens in further investigations of diagnosis, prevalence, product allergenicity, and therapies. PMID:25700348

  10. Evaluation of reduced allergenicity of irradiated peanut extract using splenocytes from peanut-sensitized mice

    NASA Astrophysics Data System (ADS)

    Oh, Sejo; Jang, Da-In; Lee, Ju-Woon; Kim, Jae-Hun; Byun, Myung-Woo; Lee, Soo-Young

    2009-07-01

    Peanut (PN) allergy is one of the most serious forms of IgE-mediated food hypersensitivity. Gamma irradiation has been widely used for the preservation of food. The results of our previous studies showed that the IgE-binding capacity to several antigens were profoundly reduced after gamma irradiation. In this study, we evaluated the changes of allergenecity and cytokine production profiles after exposure of irradiated PN extract in a PN-allergy mouse model. Mice were sensitized to PN extract by intragastric administration on days 0, 1, 2, and 7, and then challenged on day 21. Four weeks later, we evaluated the cytokine production patterns and proliferation responses of splenocytes that were stimulated with intact PN extract, compared to 10 and 50 kGy irradiated PN extract. When the cells were stimulated with 10 kGy of irradiated PN extract, a higher level of production of IFN-γ and IL-10 cytokines was observed. However, stimulation with 50 kGy of irradiated PN extract resulted in a higher level of production of only IFN-γ cytokines. In addition, the Th1/Th2 ratio increased in response to treatment with gamma-irradiated PNs. The results of this study show that the allergenicity of PN extracts could be reduced by gamma irradiation which caused downregulation of Th2 lymphocyte activity in the PN-sensitized mice.

  11. The innate response to peanut extract in ovine afferent lymph and its correlation with allergen sensitisation.

    PubMed

    Van Gramberg, Jenna L; Bischof, Robert J; O'Hehir, Robyn E; de Veer, Michael J; Meeusen, Els N

    2015-07-01

    The innate response generated after initial allergen exposure is crucial for polarising adaptive immunity, but little is known about how it drives an atopic or type-2 immune response. The present study characterises the response of skin-draining afferent lymph in sheep following injection with peanut (PN) extract in the presence or absence of aluminium hydroxide (AlOH) adjuvant. Lymph was collected and innate cell populations characterised over an 84 h time period. The innate response to PN extract in afferent lymph displayed an early increase in neutrophils and monocytes without any changes in the dendritic cell (DC) population. PN antigen was transported by neutrophils and monocytes for the first 36 h, after which time DCs were the major antigen trafficking cells. AlOH adjuvant gradually increased antigen uptake by DCs at the later time points. Following lymphatic characterisation, sheep were sensitised with PN extract by three subcutaneous injections of PN in AlOH, and the level of PN-specific immunoglobulin E (IgE) was determined. Sheep with higher levels of steady-state DCs in afferent lymph showed increased monocytic recruitment in afferent lymph and reduced PN-specific IgE following sensitisation. In addition, DCs from afferent lymph that had ingested PN antigen increased the expression of monocyte chemoattractant mRNA. The results of this study show that the innate response to PN extract involves a dynamic change in cell populations in the afferent lymph over time. In addition, DCs may determine the strength of the initial inflammatory cell response, which in turn may determine the nature of the antigen-specific adaptive response. PMID:25666095

  12. 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

  13. Allergens in veterinary medicine.

    PubMed

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

    2016-01-01

    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

  14. Allergen Quantification by Use of Electrostatic Dust Collectors (EDCs): Influence of Deployment Time, Extraction Buffer, and Storage Conditions on the Results.

    PubMed

    Sander, Ingrid; Lotz, Anne; Zahradnik, Eva; Raulf, Monika

    2016-08-01

    Sampling of endotoxin, beta-glucan, or allergens on electrostatic dust collectors (EDCs) is a convenient method for exposure assessment. However, especially for allergens few experiments on validation of this method concerning deployment time or storage and extraction procedure have been performed. The aim of study was to optimize the EDC procedure for sampling of allergens in indoor environments. EDCs were placed in households or day-care centers and after extraction, allergens were quantified by six immunoassays detecting mite antigens (Domestic mites DM, Dermatophagoides pteronyssinus Dp, Tyrophagus putrescentiae Tp) or the main allergens from cat (Fel d 1), dog (Can f 1) and mouse (Mus m 1). For 20 EDC holders, deployment times of cloths were varied between 7 and 28 days, 36 EDCs were used to test reproducibility, and for 28 EDCs extraction buffers were varied (with or without 0.05% Tween 20, borate, or phosphate buffer). The influence of storage of cloths at room temperature (2-629 days) or extracts at -80°C (7-639 days), and variation of extract storage temperature (-20°C and -80°C) for long time storage (1.5 years) on the outcome of allergen quantification were tested for about 150 EDCs. The allergens on EDC cloths increased proportionally with deployment time, and allergen loads on parallel sampled tissues were significantly correlated (P < 0.0001, Pearson of log-transformed values 0.91-0.99). Extraction without Tween reduced all results (P < 0.0001, -51% DM, -85% Dp, -60% Tp, -99% Fel d 1, -86% Can f 1, -52% Mus m 1), and borate buffer resulted in lower yields of Mus m 1 (-53%), DP (-45%), and Tp (-27%) than phosphate buffer. Storage of cloths at room temperature significantly decreased Can f 1 levels (P < 0.0001, -4.8% loss for every 30 days), whereas storage of extracts at -80°C decreased DM results (P < 0.0001, -1.2% loss for every 30 days). Extracts stored at -20°C gave at mean 12% higher DM results compared to extracts stored at -80°C for 1

  15. 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

  16. Computational detection of allergenic proteins attains a new level of accuracy with in silico variable-length peptide extraction and machine learning

    PubMed Central

    Soeria-Atmadja, D.; Lundell, T.; Gustafsson, M. G.; Hammerling, U.

    2006-01-01

    The placing of novel or new-in-the-context proteins on the market, appearing in genetically modified foods, certain bio-pharmaceuticals and some household products leads to human exposure to proteins that may elicit allergic responses. Accurate methods to detect allergens are therefore necessary to ensure consumer/patient safety. We demonstrate that it is possible to reach a new level of accuracy in computational detection of allergenic proteins by presenting a novel detector, Detection based on Filtered Length-adjusted Allergen Peptides (DFLAP). The DFLAP algorithm extracts variable length allergen sequence fragments and employs modern machine learning techniques in the form of a support vector machine. In particular, this new detector shows hitherto unmatched specificity when challenged to the Swiss-Prot repository without appreciable loss of sensitivity. DFLAP is also the first reported detector that successfully discriminates between allergens and non-allergens occurring in protein families known to hold both categories. Allergenicity assessment for specific protein sequences of interest using DFLAP is possible via ulfh@slv.se. PMID:16977698

  17. Transition of recombinant allergens from bench to clinical application.

    PubMed

    Cromwell, Oliver; Suck, Roland; Kahlert, Helga; Nandy, Andreas; Weber, Bernhard; Fiebig, Helmut

    2004-03-01

    The cloning and production of an increasing number of allergens through the use of DNA technology has provided the opportunity to use these proteins instead of natural allergen extracts for the diagnosis and therapy of IgE-mediated allergic disease. For diagnostic purposes, it is essential that the molecules exhibit IgE-reactivity comparable with that of the natural wild-type molecules, whereas T cell reactivity and immunogenic activity may be more important for allergen-specific immunotherapy. In relation to the latter, the development of hypoallergenic recombinant allergen variants is an approach which shows great promise. Clinical application of the proteins requires that they must be produced under conditions of Good Manufacturing Practice and meet the specifications set down in the appropriate Regulatory Guidelines, principally the ICH-Guidelines. Special consideration has to be given to the choice of expression system, the design of the expression vectors, and the purification strategy to obtain a pure product free from toxins and contamination. The availability of the pure recombinant molecules provides the opportunity to formulate preparations that are free from the non-allergenic ballast proteins present in natural allergen extracts and which contain relative concentrations of the allergens in clinically appropriate proportions. PMID:14962765

  18. Treatment of cashew extracts with Aspergillopepsin reduces IgE binding to cashew allergens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cashew nuts can cause serious and sometimes life threatening reactions in people that suffer from food allergies. These reactions are mediated by immunoglobulin E binding (IgE) to allergenic cashew proteins. Enzymes from Aspergillus fungal species are used in many industrial and pharmaceutical appli...

  19. 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...

  20. Reducing the allergenic capacity of peanut extracts and liquid peanut butter by phenolic compounds.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phenolic compounds are known to form soluble and insoluble complexes with proteins. The objective of this study was to determine if phenolics, such as, caffeic, chlorogenic, and ferulic acids form insoluble and irreversible complexes with major peanut allergens. We also tested whether such complexat...

  1. Effect of phenolic compounds on the allergenic properties of peanut extracts and peanut butter slurries.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phenolic compounds (PCs) are phytochemicals and antioxidants with known health benefits. They are known to bind to proteins as soluble and insoluble complexes. As soluble complexes, with major peanut allergens formed in the presence of polyphenol oxidase (PPO), PCs have been shown to be able to redu...

  2. Vaccine development for allergen-specific immunotherapy based on recombinant allergens and synthetic allergen peptides: Lessons from the past and novel mechanisms of action for the future

    PubMed Central

    Valenta, Rudolf; Campana, Raffaela; Focke-Tejkl, Margit; Niederberger, Verena

    2016-01-01

    In the past, the development of more effective, safe, convenient, broadly applicable, and easy to manufacture vaccines for allergen-specific immunotherapy (AIT) has been limited by the poor quality of natural allergen extracts. Progress made in the field of molecular allergen characterization has now made it possible to produce defined vaccines for AIT and eventually for preventive allergy vaccination based on recombinant DNA technology and synthetic peptide chemistry. Here we review the characteristics of recombinant and synthetic allergy vaccines that have reached clinical evaluation and discuss how molecular vaccine approaches can make AIT more safe and effective and thus more convenient. Furthermore, we discuss how new technologies can facilitate the reproducible manufacturing of vaccines of pharmaceutical grade for inhalant, food, and venom allergens. Allergy vaccines in clinical trials based on recombinant allergens, recombinant allergen derivatives, and synthetic peptides allow us to target selectively different immune mechanisms, and certain of those show features that might make them applicable not only for therapeutic but also for prophylactic vaccination. PMID:26853127

  3. Vaccine development for allergen-specific immunotherapy based on recombinant allergens and synthetic allergen peptides: Lessons from the past and novel mechanisms of action for the future.

    PubMed

    Valenta, Rudolf; Campana, Raffaela; Focke-Tejkl, Margit; Niederberger, Verena

    2016-02-01

    In the past, the development of more effective, safe, convenient, broadly applicable, and easy to manufacture vaccines for allergen-specific immunotherapy (AIT) has been limited by the poor quality of natural allergen extracts. Progress made in the field of molecular allergen characterization has now made it possible to produce defined vaccines for AIT and eventually for preventive allergy vaccination based on recombinant DNA technology and synthetic peptide chemistry. Here we review the characteristics of recombinant and synthetic allergy vaccines that have reached clinical evaluation and discuss how molecular vaccine approaches can make AIT more safe and effective and thus more convenient. Furthermore, we discuss how new technologies can facilitate the reproducible manufacturing of vaccines of pharmaceutical grade for inhalant, food, and venom allergens. Allergy vaccines in clinical trials based on recombinant allergens, recombinant allergen derivatives, and synthetic peptides allow us to target selectively different immune mechanisms, and certain of those show features that might make them applicable not only for therapeutic but also for prophylactic vaccination. PMID:26853127

  4. Pressurized liquid extraction-gas chromatography-mass spectrometry analysis of fragrance allergens, musks, phthalates and preservatives in baby wipes.

    PubMed

    Celeiro, Maria; Lamas, J Pablo; Garcia-Jares, Carmen; Llompart, Maria

    2015-03-01

    Baby wipes and wet toilet paper are specific hygiene care daily products used on newborn and children skin. These products may contain complexes mixtures of harmful chemicals. A method based on pressurized liquid extraction (PLE) followed by gas chromatography-mass spectrometry (GC-MS) has been developed for the simultaneous determination of sixty-five chemical compounds (fragrance allergens, preservatives, musks, and phthalates) in wipes and wet toilet paper for children. These compounds are legislated in Europe according Regulation EC No 1223/2009, being twelve of them banned for their use in cosmetics, and one of them, 3-iodo-2-propynyl butylcarbamate (IPBC), is banned in products intended for children under 3 years. Also, propyl-, and butylparaben will be prohibited in leave-on cosmetic products designed for application on the nappy area of children under 3 years from April 2015. PLE is a fast, simple, easily automated technique, which permits to integrate a clean-up step during the extraction process reducing analysis time and stages. The proposed PLE-based procedure was optimized on real non-spiked baby wipe samples by means of experimental design to study the influence on extraction of parameters such as extraction solvent, temperature, extraction time, and sorbent type. Under the selected conditions, the method was validated showing satisfactory linearity, and intra-day, and inter-day precision. Recoveries were between 80-115% for most of the compounds with relative standard deviations (RSD) lower than 15%. Finally, twenty real samples were analyzed. Thirty-six of the target analytes were detected, highlighting the presence of phenoxyethanol in all analyzed samples at high concentration levels (up to 0.8%, 800μgg(-1)). Methyl paraben (MeP), and ethyl paraben (EtP) were found in 40-50% of the samples, and the recently banned isobutyl paraben (iBuP) and isopropyl paraben (iPrP), were detected in one and seven samples, respectively, at concentrations between

  5. 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

  6. Blomia tropicalis Blo t 5 and Blo t 21 recombinant allergens might confer higher specificity to serodiagnostic assays than whole mite extract

    PubMed Central

    2013-01-01

    Background Blomia tropicalis is a dust mite and an important source of allergens in tropical regions. Up to now, the assays to diagnose atopy to this mite use whole body extract as antigens. However, anti-B. tropicalis IgE antibodies cross-react with Ascaris lumbricoides antigens, hindering the diagnosis of allergy to this mite. In this study, B. tropicalis recombinant allergens were evaluated with the purpose of developing an immunodiagnostic assay for allergy to this mite with greater specificity than those commercially available. Methods Two B. tropicalis allergens (Blo t 5 and Blo t 21) were cloned into a plasmidial expression vector, expressed in Escherichia coli and purified by affinity chromatography. Sixty-three sera containing anti-B. tropicalis extract (BtE) IgE antibodies were used to investigate IgE reactivity to the recombinant Blot 5 and 21 allergens. Inhibition assays with 20 sera pre-adsorbed with A. lumbricoides extract were performed using rBlo t 5, rBlo t 21, and BtE as antigens. All the assays were carried using indirect ELISA. Results Eighty-two point nine percent and 80.0% of the sera with anti-BtE antibodies from 35 children reacted with rBlo t 5 and rBlo t 21, respectively, whereas 92.8% and 89.3% of the 28 sera with anti-BtE antibodies from adult asthma patients reacted with the same allergens, and 96.4% of these sera reacted with a mixture of rBlo t 5 and rBlo t 21. In an inhibition ELISA, the absorption of sera by A. lumbricoides extract affected less the reaction with rBlo t 5 and rBlo t 21 than with BtE. Conclusions The rBlo t 5 and rBlo t 21 allergens contain important epitopes recognized by IgE antibodies of individuals allergic to B. tropicalis antigens. Moreover, the assays using the recombinant allergens had lower IgE cross-reactivity with A. lumbricoides antigens, a fact which would confers higher specificity to serodiagnostic assays than the crude mite extract. However, additional recombinant allergens should be evaluated in

  7. Pimecrolimus Is a Potent Inhibitor of Allergic Reactions to Hymenopteran Venom Extracts and Birch Pollen Allergen In Vitro.

    PubMed

    Heneberg, Petr; Riegerová, Kamila; Kučera, Petr

    2015-01-01

    Pimecrolimus (Elidel, SDZ ASM 981) is an anti-inflammatory and immunomodulatory 33-epichloro-derivative of macrolactam ascomycin, with low potential for affecting systemic immune responses compared with other calcineurin inhibitors, cyclosporin A and tacrolimus. Despite numerous studies focused on the mechanism of pimecrolimus action on mast cells, only the single report has addressed pimecrolimus effects on other typical FcεRI-expressing cells, the basophils. Patients allergic to birch pollen (n = 20), hymenopteran venoms (n = 23) and 10 non-allergic volunteers were examined. Primary human basophils pre-treated or not with 0.5-50 μMol pimecrolimus were exposed to various concentrations of recombinant Bet v 1a allergen, bee or wasp venom extracts and anti-IgE for 20 min, and then examined for the expression of CD45, CD193, CD203c, CD63 and CD164 using flow cytometry. The externalization of basophil activation markers (CD63 and CD164) was equally inhibited through pimecrolimus in cells activated by recombinant pollen allergen, hymenopteran venom extracts and anti-IgE. Although the individual response rate was subject to strong variation, importantly, pre-treatment with pimecrolimus lowered the number of activated basophils in response to any of the stimuli in the basophils from all patients. The inhibition was concentration-dependent; approximately half of the basophils were inhibited in the presence of 2.5 mMol pimecrolimus. Pimecrolimus is a valuable new tool for the inhibition of hyper-reactive basophils in patients with pollen allergy and a history of anaphylactic reactions to bee or wasp venoms. Further research should address short-term use of pimecrolimus in vivo in a wide spectrum of allergic diseases. PMID:26562153

  8. Pimecrolimus Is a Potent Inhibitor of Allergic Reactions to Hymenopteran Venom Extracts and Birch Pollen Allergen In Vitro

    PubMed Central

    Heneberg, Petr; Riegerová, Kamila; Kučera, Petr

    2015-01-01

    Pimecrolimus (Elidel, SDZ ASM 981) is an anti-inflammatory and immunomodulatory 33-epichloro-derivative of macrolactam ascomycin, with low potential for affecting systemic immune responses compared with other calcineurin inhibitors, cyclosporin A and tacrolimus. Despite numerous studies focused on the mechanism of pimecrolimus action on mast cells, only the single report has addressed pimecrolimus effects on other typical FcεRI-expressing cells, the basophils. Patients allergic to birch pollen (n = 20), hymenopteran venoms (n = 23) and 10 non-allergic volunteers were examined. Primary human basophils pre-treated or not with 0.5–50 μMol pimecrolimus were exposed to various concentrations of recombinant Bet v 1a allergen, bee or wasp venom extracts and anti-IgE for 20 min, and then examined for the expression of CD45, CD193, CD203c, CD63 and CD164 using flow cytometry. The externalization of basophil activation markers (CD63 and CD164) was equally inhibited through pimecrolimus in cells activated by recombinant pollen allergen, hymenopteran venom extracts and anti-IgE. Although the individual response rate was subject to strong variation, importantly, pre-treatment with pimecrolimus lowered the number of activated basophils in response to any of the stimuli in the basophils from all patients. The inhibition was concentration-dependent; approximately half of the basophils were inhibited in the presence of 2.5 mMol pimecrolimus. Pimecrolimus is a valuable new tool for the inhibition of hyper-reactive basophils in patients with pollen allergy and a history of anaphylactic reactions to bee or wasp venoms. Further research should address short-term use of pimecrolimus in vivo in a wide spectrum of allergic diseases. PMID:26562153

  9. Quantitative and qualitative optimization of allergen extraction from peanut and selected tree nuts. Part 1. Screening of optimal extraction conditions using a D-optimal experimental design.

    PubMed

    L'Hocine, Lamia; Pitre, Mélanie

    2016-03-01

    A D-optimal design was constructed to optimize allergen extraction efficiency simultaneously from roasted, non-roasted, defatted, and non-defatted almond, hazelnut, peanut, and pistachio flours using three non-denaturing aqueous (phosphate, borate, and carbonate) buffers at various conditions of ionic strength, buffer-to-protein ratio, extraction temperature, and extraction duration. Statistical analysis showed that roasting and non-defatting significantly lowered protein recovery for all nuts. Increasing the temperature and the buffer-to-protein ratio during extraction significantly increased protein recovery, whereas increasing the extraction time had no significant impact. The impact of the three buffers on protein recovery varied significantly among the nuts. Depending on the extraction conditions, protein recovery varied from 19% to 95% for peanut, 31% to 73% for almond, 17% to 64% for pistachio, and 27% to 88% for hazelnut. A modulation by the buffer type and ionic strength of protein and immunoglobuline E binding profiles of extracts was evidenced, where high protein recovery levels did not always correlate with high immunoreactivity. PMID:26471618

  10. Allergic inflammatory response to short ragweed allergenic extract in HLA-DQ transgenic mice lacking CD4 gene.

    PubMed

    Chapoval, Svetlana P; Iijima, Koji; Marietta, Eric V; Smart, Michele K; Chapoval, Andrei I; Andrews, Amy G; David, Chella S

    2002-01-15

    To investigate the role of HLA-DQ molecules and/or CD4(+) T cells in the pathogenesis of allergic asthma, we generated HLA-DQ6 and HLA-DQ8 transgenic mice lacking endogenous class II (Abeta(null)) and CD4 genes and challenged them intranasally with short ragweed allergenic extract (SRW). We found that DQ6/CD4(null) mice developed a strong eosinophilic infiltration into the bronchoalveolar lavage and lung tissue, while DQ8/CD4(null) mice were normal. However, neither cytokines nor eosinophil peroxidase in the bronchoalveolar lavage of DQ6/CD4(null) mice was found. In addition, the airway reactivity to methacholine was elevated moderately in DQ6/CD4(null) mice compared with the high response in DQ/CD4(+) counterparts and was only partially augmented by CD4(+) T cell transfer. The DQ6/CD4(null) mice showed Th1/Th2-type cytokines and SRW-specific Abs in the immune sera in contrast to a direct Th2 response observed in DQ6/CD4(+) mice. The proliferative response of spleen mononuclear cells and peribronchial lymph node cells demonstrated that the response to SRW in DQ6/CD4(null) mice was mediated by HLA-DQ-restricted CD4(-)CD8(-)NK1.1(-) T cells. FACS analysis of PBMC and spleen mononuclear cells demonstrated an expansion of double-negative (DN) CD4(-)CD8(-)TCRalphabeta(+) T cells in SRW-treated DQ6/CD4(null) mice. These cells produced IL-4, IL-5, IL-13, and IFN-gamma when stimulated with immobilized anti-CD3. IL-5 ELISPOT assay revealed that DN T cells were the cellular origin of IL-5 in allergen-challenged DQ6/CD4(null) mice. Our study shows a role for HLA-DQ-restricted CD4(+) and DN T cells in the allergic response. PMID:11777987

  11. Sequential extractions: A new way for protein quantification-data from peanut allergens.

    PubMed

    Zhou, Ningling; Li, Wenying; Wu, Zhihua; Li, Xin; Yang, Anshu; Tong, Ping; Chen, Hongbing

    2015-09-01

    Quantification of certain protein contents in the matrix is essential in protein analyses. The amount of total protein in the matrix can be determined by the Kjeldahl method. However, few methods can quantify certain protein contents in the matrix without extracting all of them in solution. Extracting all of the contents is difficult for proteins, especially relatively insoluble ones. A five-step sequential extraction method was developed for the quantification of certain proteins in defatted peanut flour based on the relationship between the extracted protein contents and the extraction times. The extracted proteins (i.e., total protein, Ara h 1, and Ara h 2) were quantitatively analyzed in each extraction of the same condition. An exponential equation was obtained between the extraction times and the respective amount of extracted protein as well as both the total protein and a particular protein. In particular, the amount of protein extracted each time can be a geometric sequence. If all proteins can be extracted with sufficient extraction times, the protein contents in the peanut matrix can be calculated using a mathematical summation formula. This sum should be all proteins in the matrix. The five-step sequential extraction method can provide a means to quantify certain proteins in the matrix. PMID:26026388

  12. Advances in the risk management of unintended presence of allergenic foods in manufactured food products--an overview.

    PubMed

    Hattersley, Sue; Ward, Rachel; Baka, Athanasia; Crevel, René W R

    2014-05-01

    Food allergy is a relatively recent newcomer to the ranks of food safety issues, only being effectively recognised as such in the last 25-30 years. This recognition, allied with the near impossibility of avoiding the unintended presence of small, yet potentially dangerous residues of allergenic constituents, brought with it the need to assess and manage the resulting risk. This paper provides an overview of the development and current knowledge and thinking on risk assessment and its application to risk management of food allergens. It also discusses the associated challenges, in particular those around communicating meaningfully that risk to allergic consumers, including the use of precautionary labelling. The paper also provides an introductory context to the more detailed analyses of these issues in the following papers, based on the deliberations of a recent stakeholder workshop. The paper concludes that consistent risk management approaches using agreed quantitative action levels based on scientifically robust principles will provide optimal protection to allergic consumers. Growing amounts of data from oral food challenges along with the parallel development of risk assessment methodologies, such as probabilistic modelling, offer a realistic possibility of agreement among stakeholders on such levels in the near future. PMID:24508586

  13. Direct contact between dendritic cells and bronchial epithelial cells inhibits T cell recall responses towards mite and pollen allergen extracts in vitro.

    PubMed

    Papazian, D; Wagtmann, V R; Hansen, S; Würtzen, P A

    2015-08-01

    Airway epithelial cells (AECs) form a polarized barrier along the respiratory tract. They are the first point of contact with airborne antigens and are able to instruct resident immune cells to mount appropriate immune responses by either soluble or contact-dependent mechanisms. We hypothesize that a healthy, polarized epithelial cell layer inhibits inflammatory responses towards allergens to uphold homeostasis. Using an in-vitro co-culture model of the airway epithelium, where a polarized cell layer of bronchial epithelial cells can interact with dendritic cells (DCs), we have investigated recall T cell responses in allergic patients sensitized to house dust mite, grass and birch pollen. Using allergen extract-loaded DCs to stimulate autologous allergen-specific T cell lines, we show that AEC-imprinted DCs inhibit T cell proliferation significantly of Bet v 1-specific T cell lines as well as decrease interleukin (IL)-5 and IL-13 production, whereas inhibition of Phl p 5-specific T cells varied between different donors. Stimulating autologous CD4(+) T cells from allergic patients with AEC-imprinted DCs also inhibited proliferation significantly and decreased production of both T helper type 1 (Th1) and Th2 cytokines upon rechallenge. The inhibitory effects of AECs' contact with DCs were absent when allergen extract-loaded DCs had been exposed only to AECs supernatants, but present after direct contact with AECs. We conclude that direct contact between DCs and AECs inhibits T cell recall responses towards birch, grass and house dust mite allergens in vitro, suggesting that AECs-DC contact in vivo constitute a key element in mucosal homeostasis in relation to allergic sensitisation. PMID:25707463

  14. 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

  15. 78 FR 66011 - Allergenic Products Advisory Committee; Notice of Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-04

    ..., perennial rye, Timothy, and Kentucky bluegrass mixed pollens allergen extract tablet for sublingual use... recommendations on the safety and efficacy of Grastek, a Timothy grass pollen allergen extract tablet...

  16. 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

  17. The relative allergenicity of Stachybotrys chartarum compared to house dust mite extracts in a mouse model

    EPA Science Inventory

    A report by the Institute of Medicine suggested that more research is needed to better understand mold effects on allergic disease, particularly asthma development. The authors compared the ability of the fungus Stachybotrys chartarum (SCE) and house dust mite (HDM) extracts to i...

  18. 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

  19. Quantitative and qualitative optimization of allergen extraction from peanut and selected tree nuts. Part 2. Optimization of buffer and ionic strength using a full factorial experimental design.

    PubMed

    L'Hocine, Lamia; Pitre, Mélanie

    2016-03-01

    A full factorial design was used to assess the single and interactive effects of three non-denaturing aqueous (phosphate, borate, and carbonate) buffers at various ionic strengths (I) on allergen extractability from and immunoglobulin E (IgE) immunoreactivity of peanut, almond, hazelnut, and pistachio. The results indicated that the type and ionic strength of the buffer had different effects on protein recovery from the nuts under study. Substantial differences in protein profiles, abundance, and IgE-binding intensity with different combinations of pH and ionic strength were found. A significant interaction between pH and ionic strength was observed for pistachio and almond. The optimal buffer system conditions, which maximized the IgE-binding efficiency of allergens and provided satisfactory to superior protein recovery yield and profiles, were carbonate buffer at an ionic strength of I=0.075 for peanut, carbonate buffer at I=0.15 for almond, phosphate buffer at I=0.5 for hazelnut, and borate at I=0.15 for pistachio. The buffer type and its ionic strength could be manipulated to achieve the selective solubility of desired allergens. PMID:26471623

  20. Dermatophagoides farinae Allergens Diversity Identification by Proteomics*

    PubMed Central

    An, Su; Chen, Lingling; Long, Chengbo; Liu, Xiaoyu; Xu, Xuemei; Lu, Xingre; Rong, Mingqiang; Liu, Zhigang; Lai, Ren

    2013-01-01

    The most important indoor allergens for humans are house dust mites (HDM). Fourteen Dermatophagoides farinae allergens (Der f 1–3, 6, 7, 10, 11, 13–18, and 22) are reported although more than 30 allergens have been estimated in D. farinae. Seventeen allergens belonging to 12 different groups were identified by a procedure of proteomics combined with two-dimensional immunoblotting from D. farina extracts. Their sequences were determined by Edman degradation, mass spectrometry analysis, and cDNA cloning. Their allergenicities were assayed by enzyme-linked immunosorbent assay inhibition tests, immunoblots, basophil activation test, and skin prick tests. Eight of them are the first report as D. farinae allergens. The procedure of using a proteomic approach combined with a purely discovery approach using sera of patients with broad IgE reactivity profiles to mite allergens was an effective method to investigate a more complete repertoire of D. farinae allergens. The identification of eight new D. farinae allergens will be helpful for HDM allergy diagnosis and therapy, especially for patients without response for HDM major allergens. In addition, the current work significantly extendedthe repertoire of D. farinae allergens. PMID:23481662

  1. Allergens causing atopic diseases in canine.

    PubMed

    Youn, Hwa-Young; Kang, Hyung-Seok; Bhang, Dong-Ha; Kim, Min-Kue; Hwang, Cheol-Yong; Han, Hong-Ryul

    2002-12-01

    Canine atopic skin disease is seasonal or sometimes non-seasonal immune-mediated skin disease which occurs commonly in Korea. The definite clinical sign is systemic pruritus, especially on periocular parts, external ear, interdigit spaces and lateral flank. For diagnosis of this dermatitis, complete history taking followed by intradermal skin test and serum in vitro IgE test needs to be performed. Allergen selection for the diagnosis and treatment of atopic dermatitis should be varied geographically. In this study, with intradermal skin test(IDST) the prevalence of atopic disease and what allergens are involved in are researched. Allergens used for IDST included 26 allergen extracts from six allergen groups: grasses, trees, weeds, molds, epidermal allergens and environmental allergens. The number of allergens was 42 in which the positive and negative controls are included. The most common positive allergen reaction was the house dust mites on IDST(22/35, 63%). The other positive allergen reactions were to flea(3/35, 9%), molds(1/35, 3%), house dusts(2/35, 6%), feathers (1/35, 3%), cedar/juniper(1/35, 3%), timothy grass(1/35, 3%) and dandelion(1/35, 3%). In this study, the most prevalent allergen causing atopic dermatitis in dogs in Korea was the house dust mites followed by the flea. PMID:12819384

  2. 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

  3. 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

  4. Allergen source materials: state-of-the-art.

    PubMed

    Esch, Robert E

    2009-01-01

    A variety of positive outcomes can be realized from validation and risk management activities (see Table 4). They are dependent on the participation of multiple functional groups including the quality unit, regulatory and legal affairs, engineering and production operations, research and development, and sales and marketing. Quality risk management is receiving increased attention in the area of public health, pharmacovigilance, and pharmaceutical manufacturing. Recent examples of its regulatory use in our industry include the assessment of the potential risks of transmissible spongiform encephalopathies (TSE) agents through contaminated products], the risks of precipitates in allergenic extracts, and the revision of the potency limits for standardized dust mite and grass allergen vaccines. Its application to allergen source material process validation activities allowed for a practical strategy, especially in a complex manufacturing environment involving hundreds of products with multiple intended uses. In addition, the use of tools such as FMEA was useful in evaluating proposed changes made to manufacturing procedures and product specifications, new regulatory actions, and customer feedback or complaints. The success of such a quality assurance programs will ultimately be reflected in the elimination or reduction of product failures, improvement in the detection and prediction of potential product failures, and increased confidence in product quality. PMID:20799440

  5. [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

  6. Peanut allergen (Ara h 1) detection in foods containing chocolate.

    PubMed

    Pomés, A; Vinton, R; Chapman, M D

    2004-04-01

    Inadvertent exposure to peanut in foods poses health risks for peanut-allergic individuals that can be reduced by improving detection systems for allergen contaminants in food products and manufacturing processes. Detection of peanut in chocolate has been especially difficult. We report the optimization of conditions for measuring a major peanut allergen, Ara h 1, in chocolate with the use of a two-site monoclonal antibody sandwich enzyme-linked immunosorbent assay. Ara h 1 was extracted from peanut in the presence or absence of chocolate with phosphate buffer, salt, and three dried milks (goat, soy, or nonfat) (0 to 25% wt/vol) for 15 min at 60 degrees C or for 2.5 h at room temperature. The best conditions for Ara h 1 extraction in the presence of chocolate were 5% nonfat dry milk for 2.5 h at room temperature. Spiking experiments of chocolate with peanut confirmed improvement of the extraction: Ara h 1 was detected in extractions of 0.16 to 0.33% peanut in chocolate. Interestingly, the best conditions for Ara h 1 extraction were different for peanut alone than with chocolate, regarding time, temperature, and percentage of nonfat dry milk in the extraction buffer. In chocolate with peanut foods, the total Ara h 1 values were 10-fold higher than when products were extracted with phosphate buffer alone and could be up to 400-fold higher for individual foods. The dramatic improvement of Ara h 1 extraction should allow specific allergen monitoring in chocolate-containing food products and assessment of Ara h 1 exposure. PMID:15083733

  7. Mechanisms of allergen-specific immunotherapy.

    PubMed

    Akdis, Mübeccel; Akdis, Cezmi A

    2007-04-01

    Allergen-specific immunotherapy (SIT) has been used for almost a century as a desensitizing therapy for allergic diseases and represents the only curative and specific method of treatment. Administration of appropriate concentrations of allergen extracts has been shown to be reproducibly effective when patients are carefully selected. The mechanisms by which allergen-SIT has its effects include the modulation of T-cell and B-cell responses and related antibody isotypes as well as effector cells of allergic inflammation, such as eosinophils, basophils, and mast cells. The balance between allergen-specific T-regulatory (Treg) and T(H)2 cells appears to be decisive in the development of allergic and healthy immune responses against allergens. Treg cells consistently represent the dominant subset specific for common environmental allergens in sensitized healthy individuals. In contrast, there is a high frequency of allergen-specific T(H)2 cells in patients with allergy. The induction of a tolerant state in peripheral T cells represents an essential step in allergen-SIT. Peripheral T-cell tolerance is characterized mainly by generation of allergen-specific Treg cells leading to suppressed T-cell proliferation and T(H)1 and T(H)2 cytokine responses against the allergen. This is accompanied by a significant increase in allergen-specific IgG(4), and also IgG(1) and IgA, and a decrease in IgE in the late stage of the disease. In addition, decreased tissue infiltration of mast cells and eosinophils and their mediator release including circulating basophils takes place. Current understanding of mechanisms of allergen-SIT, particularly the role of Treg cells in peripheral tolerance, may enable novel treatment strategies. PMID:17321578

  8. Development of a design intent extraction flow for mask manufacturing

    NASA Astrophysics Data System (ADS)

    Kato, Kokoro; Endo, Masakazu; Inoue, Tadao; Yamabe, Masaki

    2010-03-01

    The cost of photomasks has been rising year by year as the process node gets finer and the mask cost is becoming one of the headaches in the semiconductor industry. For the purpose of the mask cost reduction, ASET started Mask D2I (Mask Design, Drawing and Inspection Technology) project in 2006. In earlier papers[1-4], we introduced the idea of photomask data prioritization method which is referred to as Mask Data Rank (MDR). We have built our software system to convert Design Intent (DI) to MDR with cooperation of STARC. Then we showed the results of preliminary experiments with mask data provided by STARC. In this paper we explain the software mechanism of design intent extraction flow. Then we show the experimental results with actual chip data in three semiconductor companies and address the related issues. Finally we introduce a new idea to extract design intent from analog circuits.

  9. 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

  10. Determination of fragrance allergens in indoor air by active sampling followed by ultrasound-assisted solvent extraction and gas chromatography-mass spectrometry.

    PubMed

    Lamas, J Pablo; Sanchez-Prado, Lucia; Garcia-Jares, Carmen; Llompart, Maria

    2010-03-19

    Fragrances are ubiquitous pollutants in the environment, present in the most of household products, air fresheners, insecticides and cosmetics. Commercial perfumes may contain hundreds of individual fragrance chemicals. In addition to the widespread use and exposure to fragranced products, many of the raw fragrance materials have limited available health and safety data. Because of their nature as artificial fragrances, inhalation should be considered as an important exposure pathway, especially in indoor environments. In this work, a very simple, fast, and sensitive methodology for the analysis of 24 fragrance allergens in indoor air is presented. Considered compounds include those regulated by the EU Directive, excluding limonene; methyl eugenol was also included due to its toxicity. The proposed methodology is based on the use of a very low amount of adsorbent to retain the target compounds, and the rapid ultrasound-assisted solvent extraction (UAE) using a very low volume of solvent which avoids further extract concentration. Quantification was performed by gas chromatography coupled to mass spectrometry (GC-MS). The influence of main factors involved in the UAE step (type of adsorbent and solvent, solvent volume and extraction time) was studied using an experimental design approach to account for possible factor interactions. Using the optimized procedure, 0.2 m(-3) air are sampled, analytes are retained on 25 mg Florisil, from which they are extracted by UAE (5 min) with 2 mL ethyl acetate. Linearity was demonstrated in a wide concentration range. Efficiency of the total sampling-extraction process was studied at several concentration levels (1, 5 and 125 microg m(-3)), obtaining quantitative recoveries, and good precision (RSD<10%). Method detection limits were < or =0.6 microg m(-3). Finally, the proposed method was applied to real samples collected in indoor environments in which several of the target compounds were determined. PMID:20138288

  11. Hymenoptera allergens: from venom to "venome".

    PubMed

    Spillner, Edzard; Blank, Simon; Jakob, Thilo

    2014-01-01

    In Western Europe, Hymenoptera venom allergy (HVA) primarily relates to venoms of the honeybee and the common yellow jacket. In contrast to other allergen sources, only a few major components of Hymenoptera venoms had been characterized until recently. Improved expression systems and proteomic detection strategies have allowed the identification and characterization of a wide range of additional allergens. The field of HVA research has moved rapidly from focusing on venom extract and single major allergens to a molecular understanding of the entire "venome" as a system of unique and characteristic components. An increasing number of such components has been identified, characterized regarding function, and assessed for allergenic potential. Moreover, advanced expression strategies for recombinant production of venom allergens allow selective modification of molecules and provide insight into different types of immunoglobulin E reactivities and sensitization patterns. The obtained information contributes to an increased diagnostic precision in HVA and may serve for monitoring, re-evaluation, and improvement of current therapeutic strategies. PMID:24616722

  12. 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

  13. 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

  14. Treatment with oleic acid reduces IgE binding to peanut and cashew allergens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Oleic acid (OA) is known to bind and change the bioactivities of proteins, such as a-lactalbumin and ß-lactoglobulin in vitro. The objective of this study was to determine if OA binds to allergens from a peanut extract or cashew allergen and changes their allergenic properties. Peanut extract or c...

  15. 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

  16. Contact dermatitis to a rubber allergen with both dithiocarbamate and benzothiazole structure.

    PubMed

    Bergendorff, Ola; Hansson, Christer

    2007-05-01

    Contact dermatitis to rubber products are often caused by additives used during manufacture, and diagnosed from patch test with established rubber allergen series. In these series the compounds are divided into separate groups such as thiurams, dithiocarbamates and mercaptobenzothiazoles. The objectives were to investigate the substances with allergenic structures present in a diving mask giving rise to facial dermatitis, also those substances including structures from different groups of rubber chemicals. The rubber material was analysed by high-performance liquid chromatography and diode-array detector. The patient was tested by epicutaneous tests using pure substances, extracts and authentic rubber material. 2-Benzothiazolyl-N,N-diethylthiocarbamylsulfide, was found in the diving mask and the patient showed positive reaction to the pure compound and to extracts of the diving mask. This compound has structures of both mercaptobenzothiazole and thiuram/dithiocarbamate in its formulae. Besides the established groups of rubber accelerators, uncommon allergens with structures from more than one group can be formed or added at vulcanization. Chemical analysis of the product is needed to find these allergens. PMID:17441851

  17. Allergens of the entomopathogenic fungus Beauveria bassiana

    PubMed Central

    Westwood, Greg S; Huang, Shih-Wen; Keyhani, Nemat O

    2005-01-01

    Background Beauveria bassiana is an important entomopathogenic fungus currently under development as a bio-control agent for a variety of insect pests. Although reported to be non-toxic to vertebrates, the potential allergenicity of Beauveria species has not been widely studied. Methods IgE-reactivity studies were performed using sera from patients displaying mould hypersensitivity by immunoblot and immunoblot inhibition. Skin reactivity to B. bassiana extracts was measured using intradermal skin testing. Results Immunoblots of fungal extracts with pooled as well as individual sera showed a distribution of IgE reactive proteins present in B. bassiana crude extracts. Proteinase K digestion of extracts resulted in loss of IgE reactive epitopes, whereas EndoH and PNGaseF (glycosidase) treatments resulted in minor changes in IgE reactive banding patterns as determined by Western blots. Immunoblot inhibitions experiments showed complete loss of IgE-binding using self protein, and partial inhibition using extracts from common allergenic fungi including; Alternaria alternata, Aspergillus fumigatus, Cladosporium herbarum, Candida albicans, Epicoccum purpurascens, and Penicillium notatum. Several proteins including a strongly reactive band with an approximate molecular mass of 35 kDa was uninhibited by any of the tested extracts, and may represent B. bassiana specific allergens. Intradermal skin testing confirmed the in vitro results, demonstrating allergenic reactions in a number of individuals, including those who have had occupational exposure to B. bassiana. Conclusions Beauveria bassiana possesses numerous IgE reactive proteins, some of which are cross-reactive among allergens from other fungi. A strongly reactive potential B. bassiana specific allergen (35 kDa) was identified. Intradermal skin testing confirmed the allergenic potential of B. bassiana. PMID:15644142

  18. Safety of engineered allergen-specific immunotherapy vaccines

    PubMed Central

    Focke-Tejkl, Margarete; Valenta, Rudolf

    2015-01-01

    Purpose of review The purpose of the review is to summarize and comment on recent developments regarding the safety of engineered immunotherapy vaccines. Recent findings In the last 2 years, several studies were published in which allergy vaccines were developed on the basis of chemical modification of natural allergen extracts, the engineering of allergen molecules by recombinant DNA technology and synthetic peptide chemistry, allergen genes, new application routes and conjugation with immune modulatory molecules. Several studies exemplified the general applicability of hypoallergenic vaccines on the basis of recombinant fusion proteins consisting of nonallergenic allergen-derived peptides fused to allergen-unrelated carrier molecules. These vaccines are engineered to reduce both, immunoglobulin E (IgE) as well as allergen-specific T cell epitopes in the vaccines, and thus should provoke less IgE and T-cell-mediated side-effects. They are made to induce allergen-specific IgG antibodies against the IgE-binding sites of allergens with the T-cell help of the carrier molecule. Summary Several interesting examples of allergy vaccines with potentially increased safety profiles have been published. The concept of fusion proteins consisting of allergen-derived hypoallergenic peptides fused to allergen-unrelated proteins that seems to be broadly applicable for a variety of allergens appears to be of particular interest because it promises not only to reduce side-effects but also to increase efficacy and convenience of allergy vaccines. PMID:22885888

  19. 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

  20. Allergen immunotherapy for birch pollen-allergic patients: recent advances.

    PubMed

    Moingeon, Philippe; Floch, Véronique Bordas-Le; Airouche, Sabi; Baron-Bodo, Véronique; Nony, Emmanuel; Mascarell, Laurent

    2016-05-01

    As of today, allergen immunotherapy is performed with aqueous natural allergen extracts. Recombinant allergen vaccines are not yet commercially available, although they could provide patients with well-defined and highly consistent drug substances. As Bet v 1 is the major allergen involved in birch pollen allergy, with more than 95% of patients sensitized to this allergen, pharmaceutical-grade recombinant Bet v 1-based vaccines were produced and clinically tested. Herein, we compare the clinical results and modes of action of treatments based on either a birch pollen extract or recombinant Bet v 1 expressed as hypoallergenic or natural-like molecules. We also discuss the future of allergen immunotherapy with improved drugs intended for birch pollen-allergic patients suffering from rhinoconjunctivitis. PMID:27140409

  1. Evaluation of stability of allergen extracts for sublingual immunotherapy during transport under unfavourable temperature conditions with an innovative thermal insulating packaging.

    PubMed

    Puccinelli, P; Natoli, V; Dell'albani, I; Scurati, S; Incorvaia, C; Barbieri, S; Masieri, S; Frati, F

    2013-01-01

    Many pharmaceutical and biotechnological products are temperature-sensitive and should normally be kept at a controlled temperature, particularly during transport, in order to prevent the loss of their stability and activity. Therefore, stability studies should be performed for temperature-sensitive products, considering product characteristics, typical environmental conditions, and anticipating environmental extremes that may occur during product transport in a specific country. Staloral products for sublingual immunotherapy are temperature sensitive and are labelled for maintenance under refrigerated conditions (2-8°C). Given the peculiar climatic context of Italy and the great temperature fluctuations that may occur during transport, this study was aimed at evaluating the impact of a new engineered thermal insulating packaging for Staloral. In particular, the purpose was to assess whether the new packaging could create a container condition able to preserve the stability and immunological activity of the product during the transport phase throughout Italy. The results showed that the range of temperatures that can affect the product, in the area surrounding the product packaging, may reach a peak of 63°C during transport under the most unfavourable climatic conditions, i.e. in a non-refrigerated van during the summer season, from the site of production in France to the patient's house in Catania, the city with the highest temperatures in Italy. However, the highest temperature reached inside the vaccine did not exceed 45°C over a period of about 2 h. The ELISA inhibition test on samples subjected to the extreme temperature conditions previously defined (45°C) showed an immunological activity higher than 75% of that initially measured and was comparable to those obtained with samples stored at controlled temperature (5°C). This means that, even in the worst case scenario, the structure of the allergen extracts is not influenced and the vaccine potency is

  2. 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

  3. Recent progress in allergen immunotherapy.

    PubMed

    Nouri-Aria, Kayhan T

    2008-03-01

    The efficacy of allergen immunotherapy for the treatment of allergic rhinoconjunctivitis with or without seasonal bronchial asthma and anaphylaxis caused by the sting of the hymenoptera class of insects has been clearly demonstrated in numerous well-designed, placebo-controlled trials. Immunotherapy whether by subcutaneous injection of allergen extract or by oral/sublingual routes modifies peripheral and mucosal TH2 responses in favour of TH1 responses and augments IL-10 synthesis by TRegs both locally and by peripheral T cells. Recent researches into the cellular and molecular basis of allergic reactions have advanced our understanding of the mechanisms involved in allergic diseases. They have also helped the development of innovative approaches that are likely to further improve the control of allergic responses in the future. Novel approaches to immunotherapy that are currently being explored include the use of peptide-based allergen preparations, which do not bind IgE and therefore do not activate mast cells, but reduce both Th1 and Th2-cytokine synthesis, while increasing levels of IL-10. Alternative strategies include the use of adjuvants, such as nucleotide immunostimulatory sequences derived from bacteria CpG or monophosphoryl lipid A that potentiate Th1 responses. Blocking the effects of IgE using anti-IgE such as omalizumab, a recombinant humanized monoclonal antibody that selectively binds to IgE, has been shown to be a useful strategy in the treatment of allergic asthma and rhinitis. The combination of anti-IgE-monoclonal antibody omalizumab with allergen immunotherapy has proved beneficial for the treatment of allergic diseases, offering improved efficacy, limited adverse effects, and potential immune-modifying effects. This combination may also accelerate the rapidity by which immunotherapy induces TReg cells. If allergic diseases are due to a lack of allergen-specific TReg cells, then effective therapies should target the induction and the

  4. [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

  5. [Significance of inhaled environmental allergens].

    PubMed

    Zochert, J

    1983-01-01

    Whereas the importance of pollen as inhalative allergens has been largely investigated and is generally known, the experience in the frequency and the role of the sensibilization with air-borne fungi is relatively limited. In 720 patients with Asthma bronchiale the degree of sensitization has been tested with various extracts of air-borne fungi of SSW Dresden (mould mixture, aspergillin, mucor, cladosporium and penicillium and alternaria). The most frequent and also the strongest reactions were found with alternaria and the smallest part of positive skin reactions with penicillium. An isolated sensitization with mould has been demonstrated in 20 per cent of the cases. In 60 per cent of the tested patients a manifest mould allergy was shown by means of the Inhalative Allergen Test, the most favourable correlation between Intracutaneous Test (ICT) and Inhalative Test (IAT) was found with alternaria (76%). A conformance between ICT and basophils degranulation test (BDT) was stated in 69% of the cases. The aim should be comparable tests with allergen extracts without irritative effects and qualitative measurements of air-borne fungi. PMID:6649704

  6. 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...

  7. Tannic acid as a means to remove peanut allergens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tannic acid (TA) is a polyphenol (commonly found in tea and coffee) that has been used as a treatment for toxic substances and carpet allergens. The objectives were to determine the efficacy of TA’s binding and removal of peanut allergens from peanut butter extracts as insoluble precipitates, and to...

  8. Interfaces Between Allergen Structure and Diagnosis: Know Your Epitopes

    PubMed Central

    Pomés, Anna; Chruszcz, Maksymilian; Gustchina, Alla; Wlodawer, Alexander

    2016-01-01

    Allergy diagnosis is based on the patient’s clinical history and can be strengthened by tests that confirm the origin of sensitization. In the past 25 years, these tests have evolved from the exclusive in vivo or in vitro use of allergen extracts, to complementary molecular-based diagnostics that rely on in vitro measurements of IgE reactivity to individual allergens. For this to occur, an increase in our understanding of the molecular structure of allergens, largely due to the development of technologies such as molecular cloning and expression of recombinant allergens, X-ray crystallography, or nuclear magnetic resonance (NMR), has been essential. New in vitro microarray or multiplex systems are now available to measure IgE against a selected panel of purified natural or recombinant allergens. The determination of the three-dimensional structure of allergens has facilitated detailed molecular studies, including the analysis of antigenic determinants for diagnostic purposes. PMID:25750181

  9. Mechanisms underlying allergy vaccination with recombinant hypoallergenic allergen derivatives

    PubMed Central

    Linhart, Birgit; Valenta, Rudolf

    2015-01-01

    Hundred years ago therapeutic vaccination with allergen-containing extracts has been introduced as a clinically effective, disease-modifying, allergen-specific and long-lasting form of therapy for allergy, a hypersensitivity disease affecting more than 25% of the population. Today, the structures of most of the disease-causing allergens have been elucidated and recombinant hypoallergenic allergen derivatives with reduced allergenic activity have been engineered to reduce side effects during allergen-specific immunotherapy (SIT). These recombinant hypoallergens have been characterized in vitro, in experimental animal models and in clinical trials in allergic patients. This review provides a summary of the molecular, immunological and preclinical evaluation criteria applied for this new generation of allergy vaccines. Furthermore, we summarize the mechanisms underlying SIT with recombinant hypoallergens which are thought to be responsible for their therapeutic effect. PMID:22100888

  10. Allergen-specific immunotherapy: from therapeutic vaccines to prophylactic approaches

    PubMed Central

    Valenta, R.; Campana, R.; Marth, K.; van Hage, M.

    2015-01-01

    Immunoglobulin E-mediated allergies affect more than 25% of the population. Allergen exposure induces a variety of symptoms in allergic patients, which include rhinitis, conjunctivitis, asthma, dermatitis, food allergy and life-threatening systemic anaphylaxis. At present, allergen-specific immunotherapy (SIT), which is based on the administration of the disease-causing allergens, is the only disease-modifying treatment for allergy. Current therapeutic allergy vaccines are still prepared from relatively poorly defined allergen extracts. However, with the availability of the structures of the most common allergen molecules, it has become possible to produce well-defined recombinant and synthetic allergy vaccines that allow specific targeting of the mechanisms of allergic disease. Here we provide a summary of the development and mechanisms of SIT, and then review new forms of therapeutic vaccines that are based on recombinant and synthetic molecules. Finally, we discuss possible allergen-specific strategies for prevention of allergic disease. PMID:22640224

  11. Mold Allergens in Respiratory Allergy: From Structure to Therapy

    PubMed Central

    Twaroch, Teresa E; Curin, Mirela; Swoboda, Ines

    2015-01-01

    Allergic reactions to fungi were described 300 years ago, but the importance of allergy to fungi has been underestimated for a long time. Allergens from fungi mainly cause respiratory and skin symptoms in sensitized patients. In this review, we will focus on fungi and fungal allergens involved in respiratory forms of allergy, such as allergic rhinitis and asthma. Fungi can act as indoor and outdoor respiratory allergen sources, and depending on climate conditions, the rates of sensitization in individuals attending allergy clinics range from 5% to 20%. Due to the poor quality of natural fungal allergen extracts, diagnosis of fungal allergy is hampered, and allergen-specific immunotherapy is rarely given. Several factors are responsible for the poor quality of natural fungal extracts, among which the influence of culture conditions on allergen contents. However, molecular cloning techniques have allowed us to isolate DNAs coding for fungal allergens and to produce a continuously growing panel of recombinant allergens for the diagnosis of fungal allergy. Moreover, technologies are now available for the preparation of recombinant and synthetic fungal allergen derivatives which can be used to develop safe vaccines for the treatment of fungal allergy. PMID:25840710

  12. Measurement of Horse Allergen (Equ cx) in Schools.

    PubMed

    Merritt, Anne-Sophie; Emenius, Gunnel; Elfman, Lena; Smedje, Greta

    2011-01-01

    Background. The presence of horse allergen in public places is not well-known, unlike for instance cat and dog allergens, which have been studied extensively. The aim was to investigate the presence of horse allergen in schools and to what extent the influence of number of children with regular horse contact have on indoor allergen levels. Methods. Petri dishes were used to collect airborne dust samples during one week in classrooms. In some cases, vacuumed dust samples were also collected. All samples were extracted, frozen and analysed for Equ cx content shortly after sampling, and some were re-analysed six years later with a more sensitive ELISA assay. Results. Horse allergen levels were significantly higher in classrooms, in which many children had horse contact, regardless of sampling method. Allergen levels in extracts from Petri dish samples, which had been kept frozen, dropped about 53% over a six-year period. Conclusion. Horse allergen was present in classrooms and levels were higher in classrooms where many children had regular horse contact in their leisure time. This suggests that transfer of allergens takes place via contaminated clothing. Measures should be taken to minimize possible transfer and deposition of allergens in pet-free environments, such as schools. PMID:23724238

  13. Mold allergens in respiratory allergy: from structure to therapy.

    PubMed

    Twaroch, Teresa E; Curin, Mirela; Valenta, Rudolf; Swoboda, Ines

    2015-05-01

    Allergic reactions to fungi were described 300 years ago, but the importance of allergy to fungi has been underestimated for a long time. Allergens from fungi mainly cause respiratory and skin symptoms in sensitized patients. In this review, we will focus on fungi and fungal allergens involved in respiratory forms of allergy, such as allergic rhinitis and asthma. Fungi can act as indoor and outdoor respiratory allergen sources, and depending on climate conditions, the rates of sensitization in individuals attending allergy clinics range from 5% to 20%. Due to the poor quality of natural fungal allergen extracts, diagnosis of fungal allergy is hampered, and allergen-specific immunotherapy is rarely given. Several factors are responsible for the poor quality of natural fungal extracts, among which the influence of culture conditions on allergen contents. However, molecular cloning techniques have allowed us to isolate DNAs coding for fungal allergens and to produce a continuously growing panel of recombinant allergens for the diagnosis of fungal allergy. Moreover, technologies are now available for the preparation of recombinant and synthetic fungal allergen derivatives which can be used to develop safe vaccines for the treatment of fungal allergy. PMID:25840710

  14. 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. PMID:25778347

  15. Cultivation practices and manufacturing processes to produce Hoodia gordonii extract for weight management products.

    PubMed

    Knight, Tamsin L; Swindells, Chris M; Craddock, Andrew M; Maharaj, Vinesh J; Buchwald-Werner, Sybille; Ismaili, Smail Alaoui; McWilliam, Simon C

    2012-01-01

    Hoodia gordonii (Masson) Sweet ex Decne., is a succulent shrub, indigenous to the arid regions of southern Africa. Indigenous people have historically utilised certain species of Hoodia, including H. gordonii, as a source of food and water. Studies by the Council for Scientific and Industrial Research (CSIR, South Africa) identified that extracts of H. gordonii had appetite suppressant activity associated with specific steroid glycosides. A programme to develop weight management products based around this discovery was implemented in 1998. An agronomy programme was established which demonstrated that it was possible to cultivate this novel crop on a commercial scale (in excess of 70 ha). In parallel, a food grade manufacturing process was developed consisting of four main steps: harvesting of H. gordonii plant stems, comminution, drying under controlled conditions and extraction using food grade solvents. Appropriate Quality Control (QC) procedures were developed. The extraction process is capable of delivering a consistent composition despite natural variations in the composition of the dried H. gordonii. Specifications were developed for the resulting extract. The intended use of the standardised H. gordonii extract was as a functional food ingredient for weight management products. Other development studies on characterisation, toxicology and pharmacology are reported separately. PMID:22410259

  16. Factors influencing the Zn and Mn extraction from pyrometallurgical sludge in the steel manufacturing industry.

    PubMed

    Mocellin, J; Mercier, G; Morel, J L; Blais, J F; Simonnot, M O

    2015-08-01

    In this laboratory study, a process has been developed for selectively leaching zinc and manganese from pyrometallurgical sludge produced in the steel manufacturing industry. In the first part, the yield of Zn extraction was studied using four factors and four levels of the Box-Behnken response surface design. The optimum conditions for the step of Zn leaching were determined to be a sulfuric acid concentration of 0.25 mol/L, a pulp density of 10%, an extraction temperature of 20 °C, and three stages of leaching. Under such conditions, 75% of the Zn should be leached. For Mn leaching, the optimum conditions were determined to be a sulfuric acid concentration of 0.25 mol/L, a Na2S2O5/Mn stoichiometry of 1, a leaching time of 120 min and two leaching steps. In this case, 100% of the Mn should be leached. PMID:25958078

  17. 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

  18. PEANUT ALLERGENS AND PROCESSING

    Technology Transfer Automated Retrieval System (TEKTRAN)

    It has been suggested that boiling or frying peanuts leads to less allergenic products than roasting. In this study, we have compared the fate of the major peanut allergens in the context of peanuts subjected to boiling, frying, or roasting. As opposed to previous work, both the soluble and insolubl...

