Fungal protein MGL_1304 in sweat is an allergen for atopic dermatitis patients Takaaki Hiragun, MD, PhD, Kaori Ishii, MSc, Makiko Hiragun, MD, Hidenori.

Slides:

Advertisements

Similar presentations

Secretory IgA induces antigen-independent eosinophil survival and cytokine production without inducing effector functions Kathleen R. Bartemes, BA, Kate.

Advertisements

Napins, 2S albumins, are major allergens in oilseed rape and turnip rape Tuija J. Puumalainen, MSc, Sanna Poikonen, MD, Annika Kotovuori, MSc, Kirsi Vaali,

Cor a 1–reactive T cells and IgE are predominantly cross-reactive to Bet v 1 in patients with birch pollen–associated food allergy to hazelnut Claudia.

Kuan-Wei Chen, PhD, Katharina Blatt, MSc, Wayne R

Identification and isolation of a Fel d 1–like molecule as a major rabbit allergen Christiane Hilger, PhD, Stéphanie Kler, MSc, Karthik Arumugam, PhD,

Exploring the repertoire of IgE-binding self-antigens associated with atopic eczema Sabine Zeller, MSc, Claudio Rhyner, PhD, Norbert Meyer, MD, Peter.

Liping Xie, MD, John T. Schroeder, PhD, Jacqueline M

Identification and gene cloning of a new major allergen Cha o 3 from Chamaecyparis obtusa (Japanese cypress) pollen Toshihiro Osada, MSc, Takafumi Harada,

Reduced IFN-γ receptor expression and attenuated IFN-γ response by dendritic cells in patients with atopic dermatitis Eva Gros, MSc, Susanne Petzold,

Differential cytokine induction by the human skin–associated autoallergen thioredoxin in sensitized patients with atopic dermatitis and healthy control.

Cor a 1–reactive T cells and IgE are predominantly cross-reactive to Bet v 1 in patients with birch pollen–associated food allergy to hazelnut Claudia.

Development and characterization of a recombinant, hypoallergenic, peptide-based vaccine for grass pollen allergy Margarete Focke-Tejkl, PhD, Milena.

Blocking antibodies induced by immunization with a hypoallergenic parvalbumin mutant reduce allergic symptoms in a mouse model of fish allergy Raphaela.

Skin test diagnosis of grass pollen allergy with a recombinant hybrid molecule Carine Metz-Favre, MD, Birgit Linhart, PhD, Margarete Focke-Tejkl, PhD,

Increased expression of nuclear factor of activated T cells 1 drives IL-9–mediated allergic asthma Sonja Koch, PhD, Anna Graser, PhD, Hooman Mirzakhani,

Identification of the cysteine protease Amb a 11 as a novel major allergen from short ragweed Julien Bouley, PhD, Rachel Groeme, MSc, Maxime Le Mignon,

Act d 12 and Act d 13: Two novel, masked, relevant allergens in kiwifruit seeds Sofía Sirvent, PhD, Belén Cantó, BSc, Javier Cuesta-Herranz, MD, PhD,

Assessing basophil activation by using flow cytometry and mass cytometry in blood stored 24 hours before analysis Kaori Mukai, PhD, Nicolas Gaudenzio,

Correlation of IgE/IgG4 milk epitopes and affinity of milk-specific IgE antibodies with different phenotypes of clinical milk allergy Julie Wang, MD,

Molecular cloning, expression, and characterization of a major 38-kd cochineal allergen Yoko Ohgiya, MS, Fumihiro Arakawa, MS, Hiroshi Akiyama, PhD, Yasuo.

Julie Wang, MD, James H. Godbold, PhD, Hugh A. Sampson, MD

Allergen-induced IgE-dependent gut inflammation in a human PBMC–engrafted murine model of allergy Benno Weigmann, PhD, Nadja Schughart, MSc, Christian.

Malassezia sympodialis thioredoxin–specific T cells are highly cross-reactive to human thioredoxin in atopic dermatitis Hari Balaji, MSc, Annice Heratizadeh,

Birch pollen immunotherapy results in long-term loss of Bet v 1–specific TH2 responses, transient TR1 activation, and synthesis of IgE-blocking antibodies

Rapid and comprehensive discovery of unreported shellfish allergens using large-scale transcriptomic and proteomic resources Roni Nugraha, MSc, Sandip.

Functional analysis of protective IL1RL1 variants associated with asthma risk Vladimir Ramirez-Carrozzi, PhD, Amy Dressen, MS, Patrick Lupardus, PhD,

In vivo allergenic activity of a hypoallergenic mutant of the major fish allergen Cyp c 1 evaluated by means of skin testing Nikolaos Douladiris, MD,

Toll-like receptor 9 suppression in plasmacytoid dendritic cells after IgE-dependent activation is mediated by autocrine TNF-α John T. Schroeder, PhD,

Staphylococcus aureus fibronectin-binding protein specifically binds IgE from patients with atopic dermatitis and requires antigen presentation for cellular.