  19. Peanut Allergens and Processing

    Technology Transfer Automated Retrieval System (TEKTRAN)

    It has been suggested that boiling or frying of peanuts lead to less allergenic products than roasting. In this study, we have compared the fate of the major peanut allergens in the context of peanuts subjected to boiling, frying, or roasting. As opposed to previous work, both the soluble and insolu...

  20. 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

  1. 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

  2. LC/MS analysis of proteolytic peptides in wheat extracts for determining the content of the allergen amylase/trypsin inhibitor CM3: influence of growing area and variety.

    PubMed

    Prandi, Barbara; Faccini, Andrea; Tedeschi, Tullia; Galaverna, Gianni; Sforza, Stefano

    2013-09-01

    Food allergy from wheat is triggered by several protein classes, such as LTPs, ω5-gliadins and α-amylase/trypsin inhibitors. The latter proteins, belonging to the prolamin superfamily, are mostly involved in baker's asthma, a form of occupational allergy in which the sensitization occurs through the respiratory tract. α-Amylase/trypsin inhibitors were also found to be involved in wheat-related atopic dermatitis. In this work, the allergen Tri a 30 (the CM3 α-amylase/trypsin inhibitor) was quantified in durum wheat salt soluble extracts using a peptidomic approach. CM3 protein identification was confirmed by using LTQ-OrbiTrap analysis on peptides obtained from the enzymatically digested protein separated by gel electrophoresis. Then, marker peptides derived from the protein after enzymatic cleavage of the full wheat extracts were identified by LC-MS/MS. One of them was used as marker for quantitative determination on an UPLC/ESI-MS system by using its isotopically labelled analogue as internal standard, allowing to assess the protein content in the different samples. The CM3 allergenic proteins were found to greatly vary among different cultivation areas. PMID:23578625

  3. Hierarchy and molecular properties of house dust mite allergens.

    PubMed

    Thomas, Wayne R

    2015-10-01

    The allergenic load of house dust mite allergy is largely constituted by a few proteins with a hierarchical pattern of allergenicity. The serodominant specificities are the group 1&2 and the group 23 faecal allergens. The collective IgE binding to the group 1&2 allergens can measure unequivocal HDM sensitisation better than HDM extracts although discrepancies have been found in regions with complex acarofauna suggesting a need to investigate the specificity with allergen components. The group 4, 5, 7&21 allergens that each induce responses in about 40% of subjects are mid-tier allergens accounting for most of the remaining IgE binding. Their titres are proportional to the concomitant responses to Der p1&2. Group 2 allergen variants have different antibody binding. Body proteins only occasionally induce sensitisation although a higher prevalence of binding by atopic dermatitis patients provides a new avenue of research. A broad spectrum of IgE binding has been associated with diverse symptoms but not with the severity of asthma which is associated with low IgG antibody. Some allergens such as the group 14 large lipid binding proteins and the recently described proteins Der f 24-33, need further investigation but with the cognoscence that other denominated allergens have been found to be minor sensitisers by comparative quantitative analyses. Scabies is a confounder for diagnosis with extracts, inducing cross-reactive antibodies with Der p 4&20 as is seafood allergy with cross reactivity to Der p 10 a minor HDM allergen. The HDM genome sequence can now be used to verify allelic and paralogous variations. PMID:26433526

  4. 78 FR 54658 - Allergenic Products Advisory Committee; Notice of Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-09-05

    ... and efficacy of Oralair, a Sweet Vernal Grass, Perennial Ryegrass, Timothy Grass, Orchard Grass, and... the safety and efficacy of Grastek, a Timothy Grass Pollen Allergen Extract tablet for sublingual...

  5. Inhaled allergen bronchoprovocation tests.

    PubMed

    Diamant, Zuzana; Gauvreau, Gail M; Cockcroft, Don W; Boulet, Louis-Philippe; Sterk, Peter J; de Jongh, Frans H C; Dahlén, Barbro; O'Byrne, Paul M

    2013-11-01

    The allergen bronchoprovocation test is a long-standing exacerbation model of allergic asthma that can induce several clinical and pathophysiologic features of asthma in sensitized subjects. Standardized allergen challenge is primarily a research tool, and when properly conducted by qualified and experienced investigators, it is safe and highly reproducible. In combination with validated airway sampling and sensitive detection techniques, allergen challenge allows the study of several features of the physiology of mainly TH2 cell-driven asthma in relation to the kinetics of the underlying airway pathology occurring during the allergen-induced late response. Furthermore, given the small within-subject variability in allergen-induced airway responses, allergen challenge offers an adequate disease model for the evaluation of new (targeted) controller therapies for asthma in a limited number of subjects. In proof-of-efficacy studies thus far, allergen challenge showed a fair positive predicted value and an excellent negative predictive value for the actual clinical efficacy of new antiasthma therapies, underscoring its important role in early drug development. In this review we provide recommendations on challenge methods, response measurements, sample size, safety, and harmonization for future applications. PMID:24119772

  6. 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

  7. Treatment with oleic acid reduces IgE binding to peanut and cashew allergens.

    PubMed

    Chung, Si-Yin; Mattison, Christopher P; Reed, Shawndrika; Wasserman, Richard L; Desormeaux, Wendy A

    2015-08-01

    Oleic acid (OA) is known to bind and change the bioactivities of proteins, such as α-lactalbumin and β-lactoglobulin in vitro. The objective of this study was to determine if OA binds to allergens from a peanut extract or cashew allergen and changes their allergenic properties. Peanut extract or cashew allergen (Ana o 2) was treated with or without 5mM sodium oleate at 70°C for 60 min (T1) or under the same conditions with an additional overnight incubation at 37°C (T2). After treatment, the samples were dialyzed and analyzed by SDS-PAGE and for OA content. IgE binding was evaluated by ELISA and western blot, using a pooled serum or plasma from individuals with peanut or cashew allergies. Results showed that OA at a concentration of 5mM reduced IgE binding to the allergens. Peanut sample T2 exhibited a lower IgE binding and a higher OA content (protein-bound) than T1. Cashew allergen T2 also showed a reduction in IgE binding. We conclude that OA reduces the allergenic properties of peanut extract and cashew allergen by binding to the allergens. Our findings indicate that OA in the form of sodium oleate may be potentially useful as a coating to reduce the allergenic properties of peanut and cashew allergens. PMID:25766831

  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. PMID:26593515

  9. 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)

  10. 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

  11. Multiplex detection of food allergens and gluten.

    PubMed

    Cho, Chung Y; Nowatzke, William; Oliver, Kerry; Garber, Eric A E

    2015-05-01

    To help safeguard the food supply and detect the presence of undeclared food allergens and gluten, most producers and regulatory agencies rely on commercial test kits. Most of these are ELISAs with a few being PCR-based. These methods are very sensitive and analyte specific, requiring different assays to detect each of the different food allergens. Mass spectrometry offers an alternative approach whereby multiple allergens may be detected simultaneously. However, mass spectrometry requires expensive equipment, highly trained analysts, and several years before a quantitative approach can be achieved. Using multianalyte profiling (xMAP®) technology, a commercial multiplex test kit based on the use of established antibodies was developed for the simultaneous detection of up to 14 different food allergens plus gluten. The assay simultaneously detects crustacean seafood, egg, gluten, milk, peanut, soy, and nine tree nuts (almond, Brazil nut, cashew, coconut, hazelnut, macadamia, pine nut, pistachio, and walnut). By simultaneously performing multiple tests (typically two) for each analyte, this magnetic bead-based assay offers built-in confirmatory analyses without the need for additional resources. Twenty-five of the assays were performed on buffer extracted samples, while five were conducted on samples extracted using reduced-denatured conditions. Thus, complete analysis for all 14 allergens and gluten requires only two wells of a 96-well microtiter plate. This makes it possible to include in a single analytical run up to 48 samples. All 30 bead sets in this multiplex assay detected 5 ng/mL of food allergen and gluten with responses greater than background. In addition, 26 of the bead sets displayed signal/noise ratios of five or greater. The bead-based design makes this 30-plex assay expandable to incorporate new antibodies and capture/detector methodologies by ascribing these new detectors to any of the unassigned bead sets that are commercially available. PMID

  12. [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

  13. Clinico-Immunological Analysis of Eggplant (Solanum melongena) Allergy Indicates Preponderance of Allergens in the Peel

    PubMed Central

    2009-01-01

    Background Eggplant (Solanum melongena L.) is known to cause food allergy in some Asian countries but detailed studies on eggplant allergy are lacking. Objective The objective is to investigate sensitization to different parts of eggplant fruit, and detection of the allergens. Methods Six eggplant-allergic subjects were assessed for sensitization to eggplant (peel/pulp, and raw/cooked) by skin prick test, allergen-specific IgE, and immunoblots. Allergens were analyzed for glycoprotein nature by staining/lectinoblots, and in vitro stability in simulated gastric fluid. Results All the eggplant-sensitized subjects showed positive skin prick test with peel, pulp, raw, and cooked eggplant extracts; allergen-specific IgE to all these was positive. Raw eggplant contains 5 allergens in the range 36-71 kD. Most allergens are localized in the eggplant peel (9 allergens; 26-71 kD range) than the pulp (3 allergens; 52-71 kD); among these, the 26, 28, 36, and 71 kD allergens seem to be heat-stable. The 43, 45, 64, and 71 kD allergens are detected as glycoproteins; the 26, 64, and 71 kD allergens are stable displaying retention of IgE-binding ability in simulated gastric fluid digestion. Conclusions Eggplant is a multiallergenic vegetable in the context of presence of allergens in all edible parts of eggplant having preponderance in the peel. PMID:23283148

  14. Allergenic Characterization of a Novel Allergen, Homologous to Chymotrypsin, from German Cockroach

    PubMed Central

    Jeong, Kyoung Yong; Son, Mina; Lee, Jae-Hyun; Hong, Chein-Soo

    2015-01-01

    Purpose Cockroach feces are known to be rich in IgE-reactive components. Various protease allergens were identified by proteomic analysis of German cockroach fecal extract in a previous study. In this study, we characterized a novel allergen, a chymotrypsin-like serine protease. Methods A cDNA sequence homologous to chymotrypsin was obtained by analysis of German cockroach expressed sequence tag (EST) clones. The recombinant chymotrypsins from the German cockroach and house dust mite (Der f 6) were expressed in Escherichia coli using the pEXP5NT/TOPO vector system, and their allergenicity was investigated by ELISA. Results The deduced amino acid sequence of German cockroach chymotrypsin showed 32.7 to 43.1% identity with mite group 3 (trypsin) and group 6 (chymotrypsin) allergens. Sera from 8 of 28 German cockroach allergy subjects (28.6%) showed IgE binding to the recombinant protein. IgE binding to the recombinant cockroach chymotrypsin was inhibited by house dust mite chymotrypsin Der f 6, while it minimally inhibited the German cockroach whole body extract. Conclusions A novel allergen homologous to chymotrypsin was identified from the German cockroach and was cross-reactive with Der f 6. PMID:25749759

  15. Mammalian lipocalin allergens--insights into their enigmatic allergenicity.

    PubMed

    Virtanen, T; Kinnunen, T; Rytkönen-Nissinen, M

    2012-04-01

    Most of the important mammal-derived respiratory allergens, as well as a milk allergen and a few insect allergens, belong to the lipocalin protein family. As mammalian lipocalin allergens are found in dander, saliva and urine, they disperse effectively and are widely present in the indoor environments. Initially, lipocalins were characterized as transport proteins for small, principally hydrophobic molecules, but now they are known to be involved in many other biological functions. Although the amino acid identity between lipocalins is generally at the level of 20-30%, it can be considerably higher. Lipocalin allergens do not exhibit any known physicochemical, functional or structural property that would account for their allergenicity, that is, the capacity to induce T-helper type 2 immunity against them. A distinctive feature of mammalian lipocalin allergens is their poor capacity to stimulate the cellular arm of the human or murine immune system. Nevertheless, they induce IgE production in a large proportion of atopic individuals exposed to the allergen source. The poor capacity of mammalian lipocalin allergens to stimulate the cellular immune system does not appear to result from the function of regulatory T cells. Instead, the T cell epitopes of mammalian lipocalin allergens are few and those examined have proved to be suboptimal. Moreover, the frequency of mammalian lipocalin allergen-specific CD4(+) T cells is very low in the peripheral blood. Importantly, recent research suggests that the lipocalin allergen-specific T cell repertoires differ considerably between allergic and healthy subjects. These observations are compatible with our hypothesis that the way CD4(+) T-helper cells recognize the epitopes of mammalian lipocalin allergens may be implicated in their allergenicity. Indeed, as several lipocalins exhibit homologies of 40-60% over species, mammalian lipocalin allergens may be immunologically at the borderline of self and non-self, which would not

  16. 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 ...

  17. [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. PMID:21901563

  18. Modification of house dust mite allergens by monomethoxypolyethylene glycol. Allergenicity measured by in vitro and in vivo methods.

    PubMed

    Mosbech, H; Dreborg, S; Påhlman, I; Stahl Skov, P; Steringer, I; Weeke, B

    1988-01-01

    In animal models, allergen modification by coupling to monomethoxypolyethylene glycol (mPEG) molecules can reduce allergenicity of the extract and makes the allergen capable of suppressing boosted IgE response. To investigate in a human system the degree of attenuation implied by a mPEG modification of a house dust mite (Dermatophagoides pteronyssinus) extract, 55 adults with asthma caused by house dust mites were tested by skin prick test (SPT) and histamine release assay (HR). RAST inhibition was performed on sera from 6 additional patients. Modified extract containing 0.42 mmol mPEG/g protein was used for the analyses. In order to get the same response of the two extracts when assessed by HR and SPT, a median increase in concentration of 10-fold of the mPEG-modified extract compared to the unmodified extract was needed. Interindividual variation was limited. Sixty-four to 72% needed a dose increase within +/- half a decade from this value. In 42-49% of the patients, results from SPT and HR deviated less than half a decade. The relative potency of the modified extract as measured by RAST inhibition was reduced to 17-78% (mean 39%). Reduced allergenicity would by itself mean less side effects in immunotherapy. When planning such therapy it is important to know that mPEG modification reduces the allergenicity to a similar extent in a majority of patients. PMID:2448247

  19. [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. PMID:12134645

  20. The FEMA GRAS assessment of furfural used as a flavour ingredient. Flavor and Extract Manufacturers' Association.

    PubMed

    Adams, T B; Doull, J; Goodman, J I; Munro, I C; Newberne, P; Portoghese, P S; Smith, R L; Wagner, B M; Weil, C S; Woods, L A; Ford, R A

    1997-08-01

    The Expert Panel of the Flavor and Extract Manufacturers' Association (FEMA) has assessed the safety of furfural for its continued use as a flavour ingredient. The safety assessment takes into account the current scientific information on exposure, metabolism, pharmacokinetics, toxicology, carcinogenicity and genotoxicity. Furfural was reaffirmed as GRAS (GRASr) as a flavour ingredient under conditions of intended use based on: (1) its mode of metabolic detoxication in humans; (2) its low level of flavour use compared with higher intake levels as a naturally occurring component of food; (3) the safety factor calculated from results of subchronic and chronic studies, (4) the lack of reactivity with DNA; and (5) the conclusion that the only statistically significant finding in the 2-year NTP bioassays, an increased incidence of hepatocellular adenomas and carcinomas in the high-dose group of male mice, was secondary to pronounced hepatotoxicity. Taken together, these data do not indicate any risk to human health under conditions of use as a flavour ingredient. This evidence of safety is supported by the occurrence of furfural as a natural component of traditional foods, at concentrations in the diet resulting in a 'natural intake' that is at least 100 times higher than the intake of furfural from use as a flavour ingredient. PMID:9350219

  1. Immunological aspects of the immune response induced by mosquito allergens.

    PubMed

    Cantillo, José Fernando; Fernández-Caldas, Enrique; Puerta, Leonardo

    2014-01-01

    Allergies caused by mosquito bites may produce local or systemic reactions. The inhalation of mosquito allergens may also cause asthma and/or allergic rhinoconjunctivitis in sensitized individuals. The mechanisms implicated in the development of these immune responses involve IgE antibodies, different subtypes of IgG and proinflammatory cytokines as well as basophils, eosinophils and mast cells. Several allergenic components have been identified in the saliva and bodies of mosquitoes and some of these are present in different mosquito species. The most common species implicated in allergic reactions belong to the genera Aedes, Culex and Anopheles. Several Aedes aegypti allergens have been cloned and sequenced. The recombinant molecules show IgE reactivity similar to that of the native allergens, making them good candidates for the diagnosis of mosquito allergies. Allergen-specific immunotherapy with mosquito extracts induces a protective response characterized by a decreased production of IgE antibodies, increased IgG levels, a reduction in the severity of cutaneous and respiratory symptoms and the need for medication. The aims of this review are to summarize the progress made in the characterization of mosquito allergens and discuss the types of immune responses induced by mosquito bites and the inhalation of mosquito allergens in atopic individuals. PMID:25661054

  2. Mechanisms of subcutaneous allergen immunotherapy.

    PubMed

    Soyer, Ozge U; Akdis, Mubeccel; Akdis, Cezmi A

    2011-05-01

    Allergen-specific immunotherapy (SIT) is the only curative approach in the treatment of allergic diseases defined up-to-date. Peripheral T-cell tolerance to allergens, the goal of successful allergen-SIT, is the primary mechanism in healthy immune responses to allergens. By repeated administration of increased doses of the causative allergen, allergen-SIT induces a state of immune tolerance to allergens through the constitution of T regulatory (Treg) cells, including allergen-specific interleukin (IL)-10-secreting Treg type 1 cells and CD4(+)CD25(+)Treg cells; induction of suppressive cytokines, such as IL-10 and transforming growth factor β; suppression of allergen-specific IgE and induction of IgG4 and IgA; and suppression of mast cells, basophils, eosinophils, and inflammatory dendritic cells. This review summarizes the current knowledge on the mechanisms of allergen-SIT with emphasis on the roles of Treg cells in allergen-SIT. PMID:21530813

  3. 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...

  4. 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…

  5. Allergies associated with medical gloves. Manufacturing issues.

    PubMed

    Hamann, C P; Kick, S A

    1994-07-01

    The increase in glove usage that followed the advent of Universal Precautions has been associated with a concomitant increase in glove-related allergic reactions, many of which are potentially debilitating. Manufacturing issues that can affect the allergenicity of natural rubber latex, thermoplastic elastomer, and polyvinylchloride medical gloves are therefore examined. This information can enhance the ability of the occupational dermatologist to diagnose specific allergens and to recommend appropriate treatment based on knowledge of a product's allergenic ingredients. PMID:7923952

  6. 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. PMID:24993820

  7. Octyl-modified magnetic graphene as a sorbent for the extraction and simultaneous determination of fragrance allergens, musks, and phthalates in aqueous samples by gas chromatography with mass spectrometry.

    PubMed

    Maidatsi, Katerina V; Chatzimitakos, Theodoros G; Sakkas, Vassilios A; Stalikas, Constantine D

    2015-11-01

    An effective, simple, and low-cost sample preparation method based on dispersive SPE followed by GC with MS is developed for the multianalyte determination of fragrance allergens, musks, and phthalates, at sub-ppb levels. The extraction procedure is based on a novel magnetic graphene sorbent, which is functionalized with octylamine, taking advantage of the functionalization's hydrophobic properties and π-π interactions with the analytes. Two alkyl amines, the octylamine and octadecylamine are studied to introduce alkyl chains in the basal plane of graphene. Magnetic graphene- octadecylamine is proved to be highly hydrophobic to such a degree that is hard to disperse in the bulk aqueous matrixes. Because of this behavior, its extraction efficiency for the target analytes is low. The synthesis and applicability of the magnetic graphene-octylamine as more favored sorbent are optimized in terms of the most determining experimental conditions. The detection and quantification limits, which are calculated based on S/N ratio of 3 and 10, respectively, ranged from 0.29 to 3.2 ng L(-1) and from 0.89 to 9.6, respectively. The dispersive SPE is successfully applied to routine analysis for the determination of the target analytes in samples from municipal treatment plant of Ioannina (Greece), from Pamvotis Lake and baby bathwater. The reproducibility of the spiked biological treatment plant water sample is evaluated and the relative standard deviation values range between 2.1 and 9.4%. PMID:26311498

  8. 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.

  9. Alternative bio-based solvents for extraction of fat and oils: solubility prediction, global yield, extraction kinetics, chemical composition and cost of manufacturing.

    PubMed

    Sicaire, Anne-Gaëlle; Vian, Maryline; Fine, Frédéric; Joffre, Florent; Carré, Patrick; Tostain, Sylvain; Chemat, Farid

    2015-01-01

    The present study was designed to evaluate the performance of alternative bio-based solvents, more especially 2-methyltetrahydrofuran, obtained from crop's byproducts for the substitution of petroleum solvents such as hexane in the extraction of fat and oils for food (edible oil) and non-food (bio fuel) applications. First a solvent selection as well as an evaluation of the performance was made with Hansen Solubility Parameters and the COnductor-like Screening MOdel for Realistic Solvation (COSMO-RS) simulations. Experiments were performed on rapeseed oil extraction at laboratory and pilot plant scale for the determination of lipid yields, extraction kinetics, diffusion modeling, and complete lipid composition in term of fatty acids and micronutrients (sterols, tocopherols and tocotrienols). Finally, economic and energetic evaluations of the process were conducted to estimate the cost of manufacturing using 2-methyltetrahydrofuran (MeTHF) as alternative solvent compared to hexane as petroleum solvent. PMID:25884332

  10. Alternative Bio-Based Solvents for Extraction of Fat and Oils: Solubility Prediction, Global Yield, Extraction Kinetics, Chemical Composition and Cost of Manufacturing

    PubMed Central

    Sicaire, Anne-Gaëlle; Vian, Maryline; Fine, Frédéric; Joffre, Florent; Carré, Patrick; Tostain, Sylvain; Chemat, Farid

    2015-01-01

    The present study was designed to evaluate the performance of alternative bio-based solvents, more especially 2-methyltetrahydrofuran, obtained from crop’s byproducts for the substitution of petroleum solvents such as hexane in the extraction of fat and oils for food (edible oil) and non-food (bio fuel) applications. First a solvent selection as well as an evaluation of the performance was made with Hansen Solubility Parameters and the COnductor-like Screening MOdel for Realistic Solvation (COSMO-RS) simulations. Experiments were performed on rapeseed oil extraction at laboratory and pilot plant scale for the determination of lipid yields, extraction kinetics, diffusion modeling, and complete lipid composition in term of fatty acids and micronutrients (sterols, tocopherols and tocotrienols). Finally, economic and energetic evaluations of the process were conducted to estimate the cost of manufacturing using 2-methyltetrahydrofuran (MeTHF) as alternative solvent compared to hexane as petroleum solvent. PMID:25884332

  11. Allergen profiles of natural rubber latex (NRL) proteins on gloves and glove powders.

    PubMed

    Tomazic-Jezic, Vesna J; Sanchez, B A

    2005-01-01

    The contributing role of glove powder in sensitization to natural rubber latex (NRL) proteins has been well documented in laboratory studies and through clinical evaluations. However, the quantitative relationship of the respiratory and topical exposures in the sensitization process remains unknown because the relative levels of protein on the glove powders in relation to the total levels of protein on NRL gloves have not been determined. In NRL allergens--Hev b 1, Hev b 3, Hev b 5, and Hev b 6.02--on randomly selected surgical and examination NRL gloves. We also examined the binding pattern of the four allergens to several glove powders that showed a different affinity to NRL proteins. The level of powder-bound protein was determined by the ELISA Inhibition Assay (ASTM D6499 standard method). Two cross-linked corn starch powders, one sample of cooking corn starch and one oat starch sample, were exposed to ammoniated (AL) or nonammoniated (NAL) raw NRL protein extracts. The levels of individual allergens were determined using the NRL allergen kit. In the NRL glove extracts we observed a wide range in the total allergen levels and a great diversity in the proportion of the four allergens. On the other hand, the evaluated starches had similar ratios of four individual allergens, regardless of the differences in their total allergen levels. The exposure of starches to NRL proteins with different allergen profiles did not affect the allergen ratio. All samples demonstrated a selective affinity for binding Hev b 1 and Hev b 5 allergens and a lesser affinity for the Hev b 6.02 allergen. Allergen Hev b 6.02 made up about 60% of the total allergen in the NAL extract, but only 12-30% of Hev b 6.02 was bound to starches. In contrast, there was only 3-7% of Hev b 1 allergen in the NAL extract, but powders had 35-45% of Hev b 1. These findings indicate that allergenic properties of NRL gloves and respective glove powders may be different. PMID:15777165

  12. Precautionary labelling of foods for allergen content: are we ready for a global framework?

    PubMed Central

    2014-01-01

    Food allergy appears to be on the rise with the current mainstay of treatment centred on allergen avoidance. Mandatory allergen labelling has improved the safety of food for allergic consumers. However an additional form of voluntary labelling (termed precautionary allergen labelling) has evolved on a wide range of packaged goods, in a bid by manufacturers to minimise risk to customers, and the negative impact on business that might result from exposure to trace amounts of food allergen present during cross-contamination during production. This has resulted in near ubiquitous utilisation of a multitude of different precautionary allergen labels with subsequent confusion amongst many consumers as to their significance. The global nature of food production and manufacturing makes harmonisation of allergen labelling regulations across the world a matter of increasing importance. Addressing inconsistencies across countries with regards to labelling legislation, as well as improvement or even banning of precautionary allergy labelling are both likely to be significant steps forward in improved food safety for allergic families. This article outlines the current status of allergen labelling legislation around the world and reviews the value of current existing precautionary allergen labelling for the allergic consumer. We strongly urge for an international framework to be considered to help roadmap a solution to the weaknesses of the current systems, and discuss the role of legislation in facilitating this. PMID:24791183

  13. Food Production and Processing Considerations of Allergenic Food Ingredients: A Review

    PubMed Central

    Alvarez, Pedro A.; Boye, Joyce I.

    2012-01-01

    Although most consumers show no adverse symptoms to food allergens, health consequences for sensitized individuals can be very serious. As a result, the Codex General Standard for the Labelling of Prepackaged Foods has specified a series of allergenic ingredients/substances requiring mandatory declaration when present in processed prepackaged food products. Countries adhering to international standards are required to observe this minimum of eight substances, but additional priority allergens are included in the list in some countries. Enforcement agencies have traditionally focused their effort on surveillance of prepackaged goods, but there is a growing need to apply a bottom-up approach to allergen risk management in food manufacturing starting from primary food processing operations in order to minimize the possibility of allergen contamination in finished products. The present paper aims to review food production considerations that impact allergen risk management, and it is directed mainly to food manufacturers and policy makers. Furthermore, a series of food ingredients and the allergenic fractions identified from them, as well as the current methodology used for detection of these allergenic foods, is provided. PMID:22187573

  14. Extractable low mass proteins <30kDa from peanut display elevated antigenicity (IgG-binding) and allergenicity (IgE-binding) in vitro and are attenuated by thermal reactivity with non-peanut food ingredients.

    PubMed

    Bennett, Louise; Lee, Alvin

    2016-03-01

    Human allergic reactions to peanut proteins and the associated risk of life-threatening anaphylaxis requires vigilant management of peanuts in food processing. Processed forms of peanuts with attenuated antigenicity and less severe immunogenic responses may lower the risk. Molecular subfractions of raw (UP), blanched (BP) and roasted (RP) peanuts were prepared including water-insoluble (P1), water-soluble high mass (>30kDa, P2) and water-soluble low mass (<30kDa, P3) fractions. Products were screened by measuring binding to IgG (polyclonal antibody against peanut allergen) and IgE (sera from peanut-allergic donors, RAST>3). The results showed that IgE titres were highest for total extracts of RP, particularly for P3 fractions of UP and RP, and were affected by further heating. Antigenicity was also modulated by heating in the presence of either peanut oil or non-peanut food ingredients (lactose, coconut oil). Results support several alternative methods for regulating peanut antigenicity using food processing approaches but require further substantiation in larger numbers of allergic and control donor sera. PMID:26471622

  15. 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

  16. Identification of autoclave-resistant Anisakis simplex allergens.

    PubMed

    Carballeda-Sangiao, Noelia; Olivares, Fabiola; Rodriguez-Mahillo, Ana I; Careche, Mercedes; Tejada, Margarita; Moneo, Ignacio; González-Muñoz, Miguel

    2014-04-01

    Anisakis simplex is a fish parasite able to induce allergic reactions in humans infected when eating raw or undercooked fish parasitized with viable third-stage larvae. Some authors claim that exposure to nonviable Anisakis material can result in allergic symptoms in previously sensitized patients, indicating that parasite allergens are resistant to the thermal treatments of usual cooking procedures. Furthermore, some patients report symptoms after eating canned fish. The aim of this work was the analysis of parasite allergen stability in heating to 121 °C in an autoclave to simulate the thermal process applied to canned fish. Third-stage larvae were subjected to autoclaving for 20, 40, and 80 min, and parasite crude extracts were analyzed by electrophoresis, immunoblotting, and a flow-cytometric basophil activation test. Allergens resistant to autoclaving were separated by reversed-phase high-performance liquid chromatography and identified by ion trap mass spectrometry. Protein analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that autoclaving considerably reduced the number and intensity of identifiable protein bands in a time-dependent manner. Several allergens were detected by immunoblotting with a pool of A. simplex allergic patients' sera after autoclaving. Allergens of 9 and 14 kDa resistant to autoclaving were identified as Ani s 4 and Ani s 1 allergens, respectively. Functional analysis showed that allergens retain their capacity to activate basophils even after autoclaving for 80 min. In conclusion, some relevant A. simplex allergens retain their capacity to bind immunoglobulin E and activate basophils after being subjected to autoclaving, which is a method equivalent to that used in industrial canning processes. PMID:24680072

  17. Immunochemical characterization of acacia pollen allergens and evaluation of cross-reactivity pattern with the common allergenic pollens.

    PubMed

    Shamsbiranvand, Mohammad-Hosein; Khodadadi, Ali; Assarehzadegan, Mohammad-Ali; Borsi, Seyed Hamid; Amini, Akram

    2014-01-01

    Pollen from the Acacia has been reported as an important source of pollinosis in tropical and subtropical regions of the world. The aim of this study was to characterize the IgE binding protein of Acacia farnesiana pollen extract and evaluate cross-reactivity with the most allergenic pollens. In this study, pollen extract was fractionated by SDS-PAGE and the allergenic profile was determined by IgE-immunoblotting and specific ELISA using forty-two Acacia allergic patients. Potential cross-reactivity among Acacia and selected allergenic plants was evaluated with ELISA and immunoblotting inhibition experiments. There were several resolved protein fractions on SDS-PAGE which ranged from 12 to 85 kDa. Several allergenic protein bands with molecular weights approximately between 12 and 85 kDa were recognized by IgE-specific antibodies from Acacia allergic patients in the immunoblot assay. The inhibition by the Prosopis juliflora pollen extract was more than those by other pollen extracts. Moreover, the wheal diameters generated by the Acacia pollen extract were highly correlated with those of P. juliflora pollen extracts. The findings suggest that several proteins such as 15, 23, 45, and 50 kDa proteins could be used as diagnostic and therapeutic reagents for patients allergic to A. farnesiana and P. juliflora. PMID:24949020

  18. Immunochemical Characterization of Acacia Pollen Allergens and Evaluation of Cross-Reactivity Pattern with the Common Allergenic Pollens

    PubMed Central

    Shamsbiranvand, Mohammad-Hosein; Khodadadi, Ali; Assarehzadegan, Mohammad-Ali; Borsi, Seyed Hamid; Amini, Akram

    2014-01-01

    Pollen from the Acacia has been reported as an important source of pollinosis in tropical and subtropical regions of the world. The aim of this study was to characterize the IgE binding protein of Acacia farnesiana pollen extract and evaluate cross-reactivity with the most allergenic pollens. In this study, pollen extract was fractionated by SDS-PAGE and the allergenic profile was determined by IgE-immunoblotting and specific ELISA using forty-two Acacia allergic patients. Potential cross-reactivity among Acacia and selected allergenic plants was evaluated with ELISA and immunoblotting inhibition experiments. There were several resolved protein fractions on SDS-PAGE which ranged from 12 to 85 kDa. Several allergenic protein bands with molecular weights approximately between 12 and 85 kDa were recognized by IgE-specific antibodies from Acacia allergic patients in the immunoblot assay. The inhibition by the Prosopis juliflora pollen extract was more than those by other pollen extracts. Moreover, the wheal diameters generated by the Acacia pollen extract were highly correlated with those of P. juliflora pollen extracts. The findings suggest that several proteins such as 15, 23, 45, and 50 kDa proteins could be used as diagnostic and therapeutic reagents for patients allergic to A. farnesiana and P. juliflora. PMID:24949020

  19. Diagnosis of Chenopodium album allergy with a cocktail of recombinant allergens as a tool for component-resolved diagnosis.

    PubMed

    Nouri, Hamid Reza; Sankian, Mojtaba; Vahedi, Fatemeh; Afsharzadeh, Danial; Rouzbeh, Leila; Moghadam, Maliheh; Varasteh, Abdolreza

    2012-03-01

    Chenopodium album pollen is one of the main sources of pollen allergy in desert and semi-desert areas and contains three identified allergens, so the aim of this study is comparison of the diagnostic potential of C. album recombinant allergens in an allergenic cocktail and C. album pollen extract. Diagnostic potential of the allergenic cocktail was investigated in 32 individuals using skin prick test and obtained results were compared with the acquired results from C. album pollen extract. Specific IgE reactivity against the pollen extract and allergenic cocktail was determined by ELISA and western blotting tests. Inhibition assays were performed for the allergenic cocktail characterization. The exact sensitization profile of all patients was identified which showed that 72, 81 and 46% of allergic patients had IgE reactivity to rChe a 1, rChe a 2 and rChe a 3, respectively. Almost all of C. album allergic patients (30/32) had specific IgE against the allergenic cocktail. In addition, there was a high correlation between IgE levels against the allergenic cocktail and IgE levels against the pollen extract. The allergenic cocktail was able to completely inhibit IgE binding to natural Che a 1, Che a 2 and Che a 3 in C. album extract. In addition, positive skin test reactions were seen in allergic patients that tested by the allergenic cocktail. The reliable results obtained from this study confirmed that the allergenic cocktail with high diagnostic potential could be replaced with natural C. album allergen extracts in skin prick test and serologic tests. PMID:21713408

  20. 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

  1. 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

  2. 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

  3. 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

  4. Cockroach allergy and allergen-specific immunotherapy in asthma: Potential and Pitfalls

    PubMed Central

    Bassirpour, Gillian; Zoratti, Edward

    2014-01-01

    Purpose of review To provide a summary and discussion of cockroach allergy and clinical trials of cockroach allergen immunotherapy. Recent findings Cockroach allergen exposure among sensitized children is increasingly recognized as a key factor contributing to asthma morbidity. Recent trials suggest that cockroach immunotherapy has promise as a treatment strategy with studies demonstrating immunomodulatory and clinical effects. However, a few obstacles need to be overcome to realize the full potential of this treatment modality as cockroach allergic patients often exhibit complex sensitization patterns to multiple cockroach-associated proteins and an immunodominant allergen has not been identified. These factors have made it difficult to produce standardized cockroach allergen extracts that are potent and provide the broad allergen profiles needed for optimal treatment. There have been important advances in the identification and cloning of cockroach allergens and several strategies are being developed to provide therapeutic cockroach allergen products with enhanced clinical efficacy. Summary Allergen immunotherapy has the capability of modulating the immune response to cockroach allergen and has potential as a valuable treatment modality. Further studies of the clinical efficacy along with the development of improved therapeutic products are needed to advance our knowledge and realize the full potential of this promising therapy. PMID:25144264

  5. 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

  6. 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

  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. Effects of reduction and proteolysis on cashew allergens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Allergic reaction to cashew ingestion is frequently more severe than reaction to peanut ingestion, and food allergens are commonly resistant to digestive proteases. The purpose of this study was to characterize the sensitivity of cashew proteins to proteolysis. Cashew protein extracts and purified c...

  9. 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

  10. The Seed Biotinylated Protein of Soybean (Glycine max): A Boiling-Resistant New Allergen (Gly m 7) with the Capacity To Induce IgE-Mediated Allergic Responses.

    PubMed

    Riascos, John J; Weissinger, Sandra M; Weissinger, Arthur K; Kulis, Michael; Burks, A Wesley; Pons, Laurent

    2016-05-18

    Soybean is a common allergenic food; thus, a comprehensive characterization of all the proteins that cause allergy is crucial to the development of effective diagnostic and immunotherapeutic strategies. A cDNA library was constructed from seven stages of developing soybean seeds to investigate candidate allergens. We searched the library for cDNAs encoding a seed-specific biotinylated protein (SBP) based on its allergenicity in boiled lentils. A full-length cDNA clone was retrieved and expressed as a 75.6-kDa His-tagged recombinant protein (rSBP) in Escherichia coli. Western immunoblotting of boiled bacterial extracts demonstrated specific IgE binding to rSBP, which was further purified by metal affinity and anion exchange chromatographies. Of the 23 allergic sera screened by ELISA, 12 contained IgEs specific to the purified rSBP. Circular dichroism spectroscopy revealed a predominantly unordered structure consistent with SBP's heat stability. The natural homologues (nSBP) were the main proteins isolated from soybean and peanut embryos after streptavidin affinity purification, yet they remained low-abundance proteins in the seed as confirmed by LC-MS/MS. Using capture ELISAs, the soybean and peanut nSBPs were bound by IgEs in 78 and 87% of the allergic sera tested. The soybean nSBP was purified to homogeneity and treatments with different denaturing agents before immunoblotting highlighted the diversity of its IgE epitopes. In vitro activation of basophils was assessed by flow cytometry in a cohort of peanut-allergic children sensitized to soybean. Stronger and more frequent (38%) activations were induced by nSBP-soy compared to the major soybean allergen, Gly m 5. SBPs may represent a novel class of biologically active legume allergens with the structural resilience to withstand many food-manufacturing processes. PMID:27108990

  11. Perilla frutescens Leaf Extract Inhibits Mite Major Allergen Der p 2-induced Gene Expression of Pro-Allergic and Pro-Inflammatory Cytokines in Human Bronchial Epithelial Cell BEAS-2B

    PubMed Central

    Liu, Jer-Yuh; Chen, Yi-Ching; Lin, Chun-Hsiang; Kao, Shao-Hsuan

    2013-01-01

    Perilla frutescens has been used in traditional medicine for respiratory diseases due to its anti-bacterial and anti-inflammatory activity. This study aimed to investigate effects of Perilla frutescens leaf extract (PFE) on expression of pro-allergic and pro-inflammatory cytokines in airway epithelial cells exposed to mite major allergen Der p 2 (DP2) and the underlying mechanisms. Our results showed that PFE up to 100 µg/mL had no cytotoxic effect on human bronchial epithelial cell BEAS-2B. Further investigations revealed that PFE dose-dependently diminished mRNA expression of pro-allergic cytokine IL-4, IL-5, IL-13 and GM-CSF, as well as pro-inflammatory cytokine IL-6, IL-8 and MCP-1 in BEAS-2B cells treated with DP2. In parallel to mRNA, the DP-2-elevated levels of the tested cytokines were decreased. Further investigation showed that DP2-indued phosphorylation of p38 MAPK (P38) and JNK, but not Erk1/2, was also suppressed by PFE. In addition, PFE elevated cytosolic IκBα level and decreased nuclear NF-κB level in DP2-stimulated BEAS-2B cells. Taken together, these findings revealed that PFE significantly diminished both mRNA expression and protein levels of pro-allergic and pro-inflammatory cytokines in response to DP2 through inhibition of P38/JNK and NK-κB activation. These findings suggest that PFE should be beneficial to alleviate both allergic and inflammatory responses on airway epithelium in response to aeroallergens. PMID:24204835

  12. 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. PMID:16313690

  13. 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

  14. 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

  15. Review on immunotherapy in airway allergen sensitised patients.

    PubMed

    van der Valk, J P M; de Jong, N W; Gerth van Wijk, R

    2015-07-01

    Allergen immunotherapy is a more than 100-year-old treatment in particular for birch pollen, grass pollen, house dust mite and cat dander sensitised allergic patients. The mechanism of allergen-specific immunotherapy is complex. Different hypotheses have been postulated to explain the mode of action, such as a decrease of the number of tissue mast cells, eosinophils and basophils, an increase of IgG4 and IgA synthesis, a shift from Th2 to Th1 cells and an increase in the number and function of IL-10 producing T-regulatory cells (T-reg). All these immunological effects may contribute to immune tolerance and long-term changes in the immune system. The efficacy and safety of subcutaneous (SCIT) and sublingual immunotherapy (SLIT) with pollen and house dust mite have been investigated in many trials, meta-analyses and reviews. Nowadays grass pollen SLIT and SCIT, and birch pollen and house dust mite SCIT are implemented in clinical practice to treat therapy-resistant patients. However, the treatment is not effective for all patients and often not without side effects. Therefore, the development of new, safer and more effective immunotherapies is needed. These are approached along novel routes, including improved administration, combined treatment with immune response modifiers, fusion with immune response modifiers, allergen coupled to adjuvants and reconstruction of natural extracts with multiple recombinant allergens or with modified allergens. These developments are promising, but more research is required to implement them in clinical practice. PMID:26228190

  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. PMID:27002515

  17. Cockroach-allergen study: allergen patterns of three common cockroach species probed by allergic sera collected in two cities.

    PubMed

    Kang, B C; Wilson, M; Price, K H; Kambara, T

    1991-06-01

    Antigens/allergens of three common cockroach extracts, crude whole body extract of the American cockroach (CRa-A), crude whole body extract of the German cockroach (CRa-G), and crude whole body extract of the Oriental cockroach (CRa-O), were studied with crossed immunoelectrophoresis, crossed radioimmunoelectrophoresis, and Western blot analysis. Sera of cockroach-allergic patients with asthma, 10 from Chicago, Ill. (C group) and six patients from Lexington, Ky. (L group), were used; results were then compared with sera of control subjects with asthma. Qualitative differences in protein bands were noted among CRa-A, CRa-G, and CRa-O by crossed immunoelectrophoresis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Allergen bands on Western blot were analyzed for distribution by molecular weight (MW) with relative intensity scores. Results were compared by species and by geography. Two to 12 allergenic bands of variable MW (14 kd to greater than 116 kd) were identified by 13 of 16 individual sera from cockroach-allergic patients from all three extracts. CRa-A demonstrated 55 bands with an intensity score of 125; CRa-G, 58 bands with an intensity score of 100; and CRa-O, 51 bands with an intensity score of 108. Allergenic bands of CRa-A were identified by six sera of the C group and one sera of the L group, whereas bands of both CRa-G and CRa-O were noted by nine sera of the C group and four sera of the L group. All three species had an allergen band in MW range of 40 to 45 kd that reacted to most sera from cockroach-allergic patients with asthma. PMID:2045612

  18. A revisit to cockroach allergens.

    PubMed

    Sookrung, Nitat; Chaicumpa, Wanpen

    2010-01-01

    Among cockroaches (CR) that live in people's homes, two species, i.e., German CR (Blattella germanica) and American CR (Periplaneta americana) predominate in temperate and tropical areas, respectively. CR is an important source of inhalant indoor allergens that sensitize atopic subjects to (localized) type I hypersensitivity or atopy including allergic rhinitis and atopic asthma. In Thailand the predominant CR species is P. americana. CR allergens are found throughout CR infested houses; the number found in kitchens correlates with the degree of CR infestation while sensitization and reactivation of the allergic morbidity are likely to occur in the living room and bedroom. Levels of the CR allergens in homes of CR allergic Thais, measured by using locally made quantification test kits, revealed that the highest levels occur in dust samples collected from the wooden houses of urban slums and in the cool and dry season. CR allergens are proteins that may be derived from any anatomical part of the insect at any developmental stage. The allergens may be also from CR secretions, excretions, body washes or frass. The proteins may be the insect structural proteins, enzymes or hormones. They may exist as dimers/multimers and/or in different isoforms. Exposure to CR allergens in infancy leads to allergic morbidity later in life. Clinical symptoms of CR allergy are usually more severe and prolonged than those caused by other indoor allergens. The mechanisms of acute and chronic airway inflammation and airway hyper-responsiveness (AHR) have been addressed including specific IgE- and non-IgE-mediated mechanisms, i.e., role of protease-activated receptor-2 (PAR2). Participation of various allergen activated-CD4+ T cells of different sublineages, i.e., Th2, Th17, Th22, Th9, Th25, Tregs/Th3 as well as invariant NKT cells, in asthma pathogenesis have been mentioned. The diagnosis of CR allergy and the allergy intervention by CR population control are also discussed. PMID:21038777

  19. Aryl Hydrocarbon Receptor Protects Lungs from Cockroach Allergen-Induced Inflammation by Modulating Mesenchymal Stem Cells.

    PubMed

    Xu, Ting; Zhou, Yufeng; Qiu, Lipeng; Do, Danh C; Zhao, Yilin; Cui, Zhuang; Wang, Heng; Liu, Xiaopeng; Saradna, Arjun; Cao, Xu; Wan, Mei; Gao, Peisong

    2015-12-15

    Exposure to cockroach allergen leads to allergic sensitization and increased risk of developing asthma. Aryl hydrocarbon receptor (AhR), a receptor for many common environmental contaminants, can sense not only environmental pollutants but also microbial insults. Mesenchymal stem cells (MSCs) are multipotent progenitor cells with the capacity to modulate immune responses. In this study, we investigated whether AhR can sense cockroach allergens and modulate allergen-induced lung inflammation through MSCs. We found that cockroach allergen-treated AhR-deficient (AhR(-/-)) mice showed exacerbation of lung inflammation when compared with wild-type (WT) mice. In contrast, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), an AhR agonist, significantly suppressed allergen-induced mouse lung inflammation. MSCs were significantly reduced in cockroach allergen-challenged AhR(-/-) mice as compared with WT mice, but increased in cockroach allergen-challenged WT mice when treated with TCDD. Moreover, MSCs express AhR, and AhR signaling can be activated by cockroach allergen with increased expression of its downstream genes cyp1a1 and cyp1b1. Furthermore, we tracked the migration of i.v.-injected GFP(+) MSCs and found that cockroach allergen-challenged AhR(-/-) mice displayed less migration of MSCs to the lungs compared with WT. The AhR-mediated MSC migration was further verified by an in vitro Transwell migration assay. Epithelial conditioned medium prepared from cockroach extract-challenged epithelial cells significantly induced MSC migration, which was further enhanced by TCDD. The administration of MSCs significantly attenuated cockroach allergen-induced inflammation, which was abolished by TGF-β1-neutralizing Ab. These results suggest that AhR plays an important role in protecting lungs from allergen-induced inflammation by modulating MSC recruitment and their immune-suppressive activity. PMID:26561548

  20. Analysis of cross-reactivity between group 1 allergens from mites.

    PubMed

    Mary, Cruz Luz; López-Malpica, Fernando; Díaz, Ana María

    2008-06-01

    Mite allergen exposure can lead to sensitization in genetically predisposed individuals, and the development of asthma in previously sensitized individuals. The major allergens of mites belong to Dermatophagoides spp. and Blomia tropicalis (Bt). Various allergens of Bt have been cloned and sequenced. Some of them show homology sequence with purified allergens from Dermatophagoides pteronissynus (Dp). Recently, the allergen group 1 from Bt, Blo t 1, was cloned and sequenced at our laboratory. Recombinant Blo t 1 showed 35 % of identity and 50% of similarity with group 1 allergens as Der p 1 (from Dp), Der f 1 (from D. farinae) and Eur m 1 (from Euroglyphus maynei) at amino acid level. This would suggest that cross-reactivity between allergens of different mite species could exist. Here, we analyzed the crossreactivity between group 1 allergens from mites using recombinant proteins and monoclonal antibodies against them. ELISA inhibition assay showed that crossreactivity between homologous allergens from Dermatophagoides spp. is high, but it is low to moderate between mites from different species. IgE-reactivity analysis using serum samples from allergic individuals revealed a strong reactivity of rBlo t 1 for serum samples from subjects with highly positive reaction to Bt extract in skin test, but lack of reactivity of this protein with serum samples from individuals with highly positive reaction to house dust mite extract in the skin test. These results suggest that it is important to include Bt allergens in routine skin test in order to improve the diagnostic accuracy and precision of allergies. PMID:18616045

  1. A review of the materials and allergens in protective gloves.

    PubMed

    Rose, Rebecca F; Lyons, Paul; Horne, Helen; Mark Wilkinson, S

    2009-09-01

    The ingredients previously reported to cause protective glove allergy are presented and evaluated for strength of evidence. Allergens that have caused both delayed hypersensitivity and contact urticaria are considered for rubber, plastic, leather, and textile gloves. The current guidelines regarding glove manufacture are described. A list of materials confirmed by the industry to be used in glove production is presented together with a suggested series for investigating patients with delayed type hypersensitivity and contact urticaria secondary to glove use. PMID:19780770

  2. Determinants of efficacy and safety in epicutaneous allergen immunotherapy: summary of three clinical trials

    PubMed Central

    Senti, G; von Moos, S; Tay, F; Graf, N; Johansen, P; Kündig, T M

    2015-01-01

    The results of our third trial on epicutaneous allergen-specific immunotherapy (EPIT) will be presented and discussed in the context of our previous trials. This monocentric, placebo-controlled, double-blind phase I/IIa trial included 98 patients with grass pollen rhinoconjunctivitis. Prior to the pollen season 2009, patients received six patches (allergen extract: n = 48; placebo: n = 50) with weekly intervals, administered onto tape-stripped skin. Allergen EPIT produced a median symptom improvement of 48% in 2009 and 40% in the treatment-free follow-up year 2010 as compared to 10% and 15% improvement after placebo EPIT (P = 0.003). After allergen EPIT but not placebo EPIT, conjunctival allergen reactivity was significantly decreased and allergen-specific IgG4 responses were significantly elevated (P < 0.001). In conclusion, our three EPIT trials found that allergen EPIT can ameliorate hay fever symptoms. Overall, treatment efficacy appears to be determined by the allergen dose. Local side-effects are determined by the duration of patch administration, while risk of systemic allergic side-effects is related to the degree of stratum corneum disruption. PMID:25704072

  3. 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 ...

  4. Constructing a hybrid molecule with low capacity of IgE binding from Chenopodium album pollen allergens.

    PubMed

    Nouri, Hamid Reza; Varasteh, Abdolreza; Vahedi, Fatemeh; Chamani, Jamshidkhan; Afsharzadeh, Danial; Sankian, Mojtaba

    2012-05-30

    Allergen specific immunotherapy is the only remedy to prevent the progression of allergic diseases. Nowadays, using of recombinant allergens with reduced IgE-binding capacity is an ideal tool for allergen immunotherapy. Therefore, in this study we focused on a hybrid molecule (HM) production with reduced IgE reactivity from Chenopodium album pollen allergens. By means of genetic engineering, a head to tail structure of the three allergens of the C. album pollen was designed. The resulting DNA construct coding for a 46kDa HM was inserted into an expression vector and expressed as hexahistidine tagged fusion protein in Escherichia coli. IgE reactivity of the HM was evaluated by western blotting, inhibition ELISA and in vivo skin prick test and its immunogenic property was tested by proliferation assay. The recombinant HM was expressed and purified by nickel-affinity chromatography. Comparison of the recombinant HM with a mixture of three recombinant allergens, as well as natural allergens in the whole C. album pollen extract via immunological experiments revealed that it has a much lower potential of IgE reactivity. Furthermore, in vivo skin prick tests showed that it has a significantly lower potency to induce cutaneous reactions than the mixture of recombinant wild type allergens and whole extract while, it had been preserved immunogenic properties. Our results have demonstrated that assembling three allergens of C. album in a hybrid molecule can reduce its IgE reactivity. PMID:22504204

  5. Multi-allergen Quantitation and the Impact of Thermal Treatment in Industry-Processed Baked Goods by ELISA and Liquid Chromatography-Tandem Mass Spectrometry.

    PubMed

    Parker, Christine H; Khuda, Sefat E; Pereira, Marion; Ross, Mark M; Fu, Tong-Jen; Fan, Xuebin; Wu, Yan; Williams, Kristina M; DeVries, Jonathan; Pulvermacher, Brian; Bedford, Binaifer; Zhang, Xi; Jackson, Lauren S

    2015-12-16

    Undeclared food allergens account for 30-40% of food recalls in the United States. Compliance with ingredient labeling regulations and the implementation of effective manufacturing allergen control plans require the use of reliable methods for allergen detection and quantitation in complex food products. The objectives of this work were to (1) produce industry-processed model foods incurred with egg, milk, and peanut allergens, (2) compare analytical method performance for allergen quantitation in thermally processed bakery products, and (3) determine the effects of thermal treatment on allergen detection. Control and allergen-incurred cereal bars and muffins were formulated in a pilot-scale industry processing facility. Quantitation of egg, milk, and peanut in incurred baked goods was compared at various processing stages using commercial enzyme-linked immunosorbent assay (ELISA) kits and a novel multi-allergen liquid chromatography (LC)-tandem mass spectrometry (MS/MS) multiple-reaction monitoring (MRM) method. Thermal processing was determined to negatively affect the recovery and quantitation of egg, milk, and peanut to different extents depending on the allergen, matrix, and analytical test method. The Morinaga ELISA and LC-MS/MS quantitative methods reported the highest recovery across all monitored allergens, whereas the ELISA Systems, Neogen BioKits, Neogen Veratox, and R-Biopharm ELISA Kits underperformed in the determination of allergen content of industry-processed bakery products. PMID:26595064

  6. 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...

  7. 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

  8. Proteomic analysis of major and minor allergens from isolated pollen cytoplasmic granules.

    PubMed

    Abou Chakra, Oussama R; Sutra, Jean-Pierre; Demey Thomas, Emmanuelle; Vinh, Joëlle; Lacroix, Ghislaine; Poncet, Pascal; Sénéchal, Hélène

    2012-02-01

    Grass pollen is one of the most important vectors of aeroallergens. Under atmospheric conditions, pollen grains can release pollen cytoplasmic granules (PCGs). The allergens associated with these intrinsic subfractions induce, in laboratory animals as well as in asthmatic patients, allergic and inflammatory responses. The objectives of this study were to characterize the PCGs' intrinsic allergens and to compare them with those of pollen grains. The water-soluble proteins were extracted from pollen grains and their PCGs. IgE-binding proteins were analyzed and characterized through an allergomic strategy: 1- and 2-dimensional gel electrophoresis (1-DE and 2-DE), immunoblotting, using grass-pollen-sensitized patient sera, mass spectrometry (MS) analysis, and database searching. Several of the allergens listed in the IUIS nomenclature, Phl p 1, 4, 5, 6, and 12, were detected in pollen and PCG extracts, whereas Phl p 11 was found only in PCGs, and Phl p 2 as well as Phl p 13 were found only in pollen extract. Some other allergens not listed in the IUIS nomenclature were also characterized in both pollen and PCG extracts. Since the major grass pollen allergens were found in PCGs and because of their small size, these submicronic particles should be considered as very potent sensitizing and challenging respirable vectors of allergens. PMID:22188203

  9. Understanding allergic asthma from allergen inhalation tests.

    PubMed

    Cockcroft, Donald W; Hargreave, Fredrick E; O'Byrne, Paul M; Boulet, Louis-Philippe

    2007-10-01

    The allergen challenge has evolved, in less than 150 years, from a crude tool used to document the etiology of allergen-induced disease to a well-controlled tool used today to investigate the pathophysiology and pharmacotherapy of asthma. Highlights of the authors' involvement with the allergen challenge include confirmation of the immunoglobulin E-dependence of the late asthmatic response, importance of (nonallergic) airway hyper-responsiveness as a determinant of the airway response to allergen, identification of allergen-induced increase in airway hyper-responsiveness, documentation of beta(2)-agonist-induced increase in airway response to allergen (including eosinophilic inflammation), advances in understanding the pathophysiology and kinetics of allergen-induced airway responses, and development of a multicentre clinical trial group devoted to using the allergen challenge for investigating promising new therapeutic strategies for asthma. PMID:17948142

  10. 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

  11. Recombinant allergens for specific immunotherapy.

    PubMed

    Cromwell, Oliver; Häfner, Dietrich; Nandy, Andreas

    2011-04-01

    Recombinant DNA technology provides the means for producing allergens that are equivalent to their natural counterparts and also genetically engineered variants with reduced IgE-binding activity. The proteins are produced as chemically defined molecules with consistent structural and immunologic properties. Several hundred allergens have been cloned and expressed as recombinant proteins, and these provide the means for making a very detailed diagnosis of a patient's sensitization profile. Clinical development programs are now in progress to assess the suitability of recombinant allergens for both subcutaneous and sublingual immunotherapy. Recombinant hypoallergenic variants, which are developed with the aim of increasing the doses that can be administered while at the same time reducing the risks for therapy-associated side effects, are also in clinical trials for subcutaneous immunotherapy. Grass and birch pollen preparations have been shown to be clinically effective, and studies with various other allergens are in progress. Personalized or patient-tailored immunotherapy is still a very distant prospect, but the first recombinant products based on single allergens or defined mixtures could reach the market within the next 5 years. PMID:21377719

  12. 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

  13. Helminth infection alters IgE responses to allergens structurally related to parasite proteins

    PubMed Central

    Santiago, Helton da Costa; Ribeiro-Gomes, Flávia L.; Bennuru, Sasisekhar; Nutman, Thomas B.

    2014-01-01

    Immunological cross-reactivity between environmental allergens and helminth proteins has been demonstrated, though the clinically-related implications of this cross-reactivity have not been addressed. To investigate the impact of molecular similarity among allergens and cross-reactive homologous helminth proteins in IgE-based serologic assessment of allergic disorders in helminth-infected population, we performed Immunocap™ tests in filarial-infected and non-infected individuals for IgE measurements to allergen extracts that contained proteins with high levels of homology with helminth proteins and IgE against representative recombinant allergens with and without helminth homologues were performed. The impact of helminth infection on the levels and function of the IgE to these specific homologous and non-homologous allergens was corroborated in an animal model. We found that having a tissue-invasive filarial infection increased the serological prevalence of Immunocap™ identified IgE directed against house dust mite and cockroach, but not against timothy grass, the latter with few allergens with homologues in helminth infection. IgE ELISA confirmed that filaria-infected individuals had higher IgE prevalences to those recombinant allergens that had homologues in helminths. Mice infected with helminth Heligmosomoides polygyrus displayed increased levels of IgE and positive skin tests to allergens with homologues in the parasite. These results show that cross-reactivity among allergens and helminth proteins can have practical implications altering serologic approaches to allergen testing and brings a new perspective to the Hygiene Hypothesis. PMID:25404363

  14. Helminth infection alters IgE responses to allergens structurally related to parasite proteins.

    PubMed

    Santiago, Helton da Costa; Ribeiro-Gomes, Flávia L; Bennuru, Sasisekhar; Nutman, Thomas B

    2015-01-01

    Immunological cross-reactivity between environmental allergens and helminth proteins has been demonstrated, although the clinically related implications of this cross-reactivity have not been addressed. To investigate the impact of molecular similarity among allergens and cross-reactive homologous helminth proteins in IgE-based serologic assessment of allergic disorders in a helminth-infected population, we performed ImmunoCAP tests in filarial-infected and noninfected individuals for IgE measurements to allergen extracts that contained proteins with high levels of homology with helminth proteins as well as IgE against representative recombinant allergens with and without helminth homologs. The impact of helminth infection on the levels and function of the IgE to these specific homologous and nonhomologous allergens was corroborated in an animal model. We found that having a tissue-invasive filarial infection increased the serological prevalence of ImmunoCAP-identified IgE directed against house dust mite and cockroach, but not against timothy grass, the latter with few allergens with homologs in helminth infection. IgE ELISA confirmed that filaria-infected individuals had higher IgE prevalences to those recombinant allergens that had homologs in helminths. Mice infected with the helminth Heligmosomoides polygyrus displayed increased levels of IgE and positive skin tests to allergens with homologs in the parasite. These results show that cross-reactivity among allergens and helminth proteins can have practical implications, altering serologic approaches to allergen testing and bringing a new perspective to the "hygiene hypothesis." PMID:25404363

  15. 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

  16. Allergenic compounds on the inner and outer surfaces of natural latex gloves: MALDI mass spectrometry and imaging of proteinous allergens.

    PubMed

    Marchetti-Deschmann, Martina; Allmaier, Günter

    2009-01-01

    Natural latex gloves are the cause of a severe health problem to an increasing number of healthcare workers or patients due to the presence of protein allergens as Hevein or Rubber Elongation Factor (REF). One of the most challenging problems is the in situ localization of theses allergens in, e.g. gloves, to estimate the allergenic potential of the latex material. A sample preparation protocol applying a binary matrix-assisted laser desorption/ionization(MALDI) matrix containing alpha-cyano-4-hydroxy cinnamic acid (CHCA) and 2,5-dihydroxy benzoic acid (DHB) on trifluoro acetic acid (TFA) etched latex glove surfaces allowed the direct determination (exact molecular weight) of Hevein, REF and a truncated form of REF (tREF) within nine different brands of natural latex gloves by means of MALDI-TOF-MS in the linear mode. MALDI mass spectrometry demonstrated that Hevein, tREF and REF were present on the inner surfaces (in direct contact with the skin) of many, but not all, investigated gloves without any prior extraction procedure. Additionally, different isoforms of the allergen Hevein were detected (exhibiting ragged C-termini). tREF and REF could always be detected beside each other, but were not observed on every latex glove sample, which contained Hevein. It was also demonstrated that there is a significant difference in terms of proteins and polymers between inner and outer surfaces of gloves, which helps to explain the different allergenic potential of these.MALDI imaging allowed for the first time the unambiguous localization of all three allergens in parallel and showed that Hevein was present on 36% of the investigated area of a latex glove with a certain localization, whereupon, tREF and REF were only found on 25% of the investigated material. PMID:18720446

  17. 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...

  18. IgE response to two new allergen proteins of Solanum melongena L. (eggplant).

    PubMed

    Hoseini-Alfatemi, Seyedeh Mahsan; Bayry, Jagadeesh; Sharifi-Rad, Javad

    2015-12-01

    A number of allergens from eggplant (Solanum melongena L.) have been previously identified. In this study, we could detect IgE reactivity of two allergic subjects' sera towards two protein bands of molecular mass of about 35 and 15 kDa. As IgE were reactive to both raw and cooked eggplant extracts, a heat-stable nature of these novel allergens is apparent. PMID:26455782

  19. Allergenic potential of novel foods.

    PubMed

    Meredith, Clive

    2005-11-01

    Concerns have been expressed that the introduction of novel foods into the diet might lead to the development of new food allergies in consumers. Novel foods can be conveniently divided into GM and non-GM categories. Decision-tree approaches (e.g. International Life Sciences Institute-International Food Biotechnology Council and WHO/FAO) to assess the allergenic potential of GM foods were developed following the discovery, during product development, of the allergenic potential of GM soyabean expressing a gene encoding a storage protein from Brazil nut (Bertolletia excelsa). Within these decision trees considerations include: the source of the transgene; amino acid homology with known allergens; cross-reactivity with IgE from food-allergic individuals; resistance to proteolysis; prediction using animal models of food allergy. Such decision trees are under constant review as new knowledge and improved models emerge, but they provide a useful framework for the assessment of the allergenic potential of GM foods. For novel non-GM foods the assessment of allergenic potential is more subjective; some foods or food ingredients will need no assessment other than a robust protein assay to demonstrate the absence of protein. Where protein is present in the novel non-GM food, hazard and risk assessments need to be made in terms of the quantity of protein that might be consumed, the identity of individual protein components and their relationships to known food allergens. Where necessary, this assessment would extend to serum screening for potential cross-reactivities, skin-prick tests in previously-sensitised individuals and double-blind placebo-controlled food challenges. PMID:16313692

  20. 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

  1. The pollen enigma: modulation of the allergic immune response by non-allergenic, pollen-derived compounds.

    PubMed

    Gilles, Stefanie; Behrendt, Heidrun; Ring, Johannes; Traidl-Hoffmann, Claudia

    2012-01-01

    The question what makes an allergen an allergen puzzled generations of researchers. Pollen grains of anemophilous plants are the most important allergen carriers in ambient air, and pollinosis is a highly prevalent multi-organ disease in civilized countries. In the past, research on the allergenicity of pollen has mainly focused on elucidating genetic predisposing factors and on defining certain structural characteristics of pollen derived allergens. Recently, studies extended to the analysis of non-allergenic, adjuvant mediators co-released from pollen. Besides active proteases and oxidases, extracts of pollen contain low molecular weight molecules like pollen-associated lipid mediators or adenosine exhibiting a potential to stimulate and modulate cultured human immune cells. This article reviews our current knowledge on non-allergenic, protein and non-protein compounds from pollen and their in vitro and in vivo effects on the allergic immune response. To ultimately judge the physiological relevance of these compounds, a systematic approach will be needed comparing their releasability, content and activity in different, allergenic and non-allergenic, pollen species. System biology such as proteome and metabolome analysis will be a useful future approach to better understand pollen biology. PMID:22390694

  2. Use of artichoke (Cynara scolymus) flower extract as a substitute for bovine rennet in the manufacture of Gouda-type cheese: characterization of aspartic proteases.

    PubMed

    Llorente, Berta E; Obregón, Walter David; Avilés, Francesc X; Caffini, Néstor O; Vairo-Cavalli, Sandra

    2014-09-15

    Artichoke (Cynara scolymus L.) flower extract was assayed with the aim of replacing animal rennet in the manufacture of Gouda-type cheeses from bovine milk. Floral extract coagulated milk within a suitable time for use on an industrial scale, while the yield of cheese obtained was equal to that achieved with bovine abomasum. Five proteolytic fractions with milk-clotting activity were isolated in a two-step purification protocol, three belonging to the cardosin group. Cheeses made with C. scolymus proteases must be brined for a longer period (40 h) to prevent overproteolysis and avoid the development of a background flavor. The type of coagulant (bovine or vegetable) had no significant effect on the cheeses' chemical parameters analyzed throughout ripening, and no significant organoleptic differences were detected between those manufactured with C. scolymus or animal rennet. The results indicate that C. scolymus flower extract is suitable for replacing animal rennet in the production of Gouda-type cheeses. PMID:24767026

  3. Sputum RNA signature in allergic asthmatics following allergen bronchoprovocation test

    PubMed Central

    Zuiker, Rob G.J.A.; Tribouley, Catherine; Diamant, Zuzana; Boot, J. Diderik; Cohen, Adam F.; Van Dyck, K.; De Lepeleire, I.; Rivas, Veronica M.; Malkov, Vladislav A.; Burggraaf, Jacobus; Ruddy, Marcella K.

    2016-01-01

    Background Inhaled allergen challenge is a validated disease model of allergic asthma offering useful pharmacodynamic assessment of pharmacotherapeutic effects in a limited number of subjects. Objectives To evaluate whether an RNA signature can be identified from induced sputum following an inhaled allergen challenge, whether a RNA signature could be modulated by limited doses of inhaled fluticasone, and whether these gene expression profiles would correlate with the clinical endpoints measured in this study. Methods Thirteen non-smoking, allergic subjects with mild-to-moderate asthma participated in a randomised, placebo-controlled, 2-period cross-over study following a single-blind placebo run-in period. Each period consisted of three consecutive days, separated by a wash-out period of at least 3 weeks. Subjects randomly received inhaled fluticasone ((FP) MDI; 500 mcg BID×5 doses in total) or placebo. On day 2, house dust mite extract was inhaled and airway response was measured by FEV1 at predefined time points until 7 h post-allergen. Sputum was induced by NaCl 4.5%, processed and analysed at 24 h pre-allergen and 7 and 24 h post-allergen. RNA was isolated from eligible sputum cell pellets (<80% squamous of 500 cells), amplified according to NuGEN technology, and profiled on Affymetrix arrays. Gene expression changes from baseline and fluticasone treatment effects were evaluated using a mixed effects ANCOVA model at 7 and at 24 h post-allergen challenge. Results Inhaled allergen-induced statistically significant gene expression changes in sputum, which were effectively blunted by fluticasone (adjusted p<0.025). Forty-seven RNA signatures were selected from these responses for correlation analyses and further validation. This included Th2 mRNA levels for cytokines, chemokines, high-affinity IgE receptor FCER1A, histamine receptor HRH4, and enzymes and receptors in the arachidonic pathway. Individual messengers from the 47 RNA signatures correlated significantly

  4. Production and analysis of recombinant tree nut allergens.

    PubMed

    Willison, Leanna N; Sathe, Shridhar K; Roux, Kenneth H

    2014-03-01

    Allergic reactions to tree nuts are a growing global concern as the number of affected individuals continues to rise. Unlike some food allergies, tree nuts can cause severe reactions that persist throughout life. The tree nuts discussed in this review include those most commonly responsible for allergic reactions: cashew, almond, hazelnut, walnut, pecan, Brazil nut, pistachio, and chestnut. The native allergenic proteins derived from tree nuts are frequently difficult to isolate and purify and may not be adequately represented in aqueous nut protein extracts. Consequently, defined recombinant allergens have become useful reagents in a variety of immunoassays aimed at the diagnosis of tree nut allergy, assessing cross-reactivity between various nuts and other seeds, mapping of IgE binding epitopes, and analyzing the effects of the food matrix, food processing, and gastric digestion on allergenicity. This review describes the approaches that can be used for the production of recombinant tree nut allergens and addresses key issues associated with their production and downstream applications. PMID:23911839

  5. A review of allergy and allergen specific immunotherapy.

    PubMed

    Bidad, Katayoon; Nicknam, Mohammad Hossein; Farid, Reza

    2011-03-01

    Since 20th century, when allergy was defined, an ongoing attempt for discovering the mechanisms underlying it and its treatment began. Defining allergens as well as cells such as regulatory T-cells and characterizing the antibodies involved in the pathogenesis (including blocking antibodies) have helped very much towards a better understanding of the immunologic process. However, allergen specific immunotherapy (SIT), as a specific curative treatment for allergy also dates back to the beginning of the previous century and has progressed considerably during these years. SIT similar to natural immunomodulation, directs the immune response towards tolerance. New strategies in this field, such as using recombinant allergens, T- and B-cell-epitope-containing peptides, and DNA vaccination have shown promising results. Sublingual immunotherapy, although not yet FDA-approved, as an alternative strategy in SIT has demonstrated efficacy as well as safety. Furthermore, allergen extracts, their standardization and their modification have also been the focus of much research. Undoubtedly, specific immunotherapy is proven to be an efficacious method to treat allergy, so its cost-effectiveness should be estimated in developing countries in order to include it in the country's health priorities. Informing physicians about the new anti-vaccination movement is also crucial. PMID:21358009

  6. Allergen-specific immunotherapy in pediatric allergic asthma

    PubMed Central

    2016-01-01

    Allergen-specific immunotherapy (AIT) is the only curative way that can change the immunologic response to allergens and thus can modify the natural progression of allergic diseases. There are some important criteria which contributes significantly on efficacy of AIT, such as the allergen extract used for treatment, the dose and protocol, patient selection in addition to the severity and control of asthma. The initiation of AIT in allergic asthma should be considered in intermittent, mild and moderate cases which coexisting with other allergic diseases such as allergic rhinitis, and in case of unacceptable adverse effects of medications. Two important impact of AIT; steroid sparing effect and preventing from progression to asthma should be taken into account in pediatric asthma when making a decision on starting of AIT. Uncontrolled asthma remains a significant risk factor for adverse events and asthma should be controlled both before and during administration of AIT. The evidence concerning the efficacy of subcutaneous (SCIT) and sublingual immunotherapy (SLIT) for treatment of pediatric asthma suggested that SCIT decreases asthma symptoms and medication scores, whereas SLIT can ameliorate asthma symptoms. Although the effectiveness of SCIT has been shown for both seasonal and perennial allergens, the data for SLIT is less convincing for perennial allergies in pediatric asthma. PMID:27489785

  7. Allergen-specific immunotherapy in pediatric allergic asthma.

    PubMed

    Yukselen, Ayfer

    2016-07-01

    Allergen-specific immunotherapy (AIT) is the only curative way that can change the immunologic response to allergens and thus can modify the natural progression of allergic diseases. There are some important criteria which contributes significantly on efficacy of AIT, such as the allergen extract used for treatment, the dose and protocol, patient selection in addition to the severity and control of asthma. The initiation of AIT in allergic asthma should be considered in intermittent, mild and moderate cases which coexisting with other allergic diseases such as allergic rhinitis, and in case of unacceptable adverse effects of medications. Two important impact of AIT; steroid sparing effect and preventing from progression to asthma should be taken into account in pediatric asthma when making a decision on starting of AIT. Uncontrolled asthma remains a significant risk factor for adverse events and asthma should be controlled both before and during administration of AIT. The evidence concerning the efficacy of subcutaneous (SCIT) and sublingual immunotherapy (SLIT) for treatment of pediatric asthma suggested that SCIT decreases asthma symptoms and medication scores, whereas SLIT can ameliorate asthma symptoms. Although the effectiveness of SCIT has been shown for both seasonal and perennial allergens, the data for SLIT is less convincing for perennial allergies in pediatric asthma. PMID:27489785

  8. Maternal respiratory sensitization and gestational allergen exposure does not affect subsequent pup responses to homologous or heterologous allergen.

    PubMed

    Pucheu-Haston, Cherie M; Copeland, Lisa B; Haykal-Coates, Najwa; Ward, Marsha D W

    2010-03-01

    Evidence suggests that the predisposition towards atopy begins early in life. Maternal allergy has been associated with an increased risk of the development of allergic disease in offspring. Some studies suggest that the development of childhood atopy may also be influenced by prenatal allergen exposure. In this study, a respiratory allergen exposure model was used to determine the impact of maternal sensitization (with or without additional exposures during pregnancy) on subsequent pup responses to homologous or heterologous allergen. Female BALB/c mice received two intratracheal aspiration (IA) exposures to Metarhizium anisopliae crude antigen (MACA) or Hank's buffered salt solution (HBSS) prior to breeding. Some mice also received three additional exposures during pregnancy. Control mothers did not receive treatment. Young adult offspring received three IA exposures to MACA, house dust mite extract (HDM) or HBSS. Offspring sensitized as young adults to either HDM or MACA developed an airway inflammatory response, including increased bronchoalveolar lavage fluid lactate dehydrogenase activity, total protein and total and differential cell counts compared to offspring exposed to HBSS. Increased airway responsiveness to methacholine was observed in pups treated with HDM but not with MACA. Maternal sensitization status (with or without gestational allergen exposure) had no effect on offspring response to either MACA or HDM. In conclusion, this study demonstrates that IA administration of MACA or HDM extract to young adult BALB/c mice induces the development of an inflammatory airway response. In contrast to previous reports, neither maternal sensitization nor gestational allergen exposure could be demonstrated to have a clear effect on offspring sensitization. This discrepancy may be a function of the respiratory sensitization and exposure protocol used in this study, which mimics natural sensitization more closely than do parenteral routes of exposure. PMID

  9. Evidence of a novel allergenic protein Narcin in the bulbs of Narcissus tazetta

    PubMed Central

    Sinha, Mau; Singh, Amar; Shokeen, Akshita; Sharma, Pradeep; Kaushik, Sanket; Mitra, Dipendra K; Kaur, Punit; Sharma, Sujata; Singh, Tej P

    2013-01-01

    Several plant-derived allergens have been identified which result in the formation of immunoglobulin E antibodies. Primarily, these allergens belong to the protein families including seed storage proteins, structural proteins and pathogenesis-related proteins. Several allergens are also reported from flower bulbs which cause contact dermatitis. Such symptoms are highly common with the bulb growers handling different species of Narcissus. Narcissus toxicity is also reported if the bulbs are consumed accidentally. The present study aimed to characterize the protein from the bulbs of Narcissus tazetta responsible for its allergenic response. A 13 kDa novel allergenic protein, Narcin was isolated from the bulbs of Narcissus tazetta. The protein was extracted using ammonium sulfate fractionation. The protein was further purified by anion exchange chromatography followed by gel filtration chromatography. The N-terminal sequence of the first 15 amino-acid residues was determined using Edman degradation. The allergenicity of the protein was measured by cytokine production using flow cytometry in peripheral blood mononuclear cells. Further estimation of total IgE was performed by ELISA method. This novel protein was found to induce pro-inflammatory cytokines and thus induce allergy by elevating total IgE level. The novel protein, Narcin isolated from Narcissus tazetta was found to exhibit allergenic properties. PMID:23936740

  10. [Immunoproteomics of non water-soluble allergens from 4 legumes flours: peanut, soybean, sesame and lentil].

    PubMed

    Bouakkadia, Hayette; Boutebba, Aissa; Haddad, Iman; Vinh, Joëlle; Guilloux, Laurence; Sutra, Jean-Pierre; Sénéchal, Hélène; Poncet, Pascal

    2015-01-01

    Peanut, soybean, sesame and lentil are members of legumes worldwide consumed by human that can induce food allergy in genetically predisposed individuals. Several protein allergens, mainly water-soluble, have been described. We studied the non water-soluble fraction from these 4 food sources using immunoproteomics tools and techniques. Flour extracts were solubilized in detergent and chaotropes and analysed in 1 and 2 dimensional gel electrophoresis (2D). Results showed numerous proteins exhibiting wide ranges of isoelectric points and relative molecular masses. When IgE immunoreactivities of 18 food allergy patients were individually tested in 1 and 2D western-blots, a very diversified IgE repertoire was observed, reflecting extensive cross-reactivities but also co-sensitizations. Besides already well known and characterized allergens, mass spectrometry analysis allowed the identification of 22 allergens undescribed until now: 10 in peanut, 2 in soybean, 3 in sesame and 7 in lentil. Three allergens are legume storage proteins and the others belong to transport proteins, nucleotide binding proteins and proteins involved in the regulation of metabolism. Seven proteins are potentially similar to allergens described in plants and fungi and 11 are not related to any known allergen. Our results contribute to increase the repertoire of legume allergens that may improve the diagnosis, categorize patients and thus provide a better treatment of patients. PMID:26635049

  11. New insights into ragweed pollen allergens.

    PubMed

    Bordas-Le Floch, Véronique; Groeme, Rachel; Chabre, Henri; Baron-Bodo, Véronique; Nony, Emmanuel; Mascarell, Laurent; Moingeon, Philippe

    2015-11-01

    Pollen allergens from short ragweed (Ambrosia artemisiifolia) cause severe respiratory allergies in North America and Europe. To date, ten short ragweed pollen allergens belonging to eight protein families, including the recently discovered novel major allergen Amb a 11, have been recorded in the International Union of Immunological Societies (IUIS) allergen database. With evidence that other components may further contribute to short ragweed pollen allergenicity, a better understanding of the allergen repertoire is a requisite for the design of proper diagnostic tools and efficient immunotherapies. This review provides an update on both known as well as novel candidate allergens from short ragweed pollen, identified through a comprehensive characterization of the ragweed pollen transcriptome and proteome. PMID:26383916

  12. Allergenicity of Peanut Proteins is Retained Following Enzymatic Hydrolysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rationale: Hydrolysis of peanut proteins by food-grade enzymes may reduce allergenicity and could lead to safer forms of immunotherapy. Methods: Light roasted peanut flour extracts were digested with pepsin (37°C, pH 2), Alcalase (60°C pH 8), or Flavourzyme (50°C, pH 7) up to 1 hr, or sequentially w...

  13. Guidelines for the use of allergen immunotherapy. Canadian Society of Allergy and Clinical Immunology.

    PubMed Central

    1995-01-01

    OBJECTIVES: To recommend guidelines for the use of allergen immunotherapy to treat allergies in patients for whom allergen avoidance and drug therapy have not been sufficiently effective. OPTIONS: High-dose or low-dose allergen immunotherapy for the treatment of IgE-mediated allergy to insect stings, allergic rhinoconjunctivitis and asthma. OUTCOMES: Clinical evaluation of symptoms, objective measurement of reactions to nasal or bronchial allergen challenge, immunologic changes as a result of allergen immunotherapy and, among patients with anaphylactic reactions to stinging insects, clinical outcome of intentional sting challenge. EVIDENCE: A search of MEDLINE was conducted to identify articles that presented results of allergen immunotherapy. Proceedings of symposia held by international subcommittees and of consensus meetings, as well as references obtained from these sources, were reviewed. The articles were grouped according to their main subject: immunologic effects, specific allergies, the results of randomized placebo-controlled clinical trials, types of allergen extract and protocols for allergen immunotherapy, adverse effects and deficiencies of allergen immunotherapy. VALUES: Each member of the working group assessed the importance of such issues as basic immunologic effects, clinical efficacy, adverse effects and inappropriate use; the working group then arrived at a consensus. BENEFITS, HARMS AND COSTS: Implementation of these guidelines would lead to the appropriate use of allergen immunotherapy and control inappropriate treatment, which could result in adverse effects and increased costs of services for patients with allergies. RECOMMENDATIONS: Allergen immunotherapy with specific, standardized allergenic materials, administered in high-dose schedules, is effective in patients with an allergy to insect stings or allergic rhinoconjunctivitis, and in some patients with asthma, who have been correctly diagnosed through a meticulous history corroborated

  14. Tolerance and immunological changes of chemically modified allergen vaccine of Parietaria judaica in accelerated schedules.

    PubMed

    Asturias, J A; Ferrer, A; Arilla, M C; Andreu, C; Madariaga, B; Martínez, A

    2007-03-01

    The physicochemical modification of allergen vaccines provides a chance for administering higher doses in a shorter period of time. We sought to assess the safety and immunological changes of using a biologically standardized and modified Parietaria judaica pollen extract in accelerated schedules. Two accelerated schedules were tested in 45 P. judaica-allergic patients: 20 patients reached the maximum dose after two visits using two different concentrations and 25 patients reached the maximum dose after only one visit with two injections of the maximum concentration vial. The tolerance was assessed by recording all side effects related with immunotherapy. Specific antibody levels against native extract and rPar j 2 allergen were evaluated at the beginning and the end of the study. Allergenic potency determined by enzyme allergosorbent test (EAST) inhibition and skin prick test showed that modified P. judaica pollen had a 99.9% less allergenicity than native extract. After 650 doses administered, two clinically irrelevant local reactions (diameter<0 x 5 cm) and no systemic reactions were registered. Significant increases in allergen-specific IgG4 and IgG against P. judaica extract and rPar j 2 and significant decrease of specific IgE against Par j 2 were observed. The modified extract of P. judaica is safe to treat sensitive patients, even at accelerated regimens, and induces significant immunological changes. PMID:17302898

  15. Ara h 2 and Ara h 6 have similar allergenic activity1 and are substantially redundant

    PubMed Central

    Chen, Xueni; Wang, Qian; El-Mezayen, Rabab; Zhuang, Yonghua; Dreskin, Stephen. C.

    2013-01-01

    Background The moderately homologous (~60%) proteins, Ara h 2 and Ara h 6, are the most potent peanut allergens. This study was designed to define the relative individual contributions of Ara h 2 and Ara h 6 to the overall allergenic activity of a crude peanut extract (CPE). Methods Ara h 2 and Ara h 6 were removed from CPE by gel filtration chromatography. Ara h 2.01, Ara h 2.02, and Ara h 6 were further purified (>99%). The potency of each allergen and the ability of these allergens to reconstitute the allergenic activity of CPE depleted of Ara h 2 and Ara h 6 was measured with RBL SX-38 cells sensitized with IgE from sensitized peanut allergic patients. Results The potency of the native proteins were significantly different (p<0.0001) although not dramatically so, with a rank order of Ara h 2.01 > Ara h 2.02 > Ara h 6. The addition of either purified Ara h 2 or Ara h 6 independently at their original concentration to CPE depleted of both Ara h 2 and Ara h 6 restored 80–100% of the original CPE allergenic activity. Addition of both Ara h 2 and Ara h 6 consistently completely restored the allergenic activity of CPE. Conclusions These studies indicate that either Ara h 2 or Ara h 6 independently can account for most of the allergenic activity in a CPE and demonstrate important redundancy in the allergenic activity of these related molecules. PMID:23075924

  16. 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

  17. Glove-derived foreign proteins induce allergen-specific IgE in a mouse model.

    PubMed

    Busch, Marion; Schröder, Claudia; Baron, Jens-Malte; Ott, Hagen; Bruckner, Thomas; Diepgen, Thomas L; Mahler, Vera

    2008-04-01

    Currently, most medical gloves are produced with a low content of natural rubber latex (NRL) protein. However, they may be substituted by proteins of foreign origin to maintain specific properties of the material. The aim of this study was to investigate the allergenicity and immunogenicity of unexpected proteins (i.e., soy and casein) compared with NRL proteins in a murine model in BALB/c mice. All respective allergen sources (extracts from three brands of NRL gloves, soy, and casein) were able to induce significant allergen-specific IgE and IgG(1) responses. On average, the highest IgE induction occurred after immunization with NRL, followed by soy and casein. Certain individuals from each treatment group exhibited levels of specific IgE as high as due to NRL. To analyze further specific IgE responses on a single allergen level, we established a microarray based on recombinant allergens for allergen-specific murine IgE detection. Besides specific IgE against rHev b 3, -6, -7, -8, and -11, specific IgE against kappa-casein could be detected in mice immunized with NRL glove extract, indicating a sensitization potential of the contained foreign protein. The substitution of genuine latex proteins by proteins of foreign origin may lead to a shift and de novo increase in sensitization to the finished products. PMID:18049454

  18. First screening method for the simultaneous detection of seven allergens by liquid chromatography mass spectrometry.

    PubMed

    Heick, J; Fischer, M; Pöpping, B

    2011-02-18

    The development of a multi-method for the detection of seven allergens based on liquid chromatography and triple-quadrupole tandem mass spectrometry in multiple reaction mode is described. It is based on extraction of the allergenic proteins from a food matrix, followed by enzymatic digestion with trypsin. The chosen marker peptides were implemented into one method that is capable of the simultaneous detection of milk, egg, soy, hazelnut, peanut, walnut and almond. This method has been used to detect all seven allergenic commodities from incurred reference bread material, which was baked according to a standard recipe from the baking industry. Detected concentrations ranged from 10 to 1000 μg/g, demonstrating that the mass spectrometric based method is a useful tool for allergen screening. PMID:21227428

  19. Allergenic Characterization of 27-kDa Glycoprotein, a Novel Heat Stable Allergen, from the Pupa of Silkworm, Bombyx mori

    PubMed Central

    Son, Mina; Lee, June Yong

    2016-01-01

    Boiled silkworm pupa is a traditional food in Asia, and patients with silkworm pupa food allergy are common in these regions. Still now only one allergen from silkworm, arginine kinase, has been identified. The purpose of this study was to identify novel food allergens in silkworm pupa by analyzing a protein extract after heat treatment. Heat treated extracts were examined by proteomic analysis. A 27-kDa glycoprotein was identified, expressed in Escherichia coli, and purified. IgE reactivity of the recombinant protein was investigated by ELISA. High molecular weight proteins (above 100 kDa) elicited increased IgE binding after heat treatment compared to that before heat treatment. The molecular identities of these proteins, however, could not be determined. IgE reactivity toward a 27-kDa glycoprotein was also increased after heating the protein extract. The recombinant protein was recognized by IgE antibodies from allergic subjects (33.3%). Glycation or aggregation of protein by heating may create new IgE binding epitopes. Heat stable allergens are shown to be important in silkworm allergy. Sensitization to the 27-kDa glycoprotein from silkworm may contribute to elevation of IgE to silkworm. PMID:26770033

  20. Allergenic Characterization of 27-kDa Glycoprotein, a Novel Heat Stable Allergen, from the Pupa of Silkworm, Bombyx mori.

    PubMed

    Jeong, Kyoung Yong; Son, Mina; Lee, June Yong; Park, Kyung Hee; Lee, Jae-Hyun; Park, Jung-Won

    2016-01-01

    Boiled silkworm pupa is a traditional food in Asia, and patients with silkworm pupa food allergy are common in these regions. Still now only one allergen from silkworm, arginine kinase, has been identified. The purpose of this study was to identify novel food allergens in silkworm pupa by analyzing a protein extract after heat treatment. Heat treated extracts were examined by proteomic analysis. A 27-kDa glycoprotein was identified, expressed in Escherichia coli, and purified. IgE reactivity of the recombinant protein was investigated by ELISA. High molecular weight proteins (above 100 kDa) elicited increased IgE binding after heat treatment compared to that before heat treatment. The molecular identities of these proteins, however, could not be determined. IgE reactivity toward a 27-kDa glycoprotein was also increased after heating the protein extract. The recombinant protein was recognized by IgE antibodies from allergic subjects (33.3%). Glycation or aggregation of protein by heating may create new IgE binding epitopes. Heat stable allergens are shown to be important in silkworm allergy. Sensitization to the 27-kDa glycoprotein from silkworm may contribute to elevation of IgE to silkworm. PMID:26770033

  1. Commercialized rapid immunoanalytical tests for determination of allergenic food proteins: an overview.

    PubMed

    Schubert-Ullrich, Patricia; Rudolf, Judith; Ansari, Parisa; Galler, Brigitte; Führer, Manuela; Molinelli, Alexandra; Baumgartner, Sabine

    2009-09-01

    Food allergies have become an important health issue especially in industrialized countries. Undeclared allergenic ingredients or the presence of "hidden" allergens because of contamination during the food production process pose great health risks to sensitised individuals. The EU directive for food labelling lists allergenic foods that have to be declared on food products by the manufacturers. The list includes gluten-containing cereals, crustaceans, eggs, fish, peanuts, soybeans, milk, various nuts (e.g. almond, hazelnut, and walnut, etc.), celery, mustard, sesame seeds, lupin, and molluscs. Reliable methods for detection and quantification of food allergens are needed that can be applied in a fast and easy-to-use manner, are portable, and need only limited technical equipment. This review focuses on the latest developments in food allergen analysis with special emphasis on fast immunoanalytical methods such as rapid enzyme-linked immunosorbent assays (ELISA), lateral-flow immunochromatographic assays (LFA) and dipstick tests. Emerging technologies such as immunochemical microarrays and biosensors are also discussed and their application to food allergen analysis is reviewed. Finally, a comprehensive overview of rapid immunochemical test kits that are currently available commercially is given in tabular form. PMID:19308361

  2. Production of recombinant allergens in plants

    PubMed Central

    2010-01-01

    A large percentage of allergenic proteins are of plant origin. Hence, plant-based expression systems are considered ideal for the recombinant production of certain allergens. First attempts to establish production of plant-derived allergens in plants focused on transient expression in Nicotiana benthamiana infected with recombinant viral vectors. Accordingly, allergens from birch and mugwort pollen, as well as from apple have been expressed in plants. Production of house dust mite allergens has been achieved by Agrobacterium-mediated transformation of tobacco plants. Beside the use of plants as production systems, other approaches have focused on the development of edible vaccines expressing allergens or epitopes thereof, which bypasses the need of allergen purification. The potential of this approach has been convincingly demonstrated for transgenic rice seeds expressing seven dominant human T cell epitopes derived from Japanese cedar pollen allergens. Parallel to efforts in developing recombinant-based diagnostic and therapeutic reagents, different gene-silencing approaches have been used to decrease the expression of allergenic proteins in allergen sources. In this way hypoallergenic ryegrass, soybean, rice, apple, and tomato were developed. PMID:21258627

  3. Food Allergens: Is There a Correlation between Stability to Digestion and Allergenicity?

    PubMed

    Bøgh, Katrine Lindholm; Madsen, Charlotte Bernhard

    2016-07-01

    Food allergy is a major health problem in the Western countries, affecting 3-8% of the population. It has not yet been established what makes a dietary protein a food allergen. Several characteristics have been proposed to be shared by food allergens. One of these is resistance to digestion. This paper reviews data from digestibility studies on purified food allergens and evaluates the predictive value of digestibility tests on the allergenic potential. We point out that food allergens do not necessarily resist digestion. We discuss how the choice of in vitro digestibility assay condition and the method used for detection of residual intact protein as well as fragments hereof may greatly influence the outcome as well as the interpretation of results. The finding that digests from food allergens may retain allergenicity, stresses the importance of using immunological assays for evaluating the allergenic potential of food allergen digestion products. Studies assessing the allergenicity of digestion products, by either IgE-binding, elicitation or sensitizing capacity, shows that digestion may abolish, decrease, have no effect, or even increase the allergenicity of food allergens. Therefore, the predictive value of the pepsin resistance test for assessing the allergenic potential of novel proteins can be questioned. PMID:25607526

  4. Allergen immunotherapy in polysensitized patient.

    PubMed

    Hrubiško, M; Špičák, V

    2016-05-01

    Specific allergen immunotherapy (AIT) is the only therapeutic method with positive impact on natural course of allergic disease - affecting clinical development (including the progression of rhinitis to asthma) and new sensitisations. The actual problem is the increasing number of patients manifesting poly-sensitivity in allergy skin tests and / or in specific IgE tests. Usually, AIT is not recommended in such individuals. The objective we are facing is that in many patients tested as poly-reactive, we have to distinguish in which cases it is a true polysensitization, and when it is due to cross-reactivity of specific IgE antibodies induced by panallergens. This may really determine when AIT may be an appropriate course of action. The article focuses on this problem in more detail, applying the long time Czech and Slovak experience with allergy testing and allergen immunotherapy. PMID:27152601

  5. Multiplex component-based allergen microarray in recent clinical studies.

    PubMed

    Patelis, A; Borres, M P; Kober, A; Berthold, M

    2016-08-01

    During the last decades component-resolved diagnostics either as singleplex or multiplex measurements has been introduced into the field of clinical allergology, providing important information that cannot be obtained from extract-based tests. Here we review recent studies that demonstrate clinical applications of the multiplex microarray technique in the diagnosis and risk assessment of allergic patients, and its usefulness in studies of allergic diseases. The usefulness of ImmunoCAP ISAC has been validated in a wide spectrum of allergic diseases like asthma, allergic rhinoconjunctivitis, atopic dermatitis, eosinophilic esophagitis, food allergy and anaphylaxis. ISAC provides a broad picture of a patient's sensitization profile from a single test, and provides information on specific and cross-reactive sensitizations that facilitate diagnosis, risk assessment, and disease management. Furthermore, it can reveal unexpected sensitizations which may explain anaphylaxis previously categorized as idiopathic and also display for the moment clinically non-relevant sensitizations. ISAC can facilitate a better selection of relevant allergens for immunotherapy compared with extract testing. Microarray technique can visualize the allergic march and molecular spreading in the preclinical stages of allergic diseases, and may indicate that the likelihood of developing symptomatic allergy is associated with specific profiles of sensitization to allergen components. ISAC is shown to be a useful tool in routine allergy diagnostics due to its ability to improve risk assessment, to better select relevant allergens for immunotherapy as well as detecting unknown sensitization. Multiplex component testing is especially suitable for patients with complex symptomatology. PMID:27196983

  6. 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

  7. Food Allergens in Mattress Dust in Norwegian Homes - A Potentially Important Source of Allergen Exposure

    PubMed Central

    Bertelsen, Randi J.; Fæste, Christiane K.; Granum, Berit; Egaas, Eliann; London, Stephanie J.; Carlsen, Kai-Håkon; Carlsen, Karin C. Lødrup; Løvik, Martinus

    2014-01-01

    Background Sensitization to food allergens and food allergic reactions are mostly caused by ingesting the allergen, but can also occur from exposure via the respiratory tract or the skin. Little is known about exposure to food allergens in the home environment. Objective To describe the frequency of detection of allergens from fish, egg, milk, and peanut in mattress dust collected from homes of 13 year old adolescents, and secondly to identify home characteristics associated with the presence of food allergen contamination in dust. Methods Food allergens were measured by dot blot analysis in mattress dust from 143 homes in Oslo, Norway. We analyzed associations between home characteristics (collected by parental questionnaires and study technicians) and food allergens by multivariate regression models. Results Fish allergen was detected in 46%, peanut in 41%, milk in 39%, and egg allergen in 22% of the mattress dust samples; only three samples contained none of these allergens. All four food allergens were more frequently detected in mattresses in small dwellings (<100m2) than larger dwellings (≥130 m2); 63-71% of the small dwellings (n=24) had milk, peanut, and fish allergens in the samples compared to 33-44% of the larger dwellings (n=95). Milk, peanut, and egg allergens were more frequently detected in homes with bedroom and kitchen on the same floor as compared with different floors; with odds ratios of 2.5 (95% confidence interval (CI): 1.1, 5.6) for milk, 2.4 (95% CI: 1.0, 6.1) for peanut, and 3.1 (95% CI: 1.3, 7.5) for egg allergens. Conclusions and clinical relevance Food allergens occurred frequently in beds in Norwegian homes, with dwelling size and proximity of kitchen and bedroom as the most important determinants. Due to the amount of time children spend in the bedroom, mattress dust may be an important source of exposure to food allergens. PMID:24304208

  8. Identification of the major allergen of Macrobrachium rosenbergii (giant freshwater prawn)

    PubMed Central

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

    2012-01-01

    Objective To characterize the major allergens of Macrobrachium rosenbergii (giant freshwater prawn). Methods Raw and cooked extracts of the giant freshwater prawn were prepared. The IgE reactivity pattern was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting technique with the sera of 20 skin prick test (SPT) positive patients. The major allergen identified was then characterized using the proteomics approach involving a combination of two-dimensional (2-DE) electrophoresis, mass spectrometry and bioinformatics tools. Results SDS-PAGE of the raw extract showed 23 protein bands (15–250 kDa) but those ranging from 40 to 100 kDa were not found in the cooked extract. From immunoblotting experiments, raw and cooked extracts demonstrated 11 and 5 IgE-binding proteins, respectively, with a molecular mass ranging from 15 to 155 kDa. A heat-resistant 36 kDa protein was identified as the major allergen of both extracts. In addition, a 42 kDa heat-sensitive protein was shown to be a major allergen of the raw extract. The 2-DE gel fractionated the prawn proteins to more than 50 different protein spots. Of these, 10 spots showed specific IgE reactivity with patients' sera. Matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis led to identification of 2 important allergens, tropomyosin and arginine kinase. Conclusions It can be concluded that the availability of such allergens would help in component-based diagnosis and therapy of prawn allergies. PMID:23569834

  9. 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

  10. Latex allergens in tire dust and airborne particles.

    PubMed Central

    Miguel, A G; Cass, G R; Weiss, J; Glovsky, M M

    1996-01-01

    The prevalence and severity of latex allergy has increased dramatically in the last 15 years due to exposure to natural rubber products. Although historically this health risk has been elevated in hospital personnel and patients, a recent survey has indicated a significant potential risk for the general population. To obtain a wide-spread source for latex exposure, we have considered tire debris. We have searched for the presence of latex allergens in passenger car and truck tire tread, in debris deposited from the atmosphere near a freeway, and in airborne particulate matter samples representative of the entire year 1993 at two sites in the Los Angeles basin (California). After extraction of the samples with phosphate buffered saline, a modified-ELISA inhibition assay was used to measure relative allergen potency and Western blot analyses were used to identify latex allergens. The inhibition studies with the human IgE latex assay revealed inhibition by the tire tread source samples and ambient freeway dust, as well as by control latex sap and latex glove extracts. Levels of extractable latex allergen per unit of protein extracted were about two orders of magnitude lower for tire tread as compared to latex gloves. Western blot analyses using binding of human IgE from latex-sensitive patients showed a band at 34-36 kDa in all tire and ambient samples. Long Beach and Los Angeles, California, air samples showed four additional bands between 50 and 135 kDa. Alternative Western blot analyses using rabbit IgG raised against latex proteins showed a broad band at 30-50 kDa in all samples, with additional bands in the urban air samples similar to the IgE results. A latex cross-reactive material was identified in mountain cedar. In conclusion, the latex allergens or latex cross-reactive material present in sedimented and airborne particulate material, derived from tire debris, and generated by heavy urban vehicle traffic could be important factors in producing latex allergy

  11. Mechanisms of allergen-specific immunotherapy.

    PubMed

    Akdis, Cezmi A; Akdis, Mübeccel

    2011-01-01

    Allergen-specific immunotherapy has been used for 100 years as a desensitizing therapy for allergic diseases and represents the potentially curative and specific method of treatment. The mechanisms of action of allergen-specific immunotherapy include the very early desensitization effects, modulation of T-and B-cell responses and related antibody isotypes, and migration of eosinophils, basophils, and mast cells to tissues, as well as release of their mediators. Regulatory T (Treg) cells have been identified as key regulators of immunologic 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 patients undergoing allergen-specific immunotherapy. Naturally occurring forkhead box protein 3-positive CD4(+)CD25(+) Treg cells and inducible T(R)1 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 T(H)1, T(H)2, and T(H)17 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 Treg cell subsets. PMID:21211639

  12. Cockroach allergen exposure and risk of asthma.

    PubMed

    Do, D C; Zhao, Y; Gao, P

    2016-04-01

    Cockroach sensitization is an important risk factor for the development of asthma. However, its underlying immune mechanisms and the genetic etiology for differences in allergic responses remain unclear. Cockroach allergens identification and their expression as biologically active recombinant proteins have provided a basis for studying the mechanisms regarding cockroach allergen-induced allergic sensitization and asthma. Glycans in allergens may play a crucial role in the immunogenicity of allergic diseases. Protease-activated receptor (PAR)-2, Toll-like receptor (TLR), and C-type lectin receptors have been suggested to be important for the penetration of cockroach allergens through epithelial cells to mediate allergen uptake, dendritic cell maturation, antigen-presenting cell (APC) function in T-cell polarization, and cytokine production. Environmental pollutants, which often coexist with the allergen, could synergistically elicit allergic inflammation, and aryl hydrocarbon receptor (AhR) activation and signaling may serve as a link between these two elements. Genetic factors may also play an important role in conferring the susceptibility to cockroach sensitization. Several genes have been associated with cockroach sensitization and asthma-related phenotypes. In this review, we will discuss the epidemiological evidence for cockroach allergen-induced asthma, cockroach allergens, the mechanisms regarding cockroach allergen-induced innate immune responses, and the genetic basis for cockroach sensitization. PMID:26706467

  13. Allergens stimulate store-operated calcium entry and cytokine production in airway epithelial cells

    PubMed Central

    Jairaman, Amit; Maguire, Chelsea H.; Schleimer, Robert P.; Prakriya, Murali

    2016-01-01

    Aberrant immune responses to environmental allergens including insect allergens from house dust mites and cockroaches contribute to allergic inflammatory diseases such as asthma in susceptible individuals. Airway epithelial cells (AECs) play a critical role in this process by sensing the proteolytic activity of allergens via protease-activated receptors (PAR2) to initiate inflammatory and immune responses in the airway. Elevation of cytosolic Ca2+ is an important signaling event in this process, yet the fundamental mechanism by which allergens induce Ca2+ elevations in AECs remains poorly understood. Here we find that extracts from dust mite and cockroach induce sustained Ca2+ elevations in AECs through the activation of Ca2+ release-activated Ca2+ (CRAC) channels encoded by Orai1 and STIM1. CRAC channel activation occurs, at least in part, through allergen mediated stimulation of PAR2 receptors. The ensuing Ca2+ entry then activates NFAT/calcineurin signaling to induce transcriptional production of the proinflammatory cytokines IL-6 and IL-8. These findings highlight a key role for CRAC channels as regulators of allergen induced inflammatory responses in the airway. PMID:27604412

  14. Allergens stimulate store-operated calcium entry and cytokine production in airway epithelial cells.

    PubMed

    Jairaman, Amit; Maguire, Chelsea H; Schleimer, Robert P; Prakriya, Murali

    2016-01-01

    Aberrant immune responses to environmental allergens including insect allergens from house dust mites and cockroaches contribute to allergic inflammatory diseases such as asthma in susceptible individuals. Airway epithelial cells (AECs) play a critical role in this process by sensing the proteolytic activity of allergens via protease-activated receptors (PAR2) to initiate inflammatory and immune responses in the airway. Elevation of cytosolic Ca(2+) is an important signaling event in this process, yet the fundamental mechanism by which allergens induce Ca(2+) elevations in AECs remains poorly understood. Here we find that extracts from dust mite and cockroach induce sustained Ca(2+) elevations in AECs through the activation of Ca(2+) release-activated Ca(2+) (CRAC) channels encoded by Orai1 and STIM1. CRAC channel activation occurs, at least in part, through allergen mediated stimulation of PAR2 receptors. The ensuing Ca(2+) entry then activates NFAT/calcineurin signaling to induce transcriptional production of the proinflammatory cytokines IL-6 and IL-8. These findings highlight a key role for CRAC channels as regulators of allergen induced inflammatory responses in the airway. PMID:27604412

  15. Variable Expression of Pathogenesis-Related Protein Allergen in Mountain Cedar (Juniperus ashei) Pollen1

    PubMed Central

    Midoro-Horiuti, Terumi; Goldblum, Randall M.; Kurosky, Alexander; Wood, Thomas G.; Brooks, Edward G.

    2009-01-01

    Allergic diseases have been increasing in industrialized countries. The environment is thought to have both direct and indirect modulatory effects on disease pathogenesis, including alterating on the allergenicity of pollens. Certain plant proteins known as pathogenesis-related proteins appear to be up-regulated by certain environmental conditions, including pollutants, and some have emerged as important allergens. Thus, the prospect of environmentally regulated expression of plant-derived allergens becomes yet another potential environmental influence on allergic disease. We have identified a novel pathogenesis-related protein allergen, Jun a 3, from mountain cedar (Juniperus ashei) pollen. The serum IgE from patients with hypersensitivity to either mountain cedar or Japanese cedar were shown to bind to native and recombinant Jun a 3 in Western blot analysis and ELISA. Jun a 3 is homologous to members of the thaumatin-like pathogenesis-related (PR-5) plant protein family. The amounts of Jun a 3 extracted from mountain cedar pollen varied up to 5-fold in lots of pollen collected from the same region in different years and between different regions during the same year. Thus, Jun a 3 may contribute not only to the overall allergenicity of mountain cedar pollen, but variable levels of Jun a 3 may alter the allergenic potency of pollens produced under different environmental conditions. PMID:10657673

  16. Identification of Novel Short Ragweed Pollen Allergens Using Combined Transcriptomic and Immunoproteomic Approaches

    PubMed Central

    Bouley, Julien; Groeme, Rachel; Jain, Karine; Baron-Bodo, Véronique; Nony, Emmanuel; Mascarell, Laurent; Moingeon, Philippe

    2015-01-01

    Background Allergy to short ragweed (Ambrosia artemisiifolia) pollen is a serious and expanding health problem in North America and Europe. Whereas only 10 short ragweed pollen allergens are officially recorded, patterns of IgE reactivity observed in ragweed allergic patients suggest that other allergens contribute to allergenicity. The objective of the present study was to identify novel allergens following extensive characterization of the transcriptome and proteome of short ragweed pollen. Methods Following a Proteomics-Informed-by-Transcriptomics approach, a comprehensive transcriptomic data set was built up from RNA-seq analysis of short ragweed pollen. Mass spectrometry-based proteomic analyses and IgE reactivity profiling after high resolution 2D-gel electrophoresis were then combined to identify novel allergens. Results Short ragweed pollen transcripts were assembled after deep RNA sequencing and used to inform proteomic analyses, thus leading to the identification of 573 proteins in the short ragweed pollen. Patterns of IgE reactivity of individual sera from 22 allergic patients were assessed using an aqueous short ragweed pollen extract resolved over 2D-gels. Combined with information derived from the annotated pollen proteome, those analyses revealed the presence of multiple unreported IgE reactive proteins, including new Amb a 1 and Amb a 3 isoallergens as well as 7 novel candidate allergens reacting with IgEs from 20–70% of patients. The latter encompass members of the carbonic anhydrase, enolase, galactose oxidase, GDP dissociation inhibitor, pathogenesis related-17, polygalacturonase and UDP-glucose pyrophosphorylase families. Conclusions We extended the list of allergens identified in short ragweed pollen. These findings have implications for both diagnosis and allergen immunotherapy purposes. PMID:26317427

  17. Immunological cross-reactivity between olive and grass pollen: implication of major and minor allergens

    PubMed Central

    2014-01-01

    Background Grasses and olive trees are the most common sources of allergenic pollen worldwide. Although they share some allergens, there are few studies analyzing the in vitro cross-reactivity between them. The aim was to define the cross-reactivity between Olea europaea and Phleum pratense using well-characterized sera of allergic children from Madrid, Spain. Methods 66 patients (mean age 10.32+/−4.07 years) were included in the study. All suffered from rhinoconjuntivitis and/or asthma and had a positive skin test and/or specific IgE determination to olive and grass pollen. Serum sIgE to individual allergens was conducted and sIgE against different grass species and olive was also determined by ELISA. Inhibition assays were performed using two serum sources, containing, or not, sIgE to minor allergens. Mass spectrometry analysis was performed in both extracts. Results 59/66 (89.39%) children had a positive sIgE determination by ELISA to grasses and 57/66 (86.36%) to olive pollen. There was no significant correlation between sIgE levels to grass and olive. Inhibition assays demonstrated no cross-reactivity between P. pratense and olive pollen when using the pool containing mainly sIgE to major allergens, whereas minimal to moderate cross-reactivity was detected when the serum contained high sIgE titers to minor allergens. Proteomic analyses revealed the presence of 42 common proteins in grasses and olive pollens. Conclusion No in vitro cross-reactivity was observed when sIgE was mainly directed to major allergens. In our population, sensitization to olive and grasses is not due to cross-reactivity. The contribution of the major allergens seems to be determinant. PMID:24940475

  18. 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

  19. Antigenic and allergenic analysis of psyllium seed components.

    PubMed

    Arlian, L G; Vyszenski-Moher, D L; Lawrence, A T; Schrotel, K R; Ritz, H L

    1992-04-01

    The outer portions (husk) of psyllium seeds are a concentrated source of natural fiber used in some bulk-fiber laxatives and cereals. They are known to elicit respiratory allergic reactions after inhalation or ingestion among sensitized individuals. Antigenic and allergenic characterization of three psyllium-seed fractions (husk, endosperm, and embryo) was conducted with crossed immunoelectrophoresis (CIE), crossed radioimmunoelectrophoresis, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis to determine the source of psyllium allergenicity. Homologous CIE demonstrated psyllium endosperm and embryo extracts contained seven and four antigens, respectively. Husk extracts were too gelatinous to react by CIE. However, heterologous CIE profiles of endosperm or embryo extracts, reacted with antihusk antibodies, resulted in antigen-antibody precipitin peaks that matched the heavy staining precipitin lines of homologous reactions for endosperm and embryo, respectively. These results indicated that commercial-grade husk, endosperm, and embryo contained similar antigens. Extracts of all three seed components contained antigens that bound IgE antibodies in the sera of 11 psyllium RAST-positive individuals, as determined by crossed radioimmunoelectrophoresis. The few prominent husk protein/peptide bands resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were common in either embryo or endosperm. Immunoblots revealed common IgE reactive bands in all three seed fractions. Microscopic examination of the powdered commercial-grade psyllium (95% pure) revealed it contained endosperm and embryo particles. These immunologic, biochemical, and microscopic findings suggest that other contaminating seed components are primarily responsible for the allergenicity of commercial-grade psyllium-husk powder rather than the husk itself. PMID:1560169

  20. Dog saliva – an important source of dog allergens

    PubMed Central

    Polovic, N; Wadén, K; Binnmyr, J; Hamsten, C; Grönneberg, R; Palmberg, C; Milcic-Matic, N; Bergman, T; Grönlund, H; van Hage, M; Crameri, Reto

    2013-01-01

    Background Allergy to dog (Canis familiaris) is a worldwide common cause of asthma and allergic rhinitis. However, dander extract in routine diagnostics is not an optimal predictor of IgE-mediated dog allergy. Our objective was to evaluate saliva as an allergen source for improved diagnostics of allergy to dog. Methods IgE-binding proteins in dog saliva and dander extract were analysed by immunoblot and mass spectrometry (LC-MS/MS) using pooled or individual sera from dog-allergic patients (n = 13). Sera from 59 patients IgE positive to dander and 55 patients IgE negative to dander but with symptoms to dog were analysed for IgE against saliva and dander by ELISA. Basophil stimulation with dog saliva and dander extract was measured by flow cytometry among three dog-allergic patients. Additionally, IgE-binding protein profiles of saliva from different breeds were investigated by immunoblot. Results Greater number and diversity of IgE-binding proteins was found in saliva compared to dander extract and varied among dog breeds. In saliva, Can f 1, 2, 3 and 6 were identified but also four new saliva allergen candidates. The majority of the 59 dog dander–positive sera (n = 44) were IgE positive to dog saliva. Among patients IgE negative to dander, but with symptoms to dog, 20% were IgE positive to saliva. The biological activity of saliva was confirmed by basophil degranulation. Conclusions Dog saliva is an allergen source for improved diagnostics of dog allergy. The IgE-binding protein profile of saliva from different dogs varies. PMID:23464525

  1. 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...

  2. 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...

  3. Thioredoxin from the Indianmeal Moth Plodia interpunctella: Cloning and Test of the Allergenic Potential in Mice

    PubMed Central

    Hemmer, Wolfgang; Mahler, Vera; Panzani, Raphael C.; Jarisch, Reinhart; Wiedermann, Ursula; Duchêne, Michael

    2012-01-01

    Background/Objective The Indianmeal moth Plodia interpunctella is a highly prevalent food pest in human dwellings, and has been shown to contain a number of allergens. So far, only one of these, the arginine kinase (Plo i 1) has been identified. Objective The aim of this study was to identify further allergens and characterise these in comparison to Plo i 1. Method A cDNA library from whole adult P. interpunctella was screened with the serum of a patient with indoor allergy and IgE to moths, and thioredoxin was identified as an IgE-binding protein. Recombinant thioredoxin was generated in E. coli, and tested together with Plo i 1 and whole moth extracts in IgE immunoblots against a large panel of indoor allergic patients' sera. BALB/c mice were immunised with recombinant thioredoxin and Plo i 1, and antibody production, mediator release from RBL cells, T-cell proliferation and cytokine production were measured. Result For the first time a thioredoxin from an animal species was identified as allergen. About 8% of the sera from patients with IgE against moth extracts reacted with recombinant P. interpunctella thioredoxin, compared to 25% reacting with recombinant Plo i 1. In immunised BALB/c mice, the recombinant allergens both induced classical Th2-biased immune responses such as induction IgE and IgG1 antibodies, upregulation of IL-5 and IL-4 and basophil degranulation. Conclusion Thioredoxin from moths like Plo i 1 acts like a classical Type I allergen as do the thioredoxins from wheat or corn. This clearly supports the pan-allergen nature of thioredoxin. The designation Plo i 2 is suggested for the new P. interpunctella allergen. PMID:22844539

  4. 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

  5. Biomarkers of acute respiratory allergen exposure: Screening for sensitization potential

    SciTech Connect

    Pucheu-Haston, Cherie M.; Copeland, Lisa B.; Vallanat, Beena; Boykin, Elizabeth; Ward, Marsha D.W.

    2010-04-15

    Effective hazard screening will require the development of high-throughput or in vitro assays for the identification of potential sensitizers. The goal of this preliminary study was to identify potential biomarkers that differentiate the response to allergens vs non-allergens following an acute exposure in naive individuals. Female BALB/c mice received a single intratracheal aspiration exposure to Metarhizium anisopliae crude antigen (MACA) or bovine serum albumin (BSA) in Hank's Balanced Salt Solution (HBSS) or HBSS alone. Mice were terminated after 1, 3, 6, 12, 18 and 24 h. Bronchoalveolar lavage fluid (BALF) was evaluated to determine total and differential cellularity, total protein concentration and LDH activity. RNA was isolated from lung tissue for microarray analysis and qRT-PCR. MACA administration induced a rapid increase in BALF neutrophils, lymphocytes, eosinophils and total protein compared to BSA or HBSS. Microarray analysis demonstrated differential expression of genes involved in cytokine production, signaling, inflammatory cell recruitment, adhesion and activation in 3 and 12 h MACA-treated samples compared to BSA or HBSS. Further analyses allowed identification of approx 100 candidate biomarker genes. Eleven genes were selected for further assessment by qRT-PCR. Of these, 6 demonstrated persistently increased expression (Ccl17, Ccl22, Ccl7, Cxcl10, Cxcl2, Saa1), while C3ar1 increased from 6-24 h. In conclusion, a single respiratory exposure of mice to an allergenic mold extract induces an inflammatory response which is distinct in phenotype and gene transcription from the response to a control protein. Further validation of these biomarkers with additional allergens and irritants is needed. These biomarkers may facilitate improvements in screening methods.

  6. Biomarkers of acute respiratory allergen exposure: screening for sensitization potential.

    PubMed

    Pucheu-Haston, Cherie M; Copeland, Lisa B; Vallanat, Beena; Boykin, Elizabeth; Ward, Marsha D W

    2010-04-15

    Effective hazard screening will require the development of high-throughput or in vitro assays for the identification of potential sensitizers. The goal of this preliminary study was to identify potential biomarkers that differentiate the response to allergens vs non-allergens following an acute exposure in naïve individuals. Female BALB/c mice received a single intratracheal aspiration exposure to Metarhizium anisopliae crude antigen (MACA) or bovine serum albumin (BSA) in Hank's Balanced Salt Solution (HBSS) or HBSS alone. Mice were terminated after 1, 3, 6, 12, 18 and 24 h. Bronchoalveolar lavage fluid (BALF) was evaluated to determine total and differential cellularity, total protein concentration and LDH activity. RNA was isolated from lung tissue for microarray analysis and qRT-PCR. MACA administration induced a rapid increase in BALF neutrophils, lymphocytes, eosinophils and total protein compared to BSA or HBSS. Microarray analysis demonstrated differential expression of genes involved in cytokine production, signaling, inflammatory cell recruitment, adhesion and activation in 3 and 12 h MACA-treated samples compared to BSA or HBSS. Further analyses allowed identification of approximately 100 candidate biomarker genes. Eleven genes were selected for further assessment by qRT-PCR. Of these, 6 demonstrated persistently increased expression (Ccl17, Ccl22, Ccl7, Cxcl10, Cxcl2, Saa1), while C3ar1 increased from 6-24 h. In conclusion, a single respiratory exposure of mice to an allergenic mold extract induces an inflammatory response which is distinct in phenotype and gene transcription from the response to a control protein. Further validation of these biomarkers with additional allergens and irritants is needed. These biomarkers may facilitate improvements in screening methods. PMID:20045013

  7. Preparation and application of monoclonal antibodies for a sandwich enzyme-linked immunosorbent assay of the major soybean allergen, Gly m Bd 30K.

    PubMed

    Tsuji, H; Bando, N; Kimoto, M; Okada, N; Ogawa, T

    1993-08-01

    In order to obtain probes suitable for the determination of the major soybean allergen, Gly m Bd 30 K, in soybean-related processed foods, we have prepared two monoclonal antibodies, F5 of IgG2a and H6 of IgM, by the fusion of P3U1 myeloma cells with the spleen cells of BALB/c mice immunized with the reductively carboxymethylated allergen (RCM-allergen). The two monoclonal antibodies were shown to be specific to the intact allergen as well as the RCM-allergen and to recognize distinct epitopes on the allergen. Of the monoclonal antibodies, F5 was conjugated with peroxidase. H6 and the labeled F5 were applied as the fixing antibody and the labeled first antibody, respectively, for a direct sandwich enzyme-linked immunosorbent assay of the allergen. When the allergen was extracted from the soybean-related foods with Tris-HCl buffer containing sodium dodecylsulfate and mercaptoethanol, the allergen, Gly m Bd 30 K, was shown to be measured in a range of 5-500 ng in this assay. PMID:7506767

  8. Allergen Atlas: a comprehensive knowledge center and analysis resource for allergen information

    PubMed Central

    Tong, Joo Chuan; Lim, Shen Jean; Muh, Hon Cheng; Chew, Fook Tim; Tammi, Martti T.

    2009-01-01

    Summary: A variety of specialist databases have been developed to facilitate the study of allergens. However, these databases either contain different subsets of allergen data or are deficient in tools for assessing potential allergenicity of proteins. Here, we describe Allergen Atlas, a comprehensive repository of experimentally validated allergen sequences collected from in-house laboratory, online data submission, literature reports and all existing general-purpose and specialist databases. Each entry was manually verified, classified and hyperlinked to major databases including Swiss-Prot, Protein Data Bank (PDB), Gene Ontology (GO), Pfam and PubMed. The database is integrated with analysis tools that include: (i) keyword search, (ii) BLAST, (iii) position-specific iterative BLAST (PSI-BLAST), (iv) FAO/WHO criteria search, (v) graphical representation of allergen information network and (vi) online data submission. The latest version contains information of 1593 allergen sequences (496 IUIS allergens, 978 experimentally verified allergens and 119 new sequences), 56 IgE epitope sequences, 679 links to PDB structures and 155 links to Pfam domains. Availability: Allergen Atlas is freely available at http://tiger.dbs.nus.edu.sg/ATLAS/. Contact: martti@nus.edu.sg. PMID:19213741

  9. New routes for allergen immunotherapy.

    PubMed

    Johansen, Pål; von Moos, Seraina; Mohanan, Deepa; Kündig, Thomas M; Senti, Gabriela

    2012-10-01

    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

  10. 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

  11. Effects of NO2 and Ozone on Pollen Allergenicity.

    PubMed

    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

  12. Allergens from Brazil nut: immunochemical characterization.

    PubMed

    Bartolomé, B; Méndez, J D; Armentia, A; Vallverdú, A; Palacios, R

    1997-01-01

    The increase in the consumption of tropical nuts in the Northern Hemisphere during the last years, has evolved in a simultaneous enhancement of allergic IgE mediated (Hypersensitivity type 1) reported cases produced by this kind of food. The Brazil nut is the seed of the Bertholletia excelsa tree (Family Lecythidaceae) and, as in other seeds, proteins represent one of its major components making up 15-17% of its fresh weight and 50% of defatted flour. Of these, storage proteins are the most important ones, and the 12 S globulin legumin-like protein and the 2 S albumin have been described as the most representative. The 2 S protein, due to its high sulfur-rich amino acid content (3% cysteine and 18% methionine), is being studied, cloned and expressed in some important agronomic seeds (soybean, bean, oilseed rape) in order to enrich the nutritional quality of them. The case of a patient with serious clinical allergic symptoms (vomiting, diarrhoea and loss of consciousness) caused by oral contact with the Brazil nut, is presented. The patient gave a positive Skin Prick Test response to Brazil nut, kiwi and hazelnut extracts, and negative to regionally specific aeroallergens and other food extracts. The patient serum showed a high level of specific IgE by RAST to Brazil nut (> 17.5 PRU/ml, Class 4), and significative levels to hazelnut, and mustard. In vitro immunological studies (SDS-Immunoblotting and IEF-Immunoblotting) revealed IgE-binding proteins present in the extract. It was shown that not only the heavy (Mr 9) and light (Mr 4) subunits of the known allergenic 2 S albumin but also the alpha-subunits (Mr approximately 33.5 and 32) and at least one of the beta-subunits (Mr approximately 21) of the 12 S Brazil nut globulin, hitherto never involved in allergic problems, showed a strong IgE-binding capacity. PMID:9208050

  13. 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. PMID:15508178

  14. Effect of high pressure on peanut allergens in the presence of polyphenol oxidase and caffeic acid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    High pressure (HP) enhances enzymatic reactions. Because polyphenol oxidase (PPO) is an enzyme, and reduces IgE binding of peanut allergens in presence of caffeic acid (CA), we postulated that a further reduction in IgE binding can be achieved, using HP together with PPO and CA. Peanut extracts cont...

  15. Enzymatic treatment of peanut kernels to reduce allergen levels.

    PubMed

    Yu, Jianmei; Ahmedna, Mohamed; Goktepe, Ipek; Cheng, Hsiaopo; Maleki, Soheila

    2011-08-01

    This study investigated the use of enzymatic treatment to reduce peanut allergens in peanut kernels as affected by processing conditions. Two major peanut allergens, Ara h 1 and Ara h 2, were used as indicators of process effectiveness. Enzymatic treatment effectively reduced Ara h 1 and Ara h 2 in roasted peanut kernels by up to 100% under optimal conditions. For instance, treatment of roasted peanut kernels with α-chymotrypsin and trypsin for 1-3h significantly increased the solubility of peanut protein while reducing Ara h 1 and Ara h 2 in peanut kernel extracts by 100% and 98%, respectively, based on ELISA readings. Ara h 1 and Ara h 2 levels in peanut protein extracts were inversely correlated with protein solubility in roasted peanut. Blanching of kernels enhanced the effectiveness of enzyme treatment in roasted peanuts but not in raw peanuts. The optimal concentration of enzyme was determined by response surface to be in the range of 0.1-0.2%. No consistent results were obtained for raw peanut kernels since Ara h 1 and Ara h 2 increased in peanut protein extracts under some treatment conditions and decreased in others. PMID:25214091

  16. Diagnostic test allergens used for in vivo diagnosis of allergic diseases are at risk: a European Perspective.

    PubMed

    Klimek, L; Hoffmann, H J; Renz, H; Demoly, P; Werfel, T; Matricardi, P M; Muraro, A; Schmid-Grendelmeier, P; Cardona, V; Papadopoulos, N G

    2015-10-01

    In the European Union (EU), allergens used for diagnostic tests (TAs) are defined as medicinal products and have to be registered by national authorities. The current situation is not homogeneous. Existing authorizations need to be kept in the market in some EU states, while others need complete new authorizations requiring clinical trials, quality assurance methods, stability studies, and periodic safety update reports. Allergen manufacturers argue that offering a comprehensive panel of TAs may be economically disastrous. Expenses for initiation and maintenance of TA authorizations far exceed their related revenues and manufacturers may be forced to significantly limit their allergen portfolios. The availability of a wide range of high-quality TAs is very important for in vivo diagnoses of IgE-mediated allergies. Increased regulatory demands induce costs that need to be covered by public health organizations or reimbursed by health insurance companies. PMID:26105127

  17. Mesoporous silica nanoparticles synthesized from liquid crystal display manufacturing extracts as a potential candidate for a drug delivery carrier: evaluation of their safety and biocompatibility

    PubMed Central

    Lin, Yu-chih; Lin, Liang-Yi; Gao, Ming-Yi; Fang, Yi-Ping

    2013-01-01

    Mesoporous silica nanoparticles (MSNs) were synthesized as a promising drug delivery carrier due to the large surface area and porous characteristics. Our previous study successfully recycled wastes from the liquid crystal display (LCD) industry as the silica precursor. In this study, we substantiated the possibility of applying this material as a drug carrier. MSNs synthesized from the extraction of wastes from the manufacture of LCD panels were characterized as having an average diameter of 100 nm, a surface area of 788 m2/g, a uniform pore size distribution of 3.8 nm, and a pore volume of up to 1.04 cm3/g. Methotrexate and camptothecin were entrapped in MSNs at about 33.88% and 75.12%, respectively. The cell viability assay demonstrated that MSNs at 1 μg/mL had no significant influence on human lung fibroblast (WI-38) cells or ovarian cancer (ES-2) cells. A lactate dehydrogenase assay also indicated no inflammation occurred. Moreover, a hemolytic erythrocyte test indicated that the dose range of <100 μg/mL showed that 5% of erythrocytes were affected. After exposure to biofluids, the ordered structure was slightly degraded. The results revealed that MSNs synthesized from extraction of wastes from the manufacture of LCD panels had a good entrapment capacity for hydrophobic drugs and controllable safety conditions; they may be applied as a drug delivery carrier. PMID:24143088

  18. 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

  19. Characterisation of potential novel allergens in the fish parasite Anisakis simplex

    PubMed Central

    Fæste, Christiane Kruse; Jonscher, Karen R.; Dooper, Maaike M.W.B.; Egge-Jacobsen, Wolfgang; Moen, Anders; Daschner, Alvaro; Egaas, Eliann; Christians, Uwe

    2016-01-01

    The parasitic nematode Anisakis simplex occurs in fish stocks in temperate seas. A. simplex contamination of fish products is unsavoury and a health concern considering human infection with live larvae (anisakiasis) and allergic reactions to anisakid proteins in seafood. Protein extracts of A. simplex produce complex band patterns in gel electrophoresis and IgE-immunostaining. In the present study potential allergens have been characterised using sera from A. simplex-sensitised patients and proteome data obtained by mass spectrometry. A. simplex proteins were homologous to allergens in other nematodes, insects, and shellfish indicating cross-reactivity. Characteristic marker peptides for relevant A. simplex proteins were described. PMID:27110489

  20. 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.

  1. 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

  2. Identification of Two Metallothioneins as Novel Inhalative Coffee Allergens Cof a 2 and Cof a 3

    PubMed Central

    Peters, Ulrike; Frenzel, Karsten; Brettschneider, Reinhold; Oldenburg, Marcus; Bittner, Cordula

    2015-01-01

    Background Dust of green coffee beans is known to be a relevant cause for occupational allergic disorders in coffee industry workers. Recently, we described the first coffee allergen (Cof a 1) establishing an allergenic potential of green coffee dust. Objective Our aim was to identify allergenic components of green coffee in order to enhance inhalative coffee allergy diagnosis. Methods A Coffea arabica pJuFo cDNA phage display library was created and screened for IgE binding with sera from allergic coffee workers. Two further coffee allergens were identified by sequence analysis, expressed in E. coli, and evaluated by Western blots. The prevalence of sensitization to recombinant Cof a 1, Cof a 2, and Cof a 3 and to commercially available extract was investigated by ELISA (enzyme-linked immunosorbent assay) respectively CAP (capacity test) screening in 18 sera of symptomatic coffee workers. Results In addition to the previously described chitinase Cof a 1, two Coffea arabica cysteine-rich metallothioneins of 9 and 7 kDa were identified and included in the IUIS Allergen Nomenclature as Cof a 2 and Cof a 3. Serum IgE antibodies to at least one of the recombinant allergens were found in 8 out of 18 symptomatic coffee workers (44%). Only 2 of the analysed sera (11%) had reacted previously to the commercial allergy test. Conclusions In addition to the previously described Cof a 1 we have identified two further coffee proteins to be type I coffee allergens (Cof a 2 and Cof a 3) which may have a relevant potential for the specific diagnosis and/or therapy of coffee allergy. PMID:25962169

  3. 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

  4. Molecular Cloning and Expression of Pro J 1: A New Allergen of Prosopis Juliflora Pollen.

    PubMed

    Dousti, Fatemeh; Assarehzadegan, Mohammad-Ali; Morakabati, Payam; Khosravi, Gholam Reza; Akbari, Bahareh

    2016-04-01

    Pollen from mesquite (Prosopis juliflora) is one of the important causes of immediate hypersensitivity reactions in the arid and semi-arid regions of the world. The aim of present study is to produce and purify the recombinant form of allergenic Ole e 1-like protein from the pollen of this allergenic tree. Immunological and cross-inhibition assays were performed for the evaluation of IgE-binding capacity of purified recombinant protein. For molecular cloning, the coding sequence of the mesquite Ole e 1-like protein was inserted into pTZ57R/T vector and expressed in Escherichia coli using the vector pET-21b(+). After purification of the recombinant protein, its immunoreactivity was analysed by in vitro assays using sera from twenty one patients with an allergy to mesquite pollen. The purified recombinant allergen was a member of Ole e 1-like protein family and consisted of 150 amino acid residues, with a predicted molecular mass of 16.5 kDa and a calculated isoelectric point (pI) of 4.75. Twelve patients (57.14%) had significant specific IgE levels for this recombinant allergen. Immunodetection and inhibition assays indicated that the purified recombinant allergen might be the same as that in the crude extract. Herein, we introduce an important new allergen from P. juliflora pollen (Pro j 1), which is a member of the Ole e 1-like protein family and exhibits significant identity and similarity to other allergenic members of this family. PMID:27090365

  5. Characterization of Allergens Prepared from Smooth and Rough Strains of Brucella melitensis

    PubMed Central

    Jones, L. M.; Diaz, R.; Taylor, A. G.

    1973-01-01

    Protein allergens were prepared from Br. melitensis smooth strain Rev. 1 and rough strain B115 using a cold saline method of extraction. Guineapig skin tests showed the two preparations to be of similar potency and specificity. Gel filtration on Sephadex G-75 showed that the skin test reactive fractions of each preparation were composed of a mixture of proteins with molecular weights in the range of 12,000 to 50,000. Polyacrylamide gel electrophoresis and immunoelectrophoresis indicated that the preparations consisted of at least 12 similar, if not identical, protein constituents. At least 20 protein bands were revealed with polyacrylamide gel isoelectric focusing, which appeared to be almost identical in the two preparations. Smooth lipopolysaccharide antigen, tested in normal and infected guineapigs, was of a lower potency than protein allergens and the eyrthematous response occurred earlier, resembling an antibody mediated reaction rather than delayed hypersensitivity. The allergen prepared from the rough strain contained no detectable smooth lipopolysaccharide antigen and therefore did not stimulate a secondary rise in antibody to smooth cells in sensitized animals. As the presence of smooth lipopolysaccharide antigen in an allergen appears to be unnecessary for provoking delayed hypersensitivity responses, and undesirable because it may stimulate the production of antibodies which interfere with diagnosis, it is preferable to prepare allergens from rough strains of brucella. ImagesFigs. 1-2,4-5Fig. 7 PMID:4758380

  6. Kiwifruit Allergy in Children: Characterization of Main Allergens and Patterns of Recognition.

    PubMed

    Moreno Álvarez, Ana; Sexto, Leticia Vila; Bardina, Luda; Grishina, Galina; Sampson, Hugh A

    2015-01-01

    Kiwifruit allergy has been described mostly in the adult population, but immunoglobulin (Ig)E-mediated allergic reactions to kiwifruit appear to be occurring more frequently in children. To date, 13 allergens from kiwifruit have been identified. Our aim was to identify kiwifruit allergens in a kiwifruit allergic-pediatric population, describing clinical manifestations and patterns of recognition. Twenty-four children were included. Diagnosis of kiwifruit allergy was based on compatible clinical manifestations and demonstration of specific IgE by skin prick test (SPT) and/or serum-specific IgE determination. SDS-PAGE and immunoblotting were performed with kiwifruit extract, and proteins of interest were further analyzed by mass spectrometry/mass spectrometry. For component-resolved in vitro diagnosis, sera of kiwifruit-allergic patients were analyzed by an allergen microarray assay. Act d 1 and Act d 2 were bound by IgE from 15 of 24 children. Two children with systemic manifestations recognized a protein of 15 kDa, homologous to Act d 5. Act d 1 was the allergen with the highest frequency of recognition on microarray chip, followed by Act d 2 and Act d 8. Kiwifruit allergic children develop systemic reactions most frequently following ingestion compared to adults. Act d 1 and Act d 2 are major allergens in the pediatric age group. PMID:27417374

  7. Expression, purification and characterization of Der f 27, a new allergen from dermatophagoides farinae

    PubMed Central

    Lin, Jianli; Li, Meng; Liu, Yulin; Jiang, Congli; Wu, Yulan; Wang, Yuanyuan; Gao, Anjian; Liu, Zhigang; Yang, Pingchang; Liu, Xiaoyu

    2016-01-01

    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. farinaederived 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. PMID:27347356

  8. The peanut allergy epidemic: allergen molecular characterisation and prospects for specific therapy.

    PubMed

    de Leon, Maria P; Rolland, Jennifer M; O'Hehir, Robyn E

    2007-01-01

    Peanut (Arachis hypogaea) allergy is a major cause of food-induced anaphylaxis, with increasing prevalence worldwide. To date, there is no cure for peanut allergy, and, unlike many other food allergies, it usually persists through to adulthood. Prevention of exposure to peanuts is managed through strict avoidance, which can be compromised by the frequent use of peanuts and peanut products in food preparations. Conventional subcutaneous-injection allergen immunotherapy using crude peanut extract is not a recommended treatment because of the risk of severe side effects, largely as a result of specific IgE antibodies. Consequently, there is an urgent need to develop a suitable peanut allergen preparation that can induce specific clinical and immunological tolerance to peanuts in allergic individuals without adverse side effects. This requires detailed molecular and immunological characterisation of the allergenic components of peanut. This article reviews current knowledge on clinically relevant peanut allergens, in particular Ara h 1, Ara h 2 and Ara h 3, together with options for T-cell-reactive but non-IgE-binding allergen variants for specific immunotherapeutic strategies. These include T-cell-epitope peptide and hypoallergenic mutant vaccines. Alternative routes of administration such as sublingual are also considered, and appropriate adjuvants for delivering effective treatments at these sites examined. PMID:17210088

  9. Kiwifruit Allergy in Children: Characterization of Main Allergens and Patterns of Recognition

    PubMed Central

    Moreno Álvarez, Ana; Vila Sexto, Leticia; Bardina, Luda; Grishina, Galina; Sampson, Hugh. A.

    2015-01-01

    Kiwifruit allergy has been described mostly in the adult population, but immunoglobulin (Ig)E-mediated allergic reactions to kiwifruit appear to be occurring more frequently in children. To date, 13 allergens from kiwifruit have been identified. Our aim was to identify kiwifruit allergens in a kiwifruit allergic-pediatric population, describing clinical manifestations and patterns of recognition. Twenty-four children were included. Diagnosis of kiwifruit allergy was based on compatible clinical manifestations and demonstration of specific IgE by skin prick test (SPT) and/or serum-specific IgE determination. SDS-PAGE and immunoblotting were performed with kiwifruit extract, and proteins of interest were further analyzed by mass spectrometry/mass spectrometry. For component-resolved in vitro diagnosis, sera of kiwifruit-allergic patients were analyzed by an allergen microarray assay. Act d 1 and Act d 2 were bound by IgE from 15 of 24 children. Two children with systemic manifestations recognized a protein of 15 kDa, homologous to Act d 5. Act d 1 was the allergen with the highest frequency of recognition on microarray chip, followed by Act d 2 and Act d 8. Kiwifruit allergic children develop systemic reactions most frequently following ingestion compared to adults. Act d 1 and Act d 2 are major allergens in the pediatric age group. PMID:27417374

  10. Identification of olive pollen allergens using a fluorescence-based 2D multiplex method.

    PubMed

    Zienkiewicz, Krzysztof; Alché, Juan de Dios; Zienkiewicz, Agnieszka; Tormo, Alejandro; Castro, Antonio Jesús

    2015-04-01

    Olive (Olea europaea L.) pollen is a major health concern in the Mediterranean countries and some olive growing regions in America and Australia. The molecular variability of pollen allergens constitutes a handicap for commercial extract standardization, which is the base of current diagnosis and vaccination procedures. In this paper, we report a time-saving and plant material saving multiplex detection method for the rapid and simultaneous analysis of Ole e 1, Ole e 2, and Ole e 5 allergen polymorphism on a single blot. This method combines high-resolution 2DE techniques with high-sensitive fluorescence-based detection methods. Using this strategy, we were capable to identify a higher number of allergen forms compared with classical 1D approach. The use of fluorescent probes and the increased resolution of 2D blots avoided overlapping effects, and allow estimating the amount of individual allergen forms. In addition, the pattern and identity of the IgE-reactive proteins of either a population or individual patients allergic to olive pollen was also effortlessly determined in a single additional step. This flexible method might be extended to a higher number of olive allergens and cultivars, and is also applicable to other allergogenic plant species and sources. PMID:25640071

  11. Development of a Novel Strategy to Isolate Lipophilic Allergens (Oleosins) from Peanuts

    PubMed Central

    Schwager, Christian; Kull, Skadi; Krause, Susanne; Schocker, Frauke; Petersen, Arnd; Becker, Wolf-Meinhard; Jappe, Uta

    2015-01-01

    Background Peanut allergy is one of the most severe class I food allergies with increasing prevalence. Especially lipophilic allergens, such as oleosins, were found to be associated with severe symptoms, but are usually underrepresented in diagnostic extracts. Therefore, this study focused on isolation, molecular characterization and assessment of the allergenicity of peanut oleosins. Methods and Results A comprehensive method adapted for the isolation of peanut oil bodies of high purity was developed comprising a stepwise removal of seed storage proteins from oil bodies. Further separation of the oil body constituents, including the allergens Ara h 10, Ara h 11, the presumed allergen oleosin 3 and additional oleosin variants was achieved by a single run on a preparative electrophoresis cell. Protein identification realized by N-terminal sequencing, peptide mass fingerprinting and homology search revealed the presence of oleosins, steroleosins and a caleosin. Immunoblot analysis with sera of peanut-allergic individuals illustrated the IgE-binding capacity of peanut-derived oleosins. Conclusion Our method is a novel way to isolate all known immunologically distinct peanut oleosins simultaneously. Moreover, we were able to provide evidence for the allergenicity of oleosins and thus identified peanut oleosins as probable candidates for component-resolved allergy diagnosis. PMID:25860789

  12. Testing small molecule analogues of the Acanthocheilonema viteae immunomodulator ES-62 against clinically relevant allergens.

    PubMed

    Janicova, L; Rzepecka, J; Rodgers, D T; Doonan, J; Bell, K S; Lumb, F E; Suckling, C J; Harnett, M M; Harnett, W

    2016-06-01

    ES-62 is a glycoprotein secreted by the filarial nematode Acanthocheilonema viteae that protects against ovalbumin (OVA)-induced airway hyper-responsiveness in mice by virtue of covalently attached anti-inflammatory phosphorylcholine (PC) residues. We have recently generated a library of small molecule analogues (SMAs) of ES-62 based around its active PC moiety as a starting point in novel drug development for asthma and identified two compounds - termed 11a and 12b - that mirror ES-62's protective effects. In this study, we have moved away from OVA, a model allergen, to test the SMAs against two clinically relevant allergens - house dust mite (HDM) and cockroach allergen (CR) extract. We show that both SMAs offer some protection against development of lung allergic responses to CR, in particular reducing eosinophil infiltration, whereas only SMA 12b is effective in protecting against eosinophil-dependent HDM-induced allergy. These data therefore suggest that helminth molecule-induced protection against model allergens may not necessarily translate to clinically relevant allergens. Nevertheless, in this study, we have managed to demonstrate that it is possible to produce synthetic drug-like molecules based on a parasitic worm product that show therapeutic potential with respect to asthma resulting from known triggers in humans. PMID:27059010

  13. Mass spectrometry-based identification of allergens from Curvularia pallescens, a prevalent aerospore in India.

    PubMed

    Dey, Debarati; Saha, Bodhisattwa; Sircar, Gaurab; Ghosal, Kavita; Bhattacharya, Swati Gupta

    2016-07-01

    The worldwide prevalence of fungal allergy in recent years has augmented mining allergens from yet unexplored ones. Curvularia pallescens (CP) being a dominant aerospore in India and a major sensitiser on a wide range of allergic population, pose a serious threat to human health. Therefore, we aimed to identify novel allergens from CP in our present study. A cohort of 22 CP-sensitised patients was selected by positive Skin prick grade. Individual sera exhibited elevated specific IgE level and significant histamine release on a challenge with antigenic extract of CP. First gel-based profiling of CP proteome was done by 1- and 2-dimensional gel. Parallel 1- and 2-dimensional immunoblot were performed applying individual as well as pooled patient sera. Identification of the sero-reactive spots from the 2-dimensional gel was found to be challenging as CP was not previously sequenced. Hence, mass spectrometry-based proteomic workflow consisting of conventional database search was not alone sufficient. Therefore, de novo sequencing preceded homology search was implemented for further identification. Altogether 11 allergenic proteins including Brn-1, vacuolar protease, and fructose-bis-phosphate aldolase were identified with high statistical confidence (p<0.05). This is the first study to report on any allergens from CP. This kind of proteome-based analysis provided a catalogue of CP allergens that would lead an improved way of diagnosis and therapy of CP-related allergy. PMID:27003473

  14. 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...

  15. 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...

  16. 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...

  17. 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...

  18. 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...

  19. Identification and characterization of a second major peanut allergen, Ara h II, with use of the sera of patients with atopic dermatitis and positive peanut challenge.

    PubMed

    Burks, A W; Williams, L W; Connaughton, C; Cockrell, G; O'Brien, T J; Helm, R M

    1992-12-01

    Peanuts are frequently a cause of food hypersensitivity reactions in children. Serum from nine patients with atopic dermatitis and a positive double-blind, placebo-controlled, food challenge to peanut were used in the process of identification and purification of the peanut allergens. Identification of a second major peanut allergen was accomplished with use of various biochemical and molecular techniques. Anion exchange chromatography of the crude peanut extract produced several fractions that bound IgE from the serum of the patient pool with positive challenges. By measuring antipeanut specific IgE and by IgE-specific immunoblotting we have identified an allergic component that has two closely migrating bands with a mean molecular weight of 17 kd. Two-dimensional gel electrophoresis of this fraction revealed it to have a mean isoelectric point of 5.2. According to allergen nomenclature of the IUIS Subcommittee for Allergen Nomenclature this allergen is designated, Ara h II (Arachis hypogaea). PMID:1460200

  20. Next generation of food allergen quantification using mass spectrometric systems.

    PubMed

    Koeberl, Martina; Clarke, Dean; Lopata, Andreas L

    2014-08-01

    Food allergies are increasing worldwide and becoming a public health concern. Food legislation requires detailed declarations of potential allergens in food products and therefore an increased capability to analyze for the presence of food allergens. Currently, antibody-based methods are mainly utilized to quantify allergens; however, these methods have several disadvantages. Recently, mass spectrometry (MS) techniques have been developed and applied to food allergen analysis. At present, 46 allergens from 11 different food sources have been characterized using different MS approaches and some specific signature peptides have been published. However, quantification of allergens using MS is not routinely employed. This review compares the different aspects of food allergen quantification using advanced MS techniques including multiple reaction monitoring. The latter provides low limits of quantification for multiple allergens in simple or complex food matrices, while being robust and reproducible. This review provides an overview of current approaches to analyze food allergens, with specific focus on MS systems and applications. PMID:24824675

  1. 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

  2. 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

  3. Characterization of allergens in four South American snake species.

    PubMed

    Madero, Mauro F; Gámez, Cristina; Madero, Mauro A; Fernández-Nieto, Mar; Sastre, Joaquín; del Pozo, Victoria

    2009-01-01

    A 55-year-old herpetologist developed rhinitis, asthma, urticaria and anaphylaxis when handling 4 different viper snake venoms. Allergen characterizations were done using SDS-PAGE, IgE immunoblotting and IgE inhibition experiments. The most prominent immunoreactive proteins were analyzed by MALDI-TOF mass spectrometry, and peptide identity was demonstrated by homology with known peptide sequences. SDS-PAGE showed several protein bands ranging from 5 to 99 kDa in each of the 4 snake venoms. Immunoblotting demonstrated 4 IgE-binding bands in the Bothrops extract of about 60, 28, 14 and 7 kDa. The bands of 28 and 14 kDa were also present in Lachesis muta. Two IgE-binding proteins of about 50 and 35 kDa were found in Bothrops atrox and L. muta, respectively. A strong inhibition of IgE binding to immobilize Bothrops asper proteins was observed after preabsorption of sera with B. asper, B. atrox,Bothrops xanthograma and L. muta extracts. MALDI-TOF analysis showed a 14-kDa phospholipase and the 60- and 28-kDa proteins showed significant similarity with metalloproteinases. In this report we have characterized the snake venom allergens that can elicit IgE-mediated symptoms. PMID:19494529

  4. Survey of peanut levels in selected Irish food products bearing peanut allergen advisory labels.

    PubMed

    Robertson, Orla N; Hourihane, Jonathan O'B; Remington, Benjamin C; Baumert, Joseph L; Taylor, Steve L

    2013-01-01

    Peanut allergy affects up to 2% of consumers and is responsible for the majority of fatalities caused by food-induced anaphylaxis. Peanut-containing products must be clearly labelled. Manufacturers are not legally required to label peanut if its inclusion resulted from unintentional cross contact with foods manufactured in the same facility. However, the use of allergen advisory statements alerting consumers of the potential presence of peanut allergen has increased in recent years. In previous studies, the vast majority of foods with precautionary allergen statements did not contain detectable levels of peanut, but no data are available on Irish food products. Thirty-eight food products bearing peanut/nut allergen-related statements were purchased from multiple locations in the Republic of Ireland and analysed for the presence of peanut. Peanut was detected in at least one lot in 5.3% (2 of 38) of the products tested. The doses of peanut detected ranged from 0.14 mg to 0.52 mg per suggested serving size (0.035-0.13 mg peanut protein). No detectable levels of peanut were found in the products that indicated peanut/nuts as a minor ingredient. Quantitative risk assessment, based on the known distribution of individual threshold doses for peanut, indicates that only a very small percentage of the peanut-allergic population would be likely to experience an allergic reaction to those products while the majority of products with advisory labels appear safe for the peanut-allergic population. Food manufacturers should be encouraged to analyse products manufactured in shared facilities and even on shared equipment with peanuts for peanut residues to determine whether sufficient risk exists to warrant the use of advisory labelling. Although it appears that the majority of food products bearing advisory nut statements are in fact free of peanut contamination, advice to peanut allergy sufferers to avoid said foods should continue in Ireland and therefore in the wider European

  5. Soybean flour asthma: detection of allergens by immunoblotting

    SciTech Connect

    Bush, R.K.; Schroeckenstein, D.; Meier-Davis, S.; Balmes, J.; Rempel, D.

    1988-08-01

    A 43-year-old woman developed asthma 6 years after beginning work in a food-processing plant in which soybean flour was used as a protein extender. Symptoms of sneezing, coughing, and wheezing would begin within minutes of exposure to soybean flour and resolve 2 hours after exposure ceased. Skin tests were positive to a soy extract prepared from the flour. Airway hyperreactivity was confirmed by a positive bronchial challenge to methacholine. Bronchial challenge with soybean flour produced an immediate increase in specific airway resistance from 5.0 to 22.7 L. cm of H2O/L/sec. There was no response to challenge with lactose. The patient's allergic response to soy-flour extract was further characterized by several immunologic methods. IgE binding to soy-flour protein by direct RAST was 5.98 times that of a normal control serum. The soy-flour extract was separated by dodecyl sulfate-polyacrylamide gel electrophoresis. Twenty-four protein bands were detected in the crude soy-flour extract. After immunoblotting and subsequent autoradiography, nine proteins with molecular weights ranging from 54,500 to 14,875 were found. Cross-reactivity studies with other legumes demonstrated apparent immunologic identity between a component in green pea extract and a soybean protein with a molecular weight of 17,000. The clinical significance of this cross-reactivity is not known. We conclude that in this case of occupational asthma to soybean flour, multiple allergens were involved. Immunoblotting may be useful in identifying the allergens involved in occupational asthma.

  6. 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...

  7. The Role of Allergen Exposure and Avoidance in Asthma

    PubMed Central

    Baxi, Sachin N.; Phipatanakul, Wanda

    2010-01-01

    Allergy testing and avoidance of allergens plays an important role in asthma control. Increased allergen exposure, in genetically susceptible individuals, can lead to allergic sensitization. Continued allergen exposure can increase the risk of asthma and other allergic diseases. In a patient with persistent asthma, identification of indoor and outdoor allergens and subsequent avoidance can improve symptoms. Often times, a patient will have multiple allergies and the avoidance plan should target all positive allergens. Several studies have shown that successful allergen remediation includes a comprehensive approach including education, cleaning, physical barriers and maintaining these practices. PMID:20568555

  8. Allergen hybrids – next generation vaccines for Fagales pollen immunotherapy

    PubMed Central

    Pichler, U; Hauser, M; Hofer, H; Himly, M; Hoflehner, E; Steiner, M; Mutschlechner, S; Hufnagl, K; Ebner, C; Mari, A; Briza, P; Bohle, B; Wiedermann, U; Ferreira, F; Wallner, M

    2013-01-01

    Summary Background Trees belonging to the order of Fagales show a distinct geographical distribution. While alder and birch are endemic in the temperate zones of the Northern Hemisphere, hazel, hornbeam and oak prefer a warmer climate. However, specific immunotherapy of Fagales pollen-allergic patients is mainly performed using birch pollen extracts, thus limiting the success of this intervention in birch-free areas. Objectives T cells are considered key players in the modification of an allergic immune response during specific immunotherapy (SIT), therefore we thought to combine linear T cell epitope-containing stretches of the five most important Fagales allergens from birch, hazel, alder, oak and hornbeam resulting in a Fagales pollen hybrid (FPH) molecule applicable for SIT. Methods A Fagales pollen hybrid was generated by PCR-based recombination of low IgE-binding allergen epitopes. Moreover, a structural-variant FPH4 was calculated by in silico mutagenesis, rendering the protein unable to adopt the Bet v 1-like fold. Both molecules were produced in Escherichia coli, characterized physico-chemically as well as immunologically, and tested in mouse models of allergic sensitization as well as allergy prophylaxis. Results Using spectroscopic analyses, both proteins were monomeric, and the secondary structure elements of FPH resemble the ones typical for Bet v 1-like proteins, whereas FPH4 showed increased amounts of unordered structure. Both molecules displayed reduced binding capacities of Bet v 1-specific IgE antibodies. However, in a mouse model, the proteins were able to induce high IgG titres cross-reactive with all parental allergens. Moreover, prophylactic treatment with the hybrid proteins prevented pollen extract-induced allergic lung inflammation in vivo. Conclusion The hybrid molecules showed a more efficient uptake and processing by dendritic cells resulting in a modified T cell response. The proteins had a lower IgE-binding capacity compared with the

  9. Is high pressure treatment able to modify the allergenicity of the largemouth bass allergens?

    NASA Astrophysics Data System (ADS)

    Liu, Chu-Yi; Tao, Sha; Liu, Rong; Chen, Fu-Sheng; Xue, Wen-Tong

    2012-12-01

    The aim of this paper is to study the influence of high pressure treatment on the structural changes and allergenicity of largemouth bass. We treated the allergens at 100, 200, 300 and 400 MPa for 15 min and at 300 MPa for 5, 10, 15, 20 and 30 min at 20 °C. The treated samples from largemouth bass were tested for their IgE-binding properties by combining Sodium dodecyl sulfate-Polyacrylamide gel electrophoresis (SDS-PAGE) with western blotting (WB) and enzyme-linked immunosorbent assay (ELISA). Circular dichroism analysis was performed to characterize the structural change. In summary, we can determine that the greatest structure changes were found for samples treated by 400 MPa for 15 min. High pressure treatment did change the structure, subunit composition and molecular weight of largemouth bass allergens, but it did not change the allergenicity of the allergens.

  10. Reduction of pollutants in painting operation and suggestion of an optimal technique for extracting titanium dioxide from paint sludge in car manufacturing industries--case study (SAIPA).

    PubMed

    Khezri, Seyed Mostafa; Shariat, Seyed Mahmood; Tabibian, Sahar

    2012-06-01

    Paint sludge of car manufacturing industries are not disposed in landfills, since they contain hazardous materials with a high concentration of chromium, aluminum, titanium, barium, copper, Iron, magnesium, strontium, and so on. Thus, it is essential to find solutions in order to neutralize them or suggest cost-effective techniques, which are also environmentally acceptable. Because, this sludge contains considerable amounts of Ti pigments and unbaked resins, recycling these pigments--which could be used in a variety of industries such as paint factories--is an appropriate subject for further research. In this article, with the aim of identification of main pollutants in order to eliminate them and suggest a cost-effective solution to recover the sludge, a large number of tests including X-ray fluorescence spectroscopy, X ray diffraction spectroscopy, and diffusion thermal analysis are conducted to determine types and concentration of elements, and combinations of paint sludge in car manufacturing industries. As titanium dioxide (TiO₂) is widely used as the main pigment of automobile paints, an optimal technique is suggested for extracting TiO₂ with high purity percentage through adopting scientific methods such as membrane and electrolysis. PMID:21937529

  11. Expression, purification, and characterization of almond (Prunus dulcis) allergen Pru du 4

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Biochemical characterizations of food allergens are required for understanding the allergenicity of food allergens. Such studies require a relatively large amount of highly purified allergens. Profilins from numerous species are known to be allergens, including food allergens, such as almond (Prunus...

  12. Allergen immunotherapy: how to balance the different views from pulmonologists and allergists?

    PubMed

    Incorvaia, Cristoforo; Fuiano, Nicola; Frati, Franco

    2012-08-01

    Allergen immunotherapy (AIT) is the treatment characterizing the allergological approach to respiratory allergy. Unfortunately, most available data from the literature and current practice indicate that pulmonologists do no consider AIT when choosing the treatment strategy in patients with asthma. Indeed AIT, from its introduction in 1911 to nowadays, was unceasingly improved and has accumulated clear evidence on its effectiveness. Moreover, AIT has a characteristic not shared by drugs in the capacity to modify the natural history of asthma, due to its immunologic mechanisms of actions, and thus also works after the treatment withdrawal. This also makes AIT a clearly cost-effective treatment over time. It is surprising that pulmonologists, for whom asthma is a major disease to manage, do not consider AIT when choosing the optimal treatment in single patients. The insufficient information on AIT and the availability of allergen extracts with less than good quality are likely to be the most important factors influencing such an attitude. The current development of standardized, pharmaceutical-grade products for AIT seems capable of making allergen extracts comparable to drugs and to stimulate a rethinking of AIT's role in the treatment of asthma in pulmonologists. A reappraisal of the significance of the allergen-specific bronchial challenge could represent a further factor suggesting AIT as a reliable option. PMID:22947011

  13. Proteomic identification of allergenic seed proteins, napin and cruciferin, from cold-pressed rapeseed oils.

    PubMed

    Puumalainen, T J; Puustinen, A; Poikonen, S; Turjanmaa, K; Palosuo, T; Vaali, K

    2015-05-15

    In Finland and France atopic children commonly react to seeds of oilseed rape and turnip rape in skin prick tests (SPT) and open food challenges. These seeds are not as such in dietary use and therefore the routes of sensitization are unknown. Possible allergens were extracted from commercial cold-pressed and refined rapeseed oils and identified by gel-based tandem nanoflow liquid chromatography mass spectrometry (LC-MS/MS). Napin (a 2S albumin), earlier identified as a major allergen in the seeds of oilseed rape and turnip rape, and cruciferin (an 11S globulin), a new potential seed allergen, were detected in cold-pressed oils, but not in refined oils. Pooled sera from five children sensitized or allergic to oilseed rape and turnip rape seeds reacted to these proteins from cold-pressed oil preparations and individual sera from five children reacted to these proteins extracted from the seeds when examined with IgE immunoblotting. Hence cold-pressed rapeseed oil might be one possible route of sensitization for these allergens. PMID:25577095

  14. Developing a preventive immunization approach against insect bite hypersensitivity using recombinant allergens: A pilot study.

    PubMed

    Jonsdottir, Sigridur; Hamza, Eman; Janda, Jozef; Rhyner, Claudio; Meinke, Andreas; Marti, Eliane; Svansson, Vilhjalmur; Torsteinsdottir, Sigurbjorg

    2015-07-15

    Insect bite hypersensitivity (IBH) is an allergic dermatitis of horses caused by bites of midges (Culicoides spp.). IgE-mediated reactions are often involved in the pathogenesis of this disease. IBH does not occur in Iceland due to the absence of Culicoides, but it occurs with a high frequency in Icelandic horses exported to mainland Europe, where Culicoides are present. We hypothesize that immunization with the Culicoides allergens before export could reduce the incidence of IBH in exported Icelandic horses. The aim of the present study was therefore to compare intradermal and intralymphatic vaccination using four purified recombinant allergens, in combination with a Th1 focusing adjuvant. Twelve horses were vaccinated three times with 10μg of each of the four recombinant Culicoides nubeculosus allergens. Six horses were injected intralymphatically, three with and three without IC31(®), and six were injected intradermally, in the presence or absence of IC31(®). Antibody responses were measured by immunoblots and ELISA, potential sensitization in a sulfidoleukotriene release test and an intradermal test, cytokine and FoxP3 expression with real time PCR following in vitro stimulation of PBMC. Immunization with the r-allergens induced a significant increase in levels of r-allergen-specific IgG1, IgG1/3, IgG4/7, IgG5 and IgG(T). Application of the r-allergens in IC31(®) adjuvant resulted in a significantly higher IgG1, IgG1/3, IgG4/7 allergen-specific response. Intralymphatic injection was slightly more efficient than intradermal injection, but the difference did not reach significance. Testing of the blocking activity of the sera from the horses immunized intralymphatically with IC31(®) showed that the generated IgG antibodies were able to partly block binding of serum IgE from an IBH-affected horse to these r-allergens. Furthermore, IgG antibodies bound to protein bands on blots of C. nubeculosus salivary gland extract. No allergen-specific IgE was induced and

  15. Comprehensive proteomics approach in characterizing and quantifying allergenic proteins from northern shrimp: toward better occupational asthma prevention.

    PubMed

    Abdel Rahman, Anas M; Kamath, Sandip D; Gagné, Sébastien; Lopata, Andreas L; Helleur, Robert

    2013-02-01

    Occupational asthma is a major chronic health dilemma among workers involved in the seafood industry. Several proteins notoriously known to cause asthma have been reported in different seafood. This work involves the application of an allergenomics strategy to study the most potent allergens of northern shrimp. The proteins were extracted from shrimp tissue and profiled by gel electrophoresis. Allergenic proteins were identified based on their reactivity to patient sera and were structurally identified using tandem mass spectrometry. Northern shrimp tropomyosin, arginine kinase, and sarcoplasmic calcium-binding protein were found to be the most significant allergens. Multiple proteolytic enzymes enabled 100% coverage of the sequence of shrimp tropomyosin by tandem mass specrometry. Only partial sequence coverage was obtained, however, for the shrimp allergen arginine kinase. Signature peptides, for both tropomyosin and arginine kinase, were assigned and synthesized for use in developing the multiple reaction monitoring tandem mass spectrometric method. Subsequently, air samples were collected from a shrimp processing plant and two aerosolized proteins quantified using tandem mass specrometry. Allergens were detected in all areas of the plant, reaching levels as high as 375 and 480 ng/m(3) for tropomyosine and arginine kinase, respectively. Tropomyosine is much more abundant than arginine kinase in shrimp tissues, so the high levels of arginine kinase suggest it is more easily aerosolized. The present study shows that mass spectrometric analysis is a sensitive and accurate tool in identifying and quantifying aerosolized allergens. PMID:23268739

  16. Occupational exposure to allergenic mites in a Polish zoo.

    PubMed

    Solarz, Krzysztof; Szilman, Piotr; Szilman, Ewa

    2004-01-01

    The study was carried out from April 2000-March 2001. During this period 49 samples of dust, litter, debris and residues from cages and run-offs of mammals, birds and reptiles in the Silesian Zoo, were examined for the presence of mites, especially the allergenic taxa. Mites were extracted using the Berlese method and preserved in 70 % ethanol. For identification, the mites were mounted in Hoyer's medium on microscope slides. Mites were found in 44 of 49 samples analyzed (89.8 %). A total of 5,097 mites were collected, from which 60.3 % were found in samples collected in spring, whereas only 13 % in summer and 24.1 % in autumn. The remaining 2.6 % of the total mite population was found in winter. Majority of mites (82.7 %) were collected from aviaries of macaws and cockatiels (Ara ararauna and Nymphicus hollandicus). A total of 10 species of astigmatid mites were identified that belong to 4 families--Acaridae, Glycyphagidae, Anoetidae and Pyroglyphidae. Generally, the allergenic mites of the order Astigmata constituted 49.5 % of the total count. Among them Acarus farris was predominant (34 % of the total count), followed by Tyrophagus putrescentiae (4.7 %), Caloglyphus sp. (4.35 %) and Acarus immobilis (4.31 %). Dermatophagoides farinae, the house-dust-mite species, was for the first time found in this environment. D. farinae (0.05 % of the total population) was associated with parrots, canids and artiodactyls. Summarizing, it should be stressed, that cages and run-offs of different mammals, aviaries of parrots and terrariums of snakes are important sources of some allergenic mites, especially A. farris and T. putrescentiae, that might cause allergies in workers. PMID:15236495

  17. T-cell response to allergens.

    PubMed

    Ozdemir, Cevdet; Akdis, Mübeccel; Akdis, Cezmi A

    2010-01-01

    Anaphylaxis is a life-threatening IgE-dependent type 1 hypersensitivity reaction in which multiple organ systems are involved. The existence of allergen exposure and specific IgE are the major contributors to this systemic reaction. The decision of the immune system to respond to allergens is highly dependent on factors including the type and load of allergen, behavior and type of antigen-presenting cells, innate immune response stimulating substances in the same micromilieu, the tissue of exposure, interactions between T and B lymphocytes, costimulators, and genetic propensity known as atopy. Antigen-presenting cells introduce processed allergens to T-helper lymphocytes, where a decision of developing different types of T-cell immunity is given under the influence of several cytokines, chemokines, costimulatory signals and regulatory T cells. Among Th2-type cytokines, interleukin (IL)-4 and IL-13 are responsible for class switching in B cells, which results in production of allergen-specific IgE antibodies that bind to specific receptors on mast cells and basophils. After re-exposure to the sensitized allergen, this phase is followed by activation of IgE Fc receptors on mast cells and basophils resulting in biogenic mediator releases responsible for the symptoms and signs of anaphylaxis. Since the discovery of regulatory T cells, the concepts of immune regulation have substantially changed during the last decade. Peripheral T-cell tolerance is a key immunologic mechanism in healthy immune response to self antigens and non-infectious non-self antigens. Both naturally occurring CD4+CD25+ regulatory T (Treg) cells and inducible populations of allergen-specific, IL-10-secreting Treg type 1 cells inhibit allergen-specific effector cells and have been shown to play a central role in the maintenance of peripheral homeostasis and the establishment of controlled immune responses. On the other hand, Th17 cells are characterized by their IL-17 (or IL-17A), IL-17F, IL-6

  18. Computationally predicted IgE epitopes of walnut allergens contribute to cross-reactivity with peanuts

    PubMed Central

    Maleki, Soheila J.; Teuber, Suzanne S.; Cheng, Hsiaopo; Chen, Deliang; Comstock, Sarah S.; Ruan, Sanbao; Schein, Catherine H.

    2011-01-01

    Background Cross reactivity between peanuts and tree nuts implies that similar IgE epitopes are present in their proteins. Objective To determine whether walnut sequences similar to known peanut IgE binding sequences, according to the property distance (PD) scale implemented in the Structural Database of Allergenic Proteins (SDAP), react with IgE from sera of patients with allergy to walnut and/or peanut. Methods Patient sera were characterized by Western blotting for IgE-binding to nut protein extracts, and to peptides from walnut and peanut allergens, similar to known peanut epitopes as defined by low PD values, synthesized on membranes. Competitive ELISA was used to show that peanut and predicted walnut epitope sequences compete with purified Ara h 2 for binding to IgE in serum from a cross-reactive patient. Results Sequences from the vicilin walnut allergen Jug r 2 which had low PD values to epitopes of the peanut allergen Ara h 2, a 2s-albumin, bound IgE in sera from five patients who reacted to either walnut, peanut or both. A walnut epitope recognized by 6 patients mapped to a surface-exposed region on a model of the N-terminal pro-region of Jug r 2. A predicted walnut epitope competed for IgE binding to Ara h 2 in serum as well as the known IgE epitope from Ara h 2. Conclusions Sequences with low PD value (<8.5) to known IgE epitopes could contribute to cross-reactivity between allergens. This further validates the PD scoring method for predicting cross-reactive epitopes in allergens. PMID:21883278

  19. Immunological mechanisms of allergen-specific immunotherapy.

    PubMed

    Jutel, Marek; Akdis, C A

    2011-06-01

    The studies on the mechanisms of specific immunotherapy (SIT) point out its targets that decide on the efficacy of SIT and hence might be used for its further improvement. Several mechanisms have been proposed to explain the beneficial effects of immunotherapy. The knowledge of the mechanisms underlying allergic diseases and curative treatment possibilities has experienced exciting advances over the last three decades. Studies in several clinical trials in allergen-SIT have demonstrated that the induction of a tolerant state against allergens in many ways represents a key step in the development of a healthy immune response against allergens. Several cellular and molecular mechanisms have been demonstrated: allergen-specific suppressive capacities of both inducible subsets of CD4(+) CD25(+) forkhead box P3(+) T-regulatory and IL-10-secreting type 1 T-regulatory cells increase in peripheral blood; suppression of eosinophils, mast cells, and basophils; Ab isotype change from IgE to IgG4. This review aims at the better understanding of the observed immunological changes associated with allergen SIT. PMID:21466562

  20. Mechanisms of allergen-specific immunotherapy.

    PubMed

    Fujita, Hiroyuki; Soyka, Michael B; Akdis, Mübeccel; Akdis, Cezmi A

    2012-01-01

    Allergen-specific immunotherapy (allergen-SIT) is a potentially curative treatment approach in allergic diseases. It has been used for almost 100 years as a desensitizing therapy. The induction of peripheral T cell tolerance and promotion of the formation of regulatory T-cells are key mechanisms in allergen-SIT. Both FOXP3+CD4+CD25+ regulatory T (Treg) cells and inducible IL-10- and TGF-β-producing type 1 Treg (Tr1) cells may prevent the development of allergic diseases and play a role in successful allergen-SIT and healthy immune response via several mechanisms. The mechanisms of suppression of different pro-inflammatory cells, such as eosinophils, mast cells and basophils and the development of allergen tolerance also directly or indirectly involves Treg cells. Furthermore, the formation of non-inflammatory antibodies particularly IgG4 is induced by IL-10. Knowledge of these molecular basis is crucial in the understanding the regulation of immune responses and their possible therapeutic targets in allergic diseases. PMID:22409879

  1. Pollensomes as Natural Vehicles for Pollen Allergens.

    PubMed

    Prado, Noela; De Linares, Concepción; Sanz, María L; Gamboa, Pedro; Villalba, Mayte; Rodríguez, Rosalía; Batanero, Eva

    2015-07-15

    Olive (Olea europaea) pollen constitutes one of the most important allergen sources in the Mediterranean countries and some areas of the United States, South Africa, and Australia. Recently, we provided evidence that olive pollen releases nanovesicles of respirable size, named generically pollensomes, during in vitro germination. Olive pollensomes contain allergens, such as Ole e 1, Ole e 11, and Ole e 12, suggesting a possible role in allergy. The aim of this study was to assess the contribution of pollensomes to the allergic reaction. We show that pollensomes exhibit allergenic activity in terms of patients' IgE-binding capacity, human basophil activation, and positive skin reaction in sensitized patients. Furthermore, allergen-containing pollensomes have been isolated from three clinically relevant nonphylogenetically related species: birch (Betula verrucosa), pine (Pinus sylvestris), and ryegrass (Lolium perenne). Most interesting, pollensomes were isolated from aerobiological samples collected with an eight-stage cascade impactor collector, indicating that pollensomes secretion is a naturally occurring phenomenon. Our findings indicate that pollensomes may represent widespread vehicles for pollen allergens, with potential implications in the allergic reaction. PMID:26041541

  2. Immunochemical characterization of prosopis juliflora pollen allergens and evaluation of cross-reactivity pattern with the most allergenic pollens in tropical areas.

    PubMed

    Assarehzadegan, Mohammad-Ali; Khodadadi, Ali; Amini, Akram; Shakurnia, Abdol-Hosein; Marashi, Seyed Saeid; Ali-Sadeghi, Hosein; Zarinhadideh, Farnoosh; Sepahi, Najmeh

    2015-02-01

    Allergy to Prosopis juliflora (mesquite) pollen is one of the common causes of respiratory allergy in tropical countries. Mesquite is widely used as street trees in towns and ornamental shade trees in parks and gardens throughout arid and semiarid regions of Iran. The inhalation of mesquite pollen and several species of Amaranthus/Chenopodiaceae family is the most important cause of allergic respiratory symptoms in Khuzestan province. This study was designed to evaluate IgE banding proteins of mesquite pollen extract and its IgE cross-reactivity with other allergenic plants. Twenty patients with allergic symptoms and positive skin prick tests (SPT) for mesquite pollen extract participated in the study. Crude pollen extract was prepared from local mesquite trees and used for the evaluation of allergenic profiles of P. juliflora pollen extract by Sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and IgE-immunoblotting. There were several protein bands in mesquite pollen extract using SDS-PAGE with the approximate range of molecular weight of 10-85 kDa. The most frequent IgE reactive bands among the patients' sera were approximately 20 and 66 kDa. However, there were other IgE reactive protein bands among the patients' sera with molecular weights of 10, 15, 35, 45, 55 and 85 kDa. Inhibition experiments revealed high IgE cross-reactivity between mesquite and acacia. There are several IgE-binding proteins in P. juliflora pollen extract. Results of this study indicate that proteins with a molecular weight of 10 to 85 kDa are the major allergens in P. juliflora pollen extract. PMID:25530142

  3. Indoor determinants of dustborne allergens in Mexican homes

    PubMed Central

    Hernández-Cadena, Leticia; Zeldin, Darryl C.; Sever, Michelle L.; Sly, Peter D.; London, Stephanie J.; Escamilla-Nuñez, María Consuelo; Romieu, Isabelle

    2015-01-01

    Exposure to indoor allergens represents a significant risk factor for allergies and asthma in several parts of the world. In Mexico, few studies have evaluated indoor allergens, including cat, dog, and mouse allergens and the factors that predict their presence. This study evaluates the main environmental and household predictors of high prenatal allergen levels and multiple allergen exposures in a birth cohort from Mexico City. A cross-sectional study was conducted as part of a birth cohort study of 1094 infants recruited during pregnancy and followed until delivery. We collected dust samples in a subset of 264 homes and assessed environmental factors. Der p 1, Der f 1, dust mite group 2, Fel d 1, Can f 1, Rat n 1, Mus m 1, and Bla g 2 concentrations in dust samples were measured using immunoassays. To define detectable allergen levels, the lowest limits of detection for each allergen were taken as cutoff points. Overall allergen exposure was considered high when four or more allergens exceeded detectable levels in the same household. Logistic regression was used for predictive models. Eighty-five percent of homes had at least one allergen in dust over the detection limit, 52.1% had high exposure (four or more allergens above detectable limits), and 11.7% of homes had detectable levels for more than eight allergens. Der p 1, Der p 2, Mus m 1, and Fel d 1 were the most frequent allergens detected. Each allergen had both common and distinct predictors. The main predictors of a high multiple allergen index were the size of the home, pesticide use, mother's age, mother as homemaker, and season. Increased indoor environmental allergen exposure is mainly related to sociodemographic factors and household cleaning. PMID:25715241

  4. Indoor allergens in Minnesota schools and child care centers.

    PubMed

    Tranter, Daniel C; Wobbema, Amanda Teresa; Norlien, Kathleen; Dorschner, Dale F

    2009-09-01

    Elevated concentrations of allergens in the indoor environment may cause allergic sensitization and symptoms. Occupant exposure to indoor allergens in educational facilities should and can be controlled. This study (1) assessed the presence of indoor allergens in Minnesota schools and child care centers, (2) characterized the distribution of allergens in different materials, and (3) evaluated the effect of building and maintenance interventions on allergen concentrations. Settled dust samples were collected from carpet, vinyl tile floors, and upholstered furniture in six schools and seven child care centers before and after interventions. Interventions included changes to cleaning, ventilation, entry mats, furnishings, flooring, and classroom items. The amount of total dust, culturable fungi, and indoor allergens--cockroach, dust mite, cat, and dog--were quantified in the dust samples. Cockroach and dust mite allergens were generally low and below the detection limit, but one dust mite allergen was detected in some areas. Cat and dog allergens were frequently detected at elevated levels, with half the samples above the provisional sensitization risk thresholds and a few samples above the symptom thresholds. Allergen concentrations were highest in upholstered furniture, followed by carpeting and then vinyl floor tile. Cat and dog allergens were lower after the interventions. Cat and dog allergens, but not dust mite and cockroach allergens, seem to be ubiquitous in child care and elementary schools of the U.S. Midwest. These allergens may contribute to sensitization in atopic individuals and occasionally cause symptoms in sensitized allergic individuals. Fleecy materials that are not adequately cleaned, such as upholstered furniture, appear to be the most significant allergen reservoirs. Modest environmental interventions can be implemented by building staff, which should result in lower allergen concentrations. PMID:19585331

  5. Indoor determinants of dustborne allergens in Mexican homes.

    PubMed

    Hernández-Cadena, Leticia; Zeldin, Darryl C; Barraza-Villarreal, Albino; Sever, Michelle L; Sly, Peter D; London, Stephanie J; Escamilla-Nuñez, María Consuelo; Romieu, Isabelle

    2015-01-01

    Exposure to indoor allergens represents a significant risk factor for allergies and asthma in several parts of the world. In Mexico, few studies have evaluated indoor allergens, including cat, dog, and mouse allergens and the factors that predict their presence. This study evaluates the main environmental and household predictors of high prenatal allergen levels and multiple allergen exposures in a birth cohort from Mexico City. A cross-sectional study was conducted as part of a birth cohort study of 1094 infants recruited during pregnancy and followed until delivery. We collected dust samples in a subset of 264 homes and assessed environmental factors. Der p 1, Der f 1, dust mite group 2, Fel d 1, Can f 1, Rat n 1, Mus m 1, and Bla g 2 concentrations in dust samples were measured using immunoassays. To define detectable allergen levels, the lowest limits of detection for each allergen were taken as cutoff points. Overall allergen exposure was considered high when four or more allergens exceeded detectable levels in the same household. Logistic regression was used for predictive models. Eighty-five percent of homes had at least one allergen in dust over the detection limit, 52.1% had high exposure (four or more allergens above detectable limits), and 11.7% of homes had detectable levels for more than eight allergens. Der p 1, Der p 2, Mus m 1, and Fel d 1 were the most frequent allergens detected. Each allergen had both common and distinct predictors. The main predictors of a high multiple allergen index were the size of the home, pesticide use, mother's age, mother as homemaker, and season. Increased indoor environmental allergen exposure is mainly related to sociodemographic factors and household cleaning. PMID:25715241

  6. Increased allergen-specific Th2 responses in vitro in atopic subjects receiving subclinical allergen challenge.

    PubMed

    Gabrielsson, S; Paulie, S; Roquet, A; Ihre, E; Lagging, E; van Hage-Hamsten, M; Härfast, B; Troye-Blomberg, M

    1997-08-01

    The study aimed to determine whether inhalation of subclinical allergen doses-leads to a shift in the balance between T helper (Th) 1 and Th2 cells in asthmatic patients. Elevated IgE requires allergen-specific T cells producing cytokines such as interleukin (IL)-4 or IL-13. Interferon-gamma (IFN-gamma) produced by Th1 cells counteracts the effects of IL-4. In nature, allergic persons are often exposed to low levels of allergen, leading to hyperreactivity, but not to acute allergic reactions. In this study, nine allergic persons inhaled low doses of allergen or placebo in a double-blind manner over seven consecutive weekdays. During the study, the bronchial responsiveness to histamine challenge increased, but no subject exhibited asthmatic symptoms. Blood was drawn on days 0, 1, 4, and 9, and the number of IL-4- and IFN-gamma-producing cells was measured by enzyme-linked immunospot (ELISPOT) assay after in vitro stimulation with a low-dose phytohemagglutinin (PHA) mixed with the relevant allergen or with PHA alone. In three of the four subjects receiving allergen, the IL-4/IFN-gamma ratio increased during the time of the study. No increase was seen in the placebo group. No increase was seen in serum IgE levels in any of the groups. We conclude that a shift in the balance between Th1 and Th2 cells can be detected in subjects exposed to subclinical allergen doses. PMID:9284986

  7. Prevention of Birch Pollen-Related Food Allergy by Mucosal Treatment with Multi-Allergen-Chimers in Mice

    PubMed Central

    Hoflehner, Elisabeth; Hufnagl, Karin; Schabussova, Irma; Jasinska, Joanna; Hoffmann-Sommergruber, Karin; Bohle, Barbara; Maizels, Rick M.; Wiedermann, Ursula

    2012-01-01

    Background Among birch pollen allergic patients up to 70% develop allergic reactions to Bet v 1-homologue food allergens such as Api g 1 (celery) or Dau c 1 (carrot), termed as birch pollen-related food allergy. In most cases, specific immunotherapy with birch pollen extracts does not reduce allergic symptoms to the homologue food allergens. We therefore genetically engineered a multi-allergen chimer and tested if mucosal treatment with this construct could represent a novel approach for prevention of birch pollen-related food allergy. Methodology BALB/c mice were poly-sensitized with a mixture of Bet v 1, Api g 1 and Dau c 1 followed by a sublingual challenge with carrot, celery and birch pollen extracts. For prevention of allergy sensitization an allergen chimer composed of immunodominant T cell epitopes of Api g 1 and Dau c 1 linked to the whole Bet v 1 allergen, was intranasally applied prior to sensitization. Results Intranasal pretreatment with the allergen chimer led to significantly decreased antigen-specific IgE-dependent β-hexosaminidase release, but enhanced allergen-specific IgG2a and IgA antibodies. Accordingly, IL-4 levels in spleen cell cultures and IL-5 levels in restimulated spleen and cervical lymph node cell cultures were markedly reduced, while IFN-γ levels were increased. Immunomodulation was associated with increased IL-10, TGF-β and Foxp3 mRNA levels in NALT and Foxp3 in oral mucosal tissues. Treatment with anti-TGF-β, anti-IL10R or anti-CD25 antibodies abrogated the suppression of allergic responses induced by the chimer. Conclusion Our results indicate that mucosal application of the allergen chimer led to decreased Th2 immune responses against Bet v 1 and its homologue food allergens Api g 1 and Dau c 1 by regulatory and Th1-biased immune responses. These data suggest that mucosal treatment with a multi-allergen vaccine could be a promising treatment strategy to prevent birch pollen-related food allergy. PMID:22768077

  8. Cardiac urticaria caused by eucleid allergen

    PubMed Central

    Zhan, Xiaodong; Li, Chaopin; Wu, Qianwen

    2015-01-01

    Urticaria is a common allergic diseases, which involve respiratory and digestive system being suffered in some population. Yet, relatively little research has been done on the adverse effect on the heart. We did this research to examine the correlation between the abnormality of ECG in the patients with acute allergic urticaria and the antigen of eucleid. The antigen (allergen of eucleid and other allergens) was used to test the patients with acute allergic urticaria by skin prick test and electrocardiogram was employed to examine the patients with strong positive (moth & caterpillar) eucleid antigen. Strong positive eucleid antigen was identified in 84 cases with abnormal electrocardiographic pattern of diversity. So, the acute allergic skin urticaria caused by eucleid allergen may impose strong effect on the heart and thus lead to allergic cardiac urticaria. PMID:26885121

  9. Heat and pressure treatments effects on peanut allergenicity.

    PubMed

    Cabanillas, Beatriz; Maleki, Soheila J; Rodríguez, Julia; Burbano, Carmen; Muzquiz, Mercedes; Jiménez, María Aránzazu; Pedrosa, Mercedes M; Cuadrado, Carmen; Crespo, Jesús F

    2012-05-01

    Peanut allergy is recognized as one of the most severe food allergies. The aim of this study was to investigate the changes in IgE binding capacity of peanut proteins produced by thermal-processing methods, including autoclaving. Immunoreactivity to raw and thermally processed peanut extracts was evaluated by IgE immunoblot and skin prick test in patients with clinical allergy to peanut. Roasted peanut and autoclaved roasted peanut were selected for IgE ELISA experiments with individual sera, immunoblot experiments with antibodies against peanut allergens (Ara h 1, Ara h 2 and Ara h 3), digestion experiments, and circular dichroism spectroscopy. In vitro and in vivo experiments showed IgE immunoreactivity of roasted peanut proteins decreased significantly at extreme conditions of autoclaving. Circular dichroism experiments showed unfolding of proteins in autoclave treated samples, which makes them more susceptible to digestion. Autoclaving at 2.56atm, for 30min, produces a significant decrease of IgE-binding capacity of peanut allergens. PMID:26434302

  10. Profilins: mimickers of allergy or relevant allergens?

    PubMed

    Santos, Alexandra; Van Ree, Ronald

    2011-01-01

    Profilins are ubiquitous proteins, present in all eukaryotic cells and identified as allergens in pollen, latex and plant foods. The highly conserved structure justifies the cross-reactive nature of IgE antibodies against plant profilins and their designation as pan-allergens. Primary sensitization to profilin seems to arise from pollen sensitization with later development of cross-reactive IgE antibodies against plant food (and possibly latex) profilins. The role of profilin in inducing allergic symptoms needs to be evaluated and raises important issues in allergy diagnosis due to cross-reactivity. IgE cross-reactivity among profilins is associated with multiple pollen sensitization and with various pollen-food syndromes. In respiratory allergy, sensitization to pollen to which the patient has virtually no environmental exposure has been identified as a manifestation of profilin sensitization. As a food allergen, profilin usually elicits mild reactions, such as oral allergy syndrome, is not modified by processing and is especially important in allergy to some fruits, such as melon, watermelon, banana, tomato, citrus fruit and persimmon. Purified natural and recombinant profilins for in vitro and in vivo allergy tests are helpful in the diagnostic work-up. Herein we review the current state of knowledge about the allergen profilin and its implications in the diagnosis and treatment of allergic diseases. We conclude that, although its role in triggering allergic symptoms is still controversial, profilin is undoubtedly a relevant allergen. As a pan-allergen, profilin is associated with multiple pollen sensitization and pollen-food-latex syndromes that the allergist has to be aware of in order to accomplish an accurate diagnosis and successful treatment of allergic diseases. PMID:21293140

  11. Importance of albumin in cross-reactivity among cat, dog and horse allergens.

    PubMed

    Cabañas, R; López-Serrano, M C; Carreira, J; Ventas, P; Polo, F; Caballero, M T; Contreras, J; Barranco, P; Moreno-Ancillo, A

    2000-01-01

    Different allergenic proteins have been involved in cross-reactivity among animals. Albumins seem to be cross-sensitizing allergenic components. The aim of this study was to assess the importance of albumin as a cross-reactive allergen in patients sensitized to cat, dog and horse. One hundred and seventeen patients sensitized to cat were tested for IgE reactivity using skin prick tests and RAST assays with cat, dog and horse hair/dander extracts and their purified albumin extracts. RAST-inhibition studies were carried out to assess cross-reactivity among cat, dog and horse and among their purified albumins. It was found that 22% of patients exhibited specific IgE to cat albumin; 41% of patients sensitized to cat were also sensitized to dog and horse. Out of these patients, 21% had IgE to three albumins and 17% to two. Reciprocal inhibitions were observed among cat, dog and horse albumins and also among cat, dog and horse hair/dander extracts, using in the latter experiment sera from patients not sensitized to albumins. IgE binding to horse extract was inhibited 30% by its homologous albumin and IgE binding to cat and dog extracts in almost 15% by their respective albumins. It was concluded that albumins from these three animals share some epitopes that account for the cross-reactivity observed in around one-third of patients sensitized to cat, dog and horse. Nevertheless, more than 50% of specific IgE that cross-reacts among these three animals is directed to allergens other than albumin. PMID:10879993

  12. Skin exposure promotes a Th2-dependent sensitization to peanut allergens.

    PubMed

    Tordesillas, Leticia; Goswami, Ritobrata; Benedé, Sara; Grishina, Galina; Dunkin, David; Järvinen, Kirsi M; Maleki, Soheila J; Sampson, Hugh A; Berin, M Cecilia

    2014-11-01

    Sensitization to foods often occurs in infancy, without a known prior oral exposure, suggesting that alternative exposure routes contribute to food allergy. Here, we tested the hypothesis that peanut proteins activate innate immune pathways in the skin that promote sensitization. We exposed mice to peanut protein extract on undamaged areas of skin and observed that repeated topical exposure to peanut allergens led to sensitization and anaphylaxis upon rechallenge. In mice, this epicutaneous peanut exposure induced sensitization to the peanut components Ara h 1 and Ara h 2, which is also observed in human peanut allergy. Both crude peanut extract and Ara h 2 alone served as adjuvants, as both induced a bystander sensitization that was similar to that induced by the atopic dermatitis-associated staphylococcal enterotoxin B. In cultured human keratinocytes and in murine skin, peanut extract directly induced cytokine expression. Moreover, topical peanut extract application induced an alteration dependent on the IL-33 receptor ST2 in skin-draining DCs, resulting in Th2 cytokine production from T cells. Together, our data support the hypothesis that peanuts are allergenic due to inherent adjuvant activity and suggest that skin exposure to food allergens contributes to sensitization to foods in early life. PMID:25295541

  13. Skin exposure promotes a Th2-dependent sensitization to peanut allergens

    PubMed Central

    Tordesillas, Leticia; Goswami, Ritobrata; Benedé, Sara; Grishina, Galina; Dunkin, David; Järvinen, Kirsi M.; Maleki, Soheila J.; Sampson, Hugh A.; Berin, M. Cecilia

    2014-01-01

    Sensitization to foods often occurs in infancy, without a known prior oral exposure, suggesting that alternative exposure routes contribute to food allergy. Here, we tested the hypothesis that peanut proteins activate innate immune pathways in the skin that promote sensitization. We exposed mice to peanut protein extract on undamaged areas of skin and observed that repeated topical exposure to peanut allergens led to sensitization and anaphylaxis upon rechallenge. In mice, this epicutaneous peanut exposure induced sensitization to the peanut components Ara h 1 and Ara h 2, which is also observed in human peanut allergy. Both crude peanut extract and Ara h 2 alone served as adjuvants, as both induced a bystander sensitization that was similar to that induced by the atopic dermatitis-associated staphylococcal enterotoxin B. In cultured human keratinocytes and in murine skin, peanut extract directly induced cytokine expression. Moreover, topical peanut extract application induced an alteration dependent on the IL-33 receptor ST2 in skin-draining DCs, resulting in Th2 cytokine production from T cells. Together, our data support the hypothesis that peanuts are allergenic due to inherent adjuvant activity and suggest that skin exposure to food allergens contributes to sensitization to foods in early life. PMID:25295541

  14. Manufacturing Success

    ERIC Educational Resources Information Center

    Reese, Susan

    2007-01-01

    According to the National Association of Manufacturers (NAM), "manufacturing is the engine that drives American prosperity". When NAM and its research and education arm, The Manufacturing Institute, released the handbook, "The Facts About Modern Manufacturing," in October 2006, NAM President John Engler noted, that manufacturing output in America…

  15. 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. PMID:26023323

  16. 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

  17. Animal allergens and their presence in the environment.

    PubMed

    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

  18. Standardization and Regulation of Allergen Products in the European Union.

    PubMed

    Zimmer, Julia; Vieths, Stefan; Kaul, Susanne

    2016-03-01

    Product-specific standardization is of prime importance to ensure persistent quality, safety, and efficacy of allergen products. The regulatory framework in the EU has induced great advancements in the field in the last years although national implementation still remains heterogeneous. Scores of methods for quantification of individual allergen molecules are developed each year and also the challenging characterization of chemically modified allergen products is progressing. However, despite the unquestionable increase in knowledge and the subsequent improvements in control of quality parameters of allergen products, an important aim has not been reached yet, namely cross-product comparability. Still, comparison of allergen product potency, either based on total allergenic activity or individual allergen molecule content, is not possible due to a lack of standard reference preparations in conjunction with validated standard methods. This review aims at presenting the most recent developments in product-specific standardization as well as activities to facilitate cross-product comparability in the EU. PMID:26874849

  19. Allergen-Specific CD4(+) T Cells in Human Asthma.

    PubMed

    Ling, Morris F; Luster, Andrew D

    2016-03-01

    In allergic asthma, aeroallergen exposure of sensitized individuals mobilizes robust innate and adaptive airway immune responses, stimulating eosinophilic airway inflammation and the activation and infiltration of allergen-specific CD4(+) T cells into the airways. Allergen-specific CD4(+) T cells are thought to be central players in the asthmatic response as they specifically recognize the allergen and initiate and orchestrate the asthmatic inflammatory response. In this article, we briefly review the role of allergen-specific CD4(+) T cells in the pathogenesis of human allergic airway inflammation in allergic individuals, discuss the use of allergen-major histocompatibility complex class II tetramers to characterize allergen-specific CD4(+) T cells, and highlight current gaps in knowledge and directions for future research pertaining to the role of allergen-specific CD4(+) T cells in human asthma. PMID:27027948

  20. INDUCTION OF PLANT ALLERGENS BY ENVIRONMENTAL AGENTS

    EPA Science Inventory

    The specific objectives of the project and the progress toward meeting these objectives are listed below:

    Identify up to Three Environmental Exposures That Induce the Production of an Allergenic Protein in the Mountain Cedar Tree by Examining the Effects of Exposu...

  1. PROTEOMIC ANALYSIS OF ALLERGENS FROM METARHIZIUM ANISOPLIAE

    EPA Science Inventory

    Introduction

    The goal of this project is the identification and characterization of allergens from the fungus Metarhizium anisopliae, using mass spectrometry (MS). The US EPA, under the "Children at Risk" program, is currently addressing the problem of indoor fungal bioaer...

  2. PROTEOMIC ANALYSIS OF ALLERGENS FROM METARHIZIUM ANISOPLIEA

    EPA Science Inventory

    The goal of this project is the identification and characterization of allergens from the fungus M. Anisopliae, using mass spectrometry (MS). The US EPA, under the "Children at Risk" program, is currently addressing the problem of indoor fungal bioaerosol contamination. One of ...

  3. IDENTIFYING IMPORTANT ALLERGENS USING MASS SPECTROMETRY

    EPA Science Inventory

    The US EPA's mission is to protect human health and the environment. Fungal allergens are major potential source of allergy based disease like asthma which is now increasing, according to the Centers for Disease Control and Prevention, at epidemic rates. The Cooperative Agreemen...

  4. Allergenic pollen and pollen allergy in Europe.

    PubMed

    D'Amato, G; Cecchi, L; Bonini, S; Nunes, C; Annesi-Maesano, I; Behrendt, H; Liccardi, G; Popov, T; van Cauwenberge, P

    2007-09-01

    The allergenic content of the atmosphere varies according to climate, geography and vegetation. Data on the presence and prevalence of allergenic airborne pollens, obtained from both aerobiological studies and allergological investigations, make it possible to design pollen calendars with the approximate flowering period of the plants in the sampling area. In this way, even though pollen production and dispersal from year to year depend on the patterns of preseason weather and on the conditions prevailing at the time of anthesis, it is usually possible to forecast the chances of encountering high atmospheric allergenic pollen concentrations in different areas. Aerobiological and allergological studies show that the pollen map of Europe is changing also as a result of cultural factors (for example, importation of plants such as birch and cypress for urban parklands), greater international travel (e.g. colonization by ragweed in France, northern Italy, Austria, Hungary etc.) and climate change. In this regard, the higher frequency of weather extremes, like thunderstorms, and increasing episodes of long range transport of allergenic pollen represent new challenges for researchers. Furthermore, in the last few years, experimental data on pollen and subpollen-particles structure, the pathogenetic role of pollen and the interaction between pollen and air pollutants, gave new insights into the mechanisms of respiratory allergic diseases. PMID:17521313

  5. ASSESSING ALLERGENICITY OF INDOOR AIR FUNGAL CONTAMINANTS

    EPA Science Inventory

    Assessing Allergenicity of Indoor Air Fungal Contaminants
    M D W Ward1, M E Viana2, N Haykal-Coates1, L B Copeland1, S H Gavett1, and MJ K Selgrade1. 1US EPA, ORD, NHEERL, RTP, NC, USA. 2NCSU, CVM, Raleigh, NC, USA.
    Rationale: The indoor environment has increased in impor...

  6. Supercritical carbon dioxide extraction as a predictor of polycyclic aromatic hydrocarbon bioaccumulation and toxicity by earthworms in manufactured-gas plant site soils.

    PubMed

    Kreitinger, Joseph P; Quiñones-Rivera, Antonio; Neuhauser, Edward F; Alexander, Martin; Hawthorne, Steven B

    2007-09-01

    The toxicity and uptake of polycyclic aromatic hydrocarbons (PAHs) by earthworms were measured in soil samples collected from manufactured-gas plant sites having a wide range in PAH concentrations (170-42,000 mg/kg) and soil characteristics. Samples varied from vegetated soils to pure lampblack soot and had total organic carbon contents ranging from 3 to 87%. The biota-soil accumulation factors (BSAFs) observed for individual PAHs in field-collected earthworms (Aporrectodea caliginosa) were up to 50-fold lower than the BSAFs predicted using equilibrium-partitioning theory. Acute toxicity to the earthworm Eisenia fetida was unrelated to total PAH concentration: Mortality was not observed in some soils having high concentrations of total PAHs (>42,000 mg/kg), whereas 100% mortality was observed in other soils having much lower concentrations of total PAHs (1,520 mg/kg). Instead, toxicity appeared to be related to the rapidly released fraction of PAHs determined by mild supercritical CO2 extraction (SFE). The results demonstrate that soils having approximately 16,000 mg rapidly released total PAH/kg organic carbon can be acutely toxic to earthworms and that the concentration of PAHs in soil that is rapidly released by SFE can estimate toxicity to soil invertebrates. PMID:17705650

  7. Effect of chemical modifications on allergenic potency of peanut proteins

    PubMed Central

    Bencharitiwong, Ramon; van der Kleij, Hanneke P.M.; Koppelman, Stef J.

    2015-01-01

    Background: Modification of native peanut extracts could reduce adverse effects of peanut immunotherapy. Objective: We sought to compare native and chemically modified crude peanut extract (CPE) and major peanut allergens Ara h 2 and Ara h 6 in a mediator-release assay based on the rat basophilic leukemia (RBL) cell line transfected with human Fcε receptor. Methods: Native Ara h 2/6 was reduced and alkylated (RA), with or without additional glutaraldehyde treatment (RAGA). CPE was reduced and alkylated. Sera of subjects with peanut allergy (16 males; median age 7 years) were used for overnight RBL-passive sensitization. Cells were stimulated with 0.1 pg/mL to 10 μg/mL of peanut. β-N-acetylhexosaminidase release (NHR) was used as a marker of RBL degranulation, expressed as a percentage of total degranulation caused by Triton X. Results: Median peanut-specific immunoglobulin E was 233 kUA/L. Nineteen subjects were responders, NHR ≥ 10% in the mediator release assay. Responders had reduced NHR by RA and RAGA compared with the native Ara h 2/6. Modification resulted in a later onset of activation by 10- to 100-fold in concentration and a lowering of the maximum release. Modified RA-Ara h 2/6 and RAGA-Ara h 2/6 caused significantly lower maximum mediator release than native Ara h 2/6, at protein concentrations 0.1, 1, and 10 ng/mL (p < 0.001, < 0.001, and < 0.001, respectively, for RA; and < 0.001, 0.026, and 0.041, respectively, for RAGA). RA-CPE caused significantly lower maximum NHR than native CPE, at protein concentration 1 ng/mL (p < 0.001) and 10 ng/mL (p < 0.002). Responders had high rAra h 2 immunoglobulin E (mean, 61.1 kUA/L; p < 0.001) and higher NHR in mediator release assay to native Ara h 2/6 than CPE, which indicates that Ara h 2/6 were the most relevant peanut allergens in these responders. Conclusions: Chemical modification of purified native Ara h 2 and Ara h 6 reduced mediator release in an in vitro assay ∼100-fold, which indicates decreased

  8. The potential of papain and alcalase enzymes and process optimizations to reduce allergenic gliadins in wheat flour.

    PubMed

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

    2016-04-01

    The objectives of this study were to select effective enzymes that catalyze the hydrolysis of allergenic proteins, gliadins, in wheat flour and to optimize the enzymatic treatment conditions. Six proteases were tested. Hydrolyzed samples were tested for residual gliadin concentrations and in vitro allergenicity. The hydrolysis conditions of wheat protein by the effective enzymes were optimized by central composite design. Results showed that alcalase from Bacillus licheniformis, and papain from latex of papaya fruit had greater ability to reduce gliadin content of wheat flour than flavourzyme, pepsin, trypsin or α-chymotrypsin. The sequential-treatment of wheat flour by alcalase-papain was more effective in reducing gliadin content than single enzyme treatment. Under the optimal conditions of sequential enzymatic treatment, gliadin was almost completely removed, resulting in the flour extract showing lowest IgE-binding. Therefore, this could be a promising biotechnology for preparing low allergenic wheat products. PMID:26593625

  9. Technological processes to decrease the allergenicity of peach juice and nectar.

    PubMed

    Brenna, O; Pompei, C; Ortolani, C; Pravettoni, V; Farioli, L; Pastorello, E A

    2000-02-01

    Among vegetable foods peach (Prunus persica) has been recognized as a significant cause of allergy. The protein, which is considered to be the major peach allergen, has been named Pru p 1. Because peaches are consumed both as fresh fruits and after processing to obtain peach juice, nectar, jam, syrupy peach, etc., research was carried out to identify a technological process for production of hypo- or nonallergenic peach-based products. SDS-PAGE and immunoblotting analysis of extracts prepared from four commercial peach nectars showed that the Pru p 1 was not removed, and neither was its allergenic activity decreased by technological treatments carried out for nectar production. Some treatments oriented toward a removal of or, at least, a decrease in the allergenic power were assumed and verified at laboratory scale. A variable considered was heat treatment at 121 degrees C for 10 and 30 min: this treatment was not able to decrease the allergenicity of the Pru p 1 protein. Furthermore, the protein band was still present even after 60-min reaction with two different acidic proteases. The two technological treatments that were found to decrease the major allergen of peach were chemical lye peeling of fruits and ultrafiltration of juice through membranes with suitable cutoff. On the basis of the results obtained from this research, a processing flow sheet was defined to obtain hypoallergenic or probably nonallergenic limpid juices and nectars. These products may represent, besides finished foods, intermediates to obtain various products after addition of further ingredients such as pectins, sugars, and fiber. PMID:10691663

  10. Inactivation of dust mites, dust mite allergen, and mold from carpet.

    PubMed

    Ong, Kee-Hean; Lewis, Roger D; Dixit, Anupma; MacDonald, Maureen; Yang, Mingan; Qian, Zhengmin

    2014-01-01

    Carpet is known to be a reservoir for biological contaminants, such as dust mites, dust mite allergen, and mold, if it is not kept clean. The accumulation of these contaminants in carpet might trigger allergies or asthma symptoms in both children and adults. The purpose of this study is to compare methods for removal of dust mites, dust mite allergens, and mold from carpet. Carpets were artificially worn to simulate 1 to 2 years of wear in a four-person household. The worn carpets were inoculated together with a common indoor mold (Cladosporium species) and house dust mites and incubated for 6 weeks to allow time for dust mite growth on the carpet. The carpets were randomly assigned to one of the four treatment groups. Available treatment regimens for controlling carpet contaminants were evaluated through a literature review and experimentation. Four moderately low-hazard, nondestructive methods were selected as treatments: vacuuming, steam-vapor, Neem oil (a natural tree extract), and benzalkonium chloride (a quaternary ammonium compound). Steam vapor treatment demonstrated the greatest dust mite population reduction (p < 0.05) when compared to other methods. The two physical methods, steam vapor and vacuuming, have no statistically significant efficacy in inactivating dust mite allergens (p = 0.084), but have higher efficacy when compared to the chemical method on dust mite allergens (p = 0.002). There is no statistically significant difference in the efficacy for reducing mold in carpet (p > 0.05) for both physical and chemical methods. The steam-vapor treatment effectively killed dust mites and denatured dust mite allergen in the laboratory environment. PMID:24467247

  11. Novel structure of cockroach allergen Bla g 1 has implications for allergenicity and exposure assessment

    PubMed Central

    Mueller, Geoffrey A.; Pedersen, Lars C.; Lih, Fred B.; Glesner, Jill; Moon, Andrea F.; Chapman, Martin D.; Tomer, Kenneth B.; London, Robert E.; Pomés, Anna

    2013-01-01

    Background Sensitization to cockroach allergens is a major risk factor for asthma. The cockroach allergen Bla g 1 has multiple repeats of ~100 amino acids, but the fold of the protein and the biological function are unknown. Objective To determine the structure of Bla g 1, investigate the implications for allergic disease, and standardize cockroach exposure assays. Methods Natural Bla g 1 and recombinant constructs were compared by ELISA using specific murine IgG and human IgE. The structure of Bla g 1 was determined by X-ray crystallography. Mass spectrometry and NMR were utilized to examine ligand-binding properties of the allergen. Results The structure of a recombinant Bla g 1 construct with comparable IgE and IgG reactivity to the natural allergen was solved by X-ray crystallography. The Bla g 1 repeat forms a novel fold with 6 helices. Two repeats encapsulate a large and nearly spherical hydrophobic cavity, defining the basic structural unit. Lipids in the cavity varied depending on the allergen origin. Palmitic, oleic and stearic acids were associated with nBla g 1 from cockroach frass. One Unit of Bla g 1 was equivalent to 104 ng of allergen. Conclusions Bla g 1 has a novel fold with a capacity to bind various lipids, which suggests a digestive function associated with non-specific transport of lipid molecules in cockroaches. Defining the basic structural unit of Bla g 1 facilitates the standardization of assays in absolute units for the assessment of environmental allergen exposure. PMID:23915714

  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. PMID:26186076

  13. Cloning and expression of Che a 1, the major allergen of Chenopodium album in Escherichia coli.

    PubMed

    Vahedi, Fatemeh; Sankian, Mojtaba; Moghadam, Malihe; Mohaddesfar, Maryam; Ghobadi, Sirous; Varasteh, Abdol Reza

    2011-04-01

    Chenopodium album is a weedy annual plant in the genus Chenopodium. C. album pollen represents a predominant allergen source in Iran. The main C. album pollen allergens have been described as Che a 1, Che a 2, and Che a 3. The aim of this work was to clone the Che a 1 in Escherichia coli to establish a system for overproduction of the recombinant Che a 1 (rChe a 1). In order to clone this allergen, the pollens were subjected to RNA extraction. A full-length fragment encoding Che a 1 was prepared by polymerase chain reaction amplification of the first-strand cDNA synthesized from extracted RNA. Cloning was carried out by inserting the cDNA into the pET21b+ vector, thereafter the construct was transformed into E. coli Top10 cells and expression of the protein was induced by IPTG. The rChe a 1 was purified using histidine tag in recombinant protein by means of Ni-NTA affinity chromatography. IgE immunoblotting, ELISA, and inhibition ELISA were done to evaluate IgE binding of the purified protein. In conclusion, the cDNA for the major allergen of the C. album pollen, Che a 1, was successfully cloned and rChe a 1 was purified. Inhibition assays demonstrated allergic subjects sera reacted with rChe a 1 similar to natural Che a 1 in crude extract of C. album pollen. This study is the first report of using the E. coli as a prokaryotic system for Che a 1 cloning and production of rChe a 1. PMID:20872185

  14. Specific allergen immunotherapy attenuates allergic airway inflammation in a rat model of Alstonia scholaris pollen induced airway allergy.

    PubMed

    Datta, Ankur; Moitra, Saibal; Hazra, Iman; Mondal, Somnath; Das, Prasanta Kumar; Singh, Manoj Kumar; Chaudhuri, Suhnrita; Bhattacharya, Debanjan; Tripathi, Santanu Kumar; Chaudhuri, Swapna

    2016-01-01

    Pollen grains are well established to be an important cause of respiratory allergy. Current pharmacologic therapies for allergic asthma do not cure the disease. Allergen specific immunotherapy is the only treatment method which re-directs the immune system away from allergic response leading to a long lasting effect. The mechanism by which immunotherapy achieves this goal is an area of active research world-wide. The present experimental study was designed to develop an experimental model of allergic lung inflammation based on a relevant human allergen, Alstonia scholaris pollen, and to establish the immunological and cellular features of specific allergen immunotherapy using this same pollen extract. Our results revealed that Alstonia scholaris pollen sensitization and challenge causes eosinophilic airway inflammation with mucin hypersecretion. This is associated with increased total IgE, increased expression of FcɛRI on lung mast cells and increased levels of IL-4, IL-5 & IL-13 as confirmed by ELISA, in-situ immunofluorescence and FACS assay. Allergen specific immunotherapy reduced airway inflammation and also decreased total IgE level, FcɛRI expression, IL-4, IL-5 & IL-13 levels. It was further noted that the reduction of these levels was more by intra-nasal route than by intra-peritoneal route. Thus we present a novel animal model of Alstonia scholaris pollen allergic disease and specific allergen immunotherapy which will pave the way towards the development of better treatment modalities. PMID:26667977

  15. Purification, characterization and immunoreactivity of β'-component, a major allergen from the roe of large yellow croaker (Pseudosciaena crocea).

    PubMed

    Liu, Yan-Yun; Cao, Min-Jie; Zhang, Ming-Li; Hu, Jia-Wei; Zhang, Yong-Xia; Zhang, Ling-Jing; Liu, Guang-Ming

    2014-10-01

    Fish roe, a nutritious food, is favored by consumers, but has also been confirmed to be allergenic in salmonid fish. However, little information is available in other fish species. To determine the allergen in the roe of large yellow croaker (Pseudosciaena crocea), crude extracts were incubated with sera of allergic patients. The major allergen was purified by column chromatography methods, revealing a single band with 16 kDa and was confirmed as β'-component (β'-c) by mass spectrometry. The results of physicochemical characterization showed that β'-c was a glycoprotein and was relatively stable following thermal or acid/alkali treatment. Furthermore, β'-c was easily degraded by pepsin, but was resistant to trypsin and α-chymotrypsin. After treatment with different processing methods, including Maillard reaction (MR), ultraviolet radiation (UVR), ultrasound-heat (UH), and retorting (RT), the IgG-binding activity of β'-c decreased obviously by MR, but decreased slightly by UVR and UH. Cross-immunoreactivity results of the allergens in the roes of different species revealed that β'-c was a specific allergen in teleostean, and the cross-immunoreactivity between the roe of large yellow croaker and other kinds of fish roe was relatively strong. PMID:25046389

  16. Immunoproteomic characterization of Ambrosia artemisiifolia pollen allergens in canine atopic dermatitis.

    PubMed

    Ognjenovic, Jana; Milcic-Matic, Natalija; Smiljanic, Katarina; Vuckovic, Olga; Burazer, Lidija; Popovic, Nikola; Stanic-Vucinic, Dragana; Velickovic, Tanja Cirkovic

    2013-09-01

    Canine atopic dermatitis (CAD) is an immune system disorder that affects 10-15% of the canine population. Short ragweed (Ambrosia artemisiifolia) pollen represents one of the major seasonal sources of allergenic pollen proteins in Europe, particularly in the Pannonian valley of the Balkan region. In Serbia, about 66% of atopic dogs showed a positive intradermal skin test with its pollen extract, which is second to house dust mites. Therefore, characterization of Ambrosia artemisiifolia pollen components, in terms of defining major and minor allergens that induce clinically manifested allergic reaction in dogs, is important for valid diagnosis and efficient therapy. This study has, for the first time, characterized and identified major Ambrosia artemisiifolia allergens in CAD, using an immunoproteomic approach. To assess the prevalence of specific IgE in electrophoretically separated ragweed pollen proteins, individual reactivity of sera from dogs with CAD was analyzed and compared to the reactivity of sera from healthy dogs in the non-reducing conditions, which were found optimal for specific canine IgE detection. A specific IgE band (38 kDa) was recognized as the most dominant allergen in CAD, occurring in 81% of positive dog's sera. 2-D immunoblotting followed by a mass spectrometry peptide fingerprint analyses with pooled canine and human atopic sera, revealed that 38 kDa major Ambrosia atremisiifolia allergens in CAD were all five isoallergens of the Amb a 1 group (antigen E), including the previously named Amb a 2 (antigen K). In contrast to canine sera, human atopic sera also recognized lower mass allergens such as the β fragment of Amb a 1 and profilins (Amb a 8 variants). The most prominent ragweed proteins in CAD, represent, as in humans, variants of all five isoallergens of the Amb a 1 group (pectate lyase): Amb a 1.0101 and its natural variant E1XUL2, Amb a 1.0202, 1.0304, 1.0402 and the natural variant of Amb a 1.0501, E1XUM0, as well as the

  17. 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

  18. First National Survey of Lead and Allergens in Housing: survey design and methods for the allergen and endotoxin components.

    PubMed Central

    Vojta, Patrick J; Friedman, Warren; Marker, David A; Clickner, Robert; Rogers, John W; Viet, Susan M; Muilenberg, Michael L; Thorne, Peter S; Arbes, Samuel J; Zeldin, Darryl C

    2002-01-01

    From July 1998 to August 1999, the U.S. Department of Housing and Urban Development and the National Institute of Environmental Health Sciences conducted the first National Survey of Lead and Allergens in Housing. The purpose of the survey was to assess children's potential household exposure to lead, allergens, and bacterial endotoxins. We surveyed a sample of 831 homes, representing 96 million permanently occupied, noninstitutional housing units that permit resident children. We administered questionnaires to household members, made home observations, and took environmental samples. This article provides general background information on the survey, an overview of the survey design, and a description of the data collection and laboratory methods pertaining to the allergen and endotoxin components. We collected dust samples from a bed, the bedroom floor, a sofa or chair, the living room floor, the kitchen floor, and a basement floor and analyzed them for cockroach allergen Bla g 1, the dust mite allergens Der f 1 and Der p 1, the cat allergen Fel d 1, the dog allergen Can f 1, the rodent allergens Rat n 1 and mouse urinary protein, allergens of the fungus Alternaria alternata, and endotoxin. This article provides the essential context for subsequent reports that will describe the prevalence of allergens and endotoxin in U.S. households, their distribution by various housing characteristics, and their associations with allergic diseases such as asthma and rhinitis. PMID:12003758

  19. Effects of Nasal Corticosteroids on Boosts of Systemic Allergen-Specific IgE Production Induced by Nasal Allergen Exposure

    PubMed Central

    Egger, Cornelia; Lupinek, Christian; Ristl, Robin; Lemell, Patrick; Horak, Friedrich; Zieglmayer, Petra; Spitzauer, Susanne; Valenta, Rudolf; Niederberger, Verena

    2015-01-01

    Background Allergen exposure via the respiratory tract and in particular via the nasal mucosa boosts systemic allergen-specific IgE production. Intranasal corticosteroids (INCS) represent a first line treatment of allergic rhinitis but their effects on this boost of allergen-specific IgE production are unclear. Aim Here we aimed to determine in a double-blind, placebo-controlled study whether therapeutic doses of an INCS preparation, i.e., nasal fluticasone propionate, have effects on boosts of allergen-specific IgE following nasal allergen exposure. Methods Subjects (n = 48) suffering from grass and birch pollen allergy were treated with daily fluticasone propionate or placebo nasal spray for four weeks. After two weeks of treatment, subjects underwent nasal provocation with either birch pollen allergen Bet v 1 or grass pollen allergen Phl p 5. Bet v 1 and Phl p 5-specific IgE, IgG1–4, IgM and IgA levels were measured in serum samples obtained at the time of provocation and one, two, four, six and eight weeks thereafter. Results Nasal allergen provocation induced a median increase to 141.1% of serum IgE levels to allergens used for provocation but not to control allergens 4 weeks after provocation. There were no significant differences regarding the boosts of allergen-specific IgE between INCS- and placebo-treated subjects. Conclusion In conclusion, the application of fluticasone propionate had no significant effects on the boosts of systemic allergen-specific IgE production following nasal allergen exposure. Trial Registration http://clinicaltrials.gov/ NCT00755066 PMID:25705889

  20. Industrial Fungal Enzymes: An Occupational Allergen Perspective

    PubMed Central

    Green, Brett J.; Beezhold, Donald H.

    2011-01-01

    Occupational exposure to high-molecular-weight allergens is a risk factor for the development and pathogenesis of IgE-mediated respiratory disease. In some occupational environments, workers are at an increased risk of exposure to fungal enzymes used in industrial production. Fungal enzymes have been associated with adverse health effects in the work place, in particular in baking occupations. Exposure-response relationships have been demonstrated, and atopic workers directly handling fungal enzymes are at an increased risk for IgE-mediated disease and occupational asthma. The utilization of new and emerging fungal enzymes in industrial production will present new occupational exposures. The production of antibody-based immunoassays is necessary for the assessment of occupational exposure and the development of threshold limit values. Allergen avoidance strategies including personal protective equipment, engineering controls, protein encapsulation, and reduction of airborne enzyme concentrations are required to mitigate occupational exposure to fungal enzymes. PMID:21747869

  1. New Trends in Food Allergens Detection: Toward Biosensing Strategies.

    PubMed

    Alves, Rita C; Barroso, M Fátima; González-García, María Begoña; Oliveira, M Beatriz P P; Delerue-Matos, Cristina

    2016-10-25

    Food allergens are a real threat to sensitized individuals. Although food labeling is crucial to provide information to consumers with food allergies, accidental exposure to allergenic proteins may result from undeclared allergenic substances by means of food adulteration, fraud or uncontrolled cross-contamination. Allergens detection in foodstuffs can be a very hard task, due to their presence usually in trace amounts, together with the natural interference of the matrix. Methods for allergens analysis can be mainly divided in two large groups: the immunological assays and the DNA-based ones. Mass spectrometry has also been used as a confirmatory tool. Recently, biosensors appeared as innovative, sensitive, selective, environmentally friendly, cheaper and fast techniques (especially when automated and/or miniaturized), able to effectively replace the classical methodologies. In this review, we present the advances in the field of food allergens detection toward the biosensing strategies and discuss the challenges and future perspectives of this technology. PMID:25779935

  2. Inhibitory activities against heterologous α-amylases and in vitro allergenic reactivity of Einkorn wheats.

    PubMed

    Sánchez-Monge, R; García-Casado, G; Malpica, J M; Salcedo, G

    1996-10-01

    Salt extracts from seeds of 36 lines of Einkorn wheats were analyzed for their inhibitory activity towards two insect (Tenebrio molitor, Coleoptera, and Ephestia kuehniella, Lepidoptera) and one mammalian (human salivary) α-amylases. Whereas all ten T. monococcum accessions tested were active towards the lepidopteran enzyme, they had no effect on the coleopteran or the mammalian ones. More variability was found among the 21 lines of T. boeticum analyzed, although none of them inhibited human α-amylase. The five accessions of T. urartu showed even greater diversity. Among all Einkorn accessions tested, only two urartu lines affected the three α-amylases. These lines displayed inhibition patterns similar to those of T. aestivum and T. turgidum cultivars. Since several breadwheat α-amylase inhibitors are major allergens associated with baker's asthma, we also studied the in vitro allergenic activity of salt extracts from the Einkorn wheats under study. No significant differences in IgE-binding were found between these accessions and theT. aestivum or T. turgidum cultivars. Furthermore, putative allergens with molecular sizes in the range of 20-60 kDa were detected in these Einkorn wheats. PMID:24162403

  3. 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

  4. Perception and practice regarding allergen labeling: focus on food-related employees

    PubMed Central

    Park, Si-Eun; Kwon, Yong-Seok; Paik, Jin-Kyoung; Kwak, Tong-Kyung

    2016-01-01

    BACKGROUND/OBJECTIVES Most consumers are able to recognize allergenic foods. However, the frequency of checking such foods is reportedly low, resulting in higher prevalence of food-related allergic reactions in Korea compared to other countries. Thus, this study was performed to investigate the overall perception of allergenic food labeling and its practice level in food manufacturing company employees. SUBJECTS/METHODS The survey was administered to food safety employees and food development teams at food companies located in metropolitan areas. A total of 399 (93.8%) valid samples were used in the final analysis. Statistical analyses, including Frequency Analysis, t-test, Anova, PCA (Principal Component Analysis), and Pearson Correlation Analysis using SPSS ver. 21.0, were performed. RESULTS The correct answer rate in the analysis of allergy-related knowledge level ranged from 15.0% to 89.7%. Analysis of differences in allergy-related perception by knowledge level showed significant differences in introduction of a food recall system, strengthening of relevant laws and regulations, content labeling, description of substitutional food, and differentiated package by age. CONCLUSIONS It can be concluded that labeling of allergenic foods should be made easier and more convenient for checking by employees, developers, and consumers, and it is necessary to provide contents through the development of publicity, guidelines, or APP along with labeling. PMID:27478550

  5. 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

  6. Allergen Immunotherapy: Past, Present, and Future

    PubMed Central

    Kosowska, Anna; Smolinska, Sylwia

    2016-01-01

    Allergen-specific immunotherapy (AIT), although in clinical use for more than a century, is still the only causal treatment of allergic diseases. The safety and efficacy of AIT has been demonstrated in a large number of clinical trials. In addition to allergy symptom reduction AIT plays an essential role in preventing new allergies and asthma and shows long-term effects after discontinuation of treatment. Ideally, it is capable of curing allergy. However, AIT is not effective in all allergic individuals and is not equally effective in the treatment of various hypersensitivities to different allergens. For many years, the route of administration and the vaccine compositions have been evolving. Still there is a strong need for research in the field of new AIT modalities to increase its effectiveness and safety. Growing evidence on immunological effects of AIT, especially new T cell subsets involved in antigen/allergen tolerance, provides novel concepts for safer and more effective vaccination. Pharmacoeconomic studies have demonstrated a clear advantage of AIT over pharmacologic therapies. PMID:26922928

  7. Allergen Immunotherapy: Past, Present, and Future.

    PubMed

    Jutel, Marek; Kosowska, Anna; Smolinska, Sylwia

    2016-05-01

    Allergen-specific immunotherapy (AIT), although in clinical use for more than a century, is still the only causal treatment of allergic diseases. The safety and efficacy of AIT has been demonstrated in a large number of clinical trials. In addition to allergy symptom reduction AIT plays an essential role in preventing new allergies and asthma and shows long-term effects after discontinuation of treatment. Ideally, it is capable of curing allergy. However, AIT is not effective in all allergic individuals and is not equally effective in the treatment of various hypersensitivities to different allergens. For many years, the route of administration and the vaccine compositions have been evolving. Still there is a strong need for research in the field of new AIT modalities to increase its effectiveness and safety. Growing evidence on immunological effects of AIT, especially new T cell subsets involved in antigen/allergen tolerance, provides novel concepts for safer and more effective vaccination. Pharmacoeconomic studies have demonstrated a clear advantage of AIT over pharmacologic therapies. PMID:26922928

  8. Purification, crystallization and initial crystallographic characterization of brazil-nut allergen Ber e 2

    SciTech Connect

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

    2007-11-01

    The crystallization of the brazil nut allergen Ber e 2 is reported. Peanut and tree-nut allergies have attracted considerable attention because of their frequency and their lifelong persistence. Brazil-nut (Bertholletia excelsa) allergies have been well documented and the 11S legumin-like seed storage protein Ber e 2 (excelsin) is one of the two known brazil-nut allergens. In this study, Ber e 2 was extracted from brazil-nut kernels and purified to high purity by crystalline precipitation and gel-filtration chromatography. Well diffracting single crystals were obtained using the hanging-drop vapour-diffusion method. A molecular-replacement structural solution has been obtained. Refinement of the structure is currently under way.

  9. Cow’s milk allergy: From allergens to new forms of diagnosis, therapy and prevention☆

    PubMed Central

    Hochwallner, Heidrun; Schulmeister, Ulrike; Swoboda, Ines; Spitzauer, Susanne; Valenta, Rudolf

    2014-01-01

    The first adverse reactions to cow’s milk were already described 2000 years ago. However, it was only 50 years ago that several groups started with the analysis of cow’s milk allergens. Meanwhile the spectrum of allergy eliciting proteins within cow’s milk is identified and several cow’s milk allergens have been characterized regarding their biochemical properties, fold and IgE binding epitopes. The diagnosis of cow’s milk allergy is diverse ranging from fast and cheap in vitro assays to elaborate in vivo assays. Considerable effort was spent to improve the diagnosis from an extract-based into a component resolved concept. There is still no suitable therapy available against cow’s milk allergy except avoidance. Therefore research needs to focus on the development of suitable and safe immunotherapies that do not elicit severe side effect. PMID:23954566

  10. Potential changes in the allergenicity of three forms of peanut after thermal processing.

    PubMed

    Cabanillas, Beatriz; Cuadrado, Carmen; Rodriguez, Julia; Hart, Juana; Burbano, Carmen; Crespo, Jesus F; Novak, Natalija

    2015-09-15

    This study aimed to analyze the influence of thermal processing on the IgE binding properties of three forms of peanut, its effects in the content of individual allergens and IgE cross-linking capacity in effector cells of allergy. Three forms of peanut were selected and subjected to thermal processing. Immunoreactivity was evaluated by means of immunoblot or ELISA inhibition assay. Specific antibodies were used to identify changes in the content of the main allergens in peanut samples. The ability of treated peanut to cross-link IgE was evaluated in a basophil activation assay and Skin Prick Testing (SPT). The results showed that thermal/pressure treatments at specific conditions had the capacity to decrease IgE binding properties of protein extracts from peanut. This effect went along with an altered capacity to activate basophils sensitized with IgE from patients with peanut allergy and the wheal size in SPT. PMID:25863604

  11. [The NTP in allergy research : open questions regarding nasal provocation tests using allergens].

    PubMed

    Förster, U; Sperl, A; Klimek, L

    2013-10-01

    The nasal provocation test (NPT) is a simple procedure with high specificity and sensitivity that is used in the investigation of allergic and nonallergic diseases. Uniform standards are of particular importance in the clinical setting and for the comparability of clinical and basic allergy research. These standards should cover the composition, dosage and pharmacological formulation of provocative substances (e.g. allergen extracts), the necessity of titration, allergen application methods and the evaluation criteria for a positive NPT reaction. Detection of various mediators and cytokines in nasal discharge can be very useful in the late phase reactions. NPT finds specific applications in studies of local IgE secretion in the nasal mucosa, the diagnosis of analgesic intolerance and in assessments of the efficacy of specific immunotherapies. Additional parameters warranting further evaluation include provocation with cold dry air in nasal hyperreactivity patients and nasal nitric oxide formation. Determination of nasal blood flow during NPT provides an additional clinical parameter. PMID:24127046

  12. Cow's milk allergy: from allergens to new forms of diagnosis, therapy and prevention.

    PubMed

    Hochwallner, Heidrun; Schulmeister, Ulrike; Swoboda, Ines; Spitzauer, Susanne; Valenta, Rudolf

    2014-03-01

    The first adverse reactions to cow's milk were already described 2,000 years ago. However, it was only 50 years ago that several groups started with the analysis of cow's milk allergens. Meanwhile the spectrum of allergy eliciting proteins within cow's milk is identified and several cow's milk allergens have been characterized regarding their biochemical properties, fold and IgE binding epitopes. The diagnosis of cow's milk allergy is diverse ranging from fast and cheap in vitro assays to elaborate in vivo assays. Considerable effort was spent to improve the diagnosis from an extract-based into a component resolved concept. There is still no suitable therapy available against cow's milk allergy except avoidance. Therefore research needs to focus on the development of suitable and safe immunotherapies that do not elicit severe side effect. PMID:23954566

  13. Methods for allergen analysis in food: a review.

    PubMed

    Poms, R E; Klein, C L; Anklam, E

    2004-01-01

    Food allergies represent an important health problem in industrialized countries. Undeclared allergens as contaminants in food products pose a major risk for sensitized persons. A proposal to amend the European Food Labelling Directive requires that all ingredients intentionally added to food products will have to be included on the label. Reliable detection and quantification methods for food allergens are necessary to ensure compliance with food labelling and to improve consumer protection. Methods available so far are based on protein or DNA detection. This review presents an up-to-date picture of the characteristics of the major food allergens and collects published methods for the determination of food allergens or the presence of potentially allergenic constituents in food products. A summary of the current availability of commercial allergen detection kits is given. One part of the paper describes various methods that have been generally employed in the detection of allergens in food; their advantages and drawbacks are discussed in brief. The main part of this review, however, focuses on specific food allergens and appropriate methods for their detection in food products. Special emphasis is given to allergenic foods explicitly mentioned in the Amendment to the European Food Labelling Directive that pose a potential risk for allergic individuals, namely celery, cereals containing gluten (including wheat, rye and barley) crustaceans, eggs, fish, peanuts, soybeans, milk and dairy products, mustard, tree-nuts, sesame seeds, and sulphite at concentrations of at least 10 mg kg(-1). Sulphites, however, are not discussed. PMID:14744677

  14. 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. PMID:24383968

  15. Dust mite allergens and asthma: a worldwide problem*

    PubMed Central

    1988-01-01

    After the discovery of house dust mites in 1964 their association with asthma has been reported from many different parts of the world including the developing countries. Two sets of major allergens from mites of the genus Dermatophagoides are now well recognized. The Group I allergens are glycoproteins of relative molecular mass (Mr) 25 000, which show both structural homology and cross-reactivity. The allergen Der p I has been cloned and sequenced confirming the Mr and establishing its nature as a protease. The Group II allergens (Mr 15 000) show even closer homology and cross-reactivity. Specific immunoassays for Group I and Group II allergens, using monospecific antisera and monoclonal antibodies, have been standardized and are suitable for measuring allergen levels in different parts of the world. Measures for reducing the levels of mite allergens in houses include the covering of mattresses, hot washing of bedding, and removal of carpets from bedrooms as well as humidity control, vacuum cleaning, and the use of acaricides in the rest of the house. There is already evidence that these procedures can cause a major improvement in the symptoms of asthma. While provisional standards for both sensitization to mites and also mite allergen exposure can now be recommended, there is an urgent need for controlled studies using protocols demonstrated to reduce mite allergen levels by at least tenfold and for further international collaboration. PMID:3069235

  16. The effect of thermal processing on the behaviour of peanut allergen peptide targets used in multiple reaction monitoring mass spectrometry experiments.

    PubMed

    Sayers, R L; Johnson, P E; Marsh, J T; Barran, P; Brown, H; Mills, E N C

    2016-06-20

    Mass spectrometry-based methods offer an alternative means of determining allergens in foods. Whilst targeted methods are likely to offer the most robust approach for detection and quantification, little is known about how food processing may affect the behaviour of peptide targets. A systematic study has been undertaken to investigate the effects of thermal processing (boiling, roasting, frying) on the behaviour of a suite of peanut peptide targets representing the major clinically-relevant allergens. Initially the effect of thermal processing on protein extractability was investigated and a mass spectrometry-compatible buffer identified comprising 50 mM Tris-HCl, pH 8.8 containing 50 mM dithiothreitol and 0.04% (w/v) acid labile detergent which was able to extract 45-100% of protein from raw, boiled, roasted and fried peanuts using sonication at 60 °C. Eight peptide targets were identified including two peptides from each cupin allergen, Ara h1 and Ara h3 and four peptides from the prolamin superfamily allergens Ara h2, 6 and 7. AQUA peptide standards were synthesised and used to undertake multiple-reaction monitoring experiments, giving assay sensitivities of 0.1-30 amoles of peptide on-column (3 : 1 signal : noise), calculated limits of quantification between 96-1343 amoles of peptide on-column and a linear dynamic range of 4-5 orders of magnitude. Absolute quantification of individual peanut allergens in thermally processed samples showed that peptide targets in the cupin allergens were more prone to processing-induced effects than those from Ara h2, 6 and 7. Targets flanked by arginine residues showed greater thermostability. Identification of processing-stable targets, coupled with more efficient extraction procedures and a wide dynamic range, shows that targeted mass spectrometry methods have great potential as an additional method for quantifying peanut allergens in complex food matrices. PMID:27113917

  17. Simultaneous allergen inactivation and detoxification of castor bean cake by treatment with calcium compounds

    PubMed Central

    Fernandes, K.V.; Deus-de-Oliveira, N.; Godoy, M.G.; Guimarães, Z.A.S.; Nascimento, V.V.; de Melo, E.J.T.; Freire, D.M.G.; Dansa-Petretski, M.; Machado, O.L.T.

    2012-01-01

    Ricinus communis L. is of great economic importance due to the oil extracted from its seeds. Castor oil has been used for pharmaceutical and industrial applications, as a lubricant or coating agent, as a component of plastic products, as a fungicide or in the synthesis of biodiesel fuels. After oil extraction, a castor cake with a large amount of protein is obtained. However, this by-product cannot be used as animal feed due to the presence of toxic (ricin) and allergenic (2S albumin) proteins. Here, we propose two processes for detoxification and allergen inactivation of the castor cake. In addition, we establish a biological test to detect ricin and validate these detoxification processes. In this test, Vero cells were treated with ricin, and cell death was assessed by cell counting and measurement of lactate dehydrogenase activity. The limit of detection of the Vero cell assay was 10 ng/mL using a concentration of 1.6 × 105 cells/well. Solid-state fermentation (SSF) and treatment with calcium compounds were used as cake detoxification processes. For SSF, Aspergillus niger was grown using a castor cake as a substrate, and this cake was analyzed after 24, 48, 72, and 96 h of SSF. Ricin was eliminated after 24 h of SSF treatment. The cake was treated with 4 or 8% Ca(OH)2 or CaO, and both the toxicity and the allergenic properties were entirely abolished. A by-product free of toxicity and allergens was obtained. PMID:22911344

  18. Immunological cross-reactivity between four distant parvalbumins-Impact on allergen detection and diagnostics.

    PubMed

    Sharp, Michael F; Stephen, Juan N; Kraft, Lukas; Weiss, Thomas; Kamath, Sandip D; Lopata, Andreas L

    2015-02-01

    Fish are the largest and most diverse group of vertebrates. Fish are also a part of the eight food groups that cause the majority of IgE mediated food reactions. Detection tools for fish allergens are however limited due to the great diversity of fish species, despite fish allergy and its major allergen parvalbumin being well documented. The most commonly studied fish are frequently consumed in North America and Europe. However, much less is known about fish allergens in the Australasian region although fish is widely consumed in this region. A comprehensive phylogenetic analysis was performed of known parvalbumin amino acid sequences to determine possible candidate antigens for new cross-reactive antibodies to be used to detect most fish parvalbumins. Polyclonal rabbit antibodies were raised against parvalbumins from frequently consumed barramundi (Lates calcarifer), basa (Pangasius bocourti), pilchard (Sardinops sagax) and Atlantic salmon (Salmo salar). These were evaluated for cross-reactivity against a panel of 45 fish extracts (raw, heated and canned fish). Anti-barramundi parvalbumin proved to be the most cross-reactive antibody, detecting 87.5% of the 40 species analyzed, followed by anti-pilchard and anti-basa antibody. In contrast the anti-salmon antibody was very specific and only reacted to salmonidae and a few other fish. All analyzed fish species, except mahi mahi, swordfish, yellowfin tuna and all 5 canned fish had parvalbumin detected in raw extracts. However antibody reactivity to many fish was heat liable or susceptible to denaturation, demonstrating that some parvalbumins have most likely conformational epitopes, which lose antibody reactivity after heat treatment. We have demonstrated the generation of highly cross-reactive anti-parvalbumin antibodies that could be used for the detection of allergenic fish parvalbumin in contaminated food products. This cross-reactivity study thus shows processing of fish, especially canning, can have on impact

  19. Simultaneous allergen inactivation and detoxification of castor bean cake by treatment with calcium compounds.

    PubMed

    Fernandes, K V; Deus-de-Oliveira, N; Godoy, M G; Guimarães, Z A S; Nascimento, V V; Melo, E J T de; Freire, D M G; Dansa-Petretski, M; Machado, O L T

    2012-11-01

    Ricinus communis L. is of great economic importance due to the oil extracted from its seeds. Castor oil has been used for pharmaceutical and industrial applications, as a lubricant or coating agent, as a component of plastic products, as a fungicide or in the synthesis of biodiesel fuels. After oil extraction, a castor cake with a large amount of protein is obtained. However, this by-product cannot be used as animal feed due to the presence of toxic (ricin) and allergenic (2S albumin) proteins. Here, we propose two processes for detoxification and allergen inactivation of the castor cake. In addition, we establish a biological test to detect ricin and validate these detoxification processes. In this test, Vero cells were treated with ricin, and cell death was assessed by cell counting and measurement of lactate dehydrogenase activity. The limit of detection of the Vero cell assay was 10 ng/mL using a concentration of 1.6 x 10(5) cells/well. Solid-state fermentation (SSF) and treatment with calcium compounds were used as cake detoxification processes. For SSF, Aspergillus niger was grown using a castor cake as a substrate, and this cake was analyzed after 24, 48, 72, and 96 h of SSF. Ricin was eliminated after 24 h of SSF treatment. The cake was treated with 4 or 8% Ca(OH)2 or CaO, and both the toxicity and the allergenic properties were entirely abolished. A by-product free of toxicity and allergens was obtained. PMID:22911344

  20. 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

  1. Allergenic components in modified and unmodified rosin. Chemical characterization and studies of allergenic activity.

    PubMed

    Gäfvert, E

    1994-01-01

    Gäfvert, E. 1994. Allergenic components in modified and unmodified rosin. Chemical characterization and studies of allergenic activity. Acta Dermato-Venereologica. Suppl. 184. 36pp. Uppsala. Unmodified rosin (colophony) is a well-known cause of contact allergy (delayed type hypersensitivity). Rosin is obtained from coniferous trees and consists mainly of diterpenoid resin acids. Most rosin used in technical products is chemically modified. In the common modification of rosin with maleic anhydride, the major product formed is maleopimaric acid (MPA). MPA was identified in experimental sensitization studies as a potent contact allergen. MPA is also formed when rosin is modified with fumaric acid at high temperature and with prolonged heating. The amounts of MPA in technical quality rosins modified with maleic anhydride or fumaric acid might be enough to sensitize individuals handling these rosins. The major product of the modification of rosin with fumaric acid, fumaropimaric acid (FPA), did not elicit any reactions in the animals tested. In another common rosin modification, glycerol esterification, the major product formed was identified as glyceryl triabietate (GTA). In an experimental sensitization study none of the animals reacted to GTA. However, a minor product formed, glyceryl 1-monoabietate (GMA) showed sensitizing capacity. The presence of new contact allergens due to the modification, together with remaining unmodified material, contributes to the risk of developing allergy from contact with these types of rosin. A new main contact allergen in unmodified rosin was identified; 13,14(beta)-epoxyabietic acid. The allergenicity of this epoxide was comparable to that of an earlier identified rosin allergen, 15-hydroperoxyabietic acid (15-HPA). The allergens were detected as their methyl esters. Experimental sensitization and cross-reactivity of oxidation products of resin acids were studied. A pattern of cross-reactivity was observed which indicates that the

  2. 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

  3. Immunoproteomic tools are used to identify masked allergens: Ole e 12, an allergenic isoflavone reductase from olive (Olea europaea) pollen.

    PubMed

    Castro, Lourdes; Crespo, Jesús F; Rodríguez, Julia; Rodríguez, Rosalía; Villalba, Mayte

    2015-12-01

    Proteins performing important biochemical activities in the olive tree (Olea europaea) pollen have been identified as allergens. One novel 37-kDa protein seems to be associated to the IgE-binding profile of a group of patients suffering allergy to peach and olive pollen. Three previously described olive pollen allergens exhibit very similar molecular mass. Our objective was to identify this allergen by using immunoproteomic approaches. After 2D-electrophoresis and mass spectrometry, peptide sequences from several IgE-binding spots, allowed identifying this new allergen, as well as cloning and DNA sequencing of the corresponding gene. The allergen, named Ole e 12, is a polymorphic isoflavone reductase-like protein of 308 amino acids showing 80% and 74% identity with birch and pear allergens, Bet v 6 and Pyr c 5, respectively. A prevalence of 33% in the selected population is in contrast to 4%-10% in groups of subjects suffering from pollinosis. Recombinant allergen was produced in Escherichia coli, and deeply characterised. Immunoblotting and ELISA detection as well as inhibition experiments were performed with polyclonal antisera and allergic patients' sera. The recombinant allergen retains the IgE reactivity of its natural counterpart. Close structural and immunological relationships between members of this protein family were supported by their IgG recognition in vegetable species. In summary, Ole e 12 is a minor olive pollen allergen, which gains relevance in patients allergic to peach with olive pollinosis. Proteomic approaches used to analyse this allergen provide useful tools to identify hidden allergens, relevant for several allergic populations and thus complete allergenic panels. PMID:26391288

  4. Amino acid composition, antinutrients and allergens in the peanut protein fraction obtained by an aqueous enzymatic process.

    PubMed

    Latif, S; Pfannstiel, J; Makkar, H P S; Becker, K

    2013-01-01

    Enzyme-assisted aqueous extraction (EAE) of peanut kernel was used to extract oil and protein. The aqueous fraction (AF) obtained by EAE had a better essential amino acid profile than the residues obtained by solvent extraction (Rs) and cold pressing (Rc). No major difference in the trypsin inhibitor activity among AF, Rs and Rc was observed; however, the trypsin inhibitor activity was drastically reduced in the residue obtained after EAE. AF was subjected to MALDI-TOF/MS, revealing it to be rich in peptides (107) with molecular masses from m/z 700 to 2369Da. AF had an extremely low phytate content and was rich in peptides, which could be used as a food supplement. ESI-MS/MS data were used for the identification of major peanut allergens, viz., Ara h1, h3, h6-8. Their allergenic potential needs to be established. PMID:23017415

  5. Allergenic Properties of Enzymatically Hydrolyzed Peanut Flour Extracts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Peanut flour is a high protein, low oil, powdered material prepared from roasted 21 peanut seed. In addition to being a well-established food ingredient, peanut flour is also the 22 active ingredient in peanut oral immunotherapy trials. Enzymatic hydrolysis was evaluated as a 23 processing strategy ...

  6. Biological activity of recombinant Der p 2, Der p 5 and Der p 7 allergens of the house-dust mite Dermatophagoides pteronyssinus.

    PubMed

    Lynch, N R; Thomas, W R; Garcia, N M; Di Prisco, M C; Puccio, F A; L'opez, R I; Hazell, L A; Shen, H D; Lin, K L; Chua, K Y

    1997-09-01

    Der p 2, Der p 5 and Der p 7 are three allergens of the house-dust mite Dermatophagoides pteronyssinus that have been cloned and expressed in Escherichia coli as fusion proteins with glutathione-S-transferase (GST). We showed that these recombinant allergens produced immediate hypersensitivity skin-test reactions in 70, 60 and 52% respectively of a group of mite-sensitive allergic patients who were strongly positive to whole mite extract (WME). Comparable positivities were found for serum levels of specific IgE antibody against these allergens, as measured by the radioallergosorbant test (RAST). Overall, for the group of allergic patients that we evaluated, the serum IgE antibody concentrations against Der p 2, 5 and 7 were calculated to represent about one third, one quarter and one fifth respectively of the levels measured against the WME. However, for some patients the activity determined against the separate allergens was far higher than that detected against the WME, thus indicating that the concentration of these can be limiting in the WME. We found no significant correlations between the RAST levels against Derp 2 and either Derp 5 or 7, and RAST-inhibition tests indicated a lack of cross-reactivity between Der p 2 and the other two allergens. In contrast, the RAST results revealed the existence of a significant immunological relationship between Der p 5 and 7. Although a certain degree of reactivity against the GST fusion partner was found in the allergic patients studied, this was not a significant influence in determining the positivity against the recombinant allergens. These results confirm the in vivo biological activity of recombinant Der p 2, 5 and 7, and indicate that whilst Der p 2 is undoubtedly a major mite allergen, both Der p 5 and 7 make important contributions toward the overall allergenic activity of house-dust mites. PMID:9303332

  7. Early Exposure to Respiratory Allergens by Placental Transfer and Breastfeeding

    PubMed Central

    Macchiaverni, Patricia; Ynoue, Leandro H.; Arslanian, Christina; Verhasselt, Valérie; Condino-Neto, Antonio

    2015-01-01

    The relationship between allergen exposure and the onset of or protection from allergic diseases remains unclear. Many factors could be related to immunological responses, such as the age when the exposure occurs, type of allergen, timing, dose, and allergen route. In this study, we investigated whether exposure to respiratory allergens could occur in pregnancy or early life. In particular, we assessed whether Der p 1 and Blo t 5, as well as specific antibodies against these allergens, could be detected in 90 paired cord blood and colostrum samples. Der p 1 was detected in 58.6% of colostrum and 29% of cord blood samples, whereas Blot 5 was positive in 41.3% and 9.6% of the samples, respectively. Similar to specific IgA, which could be detected in all samples for both mites, specific IgG was found in a high number of colostrum samples, 93.5% and 94.8% for Dp and Bt, respectively. Although allergens were not detected in all cord blood samples, a high percentage of them (≥95%) were positive for specific IgM to both mites in cord blood samples, suggesting that neonates can be exposed and sensitized to airborne allergens during pregnancy. Many studies have attempted to correlate allergen exposure or its prevention in early infancy with the onset of or protection from allergic diseases. However, conflicting and inconsistent data do not show a clear correlation with or suggest a way to prevent allergen sensitization. Nevertheless, these unconvincing results could be better understood if the relationship with many aspects of allergen exposure after pregnancy could be clarified. Thus, it is necessary to address basic issues related to allergen exposure, including the development of reproducible, standardized and reliable methods, and to determine how and where the exposure occurs. PMID:26398234

  8. Manufacturing technologies

    NASA Astrophysics Data System (ADS)

    The Manufacturing Technologies Center is at the core of Sandia National Laboratories' advanced manufacturing effort which spans the entire product realization process. The center's capabilities in product and process development are summarized in the following disciplines: (1) mechanical - rapid prototyping, manufacturing engineering, machining and computer-aided manufacturing, measurement and calibration, and mechanical and electronic manufacturing liaison; (2) electronics - advanced packaging for microelectronics, printed circuits, and electronic fabrication; and (3) materials - ceramics, glass, thin films, vacuum technology, brazing, polymers, adhesives, composite materials, and process analysis.

  9. cDNA cloning and expression of Blo t 11, the Blomia tropicalis allergen homologous to paramyosin.

    PubMed

    Ramos, J D; Cheong, N; Lee, B W; Chua, K Y; Nge, C; Wah, L B; Yan, C K

    2001-12-01

    Blomia tropicalis is an important mite species in many parts of the world and the most predominant mite species in tropical countries. The prevalence of sensitization to this species has probably been underestimated because commercial extracts are largely unavailable. Identification and characterization of B. tropicalis allergens is an important step toward understanding the role of this species in allergic sensitization and could provide appropriate reagents for diagnostic and therapeutic procedures. This paper describes the isolation, sequence analysis, expression and allergenicity of a cDNA gene coding for a B. tropicalis allergen with homology to paramyosin, a high-molecular-weight allergen previously identified in Dermatophagoides farinae. The full-length Blo t 11 cDNA gene was isolated by cDNA library screening, 5'-rapid amplification of cDNA ends and long-distance PCR. Sequence analysis was performed with a combination of CLUSTAL W, CGC and BLAST program packages. The cDNA gene was expressed as a GST fusion protein in Escherichia coli and purified by affinity chromatography using the glutathione Sepharose column. Allergenicity of the rBlo t 11 was tested by human IgE dot blot immunoassay. Blo t 11 is a 3,111-bp cDNA gene with a 2,625-bp open reading frame coding for an 875-amino acid protein, exhibiting significant homology with different invertebrate paramyosins. The human IgE dot blot immunoassay showed that the rBlo t 11 reacted positively to 52% (33/63) of sera from asthmatic patients. Blo t 11 is the homolog of Der f 11 exhibiting potentially important allergenic activity. PMID:11815735

  10. Induction of Allergic Responses to Peanut Allergen in Sheep

    PubMed Central

    Van Gramberg, Jenna L.; de Veer, Michael J.; O'Hehir, Robyn E.; Meeusen, Els N. T.; Bischof, Robert J.

    2012-01-01

    Peanut allergy is the leading cause of deaths due to food-induced anaphylaxis but despite continued research, there are currently no specific treatments available. Challenge testing is limited in patients due to the high risk of adverse reactions, emphasising the need for an appropriate animal model. In the present study we examine the induction of allergic responses in a sheep model for peanut allergy. Sheep were sensitised with peanut (PN) extract and in separate injections with ovalbumin (OVA) or house dust mite (HDM) extract. Serum PN-specific IgE responses were detected in 40–50% of immunised sheep, while only 10% (1 of 10 sheep) showed detectable OVA-specific IgE. All PN-allergic sheep tested showed an Ara h 1-specific IgE response, while four out of five allergic sheep showed an Ara h 2-specific IgE response. Animals with high serum IgE levels to HDM were also PN IgE-positive. Of the PN-sensitised animals with high PN-specific IgE, 80% also showed an immediate hypersensitivity reaction following an intradermal PN injection. This new large animal model of peanut allergy may provide a useful tool for future investigations of allergen-associated immune mechanisms and specific immunotherapy. PMID:23284686

  11. Allergen immunotherapy with nanoparticles containing lipopolysaccharide from Brucella ovis.

    PubMed

    Gómez, Sara; Gamazo, Carlos; San Roman, Beatriz; Ferrer, Marta; Sanz, Maria Luisa; Espuelas, Socorro; Irache, Juan M

    2008-11-01

    The adjuvant and protective capacity against anaphylactic shock of the association between rough lipopolysaccharide of Brucella ovis (LPS) coencapsulated with ovalbumin (OVA), as a model allergen, in Gantrez AN nanoparticles was investigated. Several strategies were performed in order to study the adjuvant effect of the LPS either encapsulated or coating the nanoparticles. OVA, as well as LPS, was incorporated either during the manufacturing process (OVA-encapsulated or LPS-encapsulated nanoparticles, respectively) or after the preparation (OVA-coated or LPS-coated nanoparticles, respectively). After the administration of 10 microg of OVA incorporated in the different formulations, all the nanoparticles, with or without LPS, were capable of amplifying the immune response (IgG(1) and IgG(2a)). However, in a model of sensitized mice to OVA, the formulation with OVA and LPS-entrapped inside the nanoparticles administered intradermally in three doses of 3 microg of OVA each was the only treatment that totally protected the mice from death after a challenge with an intraperitoneal injection of OVA. In contrast, the control group administered with OVA adsorbed onto a commercial alhydrogel adjuvant showed 80% mortality. These results are highly suggestive for the valuable use of Gantrez nanoparticles combined with rough LPS of B. ovis in immunotherapy. PMID:18582571

  12. Time-Based Measurement of Personal Mite Allergen Bioaerosol Exposure over 24 Hour Periods.

    PubMed

    Tovey, Euan R; Liu-Brennan, Damien; Garden, Frances L; Oliver, Brian G; Perzanowski, Matthew S; Marks, Guy B

    2016-01-01

    Allergic diseases such as asthma and rhinitis are common in many countries. Globally the most common allergen associated with symptoms is produced by house dust mites. Although the bed has often been cited as the main site of exposure to mite allergens, surprisingly this has not yet been directly established by measurement due to a lack of suitable methods. Here we report on the development of novel methods to determine the pattern of personal exposure to mite allergen bioaerosols over 24-hour periods and applied this in a small field study using 10 normal adults. Air was sampled using a miniature time-based air-sampler of in-house design located close to the breathing zone of the participants, co-located with a miniature time-lapse camera. Airborne particles, drawn into the sampler at 2L/min via a narrow slot, were impacted onto the peripheral surface of a disk mounted on the hour-hand of either a 12 or 24 hour clock motor. The impaction surface was either an electret cloth, or an adhesive film; both novel for these purposes. Following a review of the time-lapse images, disks were post-hoc cut into subsamples corresponding to eight predetermined categories of indoor or outdoor location, extracted and analysed for mite allergen Der p 1 by an amplified ELISA. Allergen was detected in 57.2% of the total of 353 subsamples collected during 20 days of sampling. Exposure patterns varied over time. Higher concentrations of airborne mite allergen were typically measured in samples collected from domestic locations in the day and evening. Indoor domestic Der p 1 exposures accounted for 59.5% of total exposure, whereas total in-bed-asleep exposure, which varied 80 fold between individuals, accounted overall for 9.85% of total exposure, suggesting beds are not often the main site of exposure. This study establishes the feasibility of novel methods for determining the time-geography of personal exposure to many bioaerosols and identifies new areas for future technical

  13. Time-Based Measurement of Personal Mite Allergen Bioaerosol Exposure over 24 Hour Periods

    PubMed Central

    Tovey, Euan R.; Liu-Brennan, Damien; Garden, Frances L.; Oliver, Brian G.; Perzanowski, Matthew S.; Marks, Guy B.

    2016-01-01

    Allergic diseases such as asthma and rhinitis are common in many countries. Globally the most common allergen associated with symptoms is produced by house dust mites. Although the bed has often been cited as the main site of exposure to mite allergens, surprisingly this has not yet been directly established by measurement due to a lack of suitable methods. Here we report on the development of novel methods to determine the pattern of personal exposure to mite allergen bioaerosols over 24-hour periods and applied this in a small field study using 10 normal adults. Air was sampled using a miniature time-based air-sampler of in-house design located close to the breathing zone of the participants, co-located with a miniature time-lapse camera. Airborne particles, drawn into the sampler at 2L/min via a narrow slot, were impacted onto the peripheral surface of a disk mounted on the hour-hand of either a 12 or 24 hour clock motor. The impaction surface was either an electret cloth, or an adhesive film; both novel for these purposes. Following a review of the time-lapse images, disks were post-hoc cut into subsamples corresponding to eight predetermined categories of indoor or outdoor location, extracted and analysed for mite allergen Der p 1 by an amplified ELISA. Allergen was detected in 57.2% of the total of 353 subsamples collected during 20 days of sampling. Exposure patterns varied over time. Higher concentrations of airborne mite allergen were typically measured in samples collected from domestic locations in the day and evening. Indoor domestic Der p 1 exposures accounted for 59.5% of total exposure, whereas total in-bed-asleep exposure, which varied 80 fold between individuals, accounted overall for 9.85% of total exposure, suggesting beds are not often the main site of exposure. This study establishes the feasibility of novel methods for determining the time-geography of personal exposure to many bioaerosols and identifies new areas for future technical

  14. Proteomic and immunochemical characterization of glutathione transferase as a new allergen of the nematode Ascaris lumbricoides.

    PubMed

    Acevedo, Nathalie; Mohr, Jens; Zakzuk, Josefina; Samonig, Martin; Briza, Peter; Erler, Anja; Pomés, Anna; Huber, Christian G; Ferreira, Fatima; Caraballo, Luis

    2013-01-01

    Helminth infections and allergy have evolutionary and clinical links. Infection with the nematode Ascaris lumbricoides induces IgE against several molecules including invertebrate pan-allergens. These antibodies influence the pathogenesis and diagnosis of allergy; therefore, studying parasitic and non-parasitic allergens is essential to understand both helminth immunity and allergy. Glutathione transferases (GSTs) from cockroach and house dust mites are clinically relevant allergens and comparative studies between them and the GST from A. lumbricoides (GSTA) are necessary to evaluate their allergenicity. We sought to analyze the allergenic potential of GSTA in connection with the IgE response to non-parasitic GSTs. IgE to purified GSTs from Ascaris (nGSTA and rGSTA), house dust mites (rDer p 8, nBlo t 8 and rBlo t 8), and cockroach (rBla g 5) was measured by ELISA in subjects from Cartagena, Colombia. Also, multidimensional proteomic approaches were used to study the extract of A. lumbricoides and investigate the existence of GST isoforms. We found that among asthmatics, the strength of IgE levels to GSTA was significantly higher than to mite and cockroach GSTs, and there was a strong positive correlation between IgE levels to these molecules. Specific IgE to GSTA was found in 13.2% of controls and 19.5% of asthmatics. In addition nGSTA induced wheal and flare in skin of sensitized asthmatics indicating that it might be of clinical relevance for some patients. Frequency and IgE levels to GSTA were higher in childhood and declined with age. At least six GST isoforms in A. lumbricoides bind human IgE. Four isoforms were the most abundant and several amino acid substitutions were found, mainly on the N-terminal domain. In conclusion, a new allergenic component of Ascaris has been discovered; it could have clinical impact in allergic patients and influence the diagnosis of mite and cockroach allergy in tropical environments. PMID:24223794

  15. Proteomic and Immunochemical Characterization of Glutathione Transferase as a New Allergen of the Nematode Ascaris lumbricoides

    PubMed Central

    Acevedo, Nathalie; Mohr, Jens; Zakzuk, Josefina; Samonig, Martin; Briza, Peter; Erler, Anja; Pomés, Anna; Huber, Christian G.; Ferreira, Fatima; Caraballo, Luis

    2013-01-01

    Helminth infections and allergy have evolutionary and clinical links. Infection with the nematode Ascaris lumbricoides induces IgE against several molecules including invertebrate pan-allergens. These antibodies influence the pathogenesis and diagnosis of allergy; therefore, studying parasitic and non-parasitic allergens is essential to understand both helminth immunity and allergy. Glutathione transferases (GSTs) from cockroach and house dust mites are clinically relevant allergens and comparative studies between them and the GST from A. lumbricoides (GSTA) are necessary to evaluate their allergenicity. We sought to analyze the allergenic potential of GSTA in connection with the IgE response to non-parasitic GSTs. IgE to purified GSTs from Ascaris (nGSTA and rGSTA), house dust mites (rDer p 8, nBlo t 8 and rBlo t 8), and cockroach (rBla g 5) was measured by ELISA in subjects from Cartagena, Colombia. Also, multidimensional proteomic approaches were used to study the extract of A. lumbricoides and investigate the existence of GST isoforms. We found that among asthmatics, the strength of IgE levels to GSTA was significantly higher than to mite and cockroach GSTs, and there was a strong positive correlation between IgE levels to these molecules. Specific IgE to GSTA was found in 13.2% of controls and 19.5% of asthmatics. In addition nGSTA induced wheal and flare in skin of sensitized asthmatics indicating that it might be of clinical relevance for some patients. Frequency and IgE levels to GSTA were higher in childhood and declined with age. At least six GST isoforms in A. lumbricoides bind human IgE. Four isoforms were the most abundant and several amino acid substitutions were found, mainly on the N-terminal domain. In conclusion, a new allergenic component of Ascaris has been discovered; it could have clinical impact in allergic patients and influence the diagnosis of mite and cockroach allergy in tropical environments. PMID:24223794

  16. Two Allergen Model Reveals Complex Relationship Between IgE Cross-Linking and Degranulation

    PubMed Central

    Handlogten, Michael W.; Deak, Peter E.; Bilgicer, Basar

    2014-01-01

    Summary Allergy is an immune response to complex mixtures of multiple allergens yet current models use a single synthetic allergen. Multiple allergens were modeled using two well-defined tetravalent allergens each specific for a distinct IgE thus enabling a systematic approach to evaluate the effect of each allergen and percent of allergen specific IgE on mast cell degranulation. We found the overall degranulation response caused by two allergens is additive for low allergen concentrations or low percent specific IgE, does not change for moderate allergen concentrations with moderate to high percent specific IgE, and is reduced for high allergen concentrations with moderate to high percent specific IgE. These results provide further evidence that supra-optimal IgE cross-linking decreases the degranulation response and establishes the two allergen model as a relevant experimental system to elucidate mast cell degranulation mechanisms. PMID:25308278

  17. CHARACTERIZATION OF MAJOR ALLERGEN PROTEINS OF SOYBEAN BY PROTEOMIC TOOLS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We investigated profiles of three major allergen proteins (Gly m Bd 60K, Gly m Bd 30K and Gly m Bd 28K) in sixteen different soybean genotypes that included four groups; wild, cultivated, modern (elite), and ancestor. Four genotypes were studied within each group. All allergen proteins were well s...

  18. 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

  19. *Biomarkers of acute respiratory allergen exposure: Screening for sensitization potential

    EPA Science Inventory

    Effective hazard screening will require the development of high-throughput or in vitro assays for the identification of potential sensitizers. The goal of this preliminary study was to identify potential biomarkers that differentiate the response to allergens vs non-allergens fol...

  20. 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...

  1. Nasal mucosal blood flow after intranasal allergen challenge

    SciTech Connect

    Holmberg, K.; Bake, B.; Pipkorn, U.

    1988-03-01

    The nasal mucosal blood flow in patients with allergic rhinitis was determined at nasal allergen challenges with the /sup 133/Xenon washout method. Determinations were made in 12 subjects before and 15 minutes after challenge with diluent and increasing doses of allergen. The time course was followed in eight subjects by means of repeated measurements during 1 hour after a single allergen dose. Finally, the blood flow was measured after unilateral allergen challenge in the contralateral nasal cavity. A dose-dependent decrease in blood flow was found after nasal challenge with increasing doses of allergens, whereas challenge with diluent alone did not induce any changes. The highest allergen dose, which also induced pronounced nasal symptoms, resulted in a decrease in blood flow of 25% (p less than 0.001). The time-course study demonstrated a maximum decrease in blood flow 10 to 20 minutes after challenge and then a gradual return to baseline. Unilateral allergen challenge resulted in a decrease in blood flow in the contralateral, unchallenged nasal cavity, suggesting that part of the allergen-induced changes in blood flow were reflex mediated.

  2. Dust Allergens within Rural Northern Rocky Mountain Residences

    PubMed Central

    Weiler, Emily; Semmens, Erin; Noonan, Curtis; Cady, Carol; Ward, Tony

    2015-01-01

    To date, few studies have characterized allergens within residences located in rural areas of the northern Rocky Mountain region. In this study, we collected dust samples from 57 homes located throughout western Montana and northern Idaho. Dust samples were collected and later analyzed for dust mite allergens Der f 1 and Der p 1, Group 2 mite allergens (Der p 2 and Der f 2), domestic feline (Fel d 1), and canine (Can f 1). Indoor temperature and humidity levels were also measured during the sampling program, as were basic characteristics of each home. Dog (96%) and cat (82%) allergens were the most prevalent allergens found in these homes (even when a feline or canine did not reside in the home). Results also revealed the presence of dust mites. Seven percent (7%) of homes tested positive for Der p 1, 19% of homes were positive for Der f 1, and 5% of homes were positive for the Group 2 mite allergens. Indoor relative humidity averaged 27.0 ± 7.6% within the homes. Overall, humidity was not significantly associated with dust mite presence, nor was any of the other measured home characteristics. This study provides a descriptive assessment of indoor allergen presence (including dust mites) in rural areas of the northern Rocky Mountains, and provides new information to assist regional patients with reducing allergen exposure using in-home intervention strategies. PMID:25859563

  3. 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

  4. Controlling allergens in animal rooms by using curtains.

    PubMed

    Krohn, Thomas C; Itter, Gabi; Fosse, Richard; Hansen, Axel K

    2006-05-01

    The reduction and control of allergens in the animal facility is important for staff working with laboratory animals. This study was designed to evaluate the efficiency of perforated Makrolon curtains in front of racks as a method to reduce the amount of allergen in the animal room. The experimental situation we studied provides some information regarding allergen disposition in animal rooms but is clearly artificial and does not reflect a typical, 'real-world' environment in terms of preventing exposure of workers to allergens. Plastic curtains with holes were placed in front of racks, and a corridor between the racks and a curtain was present. The room was ventilated with air, which was blown into the room through the middle of the corridor, flowing downstream and passing through the holes in the curtain. This set-up resulted in air flow from the corridor through the curtain. Air samples were collected from sites in the corridor and behind the curtain. The samples were analyzed for the allergen Mus m1, and the amount of allergen was calculated. The results show air flow from the aisle through the holes in the curtains and through the racks behind the curtains, and this flow keeps allergen behind the curtains and prevents its spread from the cages into the aisle. The present study shows that the use of curtains in front of the cage racks is an efficient way to prevent spread of allergens from rodent cages to the entire animal room. PMID:16642971

  5. Biomarkers of Acute Respiratory Allergen Exposure: Screening For Sensitization Potential

    EPA Science Inventory

    Rationale: An in vitro assay to identify respiratory sensitizers will provide a rapid screen and reduce animal use. The study goal was to identify biomarkers that differentiate allergen versus non-allergen responses following an acute exposure. Methods: Female BALB/c mice rec...

  6. An overview of fruit allergy and the causative allergens.

    PubMed

    Hassan, A K G; Venkatesh, Y P

    2015-11-01

    Plant allergens, being one of the most widespread allergenic substances, are hard to avoid. Hence, their identification and characterization are of prime importance for the diagnosis and treatment of food allergy. The reported allergies to fruits mainly evoke oral allergy syndrome caused by the presence of cross-reactive IgE to certain pollens and thus, allergy to fruits has also been linked to particular pollens. Many fruit allergies are being studied for their causative allergens, and are being characterized. Some tropical or exotic fruits are responsible for region-specific allergies for which only limited information is available, and generally lack allergen characterization. From a survey of the literature on fruit allergy, it is clear that some common fruits (apple, peach, musk melon, kiwi fruit, cherry, grape, strawberry, banana, custard apple, mango and pomegranate) and their allergens appear to be at the center of current research on food allergy. The present review focuses on common fruits reported as allergenic and their identified allergens; a brief description of allergens from six rare/tropical fruits is also covered. PMID:26549334

  7. Allergen and irritant control: importance and implementation.

    PubMed

    Nelson, H S

    1998-01-01

    The Expert Panel Report 2. Guidelines for the Diagnosis and Management of Asthma (1) begins its section on controlling factors that precipitate or worsen asthma with the statement: "For successful long-term asthma management, it is essential to identify and reduce exposures to relevant allergens and irritants and to control other factors that have been shown to increase asthma symptoms and/or precipitate asthma exacerbations." The presence of allergy to indoor allergens and certain seasonal fungal spores has been found to be a risk factor for asthma in epidemiologic studies around the world. Generally between 70% and 85% of asthmatic populations studied have been reported to have positive skin-prick tests. Exposure of allergic patients to inhalant allergens increases airway inflammation, airway hyper-responsiveness, asthma symptoms, need for medication, severe attacks, and even death due to asthma. Environmental tobacco smoke exposure has been shown to increase the prevalence of childhood asthma and to increase asthma symptoms and bronchial hyperresponsiveness while reducing pulmonary function in children chronically exposed. Exposure to other indoor irritants, largely products of unvented combustion, has also been found to increase asthma symptoms. Outdoor air pollution increases asthma symptoms; levels of specific pollutants correlate with emergency room visits and hospitalization for asthma. Rhinitis/sinusitis and gastroesophageal reflux are commonly associated with asthma, and treatment of these conditions has been shown to improve asthma. In patients sensitive to aspirin and nonsteroidal anti-inflammatory drugs or metabisulfites, exposure to these agents can precipitate severe attacks of asthma. Viral infections are common causes for exacerbations of asthma. Infections with Mycoplasma pneumoniae and Chlamydia pneumoniae contribute to acute exacerbations and perhaps to long-term morbidity, as well. This chapter will discuss preventive and therapeutic measures

  8. Protease inhibitor reduces airway response and underlying inflammation in cockroach allergen-induced murine model.

    PubMed

    Saw, Sanjay; Arora, Naveen

    2015-04-01

    Protease(s) enhances airway inflammation and allergic cascade. In the present study, effect of a serine protease inhibitor was evaluated in mouse model of airway disease. Mice were sensitized with cockroach extract (CE) or Per a 10 and treated with 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF) 1 h before or after challenge to measure airway response. Mice were euthanized to collect bronchoalveolar lavage fluid (BALF), blood, and lung to evaluate inflammation. AEBSF treatment significantly reduced the AHR in allergen-challenged mice in dose-dependent manner (p≤ 0.01). IgE (p≤0.05) and Th2 cytokines (p≤0.05) were significantly reduced in treated mice. AEBSF treatment lowered total cell (p≤0.05), eosinophil (p≤0.05), and neutrophil (p≤0.05) in BALF and lung tissue. Oxidative stress parameters were impaired on treatment in allergen-challenged mice (p≤0.05). AEBSF had therapeutic effect in allergen-induced airway resistance and underling inflammation and had potential for combination or as add-on therapy for respiratory diseases. PMID:25052477

  9. Cloning and Expression of Ama r 1, as a Novel Allergen of Amaranthus retroflexus Pollen.

    PubMed

    Morakabati, Payam; Assarehzadegan, Mohammad-Ali; Khosravi, Gholam Reza; Akbari, Bahareh; Dousti, Fatemeh

    2016-01-01

    Sensitisation to Amaranthus retroflexus pollen is very common in tropical and subtropical countries. In this study we aimed to produce a recombinant allergenic Ole e 1-like protein from the pollen of this weed. To predict cross-reactivity of this allergen (Ama r 1) with other members of the Ole e 1-like protein family, the nucleotide sequence homology of the Ama r 1 was investigated. The expression of Ama r 1 in Escherichia coli was performed by using a pET-21b(+) vector. The IgE-binding potential of recombinant Ama r 1 (rAma r 1) was evaluated by immunodetection and inhibition assays using 26 patients' sera sensitised to A. retroflexus pollen. The coding sequence of the Ama r 1 cDNA indicated an open reading frame of 507 bp encoding for 168 amino acid residues which belonged to the Ole e 1-like protein family. Of the 26 serum samples, 10 (38.46%) had significant specific IgE levels for rAma r 1. Immunodetection and inhibition assays revealed that the purified rAma r 1 might be the same as that in the crude extract. Ama r 1, the second allergen from the A. retroflexus pollen, was identified as a member of the family of Ole e 1-like protein. PMID:26925110

  10. Cloning and Expression of Ama r 1, as a Novel Allergen of Amaranthus retroflexus Pollen

    PubMed Central

    Morakabati, Payam; Assarehzadegan, Mohammad-Ali; Khosravi, Gholam Reza; Akbari, Bahareh; Dousti, Fatemeh

    2016-01-01

    Sensitisation to Amaranthus retroflexus pollen is very common in tropical and subtropical countries. In this study we aimed to produce a recombinant allergenic Ole e 1-like protein from the pollen of this weed. To predict cross-reactivity of this allergen (Ama r 1) with other members of the Ole e 1-like protein family, the nucleotide sequence homology of the Ama r 1 was investigated. The expression of Ama r 1 in Escherichia coli was performed by using a pET-21b(+) vector. The IgE-binding potential of recombinant Ama r 1 (rAma r 1) was evaluated by immunodetection and inhibition assays using 26 patients' sera sensitised to A. retroflexus pollen. The coding sequence of the Ama r 1 cDNA indicated an open reading frame of 507 bp encoding for 168 amino acid residues which belonged to the Ole e 1-like protein family. Of the 26 serum samples, 10 (38.46%) had significant specific IgE levels for rAma r 1. Immunodetection and inhibition assays revealed that the purified rAma r 1 might be the same as that in the crude extract. Ama r 1, the second allergen from the A. retroflexus pollen, was identified as a member of the family of Ole e 1-like protein. PMID:26925110

  11. Characterization of arginine kinase, anovel allergen of dermatophagoides farinae (Der f 20)

    PubMed Central

    Xing, Peng; Yu, Haiqiong; Li, Meng; Xiao, Xiaojun; Jiang, Congli; Mo, Lihua; Zhang, Min; Yang, Pingchang; Liu, Zhigang

    2015-01-01

    Objective: To characterize a novel allergen, the Dermatophagoides farinae-derived arginine kinase (Der f 20). Methods: The protein of Der f 20 was synthesized by genetic engineering approaches. The allergenicity of Der f 20 was tested by enzyme-linked immunosorbent assay and an airway allergy mouse model. Results: The Der f 20 gene was cloned andpresented in the Gene Bank with an accession number of AAP57094. The Der f 20 is an arginine kinase (AK), whichshowed a close relationship with D. pteronyssinus AK and Aleuroglyphusovatus AK. Western-blot and ELISA studies showed the IgE binding capacity of Der f 20 was 66.7% in the sera from 6 dust mite allergic patients. Immune inhibition assayresults showed the IgE cross-reactivity between Der f 20 and DME (Dust mite extract). Positive responses to Der f 20 were 41.2% as shown by skin prick tests in 17 DME-allergic patients. In vitro experimental results showed that Der f 20 induced Th2 cell differentiation and the expression of T cell Ig mucin domain molecule-4 (TIM4) in DCs. Conclusions; The Der f 20 protein is a novel subtype of thedust mite allergen. PMID:26885278

  12. Isolation and full characterisation of a potentially allergenic lipid transfer protein (LTP) in almond.

    PubMed

    Buhler, Sofie; Tedeschi, Tullia; Faccini, Andrea; Garino, Cristiano; Arlorio, Marco; Dossena, Arnaldo; Sforza, Stefano

    2015-01-01

    Non-specific lipid transfer proteins (nsLTP) were shown to be among the most significant allergens, in particular in several fruits belonging to the Rosaceae family. The molecular features of LTPs, such as the presence of eight cysteine residues forming four disulfide bridges, confer a compact structure, decreasing the probability of degradation due to cooking or digestion, thereby increasing the chance of systemic absorption and severe allergic reactions. Few studies on LTP-induced allergies regarding almond (Prunus dulcis L) are available in the literature. In the present work, we describe for the first time the extraction and purification of an almond LTP, achieving its full characterisation by using liquid chromatography and exact mass spectrometry; the full sequence was identified by means of LC-ESI-Orbitrap-MS applying a bottom-up approach. The characterised protein consists of 92 amino acids and has a calculated exact MW of 9579.0. The presence of four disulfide bridges was confirmed after reduction, as shown by a mass increment of 8 Da. Finally, its potential allergenicity was confirmed via an in silico approach. The results presented here demonstrate the enormous potential of advanced MS techniques for obtaining high-quality structural and functional data of allergenic proteins in a short time. PMID:25658292

  13. Occurrence of indoor allergens in Saudi Arabia

    SciTech Connect

    Sorensen, H.; Gravesen, S.; Lind, P.; Schwartz, B.; Ashoor, A.A.; Maglad, S.

    1985-06-01

    Investigations on indoor airborne allergens in Saudi Arabia were performed by mold cultures and dust analyses by counter-current immunoelectrophoresis. Twenty fungal genera were isolated, with Aspergillus as the most often encountered. Most of the dust-bound fungi found are ubiquitous and common. Antibodies against Dermatophagoides pteronyssinus, cat- cow- and rat dander, and Cynodon dactylon pollen were used in the dust analyses. Animal antigens were found in five of the ten dust samples. House dust mites were extraordinarily rare. Pollen of Cynodon dactylon (Bermuda grass) was present in nearly all the samples, and in a concurrent clinical study this antigen was found to be the most common cause of perennial rhinitis.

  14. 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

  15. Particle size of airborne mouse crude and defined allergens.

    PubMed

    Sakaguchi, M; Inouye, S; Miyazawa, H; Kamimura, H; Kimura, M; Yamazaki, S

    1989-05-01

    Laboratory animal allergy is a serious occupational diseases of many workers and scientists engaged in animal experimentation. Control measures depend upon characterization of allergens including airborne particles. This study measured the particle size of crude mouse urine and pelt aeroallergens generated in mouse housing rooms and compared them with mouse serum albumin, a defined major allergen. Allergens were detected by specific immunological methods. Most crude and defined allergens (74.5-86.4%) concentrated on a filter with a retention size greater than 7 microns. In distrubed air, allergen concentration increased 1.4 (albumin) to 5 (crude) fold and the proportion of small particles increased from 1.4% in calm air to 4.5% in distrubed air. This information on the generation and size distribution of aeroallergens will be important in the development of effective counter measures. PMID:2724924

  16. Aspects of food processing and its effect on allergen structure.

    PubMed

    Paschke, Angelika

    2009-08-01

    The article summarizes current physical and chemical methods in food processing as storage, preparation, separation, isolation or purification and thermal application on the one hand as well as enzymatic treatment on the other and their impact on the properties of food proteins. Novel methods of food processing like high pressure, electric field application or irradiation and their impact on food allergens are presented. The EU project REDALL (Reduced Allergenicity of Processed Foods, Containing Animal Allergens: QLK1-CT-2002-02687) showed that by a combination of enzyme and heat treatment the allergic potential of hen's egg decreased about 100 fold. Clinical reactions do not appear anymore. An AiF-FV 12024 N project worked with fruits like mango, lychee and apple. Processed mango and lychee had no change in allergenic potential during heating while e. g. canning. Apple almost lost its allergenic potential after pasteurization in juice production. PMID:19557818

  17. Allergic contact dermatitis in dermatologic surgery: review of common allergens.

    PubMed

    Butler, Lara; Mowad, Christen

    2013-01-01

    With the growing number of dermatologic surgeries performed each year comes an increased potential for patient exposure and sensitization to allergens. Patients are exposed to many well-documented allergens in the preoperative, intraoperative, and postoperative settings during surgery. Postoperative skin complications of allergic contact dermatitis increase health care costs and cause patient suffering. Early recognition, diagnosis, and treatment by dermatologic surgeons are essential to decrease morbidity related to medically necessary and elective cutaneous surgeries. While a specific standard screening panel for cutaneous surgery-related allergens is not well established, we propose several categories of allergens be strongly considered and tested if a patient is suspected of having allergic contact dermatitis in an attempt to reveal pertinent allergens and prevent future exposures. PMID:24030369

  18. Novel developments in the mechanisms of immune tolerance to allergens.

    PubMed

    Eiwegger, Thomas; Gruber, Saskia; Szépfalusi, Zsolt; Akdis, Cezmi A

    2012-10-01

    Allergy is the result of a disbalanced immune response to environmental innocuous antigens. Despite of accumulating data to define the pathomechanisms that take place in case of allergic diseases a detailed understanding of sequence of events that lead to the "normal" scenario of tolerance development are still under debate. Allergen-specific immunotherapy is the only causal treatment of allergic diseases. It modifies the immune response to a particular antigen to achieve tolerance against the symptom-causing allergen. This process is considered to mirror physiological peripheral tolerance induction. A number of immunological changes have been described to occur under allergen immunotherapy, including the generation of allergen-specific regulatory T cells, the induction of allergen-specific IgG4, an increase in the Th1/Th2 cytokine ratio and decreased activation and function of effector cells such as mast cells, basophils and eosinophils. PMID:23095863

  19. Cable manufacture

    NASA Technical Reports Server (NTRS)

    Gamble, P.

    1972-01-01

    A survey is presented of flat electrical cable manufacturing, with particular reference to patented processes. The economics of manufacture based on an analysis of material and operating costs is considered for the various methods. Attention is given to the competitive advantages of the several processes and their resulting products. The historical area of flat cable manufacture is presented to give a frame of reference for the survey.

  20. Molecular Cloning and Expression of a New Allergen of Acacia farnesiana (Aca f 2).

    PubMed

    Sepahi, Najmeh; Khodadadi, Ali; Assarehzadegan, Mohammad-Ali; Amini, Akram; Zarinhadideh, Farnoosh; Ali-Sadeghi, Hosein

    2015-08-01

    Inhalation of pollens from different species of Acacia is a common cause of respiratory allergy in tropical areas of the world. Acacia farnesiana is commonly used as street trees in towns and ornamental shade trees in parks and gardens throughout arid and semi-arid regions of Asia. This study aimed to produce and purify the A. farnesiana pollen profilin (Aca f 2) and evaluate its nucleotide sequence homology with profilins of common allergenic plants to predict allergenic cross-reactivity. Thirty-nine patients who were allergic to Acacia pollens were included in the study. Cloning of Acacia profilin-coding sequence was performed by polymerase chain reaction using primers from Acacia pollen RNA. The cDNA of Acacia pollen profilin was then expressed in Escherichia coli using pET-21b(+) vector and purified by metal affinity chromatography. Immunoreactivity of the recombinant Acacia profilin (rAca f 2) was evaluated by specific ELISA, immunoblotting, and inhibition assays. The coding sequence of the Acacia profilin cDNA was recognized as a 399-bp open reading frame encoding 133 amino acid residues. Eighteen patients (18/39, 46.15%) had significant specific IgE levels against Aca f 2. Immunodetection and inhibition assays indicated that purified Aca f 2 might be the same as that in the crude extract. Aca f2, the first allergen from A. farnesiana pollen, was identified as belonging to the family of profilins. The amino acid sequence homology analysis showed high cross-reactivity between Aca f 2 and other profilins from botanically unrelated common allergenic plants. PMID:26547704

  1. Attenuated Allergenic Activity of Ovomucoid After Electrolysis

    PubMed Central

    Kido, Jun

    2015-01-01

    Ovomucoid (OMC) is the most prominent allergen causing hen's egg allergy, containing disulfide (S-S) bonds that may be responsible for its allergic action. As S-S bonds may be reduced during electrolysis, this study was undertaken to evaluate modulation of the allergic action of OMC after electrolysis. Electrolysis was carried out for 1% OMC containing 1% sodium chloride for 30 minutes with a voltage difference of 90 V, 0.23 A (30 mA/cm2). Protein assays, amino acid measurement, and mass spectrometry in untreated OMC and OMC on both the anode and cathode sides after electrolysis were performed. Moreover, 21 patients with IgE-mediated hen's egg allergy were evaluated by using the skin prick test (SPT) for untreated OMC and OMC after electrolysis. The allergic action of OMC was reduced after electrolysis on both the anode and cathode sides when evaluated by the SPT. The modifications of OMC on electrolysis caused the loss of 2 distinct peptide fragments (57E-63K and 123H-128R) as seen on matrix-associated laser desorption/ionization time-of-flight mass spectrometry. The total free SH groups in OMC were increased on the cathode side. Although the regions of S-S broken bonds were not determined in this study, the change in S-S bonds in OMC on both the anode and cathode sides may reduce the allergenic activity. PMID:26333707

  2. Attenuated Allergenic Activity of Ovomucoid After Electrolysis.

    PubMed

    Kido, Jun; Matsumoto, Tomoaki

    2015-11-01

    Ovomucoid (OMC) is the most prominent allergen causing hen's egg allergy, containing disulfide (S-S) bonds that may be responsible for its allergic action. As S-S bonds may be reduced during electrolysis, this study was undertaken to evaluate modulation of the allergic action of OMC after electrolysis. Electrolysis was carried out for 1% OMC containing 1% sodium chloride for 30 minutes with a voltage difference of 90 V, 0.23 A (30 mA/cm²). Protein assays, amino acid measurement, and mass spectrometry in untreated OMC and OMC on both the anode and cathode sides after electrolysis were performed. Moreover, 21 patients with IgE-mediated hen's egg allergy were evaluated by using the skin prick test (SPT) for untreated OMC and OMC after electrolysis. The allergic action of OMC was reduced after electrolysis on both the anode and cathode sides when evaluated by the SPT. The modifications of OMC on electrolysis caused the loss of 2 distinct peptide fragments (57E-63K and 123H-128R) as seen on matrix-associated laser desorption/ionization time-of-flight mass spectrometry. The total free SH groups in OMC were increased on the cathode side. Although the regions of S-S broken bonds were not determined in this study, the change in S-S bonds in OMC on both the anode and cathode sides may reduce the allergenic activity. PMID:26333707

  3. Making peanut allergens indigestible: a model system for reducing or preventing an allergic reaction

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Peanut allergens are not totally resistant to digestion as previously known. Creating peanut allergen conjugates that are more resistant to digestion may prevent absorption of the allergens into the bloodstream, and thereby, an allergic reaction. Peanut allergen conjugates were prepared by covalen...

  4. Cross-reactivity among non-specific lipid-transfer proteins from food and pollen allergenic sources.

    PubMed

    Morales, María; López-Matas, M Ángeles; Moya, Raquel; Carnés, Jerónimo

    2014-12-15

    Non-specific lipid-transfer proteins (nsLTPs) are a family of pan-allergens present in foods and pollen. However, sequence homology among them is limited. The objective of this study was to evaluate the IgE-mediated cross-reactivity between nsLTPs from different sources and evaluate the allergenic properties of LTPs from peach (Pru p 3) and pellitory (Par j 1/Par j 2), major fruit and pollen allergens. Both proteins were purified and characterised. Cross-reactivity studies among nsLTPs from different foods and pollens were performed by immunoblot inhibition using sera specific to peach or pellitory pollen. Cross-reactivity with Pru p 3 was observed in hazelnut, onion, corn, peanut and apple while in pollens, none of the extracts was inhibited with Par j 1/2. In conclusion, Pru p 3 did not inhibit LTPs from most fruits. Therefore, although Pru p 3 covers the largest number of epitopes, diagnosis with only this allergen may not detect all LTP sensitivities. PMID:25038692

  5. Manufacturing technologies

    SciTech Connect

    1995-09-01

    The Manufacturing Technologies Center is an integral part of Sandia National Laboratories, a multiprogram engineering and science laboratory, operated for the Department of Energy (DOE) with major facilities at Albuquerque, New Mexico, and Livermore, California. Our Center is at the core of Sandia`s Advanced Manufacturing effort which spans the entire product realization process.

  6. Manufacturing Technology.

    ERIC Educational Resources Information Center

    Barnes, James L.

    This curriculum guide is designed to assist junior high school industrial arts teachers in planning new courses and revising existing courses in manufacturing technology. Addressed in the individual units of the guide are the following topics: introduction to manufacturing, materials processing, personnel management, production management,…

  7. 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...

  8. Purification, thermal stability, and antigenicity of the immunodominant soybean allergen P34 in soy cultivars, ingredients, and products.

    PubMed

    Wilson, S; Martinez-Villaluenga, C; De Mejia, E G

    2008-08-01

    Protein P34 (Gly m Bd 30K) is the immunodominant allergen in soybean (Glycine max L.). The objectives of this study were (1) to study the effect of thermal treatment on P34 antigenicity and secondary structure after isolation and purification of P34 from soybean by chromatographic techniques; (2) to identify the variability of P34 allergen within 138 accessions from a diverse USDA soybean germplasm collection by ELISA; and (3) to quantify P34 immunoreactivity in various commercial soy ingredients and products. Thermal processing decreased P34 antigenicity. Soybean accessions with the highest P34 content were ancestral (12 mg/g defatted flour) followed by modern (10 mg/g defatted flour) and exotic (8 mg/g defatted flour). The cultivar that emerged as the lowest-expressing P34 accession was PI548657 (2.3 mg/g defatted flour). Among commercial soy ingredients, soy flour yielded the highest P34 antigenicity (32 mg/g extracted protein) followed by soy protein isolate (29 mg/g extracted protein) and soy protein concentrate (24 mg/g extracted protein). Among soy consumer products, soymilk presented the highest P34 antigenicity, ranging from 7 to 23 mg/g extracted protein, followed by tempeh (8 mg/g extracted protein), soy infant formula (3.4 mg/g extracted protein), soy powder (2 mg/g extracted protein), and soy cheese products (0.50 mg/g extracted protein). Korean miso, soy sauce, soy chili mix, soy nuts, soy cream cheese, soy meat patty, texturized soy protein, and soy cereal exhibited undetectable P34 antigenicity (detection limit = 0.45 ng). Selecting soybean varieties with low levels of this allergen, or via processing, could potentially make soybean products less antigenic. PMID:19241594

  9. Effect of Formaldehyde on Asthmatic Response to Inhaled Allergen Challenge

    PubMed Central

    Ezratty, Véronique; Bonay, Marcel; Neukirch, Catherine; Orset-Guillossou, Gaëlle; Dehoux, Monique; Koscielny, Serge; Cabanes, Pierre-André; Lambrozo, Jacques; Aubier, Michel

    2007-01-01

    Background Exposure to formaldehyde may lead to exacerbation of asthma. Objectives Our aim in this study was to investigate whether exposure to a low level (500 μg/m3) of formaldehyde enhances inhaled allergen responses. Methods Twelve subjects with intermittent asthma and allergy to pollen were exposed, at rest, in a double-blind crossover study to either formaldehyde or purified air for 60 min. The order of exposure to formaldehyde and air-only was randomized, and exposures were separated by 2 weeks. We also performed an allergen inhalation challenge after each exposure. Airway responsiveness to methacholine and lower airway inflammation (induced sputum) were assessed 8 hr after allergen challenge. Results The median dose of allergen producing a 15% decrease in forced expiratory volume in 1 sec (PD15FEV1) was 0.80 IR (index of reactivity) after formaldehyde exposure compared with 0.25 IR after air-only exposure (p = 0.06). Formaldehyde exposure did not affect allergen-induced increase in responsiveness to methacholine (p = 0.42). We found no formaldehyde-associated effect on the airway inflammatory response, in particular the eosinophilic inflammatory response, induced by the allergen challenge 8 hr before. Conclusion In this study, exposure to 500 μg/m3 formaldehyde had no significant deleterious effect on airway allergen responsiveness of patients with intermittent asthma; we found a trend toward a protective effect. PMID:17384766

  10. Clinical presentation, allergens, and management of wheat allergy.

    PubMed

    Quirce, Santiago; Boyano-Martínez, Teresa; Díaz-Perales, Araceli

    2016-05-01

    IgE-mediated allergy to wheat proteins can be caused by exposure through ingestion, inhalation, or skin/mucosal contact, and can affect various populations and age groups. Respiratory allergy to wheat proteins is commonly observed in adult patients occupationally exposed to flour, whereas wheat food allergy is more common in children. Wheat allergy is of growing importance for patients with recurrent anaphylaxis, especially when exercise related. The diagnosis of wheat allergy relies on a consistent clinical history, skin prick testing with well-characterized extracts and specific IgE tests. The accuracy of wheat allergy diagnosis may be improved by measuring IgE responses to several wheat components. However, a high degree of heterogeneity has been found in the recognition pattern of allergens among patient groups with different clinical profiles, as well as within each group. Thus, oral provocation with wheat or the implicated cereal is the reference test for the definitive diagnosis of ingested wheat/cereal allergy. PMID:26800201

  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 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

  13. Environmental tobacco smoke, indoor allergens, and childhood asthma.

    PubMed Central

    Gold, D R

    2000-01-01

    Both environmental tobacco smoke and indoor allergens can exacerbate already established childhood albeit primarily through quite disparate mechanisms. In infancy and childhood, environmental tobacco smoke (ETS) exposure is associated with measures of decreased flow in the airways, bronchial hyperresponsiveness, and increased respiratory infections, but the relationship between ETS and allergy is poorly understood. Indoor allergens from dust mite, cockroach, and cat can be associated with asthma exacerbation in children sensitized to the specific allergens. The precise role of either ETS or indoor allergens in the development of asthma is less well understood. The strong and consistent association between ETS and asthma development in young children may relate to both prenatal and postnatal influences on airway caliber or bronchial responsiveness. Dust mite allergen levels predict asthma in children sensitized to dust mite. The tendency to develop specific IgE antibodies to allergens (sensitization) is associated with and may be preceded by the development of a T-helper (Th)2 profile of cytokine release. The importance of either ETS or indoor allergens in the differentiation of T cells into a Th2-type profile of cytokine release or in the localization of immediate-type allergic responses to the lung is unknown. This article evaluates the strength of the evidence that ETS or indoor allergens influence asthma exacerbation and asthma development in children. We also selectively review data for the effectiveness of allergen reduction in reducing asthma symptoms and present a potential research agenda regarding these two broad areas of environmental exposure and their relationship to childhood asthma. PMID:10931782

  14. Intranasal mite allergen induces allergic asthma-like responses in NC/Nga mice.

    PubMed

    Shibamori, Masafumi; Ogino, Keiki; Kambayashi, Yasuhiro; Ishiyama, Hironobu

    2006-01-25

    Airway responses induced by intranasal administration of mite allergen without adjuvant were studied in NC/Nga mice. A crude extract of Dermatophagoides farinae (Df) was administered for 5 consecutive days and a single intranasal challenge booster dose was given 1 week after the last sensitization. 24 h after the single challenge, the airway hyperresponsiveness (AHR) was measured and the bronchoalveolar lavage fluid (BALF) was analyzed for numbers of eosinophils and neutrophils, and both cytokine and chemokine levels. There were marked increases in number of eosinophils in the BALF, AHR, Th2 cytokines (IL-5 and IL-13), and chemokine (eotaxin-1 and eotaxin-2) levels in the BALF following Df exposure. C57BL/6N, A/J, BALB/c, and CBA/JN mouse strains were also exposed to Df crude extract, but all of the measured responses were strongest in NC/Nga mice. Furthermore, Df-exposed NC/Nga mice showed the goblet cell hyperplasia, pulmonary eosinophilic inflammation, and increases in both total serum IgE and Df-specific IgG1. After intranasal exposure of NC/Nga mice to crude extract of Dermatophagoides pteronyssinus, the BALF eosinophilia and AHR were similar to responses induced by Df. None of the study parameters were increased in response to intranasal exposure to ovalbumin. These data demonstrated that NC/Nga mice developed allergic asthma-like responses after intranasal exposure to mite allergens. PMID:16229861

  15. 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. PMID:12615623

  16. Effects of atmospheric pollutants (CO, O3, SO2) on the allergenicity of Betula pendula, Ostrya carpinifolia, and Carpinus betulus pollen.

    PubMed

    Cuinica, Lázaro G; Cruz, Ana; Abreu, Ilda; da Silva, Joaquim C G Esteves

    2015-01-01

    Pollen of Betula pendula, Ostrya carpinifolia, and Carpinus betulus was exposed in vitro to relatively low levels of the air pollutants, namely carbon monoxide, ozone, and sulfur dioxide. The allergenicity of the exposed pollen was compared with that of non-exposed pollen samples to assess if air pollution exposition affects the allergenicity potential of pollen. The immunodetection assays indicated higher IgE recognition by all sera of allergic patients to the pollen protein extracts in all exposed samples in comparison to the non-exposed samples. These results show that the pollen exposition to low pollutants' levels induces increased allergic reaction to sensitized individuals. PMID:25055718

  17. 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

  18. Identification of novel allergen in edible insect, Gryllus bimaculatus and its cross-reactivity with Macrobrachium spp. allergens.

    PubMed

    Srinroch, Chutima; Srisomsap, Chantragan; Chokchaichamnankit, Daranee; Punyarit, Phaibul; Phiriyangkul, Pharima

    2015-10-01

    Edible insects have recently been promoted as a source of protein and have a high nutrition value. Identification of allergens and cross-reactivity between Macrobrachium spp. and the field cricket (Gryllus bimaculatus) is necessary for food safety control and to assist in the diagnosis and therapy of allergy symptoms. Denaturing polyacrylamide gel electrophoresis (SDS-PAGE) was used to separate proteins. Allergens were determined and identified by IgE-immunoblotting with pooled sera from prawn-allergic patients (n=16) and LC-MS/MS. Arginine kinase (AK) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were determined as the important allergens in muscle of Macrobrachium rosenbergii whereas, hemocyanin (HC) was identified as an allergen in Macrobrachium spp. The allergens in Macrobrachium lanchesteri were identified as AK and HC. In addition, hexamerin1B (HEX1B) was identified as a novel and specific allergen in G. bimaculatus. The important allergen in G. bimaculatus and Macrobrachium spp. is AK and was found to cross-react between both species. PMID:25872439

  19. Manufacturing technology

    SciTech Connect

    Blaedel, K.L.

    1997-02-01

    The specific goals of the Manufacturing Technology thrust area are to develop an understanding of fundamental fabrication processes, to construct general purpose process models that will have wide applicability, to document our findings and models in journals, to transfer technology to LLNL programs, industry, and colleagues, and to develop continuing relationships with industrial and academic communities to advance our collective understanding of fabrication processes. Advances in four projects are described here, namely Design of a Precision Saw for Manufacturing, Deposition of Boron Nitride Films via PVD, Manufacturing and Coating by Kinetic Energy Metallization, and Magnet Design and Application.

  20. Thresholds of allergenic proteins in foods

    SciTech Connect

    Hourihane, Jonathan O'B. . E-mail: J.Hourihane@soton.ac.uk; Knulst, Andre C.

    2005-09-01

    Threshold doses or Estimated Eliciting Doses (EEDs) represent an important new field of research in food allergy. Clinicians and regulators have embraced some toxicological concepts such as LOAEL and NOAEL and applied them to an area of significant clinical uncertainty and interest. The impact of intrinsic human factors (e.g., asthma and exercise) and extrinsic event factors (e.g., season, location and especially dose of allergen) on a future allergic reaction in the community needs to be considered carefully when interpreting results of clinical and research low-dose food challenges. The ongoing cooperation of food allergy research groups in medicine, food science and government will surely deliver results of the highest importance to the wider communities of allergology, food science and technology and the increasing number of allergic consumers.

  1. Smart Manufacturing.

    PubMed

    Davis, Jim; Edgar, Thomas; Graybill, Robert; Korambath, Prakashan; Schott, Brian; Swink, Denise; Wang, Jianwu; Wetzel, Jim

    2015-01-01

    Historic manufacturing enterprises based on vertically optimized companies, practices, market share, and competitiveness are giving way to enterprises that are responsive across an entire value chain to demand dynamic markets and customized product value adds; increased expectations for environmental sustainability, reduced energy usage, and zero incidents; and faster technology and product adoption. Agile innovation and manufacturing combined with radically increased productivity become engines for competitiveness and reinvestment, not simply for decreased cost. A focus on agility, productivity, energy, and environmental sustainability produces opportunities that are far beyond reducing market volatility. Agility directly impacts innovation, time-to-market, and faster, broader exploration of the trade space. These changes, the forces driving them, and new network-based information technologies offering unprecedented insights and analysis are motivating the advent of smart manufacturing and new information technology infrastructure for manufacturing. PMID:25898070

  2. House-dust mite allergy: mapping of Dermatophagoides pteronyssinus allergens for dogs by two-dimensional immunoblotting

    PubMed Central

    Marques, Andreia Grilo; Pereira, Luísa Maria Dotti Silva; Goicoa, Ana; Semião-Santos, Saul José; Bento, Ofélia Pereira

    2015-01-01

    Introduction Specific immunotherapy has shown to be very useful for allergy control in dogs, with a common success rate ranging from 65% to 70%. However, this efficacy could probably be improved and the identification of individual allergomes, with the choice of more adequate molecular allergen pools for specific immunotherapy, being the strategy. Aim To map Dermatophagoides pteronyssinus (Der p) allergens for mite-sensitized atopic dogs, for better understanding how individual allergograms may influence the response to house-dust mite immunotherapy. Material and methods To identify the Der p mite allergome for dogs, 20 individuals allergic to dust-mites and sensitized to Der p, were selected. The extract from Der p was submitted to isoelectric focusing (IEF), one-dimensional (1-D) and two-dimensional (2-D) sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Separated proteins were blotted onto polyvinylidene difluoride (PVDF) membranes and immunoblottings were performed with patient sera. Allergen-bound specific IgE was detected. Results Eleven allergens were identified from isoelectric focusing (IEF), as well as from 1-D SDS PAGE. From 2-D SDS-PAGE, 24 spots were identified. Conclusions Several similarities were found between dog and human allergograms and no absolute correlation between sensitization and allergy was observed either. As in humans, different individual allergograms do not seem to implicate different clinical patterns, but may influence the response to specific immunotherapy. The molecular epidemiology approach in veterinary allergy management, by the characterization of individual patients’ allergoms and by choosing the best molecular allergen pool for each patient could also improve the efficacy of allergy immunotherapy. PMID:26015775

  3. Expression of a codon-optimised recombinant Ara h 2.02 peanut allergen in Escherichia coli.

    PubMed

    Lew, Min Han; Lim, Renee Lay Hong

    2016-01-01

    Current diagnostic tools for peanut allergy using crude peanut extract showed low predictive value and reduced specificity for detection of peanut allergen-specific immunoglobulin E (IgE). The Ara h 2.02, an isoform of the major peanut allergen Ara h 2, contains three IgE epitope recognition sequence of 'DPYSPS' and may be a better reagent for component resolve diagnosis. This research aimed to generate a codon-optimised Ara h 2.02 gene for heterologous expression in Escherichia coli and allergenicity study of this recombinant protein. The codon-optimised gene was generated by PCR using overlapping primers and cloned into the pET-28a (+) expression vector. Moderate expression of a 22.5 kDa 6xhistidine-tagged recombinant Ara h 2.02 protein (6xHis-rAra h 2.02) in BL21 (DE3) host cells was observed upon induction with 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG). The insoluble recombinant protein was purified under denaturing condition using nickel-nitrilotriacetic acid (Ni-NTA) affinity chromatography and refolded by dialysis in decreasing urea concentration, amounting to a yield of 74 mg/l of expression culture. Matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) and immunoblot analysis confirmed the production of the recombinant 6xHis-rAra h 2.02. The refolded recombinant 6xHis-rAra h 2.02, with or without adjuvant, was able to elicit comparable level of allergen-specific IgE and IgG1 in sensitised Balb/c mice. In addition, the specific IgE antibodies raised against the recombinant protein were able to recognise the native Ara h 2 protein, demonstrating its allergenicity and potential as a reagent for diagnosis and therapeutic study. PMID:26411458

  4. Purification and Immunobiochemical Characterization of a 31 kDa Cross-Reactive Allergen from Phaseolus vulgaris (Kidney Bean)

    PubMed Central

    Kasera, Ramkrashan; Singh, Anand Bahadur; Lavasa, Shakuntala; Nagendra, Komarla; Arora, Naveen

    2013-01-01

    Background Legumes are a rich source of proteins but are also potential elicitors of IgE-mediated food allergy. This study aimed to isolate and characterize a major allergen of Phaseolus vulgaris (kidney bean) and determine its allergenicity. Methodology Kidney bean allergen was purified using Q Sepharose column (anion exchanger) and eluates with high intensity were pooled to purify protein using Superdex 75 (gel filtration) and C18 column (RP-HPLC). Patients with history of kidney bean allergy were skin prick tested (SPT) with crude kidney bean extract and the purified protein. Specific IgE was estimated in sera by enzyme-linked immunosorbent assay (ELISA). Characterization of purified protein and its cross-reactivity was investigated by immunobiochemical methods. Identification of purified protein was carried out by tandem mass spectrometry. Principal Findings Purified protein appeared as a single band at 31 kDa on SDS-PAGE and showed IgE binding to 88% patients’ sera by ELISA and immunoblotting. SPT with purified protein identified 78% hypersensitive patients of kidney bean. Significant release of histamine from sensitized basophils was observed after challenge with purified protein. PAS staining suggested it to be a glycoprotein, but no change in IgE binding was observed after periodate oxidation. The 31 kDa protein remained stable for 60 min on incubation with pepsin. The purified protein had high allergenic potential since it required only 102 ng of self protein for 50% IgE inhibition. Mass spectrometric analysis identified it as Phytohemagglutinin. It also showed hemagglutination with human RBCs. Cross-reactivity was observed with peanut and black gram with IC50 of 185 and 228 ng respectively. Conclusion/Significance A 31 kDa major allergen of kidney bean was purified and identified as phytohemagglutinin with cross-reactivity to peanut and black gram. PMID:23671655

  5. Isoform identification, recombinant production and characterization of the allergen lipid transfer protein 1 from pear (Pyr c 3).

    PubMed

    Ramazzina, Ileana; Amato, Stefano; Passera, Elisabetta; Sforza, Stefano; Mistrello, Gianni; Berni, Rodolfo; Folli, Claudia

    2012-01-10

    Non-specific lipid transfer proteins belonging to LTP1 family represent the most important allergens for non pollen-related allergies to Rosaceae fruits in the Mediterranean area. Peach LTP1 (Pru p 3) is a major allergen and is considered the prototypic allergenic LTP. On the contrary, pear allergy without pollinosis seems to be under-reported when compared to other Rosaceae fruits suggesting that the as-yet-uncharacterized pear LTP1 (Pyr c 3) has in vivo a low allergenicity. We report here on the identification of four cDNAs encoding for LTP1 in pear fruits. The two isoforms exhibiting amino acid sequences most similar to those of peach and apple homologues were obtained as recombinant proteins. Such isoforms exhibited CD spectra and lipid binding ability typical of LTP1 family. Moreover, pear LTP1 mRNA was mainly found in the peel, as previously shown for other Rosaceae fruits. By means of IgE ELISA assays a considerable immunoreactivity of these proteins to LTP-sensitive patient sera was detected, even though allergic reactions after ingestion of pear were not reported in the clinical history of the patients. Finally, the abundance of LTP1 in protein extracts from pear peel, in which LTP1 from Rosaceae fruits is mainly confined, was estimated to be much lower as compared to peach peel. Our data suggest that the two isoforms of pear LTP1 characterized in this study possess biochemical features and IgE-binding ability similar to allergenic LTPs. Their low concentrations in pear might be the cause of the low frequency of LTP-mediated pear allergy. PMID:22015956

  6. Mimotopes for Api g 5, a Relevant Cross-reactive Allergen, in the Celery-Mugwort-Birch-Spice Syndrome

    PubMed Central

    Lukschal, Anna; Wallmann, Julia; Bublin, Merima; Hofstetter, Gerlinde; Mothes-Luksch, Nadine; Breiteneder, Heimo; Pali-Schöll, Isabella

    2016-01-01

    Purpose In the celery-mugwort-birch-spice syndrome, a significant proportion of IgE is directed against high molecular weight (HMW) glycoproteins, including the celery allergen Api g 5. BIP3, a monoclonal antibody originally raised against birch pollen, recognizes HMW allergens in birch and mugwort pollens, celery, and Apiaceae spices. Our aim was to generate mimotopes using BIP3 for immunization against the HMW allergens relevant in the celery-mugwort-birch-spice cross reactivity syndrome. Methods Mimotopes were selected from a random-peptide display library by BIP3 and applied in IgE inhibition assays. The 3 phage clones with the highest inhibitory capacity were chosen for immunization of BALB/c mice. Mouse immune sera were tested for IgG binding to blotted birch pollen extract and used for inhibiting patients' IgE binding. Furthermore, sera were tested for binding to Api g 5, to horseradish peroxidase (HRP) as a second glycoprotein, or to non-glycosylated control allergen Phl p 5 in ELISA, and the specific Api g 5-specific IgG titers were determined. Results Three rounds of biopanning resulted in phage clones exhibiting 7 different sequences including 1 dominant, 1-6-cyclo-CHKLRCDKAIA. Three phage clones had the capacity to inhibit human IgE binding and induced IgG to the HMW antigen when used for immunizing BALB/c mice. The induced BIP3-mimotope IgG reached titers of 1:500 specifically to Api g 5, but hardly reacted to glycoprotein HRP, revealing a minor role of carbohydrates in their epitope. Conclusions The mimotopes characterized in this study mimic the epitope of BIP3 relevant for Api g 5, one of the cross-reactive HMW allergens relevant in the celery-mugwort-birch-spice syndrome. BIP3 mimotopes may be used in the future for hyposensitization in this clinical syndrome by virtue of good and specific immunogenicity. PMID:26739405

  7. 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. PMID:26830572

  8. 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

  9. Molecular cloning, expression and immunological characterisation of Pas n 1, the major allergen of Bahia grass Paspalum notatum pollen.

    PubMed

    Davies, Janet M; Mittag, Diana; Dang, Thanh D; Symons, Karen; Voskamp, Astrid; Rolland, Jennifer M; O'Hehir, Robyn E

    2008-12-01

    Bahia grass, Paspalum notatum, is a clinically important subtropical grass with a prolonged pollination season from spring to autumn. We aimed to clone and characterise the major Bahia grass pollen allergen, Pas n 1. Grass pollen-allergic patients presenting to a tertiary hospital allergy clinic were tested for IgE reactivity with Bahia grass pollen extract by skin prick testing, ImmunoCAP, ELISA and immunoblotting. Using primers deduced from the N-terminal peptide sequence of a group 1 allergen of Bahia grass pollen extract separated by two-dimensional gel electrophoresis, the complete Pas n 1 cDNA was obtained by rapid amplification of cDNA ends and cloned. Biological relevance of recombinant Pas n 1 expressed in Escherichia coli was assessed by serum IgE reactivity and basophil activation. Twenty-nine of 34 (85%) consecutive patients presenting with grass pollen allergy were skin prick test positive to Bahia grass pollen. The Pas n 1 cDNA has sequence homology with the beta-expansin 1 glycoprotein family and is more closely related to the maize pollen group 1 allergen (85% identity) than to ryegrass Lol p 1 or Timothy grass Phl p 1 (64 and 66% identity, respectively). rPas n 1 reacted with serum IgE in 47 of 55 (85%) Bahia grass pollen-allergic patients, activated basophils and inhibited serum IgE reactivity with the 29 kDa band of Bahia grass pollen extract. In conclusion the cDNA for the major group 1 allergen of the subtropical Bahia grass pollen, Pas n 1, was identified and cloned. rPas n 1 is immunologically active and is a valuable reagent for diagnosis and specific immunotherapy of grass pollen allergy. PMID:18817975

  10. Assessment of the Sensitizing Potential of Processed Peanut Proteins in Brown Norway Rats: Roasting Does Not Enhance Allergenicity

    PubMed Central

    Kroghsbo, Stine; Rigby, Neil M.; Johnson, Philip E.; Adel-Patient, Karine; Bøgh, Katrine L.; Salt, Louise J.; Mills, E. N. Clare; Madsen, Charlotte B.

    2014-01-01

    Background IgE-binding of process-modified foods or proteins is the most common method for examination of how food processing affects allergenicity of food allergens. How processing affects sensitization capacity is generally studied by administration of purified food proteins or food extracts and not allergens present in their natural food matrix. Objectives The aim was to investigate if thermal processing increases sensitization potential of whole peanuts via the oral route. In parallel, the effect of heating on sensitization potential of the major peanut allergen Ara h 1 was assessed via the intraperitoneal route. Methods Sensitization potential of processed peanut products and Ara h 1 was examined in Brown Norway (BN) rats by oral administration of blanched or oil-roasted peanuts or peanut butter or by intraperitoneal immunization of purified native (N-), heated (H-) or heat glycated (G-)Ara h 1. Levels of specific IgG and IgE were determined by ELISA and IgE functionality was examined by rat basophilic leukemia (RBL) cell assay. Results In rats dosed orally, roasted peanuts induced significant higher levels of specific IgE to NAra h 1 and 2 than blanched peanuts or peanut butter but with the lowest level of RBL degranulation. However, extract from roasted peanuts was found to be a superior elicitor of RBL degranulation. Process-modified Ara h 1 had similar sensitizing capacity as NAra h 1 but specific IgE reacted more readily with process-modified Ara h 1 than with native. Conclusions Peanut products induce functional specific IgE when dosed orally to BN rats. Roasted peanuts do not have a higher sensitizing capacity than blanched peanuts. In spite of this, extract from roasted peanuts is a superior elicitor of RBL cell degranulation irrespectively of the peanut product used for sensitization. The results also suggest that new epitopes are formed or disclosed by heating Ara h 1 without glucose. PMID:24805813

  11. 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.

  12. Lanolin allergy: history, epidemiology, responsible allergens, and management.

    PubMed

    Lee, Bailey; Warshaw, Erin

    2008-01-01

    Allergy to lanolin has been recognized by dermatologists for decades. This review summarizes the history, epidemiology, and allergenicity of lanolin and its derivatives. "The lanolin paradox" and the safety of pharmaceutical-grade lanolin products are also discussed. PMID:18413106

  13. Structure of allergens and structure based epitope predictions☆

    PubMed Central

    Dall’Antonia, Fabio; Pavkov-Keller, Tea; Zangger, Klaus; Keller, Walter

    2014-01-01

    The structure determination of major allergens is a prerequisite for analyzing surface exposed areas of the allergen and for mapping conformational epitopes. These may be determined by experimental methods including crystallographic and NMR-based approaches or predicted by computational methods. In this review we summarize the existing structural information on allergens and their classification in protein fold families. The currently available allergen-antibody complexes are described and the experimentally obtained epitopes compared. Furthermore we discuss established methods for linear and conformational epitope mapping, putting special emphasis on a recently developed approach, which uses the structural similarity of proteins in combination with the experimental cross-reactivity data for epitope prediction. PMID:23891546

  14. Cross-reactions between respiratory and food allergens.

    PubMed

    de Blay, F; Pauli, G; Bessot, J C

    1991-01-01

    Cross-reactions between inhaled and food allergens are usually attributed to pollen hypersensitivity associated with fruit and vegetable allergy. However, other allergens are involved in these types of cross-reactions. In a few cases, there is a complete similarity between the inhaled and food allergens (garlic, crustacea proteins). More frequently, partial similarity has been demonstrated: whole inhaled allergens are included in ingested substances. Moreover, immunological techniques can demonstrate common antigenic epitopes in organic substances without any apparent relationship. This has been demonstrated by RAST-inhibition and/or immunoblot techniques, using sera from patients cross-sensitized to (1) pollens and fruits or vegetables or (2) avian sera and eggs. Respiratory sensitization always seems to precede food allergy symptoms. PMID:1720408

  15. Bioinformatics Approaches to Classifying Allergens and Predicting Cross-Reactivity

    PubMed Central

    Schein, Catherine H.; Ivanciuc, Ovidiu; Braun, Werner

    2007-01-01

    The major advances in understanding why patients respond to several seemingly different stimuli have been through the isolation, sequencing and structural analysis of proteins that induce an IgE response. The most significant finding is that allergenic proteins from very different sources can have nearly identical sequences and structures, and that this similarity can account for clinically observed cross-reactivity. The increasing amount of information on the sequence, structure and IgE epitopes of allergens is now available in several databases and powerful bioinformatics search tools allow user access to relevant information. Here, we provide an overview of these databases and describe state-of-the art bioinformatics tools to identify the common proteins that may be at the root of multiple allergy syndromes. Progress has also been made in quantitatively defining characteristics that discriminate allergens from non-allergens. Search and software tools for this purpose have been developed and implemented in the Structural Database of Allergenic Proteins (SDAP, http://fermi.utmb.edu/SDAP/). SDAP contains information for over 800 allergens and extensive bibliographic references in a relational database with links to other publicly available databases. SDAP is freely available on the Web to clinicians and patients, and can be used to find structural and functional relations among known allergens and to identify potentially cross-reacting antigens. Here we illustrate how these bioinformatics tools can be used to group allergens, and to detect areas that may account for common patterns of IgE binding and cross-reactivity. Such results can be used to guide treatment regimens for allergy sufferers. PMID:17276876

  16. 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

  17. Characterization of Allergen Emission Sources in Urban Areas.

    PubMed

    Cariñanos, Paloma; Adinolfi, Cristiano; Díaz de la Guardia, Consuelo; De Linares, Concepción; Casares-Porcel, Manuel

    2016-01-01

    Pollen released by urban flora-a major contributor to airborne allergen content during the pollen season-has a considerable adverse impact on human health. Using aerobiological techniques to sample and characterize airborne biological particulate matter (BPM), we can identify the main species contributing to the pollen spectrum and chart variations in counts and overall pollen dynamics throughout the year. However, given the exponential increase in the number of pollen allergy sufferers in built-up areas, new strategies are required to improve the biological quality of urban air. This paper reports on a novel characterization of the potential allergenicity of the tree species most commonly used as ornamentals in Mediterranean cities. Values were assigned to each species based on a number of intrinsic features including pollination strategy, pollen season duration, and allergenic capacity as reported in the specialist literature. Findings were used to generate a database in which groups of conifers, broadleaves, and palm trees were assigned a value of between 0 and 36, enabling their allergenicity to be rated as nil, low, moderate, high, or very high. The case study presented here focuses on the city of Granada in southern Spain. The major airborne-pollen-producing species were identified and the allergenicity of species growing in urban green zones was estimated. Corrective measures are proposed to prevent high allergen levels and thus improve biological air quality. PMID:26828180

  18. Effect of processing technologies on the allergenicity of food products.

    PubMed

    Jiménez-Saiz, Rodrigo; Benedé, Sara; Molina, Elena; López-Expósito, Iván

    2015-01-01

    Heat treatment has been used since ancient times for food processing, first to ensure the safety of food and its storage, but also to transform its characteristics (in its raw form) and obtain new textures, flavors, or novel foods. However, the transformation experienced by food components when heated, or processed, can dramatically affect the allergenicity of food, either reducing or increasing it. To date, most of the articles published dealing with the changes in the potential allergenicity of food are focused on heat treatment and the Maillard reaction. However, it is also important to give prominence to other group of new technologies developed nowadays, such as high-pressure processing, microwaves and food irradiation. These techniques are not likely to replace traditional processing methods, but they are becoming attractive for the food industry due to different reasons, and it is expected in the near future to have different products on the market processed with these new technologies at an affordable cost. Moreover, other biochemical modifications, particularly enzymatic cross-linking of proteins, have attracted wide-spread attention and will be considered as well in this review, because of its great opportunities to induce protein modification and thus affect food allergenicity. Together with the effect of processing of food allergens, this review will place special attention on gastroduodenal digestion of processed allergens, which directly affects their allergenicity. PMID:24734775

  19. A personalized food allergen testing platform on a cellphone

    PubMed Central

    Coskun, Ahmet F.; Wong, Justin; Khodadadi, Delaram; Nagi, Richie; Tey, Andrew; Ozcan, Aydogan

    2013-01-01

    We demonstrate a personalized food allergen testing platform, termed iTube, running on a cellphone that images and automatically analyses colorimetric assays performed in test tubes toward sensitive and specific detection of allergens in food samples. This cost-effective and compact iTube attachment, weighing approximately 40 grams, is mechanically installed on the existing camera unit of a cellphone where the test and control tubes are inserted from the side and are vertically illuminated by two separate light-emitting-diodes. The illumination light is absorbed by the allergen assay that is activated within the tubes, causing an intensity change in the acquired images by the cellphone camera. These transmission images of the sample and control tubes are digitally processed within1 sec using a smart application running on the same cellphone for detection and quantification of allergen contamination in food products. We evaluated the performance of this cellphone based iTube platform using different types of commercially available cookies, where the existence of peanuts was accurately quantified after a sample preparation and incubation time of ~20 min per test. This automated and cost-effective personalized food allergen testing tool running on cellphones can also permit uploading of test results to secure servers to create personal and/or public spatio-temporal allergen maps, which can be useful for public health in various settings. PMID:23254910

  20. A personalized food allergen testing platform on a cellphone.

    PubMed

    Coskun, Ahmet F; Wong, Justin; Khodadadi, Delaram; Nagi, Richie; Tey, Andrew; Ozcan, Aydogan

    2013-02-21

    We demonstrate a personalized food allergen testing platform, termed iTube, running on a cellphone that images and automatically analyses colorimetric assays performed in test tubes toward sensitive and specific detection of allergens in food samples. This cost-effective and compact iTube attachment, weighing approximately 40 grams, is mechanically installed on the existing camera unit of a cellphone, where the test and control tubes are inserted from the side and are vertically illuminated by two separate light-emitting-diodes. The illumination light is absorbed by the allergen assay, which is activated within the tubes, causing an intensity change in the acquired images by the cellphone camera. These transmission images of the sample and control tubes are digitally processed within 1 s using a smart application running on the same cellphone for detection and quantification of allergen contamination in food products. We evaluated the performance of this cellphone-based iTube platform using different types of commercially available cookies, where the existence of peanuts was accurately quantified after a sample preparation and incubation time of ~20 min per test. This automated and cost-effective personalized food allergen testing tool running on cellphones can also permit uploading of test results to secure servers to create personal and/or public spatio-temporal allergen maps, which can be useful for public health in various settings. PMID:23254910

  1. Thermal processing effects on peanut allergen Ara h 2 allergenicity in mice and its antigenic epitope structure.

    PubMed

    Zhang, Wenju; Zhu, Qingqing; Zhang, Tong; Cai, Qin; Chen, Qin

    2016-12-01

    Ara h 2 was purified from peanuts that were thermally treated by various processes, including boiling, glycation, frying and roasting. The allergenicity of Ara h 2 in Balb/c mice and the influence of thermal processing on the structural characteristics, and binding capacity of three core antigenic epitopes were studied. The results demonstrated that boiling, glycation and frying induced the down-regulation of the allergenicity of Ara h 2 in Balb/c mice, the collapse of its tertiary/secondary structure, and a reduction in the core epitope binding capacity; roasting showed a comparable allergenicity and the weakest inhibitory effect on core epitope binding capacity. These results indicate that thermal processing causes alteration of the protein structure and core epitopes of Ara h 2, and may affect its allergenicity. PMID:27374581

  2. DIFFERENTIAL ALLERGIC AND NEUROTROPHIN RESPONSES TO FUNGAL COMPONENT EXTRACTS IN BALB/C MICE

    EPA Science Inventory

    Metarhizium anisopliae mycelium (MYC), conidia (CON) and inducible protease (IND) extracts were combined to produce the antigen MACA to screen for allergenic potential. Involuntary aspiration (IA) exposure to MACA in BALB/c mice has caused immune, inflammatory and physiological ...

  3. Solid-phase microextraction gas chromatography-mass spectrometry determination of fragrance allergens in baby bathwater.

    PubMed

    Lamas, J Pablo; Sanchez-Prado, Lucia; Garcia-Jares, Carmen; Llompart, Maria

    2009-07-01

    A method based on solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) has been optimized for the determination of fragrance allergens in water samples. This is the first study devoted to this family of cosmetic ingredients performed by SPME. The influence of parameters such as fibre coating, extraction and desorption temperatures, salting-out effect and sampling mode on the extraction efficiency has been studied by means of a mixed-level factorial design, which allowed the study of the main effects as well as two-factor interactions. Excluding desorption temperature, the other parameters were, in general, very important for the achievement of high response. The final procedure was based on headspace sampling at 100 degrees C, using polydimethylsiloxane/divinylbenzene fibres. The method showed good linearity and precision for all compounds, with detection limits ranging from 0.001 to 0.3 ng mL(-1). Reliability was demonstrated through the evaluation of the recoveries in different real water samples, including baby bathwater and swimming pool water. The absence of matrix effects allowed the use of external standard calibration to quantify the target compounds in the samples. The proposed procedure was applied to the determination of allergens in several real samples. All the target compounds were found in the samples, and, in some cases, at quite high concentrations. The presence and the levels of these chemicals in baby bathwater should be a matter of concern. PMID:19458938

  4. Allergens involved in occupational asthma caused by baby's breath (Gypsophila paniculata).

    PubMed

    Schroeckenstein, D C; Meier-Davis, S; Yunginger, J W; Bush, R K

    1990-08-01

    Although the floral industry deals with many potential allergens, few examples of occupational asthma exist in this industry. A 22-year-old florist experienced symptoms of rhinitis, conjunctivitis, and asthma on exposure to baby's breath. To determine the contribution of baby's breath to the patient's symptoms, an extract of baby's breath was prepared. Prick skin tests with a 1:10(-5) wt/vol concentration of the extract produced an immediate response, whereas nonexposed atopic and normal control subjects did not react. The patient's asthmatic response to baby's breath was confirmed by bronchial challenge that caused an immediate fall in FEV1 of 26.2% from baseline after inhalation of 88 breath units of the extract. With a direct RAST, the patient's serum bound 38 times the amount of IgE bound by the negative control. IgE binding in the RAST was inhibited by the baby's breath extract but not by unrelated inhibitors (ragweed and tree pollens). Immunoblotting demonstrated IgE binding to 13 protein bands in the extract with molecular weights ranging from 11.5 to 68 kd. Serum from a patient previously reported to have sensitivity to baby's breath recognized five protein bands. Three proteins with molecular weights of 27, 31, and 37 kd were recognized by both patients' sera. We conclude that this patient developed IgE-mediated sensitivity to multiple allergens in baby's breath. This study confirms the importance of this plant as a potential cause of occupational asthma in the floral industry. PMID:2384649

  5. Analyzing the influence of manufacturing conditions of reclaimed asphalt concrete on the characteristics of the asphalt binder: development of a gradual binder extraction method

    NASA Astrophysics Data System (ADS)

    Navaro, J.; Bruneau, D.; Drouadaine, I.; Pouteau, B.; Colin, J.; Dony, A.

    2012-05-01

    When asphalt concrete is manufactured incorporating a high percentage (almost 70%) of reclaimed materials from the deconstruction of road surfaces under renovation, and when the corresponding production device is designed specifically to reduce the energy input need (lowering the production temperature), the resulting manufacturing process contributes to the protection of the environment and reduces production costs. However, to meet the quality requirements of the finished product, virgin materials of appropriate quality and quantity must also be added (mineral aggregates and new asphalt binder) and control systems set up to quantify and optimize the parameters involved (thus avoiding the guess work which still often prevails today). It was for this reason that a new experimental technique described here was devised, which will ultimately be used in asphalt concrete production plants. The technique involves lixiviating reclaimed asphalt concrete using a chlorinated solvent; the resulting solute is collected gradually, then the mixture of binders (virgin and reclaimed asphalt concrete) can be characterized and their mass fractions quantified using a combination of UV and IR spectrometry. With this experimental technique we were able to assess the extent to which the reclaimed asphalt pavement binder participates in the agglomeration and cohesion of the reclaimed asphalt concrete. This assessment was made in terms of the main parameters in the production process, temperature of the materials and mixing time.

  6. 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

  7. Microgravity Manufacturing

    NASA Technical Reports Server (NTRS)

    Cooper, Ken; Munafo, Paul M. (Technical Monitor)

    2002-01-01

    Manufacturing capability in outer space remains one of the critical milestones to surpass to allow humans to conduct long-duration manned space exploration. The high cost-to-orbit for leaving the Earth's gravitational field continues to be the limiting factor in carrying sufficient hardware to maintain extended life support in microgravity or on other planets. Additive manufacturing techniques, or 'chipless' fabrication, like RP are being considered as the most promising technologies for achieving in situ or remote processing of hardware components, as well as for the repair of existing hardware. At least three RP technologies are currently being explored for use in microgravity and extraterrestrial fabrication.

  8. Extractable resources

    NASA Technical Reports Server (NTRS)

    1975-01-01

    The use of information from space systems in the operation of extractive industries, particularly in exploration for mineral and fuel resources was reviewed. Conclusions and recommendations reported are based on the fundamental premise that survival of modern industrial society requires a continuing secure flow of resources for energy, construction and manufacturing, and for use as plant foods.

  9. Validation of standard manufacturing procedure of Guḍūcī sattva (aqueous extract of Tinospora cordifolia (Willd.) Miers) and its tablets

    PubMed Central

    Sharma, Rohit; Amin, Hetal; Galib; Prajapati, P. K.

    2013-01-01

    Introduction: Guḍūci Sattva is a highly valued formulation among ayurvedic physicians, commonly recommended in conditions such as Jvara (fever), Dāha (burning sensation) and other conditions of Pitta predominance. In spite of its numerous medicinal attributes, no published work is available until date on manufacturing guidelines along with its quality control parameters. Aims and Objectives: The aim of this study is to develop the standard manufacturing procedure for preparation of Guḍūci Sattva and its tablets. Materials and Methods: A total of 15 batches of Guḍūci Sattva were prepared in the laboratory. During its preparation, pharmaceutical findings and observations were systematically recorded. To maintain quality control, Guḍūci Sattva tablets were further subjected to analysis such as shape, diameter, width, hardness, weight variation, disintegration time (DT) and friability. Qualitative analysis to detect the presence of various functional groups and high performance thin layer chromatography (HPTLC) profile were also carried out. Results and Conclusion: The average percentage of dried Sattva obtained was 3.8%. The tablets were prepared by direct compression method as per pharmacopoeal specifications. Optimum hardness, weight of tablets, DT and friability of Guḍūci Sattva tablets were found complying with official standards. Alkaloids, carbohydrates and starch were found present in Sattva tablets. Number of peaks obtained in HPTLC also corresponds to this finding. Data obtained by present study may be considered as standard for future studies. Conclusion: The average percentage of dried Sattva obtained was 3.8%. The tablets were prepared by direct compression method as per pharmacopoeal specifications. Optimum hardness, weight of tablets, DT and friability of Guḍūci Sattva tablets were found complying with official standards. Alkaloids, carbohydrates and starch were found present in Sattva tablets. Number of peaks obtained in HPTLC also

  10. Clinical and Laboratory Studies of the Fate of Intranasal Allergen

    PubMed Central

    Rimmer, Janet; Santos, Conceição; Yli-Panula, Eija; Noronha, Virginia; Viander, Markku

    2015-01-01

    Background The precise way in which allergen is handled by the nose is unknown. The objective of this study was to determine recovery of Der p 1 allergen following nasal administration and to determine whether Der p 1 can be detected in nasal biopsies after natural exposure and nasal challenge to allergen. Methods (1) 20 nonatopic non-rhinitics were challenged with Der p 1 and recovery was measured by ELISA in the nasal wash, nasal mucus and induced sputum up to 30 minutes. Particulate charcoal (<40 μm) served as control. (2) In 8 subjects (5 atopics), 30 to 60 minutes after challenge histological localisation of Der p 1 in the nasal mucosal epithelium, subepithelial mucous glands and lamina propria was performed. Co-localisation of Der p 1 with macrophages and IgE-positive cells was undertaken. Results (1) Less than 25% of total allergen was retrievable after aqueous or particulate challenge, most from the nasal mucus during 1-5 min after the challenge. The median of carbon particles recovered was 9%. (2) Prechallenge Der p 1 staining was associated with the epithelium and subepithelial mucous glands. After challenge there was a trend for greater Der p 1 deposition in atopics, but both atopics and nonatopics showed increases in the number of Der p 1 stained cells and stained tissue compartments. In atopics, increased eosinophils, macrophages and IgE positive cells co-localized with Der p 1 staining. Conclusions Der p 1 allergen is detected in nasal tissue independent of atopic status after natural exposure. After challenge the nose effectively retains allergen, which remains mucosally associated; in atopics there is greater Der p 1 deposition and inflammatory response than in nonatopics. These results support the hypothesis that nasal mucus and tissue act as a reservoir for the inhaled Der p 1 allergen leading to a persistent allergic inflammatory response in susceptible individuals. PMID:25969994

  11. Chapter 3: Allergen immunotherapy: definition, indication, and reactions.

    PubMed

    Georgy, Mary S; Saltoun, Carol A

    2012-01-01

    Specific allergen immunotherapy is the administration of increasing amounts of specific allergens to which the patient has type I immediate hypersensitivity. It is a disease modifying therapy, indicated for the treatment of allergic rhinitis, allergic asthma, and hymenoptera hypersensitivity. Specific IgE antibodies for appropriate allergens for immunotherapy must be documented. Indications for allergen immunotherapy include (1) inadequate symptom control despite pharmacotherapy and avoidance measures, (2) a desire to reduce the morbidity from allergic rhinitis and/or asthma or reduce the risk of anaphylaxis from a future insect sting, (3) when the patient experiences undesirable side effects from pharmacotherapy, and (4) when avoidance is not possible. Furthermore, patients may seek to benefit from economic savings of allergen immunotherapy compared with pharmacotherapy over time. Several studies have reported that immunotherapy in children with allergic rhinitis appears to prevent the development of new allergic sensitizations and/or new-onset asthma. Humoral, cellular, and tissue level changes occur with allergen immunotherapy including large increases in antiallergen IgG(4) antibodies, a decrease in the postseasonal rise of antiallergen IgE antibodies, reduced numbers of nasal mucosal mast cells and eosinophils, induction of Treg cells, and suppression of Th2 more than Th1 lymphocytes. There is a corresponding increase in IL-10 and transforming growth factor beta. In the United States, allergen immunotherapy is administered by the subcutaneous route in the physician's office, whereas primarily in some countries in Europe, it is administered for allergic rhinitis and asthma by the sublingual route by the patient at home. PMID:22794676

  12. Probiotics promote endocytic allergen degradation in gut epithelial cells

    SciTech Connect

    Song, Chun-Hua; Liu, Zhi-Qiang; Huang, Shelly; Zheng, Peng-Yuan; Yang, Ping-Chang

    2012-09-14

    Highlights: Black-Right-Pointing-Pointer Knockdown of A20 compromised the epithelial barrier function. Black-Right-Pointing-Pointer The fusion of endosome/lysosome was disturbed in the A20-deficient HT-29 cells. Black-Right-Pointing-Pointer Antigens transported across A20-deficient HT-29 monolayers conserved antigenicity. Black-Right-Pointing-Pointer Probiotic proteins increased the expression of A20 in HT-29 cells. -- Abstract: Background and aims: Epithelial barrier dysfunction plays a critical role in the pathogenesis of allergic diseases; the mechanism is to be further understood. The ubiquitin E3 ligase A20 (A20) plays a role in the endocytic protein degradation in the cells. This study aims to elucidate the role of A20 in the maintenance of gut epithelial barrier function. Methods: Gut epithelial cell line, HT-29 cell, was cultured into monolayers to evaluate the barrier function in transwells. RNA interference was employed to knock down the A20 gene in HT-29 cells to test the role of A20 in the maintenance of epithelial barrier function. Probiotic derived proteins were extracted from the culture supernatants using to enhance the expression of A20 in HT-29 cells. Results: The results showed that the knockdown of A20 compromised the epithelial barrier function in HT-29 monolayers, mainly increased the intracellular permeability. The fusion of endosome/lysosome was disturbed in the A20-deficient HT-29 cells. Allergens collected from the transwell basal chambers of A20-deficient HT-29 monolayers still conserved functional antigenicity. Treating with probiotic derived proteins increased the expression of A20 in HT-29 cells and promote the barrier function. Conclusion: A20 plays an important role in the maintenance of epithelial barrier function as shown by HT-29 monolayer. Probiotic derived protein increases the expression of A20 and promote the HT-29 monolayer barrier function.

  13. 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

  14. Comparative responses to nasal allergen challenge in allergic rhinitic subjects with or without asthma

    PubMed Central

    2011-01-01

    Background Nasal allergen challenge (NAC) is useful to study the pathophysiology of rhinitis, and multiple challenges may more adequately approximate natural exposure. Objective To determine the effect of 4 consecutive daily NAC, on clinical and inflammatory parameters in rhinitics with or without asthma. Methods Rhinitic subjects were recruited: 19 with mild asthma and 13 without asthma. Subjects underwent a control challenge (normal saline) followed by 4 consecutive daily NAC. Allergen challenge consisted of spraying the chosen allergen extract into each nostril until a positive nasal response occurred. Symptoms were recorded on a Likert scale, and oral peak expiratory and nasal peak inspiratory flows allowed assessment of a nasal blockage index (NBI), for a period of 7 hours. Induced sputum and nasal lavage were performed on control day and after 1 and 4 days of NAC. Results Compared with the control day, there was a significant increase in symptom scores and NBI 10 minutes after each last daily NAC in both groups (p < 0.05). Symptom scores and NBI were similar for the 2 groups, except for nasal obstruction and rhinorrhea, which were more marked in subjects with asthma and rhinitis, respectively. Nasal lavage eosinophils were increased after 4 days of challenges in both groups, but there was no change in sputum eosinophils. No cumulative effect or any late response were observed in any of the groups over the challenge period. Conclusion Multiple NAC may be a useful tool to study the pathophysiology of allergic rhinitis or its relationships with asthma. Trial registration ClinicalTrials.gov NCT01286129 PMID:21507261

  15. An adjuvant free mouse model of oral allergenic sensitization to rice seeds protein

    PubMed Central

    2011-01-01

    Background Rice is commonly known as a staple crop consumed worldwide, though with several rice proteins being reported for allergic properties in clinical studies. Thus, there is a growing need for the development of an animal model to better understand the allergenicity of rice proteins and the immunological and pathophysiological mechanisms underlying the development of food allergy. Methods Groups of BALB/c mice were sensitized daily with freshly homogenized rice flour (30 mg or 80 mg) without adjuvant by intragastric gavage. In addition, the mice were challenged with extracted rice flour proteins at several time points intragastrically. Hypersensitivity symptoms in mice were evaluated according to a scoring system. Vascular leakage, ELISA of rice protein-specific IgE, histopathology of small intestine, and passive cutaneous anaphylaxis were conducted on challenged mice. Results An adjuvant free mouse model of rice allergy was established with sensitized mice showing increased scratching behaviors and increased vascular permeability. Rice protein-specific IgE was detected after eighteen days of sensitization and from the fifth challenge onwards. Inflammatory damage to the epithelium in the small intestine of mice was observed beyond one month of sensitization. Passive cutaneous anaphylaxis results confirmed the positive rice allergy in the mouse model. Conclusions We introduced a BALB/c mouse model of rice allergy with simple oral sensitization without the use of adjuvant. This model would serve as a useful tool for further analysis on the immunopathogenic mechanisms of the various rice allergens, for the evaluation of the hypersensitivity of rice or other cereal grains, and to serve as a platform for the development of immunotherapies against rice allergens. PMID:21605393

  16. Determination of dimethyl fumarate and other potential allergens in desiccant and antimould sachets.

    PubMed

    Lamas, J Pablo; Sanchez-Prado, Lucia; Regueiro, Jorge; Llompart, Maria; Garcia-Jares, Carmen

    2009-08-01

    A method for the determination of dimethyl fumarate (DMF), benzothiazole (BT) and tert-butylphenol (TBP) in desiccant and antimould agents employed for protecting consumer products from humidity and mould has been developed. The method is based on ultrasound-assisted extraction (UAE) followed by GC-MS analysis. Parameters that could affect the extraction of the compounds have been optimised using a multivariate approach. In the final conditions, the extraction is performed using only 0.5 or 1 mL ethyl acetate and applying ultrasound energy for 5 min. Simultaneous extractions could also be carried out in 5 min without losing efficiency. The method was validated showing good linearity (R2 >0.995). Both intra- and inter-day precisions were studied at several concentration levels, being satisfactory in all cases (RSD <10%). Recovery was evaluated in four real desiccant samples at different compound concentrations, ranging between 87% and 109%. Limits of detection and quantification were in the low nanogramme per gramme level, thus allowing the determination of DMF at concentrations well below the limit established by the recent EU Directive (0.1 microg/g). The proposed procedure was applied to the determination of the target compounds in several desiccant and antimould samples. Although most of them were simply labelled as "silica gel", more than 70% of the tested samples contained high amounts of DMF, many of them at the high microgram per gramme level. Many samples also showed the presence of the other two potential allergens. These results demonstrate that the content of the "desiccant" sachets and tablets in consumer products does not usually belong with the label of the desiccant, and hence, the high risk of exposition to the powerful allergen DMF and other potentially harmful chemicals through consumer goods should be a matter of concern. PMID:19578833

  17. Effectiveness of vacuum cleaning and wet cleaning in reducing house-dust mites, fungi and mite allergen in a cotton carpet: a case study.

    PubMed

    Wassenaar, D P

    1988-02-01

    In order to evaluate the effect of continuous, thorough vacuuming on house dust organisms and mite allergen, a cotton carpet was vacuumed every other day, six times in total. Seven weeks later, the carpet was cleaned by means of spray extraction. Samples were taken before and after this wet cleaning. In total 1150 g of dust was collected, containing approximately 174,000 arthropods (dead and alive) and 9000 X 10(6) fungal spores. In the course of the experiment, the amount of dust collected at each vacuuming decreased. The number of extracted house dust mites did not decrease significantly, but that of the predator mite Cheyletus did. The number of extracted fungal spores showed a significant decrease (from 142 to 16 X 10(6) spores/m2 per minute vacuuming), as did the extracted mite allergen per m2. After 7 weeks the number of mite eggs and complete house dust mites had increased enormously. After cleaning by spray extraction another increase in the number of complete mites and mite eggs was found, while the amount of mite allergen was diminished. The population growth of the house dust mite between the 6th and the 7th vacuuming is probably due to the decrease of their most important predator, Cheyletus. After the wet cleaning a number of extra eggs hatched, probably due to the high humidity in the carpet. The procedures used in this study to combat house dust mites may have an adverse effect in the long run. PMID:3378462

  18. Engineering, purification and applications of His-tagged recombinant antibody fragments with specificity for the major birch pollen allergen, bet v1.

    PubMed

    Flicker, S; Laffer, S; Steinberger, P; Alhani, B; Zhu, Y; Laukkanen, M L; Keinänen, K; Kraft, D; Valenta, R

    2000-01-01

    Type I allergy, an immunodisorder affecting almost 20% of the population worldwide, is based on the production of IgE antibodies against per se harmless allergens. We report the expression of hexahistidine-tagged antibody fragments (Fabs) with specificity for Bet v1, the major birch pollen allergen, in Escherichia coli. The cDNA coding for the heavy chain fragment of a mouse monoclonal anti-Bet v1 antibody, Bip 1, was engineered by PCR to contain a hexahistidine-encoding 3' end. The modified Bip1 heavy chain cDNA was co-expressed in E. coli XL-1 Blue with the Bip 1 light chain cDNA using the combinatorial plasmid pComb3H. His-tagged recombinant (r) Bip 1 Fabs were isolated by nickel affinity chromatography and rBip 1 Fabs without His-tag were purified via affinity to rBet v1. rBip 1 Fabs with and without His-tag bound specifically to rBet v1 and, like Bet v1 -specific human serum IgE and rabbit-anti rBet v1 antibodies, cross-reacted with Bet v1-related allergens in other plant-species (alder, oak, hazelnut). We demonstrate the usefulness of His-tagged rBip 1 Fabs (1) for the identification of pollen samples containing Bet v 1 by particle blotting, (2) forthe detection of Bet v1-specific IgE antibodies in human serum samples by sandwich ELISA and (3) for the quantification of Bet v1 in solution. Based on these examples we suggest to use rBip 1 Fabs for the detection of Bet v1 and Bet v1-related allergens in natural allergen sources for allergy prevention, as well as for the standardization of natural allergen extracts produced for diagnosis and immunotherapy of birch pollen allergy. PMID:10722049

  19. 27 CFR 5.32b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... human health; or (2) Does not contain allergenic protein derived from one of the foods identified in § 5.32a(a)(1)(i), even though a major food allergen was used in production. (b) Decision on petition....

  20. 27 CFR 7.22b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... contain allergenic protein derived from one of the foods identified in § 7.22(a)(1)(i), even though a major food allergen was used in production. (b) Decision on petition. TTB will approve or deny...

  1. 27 CFR 5.32b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... human health; or (2) Does not contain allergenic protein derived from one of the foods identified in § 5.32a(a)(1)(i), even though a major food allergen was used in production. (b) Decision on petition....

  2. 27 CFR 5.32b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... human health; or (2) Does not contain allergenic protein derived from one of the foods identified in § 5.32a(a)(1)(i), even though a major food allergen was used in production. (b) Decision on petition....

  3. 27 CFR 7.22b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... contain allergenic protein derived from one of the foods identified in § 7.22(a)(1)(i), even though a major food allergen was used in production. (b) Decision on petition. TTB will approve or deny...

  4. 27 CFR 7.22b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... contain allergenic protein derived from one of the foods identified in § 7.22(a)(1)(i), even though a major food allergen was used in production. (b) Decision on petition. TTB will approve or deny...

  5. 27 CFR 7.22b - Petitions for exemption from major food allergen labeling.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... contain allergenic protein derived from one of the foods identified in § 7.22(a)(1)(i), even though a major food allergen was used in production. (b) Decision on petition. TTB will approve or deny...

  6. Common environmental allergens causing respiratory allergy in India.

    PubMed

    Singh, A B; Kumar, Pawan

    2002-03-01

    Respiratory allergy affects all age groups but the children are the worst affected by the respiratory allergy. Bioparticles from different biological sources are the main cause of allergy. Pollen grains, fungal spores, insect and other materials of biological origin form the most important allergen load in the air. For the efficient diagnosis of the allergy and its effective treatment it is very important to know about the prevalence, seasonal and annual variations of aeroallergens of the area. India being the climatically diversed country, there is diversity in the flora and fauna of different parts of the country. Atmospheric surveys carried out in different parts of India reveal that, Alanus nitida, Amarantus spinosus, Argemone mexicana Cocos nucifera, Betula utilis, Borasus flabellifer, Caraica papaya, Cedrus deodara, Cassia fistula, Parthenium, Chenopodium album, Dodonaea viscosa, Malotus phillipensis, Plantago ovata, Prosopis juliflora, Ricinus communis, Holoptelea intergifolia are the allergenically important pollens of the country. Among the fungal aeroallergens, Alternaria, Candida aibieans, Aspergillus versicolor, Aspergillus terreus, Aspergillus japonicus, Cladosporium cladosporoides, Fusarium roseum, Ganoderma lucidum,Neurospora sitophila Helminthosporium, Ustilago trtici, Uromyses are important allergens. Dust mites D. farinae, D.pteronyssinus are also important source of inhalant allergens particularly in the coastal areas of the country. Cockroaches, beetles, weevils, mosquitoes, house flies also contribute towards the aeroallergen load and are allergenically implicated. Avoidance of the indoor and outdoor aeroallergens is recommended for better management of respiratory allergy. PMID:12003301

  7. Signaling pathways activated by a protease allergen in basophils

    PubMed Central

    Rosenstein, Rachel K.; Bezbradica, Jelena S.; Yu, Shuang; Medzhitov, Ruslan

    2014-01-01

    Allergic diseases represent a significant burden in industrialized countries, but why and how the immune system responds to allergens remain largely unknown. Because many clinically significant allergens have proteolytic activity, and many helminths express proteases that are necessary for their life cycles, host mechanisms likely have evolved to detect the proteolytic activity of helminth proteases, which may be incidentally activated by protease allergens. A cysteine protease, papain, is a prototypic protease allergen that can directly activate basophils and mast cells, leading to the production of cytokines, including IL-4, characteristic of the type 2 immune response. The mechanism of papain’s immunogenic activity remains unknown. Here we have characterized the cellular response activated by papain in basophils. We find that papain-induced IL-4 production requires calcium flux and activation of PI3K and nuclear factor of activated T cells. Interestingly, papain-induced IL-4 production was dependent on the immunoreceptor tyrosine-based activation motif (ITAM) adaptor protein Fc receptor γ-chain, even though the canonical ITAM signaling was not activated by papain. Collectively, these data characterize the downstream signaling pathway activated by a protease allergen in basophils. PMID:25369937

  8. Allergen Immunotherapy (AIT): a prototype of Precision Medicine.

    PubMed

    Canonica, G W; Bachert, C; Hellings, P; Ryan, D; Valovirta, E; Wickman, M; De Beaumont, O; Bousquet, J

    2015-01-01

    Precision medicine is a medical model aiming to deliver customised healthcare - with medical decisions, practices, and/or products tailored to the individual patient informed but not directed by guidelines. Allergen immunotherapy has unique immunological rationale, since the approach is tailored to the specific IgE spectrum of an individual and modifies the natural course of the disease as it has a persistent efficacy after completion of treatment. In this perspective Allergen Immunotherapy - AIT has to be presently considered a prototype of Precision Medicine. Precise information and biomarkers provided by systems medicine and network medicine will address the discovery of Allergen immunotherapy biomarkers for (i) identification of the causes, (ii) stratification of eligible patients for AIT and (iii) the assessment of AIT efficacy. This area of medical technology is evolving rapidly and, compelemented by e-health, will change the way we practice medicine. It will help to monitor patients' disease control and data for (i) patient stratification, (ii) clinical trials, (iii) monitoring the efficacy and safety of targeted therapies which are critical for reaching an appropriate reimbursement. Biomarkers associated with e-health combined with a clinical decision support system (CDSS) will change the scope of Allergen immunotherapy. The cost/effectiveness of Allergen immunotherapy is a key issue for successful implementation. It should include the long-term benefits in the pharmaco-economic evaluation, since no other allergy treatment has this specific characteristic. AIT is the prototype of current and future precision medicine. PMID:26594303

  9. Apparel Manufacture

    NASA Technical Reports Server (NTRS)

    1995-01-01

    Marshall Space Flight Center teamed with the University of Alabama in Huntsville (UAH) in 1989 on a program involving development of advanced simulation software. Concurrently, the State of Alabama chartered UAH to conduct a technology advancement program in support of the state's apparel manufacturers. In 1992, under contract to Marshall, UAH developed an apparel-specific software package that allows manufacturers to design and analyze modules without making an actual investment -- it functions on ordinary PC equipment. By 1995, Marshall had responded to requests for the package from more than 400 companies in 36 states; some of which reported savings up to $2 million. The National Garment Company of Missouri, for example, uses the system to design and balance a modular line before committing to expensive hardware; for setting up sewing lines; and for determining the composition of a new team.

  10. Manufacturing technology

    SciTech Connect

    Leonard, J.A.; Floyd, H.L.; Goetsch, B.; Doran, L.

    1993-08-01

    This bulletin depicts current research on manufacturing technology at Sandia laboratories. An automated, adaptive process removes grit overspray from jet engine turbine blades. Advanced electronic ceramics are chemically prepared from solution for use in high- voltage varistors. Selective laser sintering automates wax casting pattern fabrication. Numerical modeling improves performance of photoresist stripper (simulation on Cray supercomputer reveals path to uniform plasma). And mathematical models help make dream of low- cost ceramic composites come true.

  11. Passive immunization with allergen-specific IgG antibodies for treatment and prevention of allergy

    PubMed Central

    Flicker, Sabine; Linhart, Birgit; Wild, Carmen; Wiedermann, Ursula; Valenta, Rudolf

    2013-01-01

    IgE antibody-mediated allergies affect more than 25% of the population worldwide. To investigate therapeutic and preventive effects of passive immunization with allergen-specific IgG antibodies on allergy in mouse models we used clinically relevant pollen allergens. In a treatment model, mice were sensitized to the major birch pollen allergen Bet v 1 and to the major grass pollen allergens, Phl p 1 and Phl p 5 and then received passive immunization with rabbit IgG antibodies specific for the sensitizing or an unrelated allergen. In a prevention model, mice obtained passive immunization with allergen-specific rabbit IgG before sensitization. Kinetics of the levels of administered IgG antibodies, effects of administered allergen-specific IgG on allergen-specific IgE reactivity, the development of IgE and IgG responses and on immediate allergic reactions were studied by ELISA, rat basophil leukaemia degranulation assays and skin testing, respectively. Treated mice showed an approximately 80% reduction of allergen-specific IgE binding and basophil degranulation which was associated with the levels of administered allergen-specific IgG antibodies. Preventive administration of allergen-specific IgG antibodies suppressed the development of allergen-specific IgE and IgG1 antibody responses as well as allergen-induced basophil degranulation and skin reactivity. Our results show that passive immunization with allergen-specific IgG antibodies is effective for treatment and prevention of allergy to clinically important pollen allergens in a mouse model and thus may pave the road for the clinical application of allergen-specific antibodies in humans. PMID:23182706

  12. Green Manufacturing

    SciTech Connect

    Patten, John

    2013-12-31

    Green Manufacturing Initiative (GMI): The initiative provides a conduit between the university and industry to facilitate cooperative research programs of mutual interest to support green (sustainable) goals and efforts. In addition to the operational savings that greener practices can bring, emerging market demands and governmental regulations are making the move to sustainable manufacturing a necessity for success. The funding supports collaborative activities among universities such as the University of Michigan, Michigan State University and Purdue University and among 40 companies to enhance economic and workforce development and provide the potential of technology transfer. WMU participants in the GMI activities included 20 faculty, over 25 students and many staff from across the College of Engineering and Applied Sciences; the College of Arts and Sciences' departments of Chemistry, Physics, Biology and Geology; the College of Business; the Environmental Research Institute; and the Environmental Studies Program. Many outside organizations also contribute to the GMI's success, including Southwest Michigan First; The Right Place of Grand Rapids, MI; Michigan Department of Environmental Quality; the Michigan Department of Energy, Labor and Economic Growth; and the Michigan Manufacturers Technical Center.

  13. 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. PMID:24215698

  14. Indoor mite allergens in patients with respiratory allergy living in Porto, Portugal.

    PubMed

    Plácido, J L; Cuesta, C; Delgado, L; da Silva, J P; Miranda, M; Ventas, P; Vaz, M

    1996-09-01

    We investigated the levels of mite allergens (Der p 1, Der f 1, Der 2, and Lep d 1) in dust samples from the homes of 59 patients with asthma, 36 sensitized to house-dust mites (HDM) and 23 to grass pollen (controls), living in Porto, northern Portugal. The relationship between exposure and sensitization to HDM and the influence of housing conditions on mite-allergen levels were also evaluated. Der p 1 (median 9.2 micrograms/g) and Der 2 (4.6 micrograms/g) were the main allergens, while Der f 1 and Lep d 1 levels were always < 1 microgram/g dust and undetectable in 11% and 47% of samples, respectively. All HDM-sensitized asthmatics were exposed to Der p 1 levels > 2 micrograms/g and their homes contained significantly higher levels of Der p 1 (median 12.5 vs 6.4 micrograms/g; P = 0.008) and Der 2 (6.2 vs 3.0 micrograms/g; P = 0.004) when compared to the control group. A significant correlation was observed between the exposure to Der p 1 and the wheal area at skin testing with the Dermatophagoides pteronyssinus (Dp) extract (P = 0.01) as well as with serum specific IgE levels to Dp (P = 0.03). Patients with higher levels of serum specific IgE (> or = 17.5 HRU/ml) were also more frequently exposed to Der p 1 levels > or = 10 micrograms/g (P = 0.002). Old homes, presence of carpets, and signs of dampness were conditions associated with significantly higher levels of mite allergens. In conclusion, we found high levels of Der p 1 and Der 2 particularly in the homes of HDM-sensitized patients and we confirm the relationship between exposure and sensitization to HDM, assessed by both in vivo and in vitro methods. In additional to a favorable outdoor climate, we found in our region housing conditions propitious to mite growth, suggesting that specific geographic characteristics must also be taken into account for the correct planning of mite-avoidance measures. PMID:8899115

  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. 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…

  17. Collaborative validation of the quantification method for suspected allergens and test of an automated data treatment.

    PubMed

    Chaintreau, Alain; Cicchetti, Esmeralda; David, Nathalie; Earls, Andy; Gimeno, Pascal; Grimaud, Béatrice; Joulain, Daniel; Kupfermann, Nikolai; Kuropka, Gryta; Saltron, Frédéric; Schippa, Christine

    2011-10-28

    Previous publications investigated different data treatment strategies for quantification of volatile suspected allergens by GC/MS. This publication presents the validation results obtained on "ready to inject" samples under reproducibility conditions following inter-laboratory ring-testing. The approach is based on the monitoring of three selected ions per analyte using two different GC capillary columns. To aid the analysts a decisional tree is used for guidance during the interpretation of the analytical results. The method is evaluated using a fragrance oil concentrate spiked with all suspected allergens to mimic the difficulty of a real sample extract or perfume oil. At the concentrations of 10 and 100mg/kg, imposed by Directive 76/768/EEC for labeling of leave-on and rinse-off cosmetics, the mean bias is +14% and -4%, respectively. The method is linear for all analytes, and the prediction intervals for each analyte have been determined. To speed up the analyst's task, an automated data treatment is also proposed. The method mean bias is slightly shifted towards negative values, but the method prediction intervals are close to that resulting from the decisional tree. PMID:21945622

  18. A study of cross-reactions between mango contact allergens and urushiol.

    PubMed

    Oka, Keiko; Saito, Fumio; Yasuhara, Tadashi; Sugimoto, Akiko

    2004-01-01

    The allergens causing mango dermatitis have long been suspected to be alk(en)yl catechols and/or alk(en)yl resorcinols on the basis of observed cross-sensitivity reactions to mango in patients known to be sensitive to poison ivy and oak (Toxicodendron spp.). Earlier, we reported the 3 resorcinol derivatives: heptadecadienylresorcinol (I), heptadecenylresorcinol (II) and pentadecylresorcinol (III); collectively named 'mangol', as mango allergens. In this study, we extracted the 1st 2 components (I and II) from the Philippine mango, adjusted them to 0.05% concentration in petrolatum and patch tested the components on 2 subjects with mango dermatitis. Both subjects reacted to I. 1 subject also elicited a weaker positive reaction to II. To investigate the cross-reaction between mangol and urushiol, we also patch tested the same subjects with urushiol. The subject sensitive to II reacted to urushiol. 6 subjects with a history of lacquer contact dermatitis and positive reactions to urushiol were similarly patch tested. 5 persons reacted to I. 2 subjects also exhibited a slower but positive reaction to II. This is the 1st report in which heptadec(adi)enyl resorcinols known to be present in mango have been shown to elicit positive patch test reactions in mango-sensitive patients. PMID:15606656

  19. Detection by real time PCR of walnut allergen coding sequences in processed foods.

    PubMed

    Linacero, Rosario; Ballesteros, Isabel; Sanchiz, Africa; Prieto, Nuria; Iniesto, Elisa; Martinez, Yolanda; Pedrosa, Mercedes M; Muzquiz, Mercedes; Cabanillas, Beatriz; Rovira, Mercè; Burbano, Carmen; Cuadrado, Carmen

    2016-07-01

    A quantitative real-time PCR (RT-PCR) method, employing novel primer sets designed on Jug r 1, Jug r 3, and Jug r 4 allergen-coding sequences, was set up and validated. Its specificity, sensitivity, and applicability were evaluated. The DNA extraction method based on CTAB-phenol-chloroform was best for walnut. RT-PCR allowed a specific and accurate amplification of allergen sequence, and the limit of detection was 2.5pg of walnut DNA. The method sensitivity and robustness were confirmed with spiked samples, and Jug r 3 primers detected up to 100mg/kg of raw walnut (LOD 0.01%, LOQ 0.05%). Thermal treatment combined with pressure (autoclaving) reduced yield and amplification (integrity and quality) of walnut DNA. High hydrostatic pressure (HHP) did not produce any effect on the walnut DNA amplification. This RT-PCR method showed greater sensitivity and reliability in the detection of walnut traces in commercial foodstuffs compared with ELISA assays. PMID:26920302

  20. Risk assessment of allergen metals in cosmetic products.

    PubMed

    Sipahi, Hande; Charehsaz, Mohammad; Güngör, Zerrin; Erdem, Onur; Soykut, Buğra; Akay, Cemal; Aydin, Ahmet

    2015-01-01

    Cosmetics are one of the most common reasons for hospital referrals with allergic contact dermatitis. Because of the increased use of cosmetics within the population and an increase in allergy cases, monitoring of heavy metals, especially allergen metals, is crucial. The aim of this study was to investigate the concentration of allergen metals, nickel (Ni), cobalt (Co), and chromium (Cr), in the most commonly used cosmetic products including mascara, eyeliner, eye shadow, lipstick, and nail polish. In addition, for safety assessment of cosmetic products, margin of safety of the metals was evaluated. Forty-eight makeup products were purchased randomly from local markets and large cosmetic stores in Istanbul, Turkey, and an atomic absorption spectrometer was used for metal content determination. Risk assessment of the investigated cosmetic products was performed by calculating the systemic exposure dosage (SED) using Scientific Committee on Consumer Safety guideline. According to the results of this investigation in all the samples tested, at least two of the allergen metals, Ni and/or Co and/or Cr were detected. Moreover, 97% of the Ni-detected products, 96% of Cr- and 54% of Co-detected products, contained over 1 μg/g of this metals, which is the suggested ultimate target value for sensitive population and thereby can be considered as the possible allergen. On the basis of the results of this study, SED of the metals was negligible; however, contact dermatitis caused by cosmetics is most probably due to the allergen metal content of the products. In conclusion, to assess the safety of the finished products, postmarketing vigilance and routine monitoring of allergen metals are very important to protect public health. PMID:26753435

  1. [Assessment of allergenicity of genetically modified food crops].

    PubMed

    Schauzu, M; Pöting, A; Rubin, D; Lampen, A

    2012-03-01

    The placing on the European Union's market of genetically modified crops requires authorization by the European Commission which is based on the proof that the derived foods are as safe as their conventional counterparts. The assessment of potential allergenicity is part of the necessary investigations recommended in the updated Guidance Document of the Scientific Panel on Genetically Modified Organisms (GMO) of the European Food Safety Authority (EFSA), which is based on internationally agreed recommendations. All genetically modified crops which so far have been authorized in the European Union were evaluated by the EFSA GMO Panel which considered it unlikely that their overall allergenicity has been altered. PMID:22373855

  2. Manufacturing technology

    SciTech Connect

    Blaedel, K L

    1998-01-01

    The mission of the Manufacturing Technology thrust area at Lawrence Livermore National Laboratory (LLNL) has been to have an adequate base of manufacturing technology, not necessarily resident at LLNL, to conduct their future business. The specific goals were (1) to develop an understanding of fundamental fabrication processes; (2) to construct general purpose process models that have wide applicability; (3) to document their findings and models in journals; (4) to transfer technology to LLNL programs, industry, and colleagues; and (5) to develop continuing relationships with the industrial and academic communities to advance their collective understanding of fabrication processes. In support of this mission, two projects were reported here, each of which explores a way to bring higher precision to the manufacturing challenges that we face over the next few years. The first, ''A Spatial-Frequency-Domain Approach to Designing a Precision Machine Tools,'' is an overall view of how they design machine tools and instruments to make or measure workpieces that are specified in terms of the spatial frequency content of the residual errors of the workpiece surface. This represents an improvement of an ''error budget,'' a design tool that saw significant development in the early 1980's, and has been in active use since then. The second project, ''Micro-Drilling of ICF Capsules,'' is an attempt to define the current state in commercial industry for drilling small holes, particularly laser-drilling. The report concludes that 1-{micro}m diameter holes cannot currently be drilled to high aspect ratios, and then defines the engineering challenges that will have to be overcome to machine holes small enough for NIF capsules.

  3. Lack of Detectable Allergenicity in Genetically Modified Maize Containing “Cry” Proteins as Compared to Native Maize Based on In Silico & In Vitro Analysis

    PubMed Central

    Mathur, Chandni; Kathuria, Pooran C.; Dahiya, Pushpa; Singh, Anand B.

    2015-01-01

    Background Genetically modified, (GM) crops with potential allergens must be evaluated for safety and endogenous IgE binding pattern compared to native variety, prior to market release. Objective To compare endogenous IgE binding proteins of three GM maize seeds containing Cry 1Ab,1Ac,1C transgenic proteins with non GM maize. Methods An integrated approach of in silico & in vitro methods was employed. Cry proteins were tested for presence of allergen sequence by FASTA in allergen databases. Biochemical assays for maize extracts were performed. Specific IgE (sIgE) and Immunoblot using food sensitized patients sera (n = 39) to non GM and GM maize antigens was performed. Results In silico approaches, confirmed for non sequence similarity of stated transgenic proteins in allergen databases. An insignificant (p> 0.05) variation in protein content between GM and non GM maize was observed. Simulated Gastric Fluid (SGF) revealed reduced number of stable protein fractions in GM then non GM maize which might be due to shift of constituent protein expression. Specific IgE values from patients showed insignificant difference in non GM and GM maize extracts. Five maize sensitized cases, recognized same 7 protein fractions of 88-28 kD as IgE bindng in both GM and non-GM maize, signifying absence of variation. Four of the reported IgE binding proteins were also found to be stable by SGF. Conclusion Cry proteins did not indicate any significant similarity of >35% in allergen databases. Immunoassays also did not identify appreciable differences in endogenous IgE binding in GM and non GM maize. PMID:25706412

  4. Exposure to multiple indoor allergens in US homes and relationship to asthma

    PubMed Central

    Salo, Päivi M.; Arbes, Samuel J.; Crockett, Patrick W.; Thorne, Peter S.; Cohn, Richard D.; Zeldin, Darryl C.

    2008-01-01

    Background The National Survey of Lead and Allergens in Housing was the first population-based study to measure indoor allergen levels in US homes. Objective We characterized the overall burden to multiple allergens and examined whether elevated allergen levels were associated with occupants’ asthma status. Methods This cross-sectional study surveyed a nationally representative sample of 831 housing units in 75 different locations throughout the US. Information was collected by questionnaire and environmental assessments. Allergen concentrations in dust samples were assessed by immunoassays. The following cut points were used to define elevated allergen levels: 10 μg/g for Der p 1, Der f 1, and Can f 1; 8 μg/g for Fel d 1; 8 U/g Bla g 1; 1.6 μg/g for mouse urinary protein; and 7 μg/g for Alternaria antigens. Allergen burden was considered high when 4 or more allergens exceeded elevated levels in any of the sampling locations. Results Exposure to multiple allergens was common in US homes. Of the surveyed homes, 51.5% had at least 6 detectable allergens and 45.8% had at least 3 allergens exceeding elevated levels. Occupants’ race, income, housing type, absence of children, and presence of smokers, pets, cockroaches, rodents and mold/moisture related problems were independent predictors of high allergen burden. Among atopics, high allergen burden increased the odds of having asthma symptoms (OR=1.81, 95% CI: 1.04-3.15). Conclusion Elevated allergen levels in the home are associated with asthma symptoms in allergic individuals. Clinical implication In allergic asthma, indoor allergen exposures play an important role in asthma exacerbations. PMID:18255132

  5. Parachute Manufacturing

    NASA Technical Reports Server (NTRS)

    2008-01-01

    Shown is the fabrication of the First Stage Main Parachute in support of Ares/CLV at the Pioneer Zodiac Facility in Mississippi in support of the Constellation/Ares project. This image is extracted from a high definition video file and is the highest resolution available

  6. Ole e 13 is the unique food allergen in olive: Structure-functional, substrates docking, and molecular allergenicity comparative analysis.

    PubMed

    Jimenez-Lopez, J C; Robles-Bolivar, P; Lopez-Valverde, F J; Lima-Cabello, E; Kotchoni, S O; Alché, J D

    2016-05-01

    Thaumatin-like proteins (TLPs) are enzymes with important functions in pathogens defense and in the response to biotic and abiotic stresses. Last identified olive allergen (Ole e 13) is a TLP, which may also importantly contribute to food allergy and cross-allergenicity to pollen allergen proteins. The goals of this study are the characterization of the structural-functionality of Ole e 13 with a focus in its catalytic mechanism, and its molecular allergenicity by extensive analysis using different molecular computer-aided approaches covering a) functional-regulatory motifs, b) comparative study of linear sequence, 2-D and 3D structural homology modeling, c) molecular docking with two different β-D-glucans, d) conservational and evolutionary analysis, e) catalytic mechanism modeling, and f) IgE-binding, B- and T-cell epitopes identification and comparison to other allergenic TLPs. Sequence comparison, structure-based features, and phylogenetic analysis identified Ole e 13 as a thaumatin-like protein. 3D structural characterization revealed a conserved overall folding among plants TLPs, with mayor differences in the acidic (catalytic) cleft. Molecular docking analysis using two β-(1,3)-glucans allowed to identify fundamental residues involved in the endo-1,3-β-glucanase activity, and defining E84 as one of the conserved residues of the TLPs responsible of the nucleophilic attack to initiate the enzymatic reaction and D107 as proton donor, thus proposing a catalytic mechanism for Ole e 13. Identification of IgE-binding, B- and T-cell epitopes may help designing strategies to improve diagnosis and immunotherapy to food allergy and cross-allergenic pollen TLPs. PMID:27017426

  7. Specific IgE in the identification of allergens in allergic rhinitis Malaysian patients.

    PubMed

    Choon-Kook, S; Teck-Soong, S L

    1995-06-01

    The specific serum IgE levels to 20 allergens were determined by enzyme immunoassay in 90 Malaysian patients with allergic rhinitis. Ninety-two percent of patients had elevated IgE to at least 1 of the allergens. The housedust mites D. pteronyssinus and D. farinae were the major allergens, elevated IgE to either allergen being present in 86% of the patients. Prick skin tests were carried out in some of the patients, housedust mites, cat fur, dog hair and shrimp were the allergens used. Close correspondence was found between IgE and prick skin tests to the mites. PMID:7488340

  8. Experiences on IGSCC crack manufacturing

    SciTech Connect

    Veron, P.

    1997-02-01

    The author presents his experience in manufacturing IGSCC realistic defects, mainly in INCONEL 600 MA Steam Generator Tubes. From that experience he extracts some knowledge about this cracking (influence of chemistry in the environment, stress state, crack growth rate, and occurrence in laboratory condition of break before leak).

  9. Cross-Reactivity between Schistosoma mansoni Antigens and the Latex Allergen Hev b 7: Putative Implication of Cross-Reactive Carbohydrate Determinants (CCDs)

    PubMed Central

    Doenhoff, Michael J.; El-Faham, Marwa; Liddell, Susan; Fuller, Heidi R.; Stanley, Ronald G.; Schramm, Gabriele; Igetei, Joseph E.

    2016-01-01

    IgG antibodies produced by rabbits immunized against S. mansoni antigens cross-reacted with aqueous soluble constituents of a variety of allergens. The antibody cross-reactivity was largely sensitive to degradation by treatment of the target antigens with sodium meta-periodate, suggesting the cross-reactivity was due to carbohydrate determinants that were common to both the schistosome and the allergens (CCDs). The reaction between the rabbit antibodies and a 43 kDa molecule in a rubber latex extract was analysed further: tandem mass spectrometry identified the latex molecule as allergen Hev b 7. Rabbit anti-schistosome IgG antibodies purified by acid-elution from solid-phase latex Hev b 7 reacted with the S. mansoni egg antigens IPSE/alpha-1 and kappa-5 and cercarial antigens SPO-1 and a fatty acid-binding protein. Moreover, purified anti-S. mansoni egg, latex cross-reactive antibodies reacted with antigenic constituents of some fruits, a result of potential relevance to the latex-fruit syndrome of allergic reactions. We propose that IgG anti-schistosome antibodies that cross-react with allergens may be able to block IgE-induced allergic reactions and thus provide a possible explanation for the hygiene hypothesis. PMID:27467385

  10. Cross-Reactivity between Schistosoma mansoni Antigens and the Latex Allergen Hev b 7: Putative Implication of Cross-Reactive Carbohydrate Determinants (CCDs).

    PubMed

    Doenhoff, Michael J; El-Faham, Marwa; Liddell, Susan; Fuller, Heidi R; Stanley, Ronald G; Schramm, Gabriele; Igetei, Joseph E

    2016-01-01

    IgG antibodies produced by rabbits immunized against S. mansoni antigens cross-reacted with aqueous soluble constituents of a variety of allergens. The antibody cross-reactivity was largely sensitive to degradation by treatment of the target antigens with sodium meta-periodate, suggesting the cross-reactivity was due to carbohydrate determinants that were common to both the schistosome and the allergens (CCDs). The reaction between the rabbit antibodies and a 43 kDa molecule in a rubber latex extract was analysed further: tandem mass spectrometry identified the latex molecule as allergen Hev b 7. Rabbit anti-schistosome IgG antibodies purified by acid-elution from solid-phase latex Hev b 7 reacted with the S. mansoni egg antigens IPSE/alpha-1 and kappa-5 and cercarial antigens SPO-1 and a fatty acid-binding protein. Moreover, purified anti-S. mansoni egg, latex cross-reactive antibodies reacted with antigenic constituents of some fruits, a result of potential relevance to the latex-fruit syndrome of allergic reactions. We propose that IgG anti-schistosome antibodies that cross-react with allergens may be able to block IgE-induced allergic reactions and thus provide a possible explanation for the hygiene hypothesis. PMID:27467385

  11. Characterization of mustard 2S albumin allergens by bottom-up, middle-down, and top-down proteomics: a consensus set of isoforms of Sin a 1.

    PubMed

    Hummel, Marlene; Wigger, Tina; Brockmeyer, Jens

    2015-03-01

    The mustard allergen Sin a 1 belongs to the 2S-albumin family of seed-storage proteins. Because of its high abundance in mustard seeds and the potential to elicit severe allergenic reactions, Sin a 1 is considered to be a major allergen in mustard. Eight Sin a 1 isoforms have been identified using DNA cloning and sequencing, and we aim in this study to thoroughly investigate sequence heterogeneity using a novel combination of bottom-up, middle-down, and top-down proteomics. The characterization of purified Sin a 1 extract shows that sequence diversity is far more pronounced than previously assumed. We identified in total 24 sequence polymorphisms including 17 yet unpublished point mutations. Using middle-down and top-down approaches on the subunit and protein level of Sin a 1, we were able to detect eight consensus isoforms of Sin a 1(including four novel isoforms), which we detect in the majority of the four different mustard samples included in this study. In addition, we provide for the first time data on relative abundance of the main Sin a 1 isoforms and identify phytic acid as a potential ligand of Sin a 1. Together, these data can form the basis for a more detailed investigation of the effect of Sin a 1 polymorphic sites on allergenicity of isoforms. PMID:25660635

  12. Allergen sensitization linked to climate and age, not to intermittent-persistent rhinitis in a cross-sectional cohort study in the (sub)tropics

    PubMed Central

    2014-01-01

    could be detected. Sensitization patterns vary with age (child HDM, adult pollen). Clinical implications of our findings are dual: only a few allergens –some region specific- cover the majority of sensitizations in (sub)tropical climate zones. This is of major importance for allergen manufacturers and immunotherapy planning. Secondly, patient selection in clinical trials should be based on the intermittent-persistent and severity classifications, rather than on the seasonal-perennial AR subtypes, especially when conducted in (sub)tropical countries. PMID:24976949

  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. PMID:23205714

  14. Molecular characterization of recombinant T1, a non-allergenic periwinkle (Catharanthus roseus) protein, with sequence similarity to the Bet v 1 plant allergen family.

    PubMed Central

    Laffer, Sylvia; Hamdi, Said; Lupinek, Christian; Sperr, Wolfgang R; Valent, Peter; Verdino, Petra; Keller, Walter; Grote, Monika; Hoffmann-Sommergruber, Karin; Scheiner, Otto; Kraft, Dietrich; Rideau, Marc; Valenta, Rudolf

    2003-01-01

    More than 25% of the population suffer from Type I allergy, an IgE-mediated hypersensitivity disease. Allergens with homology to the major birch ( Betula verrucosa ) pollen allergen, Bet v 1, belong to the most potent elicitors of IgE-mediated allergies. T1, a cytokinin-inducible cytoplasmic periwinkle ( Catharanthus roseus ) protein, with significant sequence similarity to members of the Bet v 1 plant allergen family, was expressed in Escherichia coli. Recombinant T1 (rT1) did not react with IgE antibodies from allergic patients, and failed to induce basophil histamine release and immediate-type skin reactions in Bet v 1-allergic patients. Antibodies raised against purified rT1 could be used for in situ localization of natural T1 by immunogold electron microscopy, but did not cross-react with most of the Bet v 1-related allergens. CD analysis showed significant differences regarding secondary structure and thermal denaturation behaviour between rT1 and recombinant Bet v 1, suggesting that these structural differences are responsible for the different allergenicity of the proteins. T1 represents a non-allergenic member of the Bet v 1 family that may be used to study structural requirements of allergenicity and to engineer hypo-allergenic plants by replacing Bet v 1-related allergens for primary prevention of allergy. PMID:12656672

  15. Role of Allergen Source-Derived Proteases in Sensitization via Airway Epithelial Cells

    PubMed Central

    Matsumura, Yasuhiro

    2012-01-01

    Protease activity is a characteristic common to many allergens. Allergen source-derived proteases interact with lung epithelial cells, which are now thought to play vital roles in both innate and adaptive immune responses. Allergen source-derived proteases act on airway epithelial cells to induce disruption of the tight junctions between epithelial cells, activation of protease-activated receptor-2, and the production of thymic stromal lymphopoietin. These facilitate allergen delivery across epithelial layers and enhance allergenicity or directly activate the immune system through a nonallergic mechanism. Furthermore, they cleave regulatory cell surface molecules involved in allergic reactions. Thus, allergen source-derived proteases are a potentially critical factor in the development of allergic sensitization and appear to be strongly associated with heightened allergenicity. PMID:22523502

  16. [Recombinant allergens for diagnosis and specific immunotherapy--value in pediatric patients].

    PubMed

    Couderc, Rémy; Just, Jocelyne

    2013-03-01

    Identification of culprit allergens is important for prophylactic measures and specific allergen immunotherapy (SIT). Since the late 1980s, the use of molecular cloning technology has led to a major improvement in our knowledge of epitopes involved in IgE-mediated allergy, and has also allowed in vitro production of recombinant allergens of interest for the diagnosis of allergenic sensitization. It has also improved our understanding of allergen cross-reactivity, which can be responsible for severe clinical manifestations, particularly in children with food allergy and allergic asthma. Better knowledge of molecular and cellular mechanisms of allergenic sensitization, based on the use of natural or modified recombinant allergens, has led to the development of effective SIT strategies which, in the foreseeable future, could provide genuine cure, therefore avoiding use of symptomatic therapeutics, starting very early in childhood. PMID:25163347

  17. RISK ASSESSMENT OF FOOD ALLERGENICITY BY A DATA BASE APPROACH

    EPA Science Inventory

    The overall goal of the proposal is the further development of our Structural Database of Allergenic Proteins (SDAP) (http://fermi.utmb.edu/SDAP/ 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

  18. CLINICALLY RELEVANT IGE-CROSS-REACTIVITY OF NUT ALLERGENS

    EPA Science Inventory

    All data resulting from this study will be catalogued in SDAP .This work will generate important information relating the structure/ physicochemical properties of cross-reactive IgE epitopes to clinical response, and model factors that underlie allergen recognition by the immu...

  19. 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...

  1. Effect of Non-Thermal Processing on Peanut Allergens.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Peanut allergy is on the rise, and the reason is still unclear. Previously, roasting by thermal method has been shown to increase the allergenic potency of peanuts. In this study, we determined if non-thermal methods, such as, pulsed electric fields (PEF) and pulsed UV lights (PUV) affect peanut all...

  2. Cross-reactivity of termite myosin; a potential allergen

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Myosin and myosin isoforms are common food allergens in crustaceans; such as, shrimp, lobster, and crab. Allergy to Shellfish is a prevalent and potentially long lasting disorder that can severely affect health and quality of life. Myosin and myosin isoforms of dust mites and cockroaches are simil...

  3. 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.

  4. THE ALLERGENIC POTENTIAL OF INDOOR AIR FUNGAL CONTAMINANTS

    EPA Science Inventory


    The Allergenic Potential of Indoor Air Fungal Contaminants
    Marsha D W Ward1, Michael E Viana2, Yongjoo Chung3, Najwa Haykal-Coates1, Lisa B Copeland1, Steven H Gavett1, and MaryJane K Selgrade1. 1US EPA, ORD, NHEERL, RTP, NC, USA. 2NCSU, CVM, Raleigh, NC, USA, 3 UNC, SPH,...

  5. Hydrophobic allergens from the bottom fraction membrane of Hevea brasiliensis.

    PubMed

    Mengumpun, Kesajee; Tayapiwatana, Chatchai; Hamilton, Robert G; Sangsupawanich, Pasuree; Wititsuwannakul, Rapepun

    2008-01-01

    Several proteins of rubber latex have been recognized as allergens causing immediate hypersensitivity in humans. In this study, a bottom fraction membrane (BFM) protein preparation from Hevea brasiliensis trees grown in southern Thailand was used to detect specific IgE in four groups of serum samples. The first group included 170 samples of latex glove factory workers (LGWs); group 2 consisted of the sera of 35 health care workers (HCWs) who were repeatedly exposed to powdered latex gloves; groups 3 and 4 were 31 positive and 22 negative sera, respectively, obtained from Johns Hopkins University School of Medicine, Baltimore, USA, tested for IgE to latex allergen. It was found that 56/170 (33%), 5/35 (14%), 11/31 (35.5%) and 1/22 (4.5%) samples of the LGWs, HCWs, CAP+ and CAP- groups had significant IgE to the BFM proteins, respectively. However, of all subjects only one subject of group 1 had experienced allergic morbidity consisting of eczema, conjunctivitis and asthma. The IgE of this subject bound to a 55 kDa component in the rubber latex BFM preparation. Thus, this protein may be regarded as a novel, although minor, latex allergen. Further investigation is needed to characterize the component and to pinpoint its allergenic role. PMID:19054931

  6. Categorisation of protein respiratory allergens: the case of Subtilisin.

    PubMed

    Kimber, Ian; Basketter, David A

    2014-04-01

    Characterisation of the relative sensitizing potency of protein and chemical allergens remains challenging, particularly for materials causing allergic sensitization of the respiratory tract. There nevertheless remains an appetite, for priority setting and risk management, to develop paradigms that distinguish between individual respiratory allergens according to perceptions of the hazards and risks posed to human health. One manifestation thereof is recent listing of certain respiratory allergens as Substances of Very High Concern (SVHC) under the provisions of REACH (Registration, Evaluation, Authorisation and restriction of Chemicals). Although priority setting is a laudable ambition, it is important the process is predicated on evidence-based criteria that are transparent, understood and owned. The danger is that in the absence of rigorous criteria unwanted precedents can be created, and confidence in the process is compromised. A default categorisation of sensitisers as SVHC requiring assessment under the authorisation process is not desirable. We therefore consider here the value and limitations of selective assignment of certain respiratory allergens as being SVHC. The difficulties of sustaining such designations in a sound and equitable way is discussed in the context of the challenges that exist with respect to assessment of potency, and information available regarding the effectiveness of exposure-based risk management. PMID:24534002

  7. Lipid transfer proteins and 2S albumins as allergens.

    PubMed

    Pastorello, E A; Pompei, C; Pravettoni, V; Brenna, O; Farioli, L; Trambaioli, C; Conti, A

    2001-01-01

    Plant lipid transfer proteins, a widespread family of proteins, have been recently identified as important food allergens. Their common structural features, such as eight conserved cysteines forming disulfide bridges, basic isoelectric point and high similarity in amino acid sequence, are the basis of allergic clinical cross-reactivity. This has been demonstrated for the LTP allergens of the Prunoideae subfamily, whose similarity is about 95% as demonstrated for the purified allergens of peach, apricot, plum and apple. A relevant aspect is the existence of sequence homology of LTPs of botanically unrelated foods, as demonstrated for LTPs of maize and peach. A class of food allergens of well recognized clinical importance is that of seed storage 2S albumins. They have been identified in the most diffused edible seeds and nuts, such as mustard, sesame, Brazil nut, walnut and peanut. In particular, a strong correlation between IgE-binding to these proteins and food-induced anaphylaxis has been demonstrated for Brazil nut and sesame seeds. PMID:11298008

  8. 78 FR 78368 - Allergenic Products Advisory Committee; Notice of Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-26

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration Allergenic Products Advisory Committee; Notice of Meeting AGENCY: Food and Drug Administration, HHS. ACTION: Notice. This notice announces a forthcoming meeting of a public advisory committee of the Food...

  9. 76 FR 14414 - Allergenic Products Advisory Committee; Notice of Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-16

    ... From the Federal Register Online via the Government Publishing Office ] DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration Allergenic Products Advisory Committee; Notice of Meeting AGENCY: Food and Drug Administration, HHS. ACTION: Notice. This notice announces a forthcoming meeting of a public advisory committee of the Food...

  10. Turbine Manufacture

    NASA Technical Reports Server (NTRS)

    1979-01-01

    The machinery pictured is a set of Turbodyne steam turbines which power a sugar mill at Bell Glade, Florida. A NASA-developed computer program called NASTRAN aided development of these and other turbines manufactured by Turbodyne Corporation's Steam Turbine Division, Wellsville, New York. An acronym for NASA Structural Analysis Program, NASTRAN is a predictive tool which advises development teams how a structural design will perform under service use conditions. Turbodyne uses NASTRAN to analyze the dynamic behavior of steam turbine components, achieving substantial savings in development costs. One of the most widely used spinoffs, NASTRAN is made available to private industry through NASA's Computer Software Management Information Center (COSMIC) at the University of Georgia.

  11. Influence of processing on the allergenic properties of pistachio nut assessed in vitro.

    PubMed

    Noorbakhsh, Reihaneh; Mortazavi, Seyed Ali; Sankian, Mojtaba; Shahidi, Fakhri; Maleki, Soheila J; Nasiraii, Leila Roozbeh; Falak, Reza; Sima, Hamid Reza; Varasteh, AbdolReza

    2010-09-22

    Pistachio (Pistacia vera) is a tree nut that has been reported to cause IgE-mediated allergic reactions. This study was undertaken to investigate the distinctions between different cultivars of pistachio nut and the influence of different processing on the IgE-binding capacity of whole pistachio protein extracts. The influence of different processes on allergenicity was investigated using competitive inhibition ELISA and Western blotting assays. The Western blotting results of extracts from pistachio cultivars showed no marked difference among them. The IgE-binding capacity was significantly lower for the protein extract prepared from steam-roasted than from raw and dry-roasted pistachio nuts. The results of sensory evaluation analysis and hedonic rating proved no significant differences in color, taste, flavor, and overall quality of raw, roasted, and steam-roasted pistachio nut treatments. The most significant finding of the present study was the successful reduction of IgE-binding by pistachio extracts using steam-roast processing without any significant changes in sensory quality of product. PMID:20735130

  12. Cross-reactivity between aeroallergens and food allergens.

    PubMed

    Popescu, Florin-Dan

    2015-06-26

    In patients with respiratory allergy, cross-reactivity between aeroallergens and foods may induce food allergy, symptoms ranging from oral allergy syndrome to severe anaphylaxis. Clinical entities due to IgE sensitization to cross-reactive aeroallergen and food allergen components are described for many sources of plant origin (pollen-food syndromes and associations, such as birch-apple, cypress-peach and celery-mugwort-spice syndromes, and mugwort-peach, mugwort-chamomile, mugwort-mustard, ragweed-melon-banana, goosefoot-melon associations), fungal origin (Alternaria-spinach syndrome), and invertebrate, mammalian or avian origin (mite-shrimp, cat-pork, and bird-egg syndromes). Clinical cases of allergic reactions to ingestion of food products containing pollen grains of specific plants, in patients with respiratory allergy to Asteraceae pollen, especially mugwort and ragweed, are also mentioned, for honey, royal jelly and bee polen dietary supplements, along with allergic reactions to foods contaminated with mites or fungi in patients with respiratory allergy to these aeroallergens. Medical history and diagnosis approach may be guided by the knowledge about the diverse cross-reacting allergens involved, and by the understanding of these clinical entities which may vary significantly or may be overlapping. The association between primary IgE sensitization with respiratory symptoms to inhaled allergens and food allergy due to cross-reactive allergen components is important to assess in allergy practice. The use of molecular-based diagnosis improves the understanding of clinically relevant IgE sensitization to cross-reactive allergen components from aeroallergen sources and foods. PMID:26140270

  13. Interactions between FLG mutations and allergens in atopic dermatitis.

    PubMed

    Li, Ming; Liu, Jiang-Bo; Liu, Qiang; Yao, Mianzhi; Cheng, Ruhong; Xue, Hui; Zhou, Hua; Yao, Zhirong

    2012-12-01

    Filaggrin gene (FLG) mutations and sensitization in patients with atopic dermatitis (AD) have been well documented. However, whether an interaction exists between these mutations and specific sensitization in AD patients is still unknown. The aim of the study was to explore the interaction between FLG mutations and specific sensitization in AD patients. A total of 249 AD outpatients were recruited in the current study. Skin prick tests were conducted to assess the patient's sensitization to specific allergens. FLG mutations were analyzed through comprehensive sequencing. Logistic regression analyses were conducted to determine the interactions between FLG mutations and sensitization present. The mean age of the patients was 3.5 years, and the mean age of onset of AD was 9.6 months. The mean SCORAD of the patients was 25.8. Fourteen types of mutations were identified in the FLG of 64 patients. A total of 24 (9.6 %) and 29 (11.6 %) cases were mutated with 3321delA and K4671X, respectively. Sensitization to at least one type of allergen was detected in 118 patients (47.4 %). Logistic regression analyses showed that FLG mutations presented an interaction with sensitization to peanut and did not interact with the other detected allergens among AD patients. Sensitization to peanut allergens would have an interaction with the mutation of K4671X and the combined mutations in FLG in patients with atopic dermatitis. However, sensitization to the other common allergens might not interact with FLG mutations in the development of atopic dermatitis. PMID:22903496

  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. Cross-reactivity between aeroallergens and food allergens

    PubMed Central

    Popescu, Florin-Dan

    2015-01-01

    In patients with respiratory allergy, cross-reactivity between aeroallergens and foods may induce food allergy, symptoms ranging from oral allergy syndrome to severe anaphylaxis. Clinical entities due to IgE sensitization to cross-reactive aeroallergen and food allergen components are described for many sources of plant origin (pollen-food syndromes and associations, such as birch-apple, cypress-peach and celery-mugwort-spice syndromes, and mugwort-peach, mugwort-chamomile, mugwort-mustard, ragweed-melon-banana, goosefoot-melon associations), fungal origin (Alternaria-spinach syndrome), and invertebrate, mammalian or avian origin (mite-shrimp, cat-pork, and bird-egg syndromes). Clinical cases of allergic reactions to ingestion of food products containing pollen grains of specific plants, in patients with respiratory allergy to Asteraceae pollen, especially mugwort and ragweed, are also mentioned, for honey, royal jelly and bee polen dietary supplements, along with allergic reactions to foods contaminated with mites or fungi in patients with respiratory allergy to these aeroallergens. Medical history and diagnosis approach may be guided by the knowledge about the diverse cross-reacting allergens involved, and by the understanding of these clinical entities which may vary significantly or may be overlapping. The association between primary IgE sensitization with respiratory symptoms to inhaled allergens and food allergy due to cross-reactive allergen components is important to assess in allergy practice. The use of molecular-based diagnosis improves the understanding of clinically relevant IgE sensitization to cross-reactive allergen components from aeroallergen sources and foods. PMID:26140270

  16. Maize pollen is an important allergen in occupationally exposed workers

    PubMed Central

    2011-01-01

    Background The work- or environmental-related type I sensitization to maize pollen is hardly investigated. We sought to determine the prevalence of sensitization to maize pollen among exposed workers and to identify the eliciting allergens. Methods In July 2010, 8 out of 11 subjects were examined who were repeatedly exposed to maize pollen by pollinating maize during their work in a biological research department. All 8 filled in a questionnaire and underwent skin prick testing (SPT) and immune-specific analyses. Results 5 out of the 8 exposed subjects had repeatedly suffered for at least several weeks from rhinitis, 4 from conjunctivitis, 4 from urticaria, and 2 from shortness of breath upon occupational exposure to maize pollen. All symptomatic workers had specific IgE antibodies against maize pollen (CAP class ≥ 1). Interestingly, 4 of the 5 maize pollen-allergic subjects, but none of the 3 asymptomatic exposed workers had IgE antibodies specific for grass pollen. All but one of the maize pollen-allergic subjects had suffered from allergic grass pollen-related symptoms for 6 to 11 years before job-related exposure to maize pollen. Lung function testing was normal in all cases. In immunoblot analyses, the allergenic components could be identified as Zea m 1 and Zea m 13. The reactivity is mostly caused by cross-reactivity to the homologous allergens in temperate grass pollen. Two sera responded to Zea m 3, but interestingly not to the corresponding timothy allergen indicating maize-specific IgE reactivity. Conclusion The present data suggest that subjects pollinating maize are at high risk of developing an allergy to maize pollen as a so far underestimated source of occupational allergens. For the screening of patients with suspected maize pollen sensitization, the determination of IgE antibodies specific for maize pollen is suitable. PMID:22165847

  17. Effect on quality of life of the mixed house dust mite/weed pollen extract immunotherapy

    PubMed Central

    Li, Lisha

    2016-01-01

    Background Although many patients with allergic rhinitis have symptoms due to sensitization to more than one kind of allergens, and mixed allergen extracts are widely used for immunotherapy, there are few published trials. Objective Our study aimed to evaluate the effect of multiple-allergen immunotherapy on improving the symptoms and quality of life of allergic rhinitis patients. Methods We performed a 1-year single-center observation study of subcutaneous immunotherapy using house dust mite extract (n = 12), weed pollen extract (n = 21), or mixed house dust mite/weed pollen extract (n = 11) in 44 allergic rhinitis patients. All the allergens responsible for the symptom of each patient were included in his immunotherapy. Symptom score, medication score, and quality of life of the patients were evaluated before and after 1-year immunotherapy. Quality of life was evaluated with the Rhinoconjunctivitis Quality of Life Questionnaire. Results In all 3 groups receiving subcutaneous immunotherapy, significant improvement of symptom score, medication score, and quality of life was found vs. baseline at 1 year, irrespective of the allergen used. In the weed pollen season, the changes of quality of life questionnaire score after 1-year treatment were not significantly different between the weed pollen group (1.55 ± 1.24) and the mixed house dust mite/weed pollen group (1.14 ± 1.01). The same happened in the nonpollen seasons, during which dust mite immunotherapy (1.23 ± 1.63) and mixed immunotherapy (0.60 ± 0.47) did not show significantly different effect on the quality of life. Conclusion The multiple-allergen immunotherapy might be effective in polysensitized allergic rhinitis patients, and could improve their quality of life. Our result did not show significant difference between the effects of multiple-allergen immunotherapy and mono-allergen immunotherapy. PMID:27489789

  18. Histamine release in skin monitored with the microdialysis technique does not correlate with the weal size induced by cow allergen.

    PubMed

    Horsmanheimo, L; Harvima, I T; Harvima, R J; Ylönen, J; Naukkarinen, A; Horsmanheimo, M

    1996-01-01

    The purpose of this study was to monitor histamine release in immediate-type hypersensitivity reactions in the skin of 10 atopic patients, sensitive to cow, by using the microdialysis technique. Three healthy subjects, without any atopic features or background, served as the control group. The probe inserted into the forearm dermal skin was perfused with isotonic saline solution. Samples were collected at 15-min intervals. After the first allergen challenge of four prick tests close to the probe with cow allergen extract, the skin was similarly repricked again in five patients and three healthy subjects, and in five other patients, 25 microliters of 10 mumol/l substance P (SP) was injected intracutaneously. The samples were analysed for histamine by radioenzyme assay. The patients were clinically evaluated for allergic symptoms, prick- and scratch-patch test reactivity and for serum cow-specific, and total, IgE levels. The baseline histamine concentration was 7.5 +/- 4.0 nmol/l (mean +/- standard deviation: SD; n = 10). After the allergen challenge, the histamine concentration in the consecutive samples was 11.9 +/- 11.0 nmol/l, 91.1 +/- 127.3 nmol/l, 61.0 +/- 94.2 nmol/l and 33.7 +/- 53.7 nmol/l. The peak concentration was detected in the 15-30 min fraction, and it varied between 0 and 406 nmol/l regardless of the weal size. The second allergen challenge was unable to induce marked additional histamine release, but SP induced extensive histamine liberation in those patients who did not exhibit histamine release during the preceding prick tests. In three healthy subjects, the baseline histamine concentration was 6.2 +/- 3.9 nmol/l. After the allergen challenge, no additional histamine liberation could be measured. Surprisingly, the histamine release was not related to the size of the cow-induced weal nor was it related to any specific allergic symptoms or IgE levels. The results suggest that, in some patients, mast cell mediators other than histamine play a

  19. Low-flow, long-term air sampling under normal domestic activity to measure house dust mite and cockroach allergens.

    PubMed

    Park, Jung Won; Kim, Cheol Woo; Kang, Dae Bong; Lee, In Young; Choi, Soo Young; Yong, Tai-Soon; Shin, Dong Cheon; Kim, Kyu Earn; Hong, Chein-Soo

    2002-01-01

    Successful applications of air sampling for the quantification of exposure to indoor allergens have been reported, but its efficiency is still controversial. We evaluated whether the low-flow, long-term air sampling in normal domestic activity conditions can quantify the exposure of house dust mites (HDM) and cockroaches (CR) allergens or not. Airborne Der f 1 and Bla g 1 were captured with a personal air sampler in 25 bedrooms during normal domestic activity. Quantification of the major allergens in the reservoir dust and the extraction of the air sampler filters were done with two-site ELISA kits. Airborne Der f 1 was measured above the threshold level of detection in 15 houses (60%). Detection rate of airborne Der f 1 was significantly higher in those houses where D. farinae was microscopically found in the reservoir dusts (76.5% vs. 25%, chi 2 = 6.0, p = 0.014). Airborne Der f 1 was more frequently detected in the houses with higher Der f 1 (> or = 10 micrograms/g dust) in bedding reservoir dust than the other group (91% vs. 35.7%, chi 2 = 7.819, p = 0.005), and the median value of airborne Der f 1 was also significantly higher in that group (14.0 pg/m3 vs. below detection limit, p = 0.002). Airborne Der f 1 was significantly correlated with Der f 1 in bedding reservoir dust (r = 0.591, p < 0.01). Airborne Bla g 1 was measured with ELISA in 16 houses (64%), and it was more frequently detected in the houses where the CRs were captured by adhesive traps (91% vs. 57%, chi 2 = 3,484, p = 0.06). The median concentration of Bla g 1 in the filter was also higher in the houses with captured CRs (0.12 vs. 0.05 mU/m3, p = 0.06), but the level of Bla g 1 did not correlate with that of the bedding dusts or the floor dusts of kitchen. These results suggested that airborne HDM or CR allergens could be measured by low-flow, long-term air sampling, and that it might be one of appropriate modalities for evaluating personal exposure to HDM and CR allergens. PMID:12926189

  20. Identification of Italian cypress (Cupressus sempervirens) pollen allergen Cup s 3 using homology and cross-reactivity

    PubMed Central

    Togawa, Akihisa; Panzani, Raphael C.; Garza, Maritza A.; Kishikawa, Reiko; Goldblum, Randall M.; Midoro-Horiuti, Terumi

    2008-01-01

    Background The prevalence of seasonal allergic diseases of the upper airways is increasing in industrialized countries. The Cupressaceae are important causes of pollinosis, particularly in Europe. Objective To determine whether the pollen from Cupressus sempervirens (Italian cypress) contains a pathogenesis-related group 5 (PR-5) protein, similar to that found in other allergenic Cupressaceae pollens. Methods Messenger RNA was purified from Italian cypress pollen, and complementary DNA (cDNA) was synthesized. cDNAs for PR-5 proteins were amplified by polymerase chain reaction and extended by rapid amplification of cDNA ends methods. Recombinant Cup s 3 was expressed in Escherichia coli as a fusion protein. Inhibition enzyme-linked immunosorbent assays