Kathleen R. Bartemes, BA, Gail M. Kephart, BS, Stephanie J

CD94/NKG2C is a killer effector molecule in patients with Stevens-Johnson syndrome and toxic epidermal necrolysis Esther Morel, PhD, Salvador Escamochero,

Interleukin receptor-associated kinase-4 deficiency impairs Toll-like receptor–dependent innate antiviral immune responses Douglas R. McDonald, MD, PhD,

Allergen microarray detects high prevalence of asymptomatic IgE sensitizations to tropical pollen-derived carbohydrates Clarissa R. Cabauatan, PhD, Christian.

Estrogen increases the severity of anaphylaxis in female mice through enhanced endothelial nitric oxide synthase expression and nitric oxide production

Fibronectin is a TH1-specific molecule in human subjects

ImmunoCAP cellulose displays cross-reactive carbohydrate determinant (CCD) epitopes and can cause false-positive test results in patients with high anti-CCD.

Protein unfolding strongly modulates the allergenicity and immunogenicity of Pru p 3, the major peach allergen Masako Toda, PhD, Gerald Reese, PhD, Gabriele.

Suppression of the basophil response to allergen during treatment with omalizumab is dependent on 2 competing factors Donald W. MacGlashan, MD, PhD,

Thymic stromal lymphopoietin–activated invariant natural killer T cells trigger an innate allergic immune response in atopic dermatitis Wen Hao Wu, PhD,

IL-17 in atopic eczema: Linking allergen-specific adaptive and microbial-triggered innate immune response Kilian Eyerich, MD, Davide Pennino, PhD, Claudia.

Fibronectin is a TH1-specific molecule in human subjects

Genetically engineered hybrid proteins from Parietaria judaica pollen for allergen- specific immunotherapy Roberto González-Rioja, PhD, Ignacio Ibarrola,

A hypoallergenic variant of Der p 1 as a candidate for mite allergy vaccines David Walgraffe, PhD, Christel Mattéotti, PhD, Mohamed el Bakkoury, PhD,

Kerstin Bauermeister, MSc, Barbara K

Individual IL-3 priming is crucial for consistent in vitro activation of donor basophils in patients with chronic urticaria Thomas Gentinetta, MSc, Tatjana.

Staphylococcal exotoxins are strong inducers of IL-22: A potential role in atopic dermatitis Margarete Niebuhr, MD, Helena Scharonow, MS, Merle Gathmann,

Peanut T-cell epitope discovery: Ara h 1

Kuan-Wei Chen, PhD, Katharina Blatt, MSc, Wayne R

Analysis of the structure and allergenicity of recombinant pro- and mature Der p 1 and Der f 1: Major conformational IgE epitopes blocked by prodomains

IL-17E upregulates the expression of proinflammatory cytokines in lung fibroblasts Séverine Létuvé, PhD, Stéphane Lajoie-Kadoch, MSc, Séverine Audusseau,

A major allergen gene-fusion protein for potential usage in allergen-specific immunotherapy Fatimah Kussebi, MD, Fariba Karamloo, PhD, Claudio Rhyner,

Structural and immunologic cross-reactivity among filarial and mite tropomyosin: Implications for the hygiene hypothesis Helton C. Santiago, MD, PhD,

Association between specific timothy grass antigens and changes in TH1- and TH2-cell responses following specific immunotherapy Véronique Schulten, PhD,

A hypoallergenic cat vaccine based on Fel d 1–derived peptides fused to hepatitis B PreS Katarzyna Niespodziana, MSc, Margarete Focke-Tejkl, PhD, Birgit.

Sara Paveglio, PhD, MS, Erin Bennett, MS, Kelly L. Hawley, PhD, Adam P

Liping Xie, MD, John T. Schroeder, PhD, Jacqueline M

Does “autoreactivity” play a role in atopic dermatitis?

Allergy to human seminal fluid: Cross-reactivity with dog dander

Expression of chemokines and chemokine receptors in lesional and nonlesional upper skin of patients with atopic dermatitis Eva Gros, MSc, Caroline Bussmann,

Cloning, sequencing, and recombinant production of Sin a 2, an allergenic 11S globulin from yellow mustard seeds Oscar Palomares, PhD, Andrea Vereda,

CCL17/thymus and activation-regulated chemokine induces calcitonin gene–related peptide in human airway epithelial cells through CCR4 Kandace Bonner,

Profilin (Che a 2) and polcalcin (Che a 3) are relevant allergens of Chenopodium album pollen: Isolation, amino acid sequences, and immunologic properties

Teresa Asensio, MD, Jesus F

Identification of wheat gliadins as an allergen family related to baker's asthma Cordula Bittner, MD, Britta Grassau, MS, Karsten Frenzel, PhD, Xaver.

Bleomycin hydrolase downregulation in lesional skin of adult atopic dermatitis patients is independent of FLG gene mutations Laurence Pellerin, PhD,

IgE sensitization profiles toward green and gold kiwifruits differ among patients allergic to kiwifruit from 3 European countries Merima Bublin, MSc,

Carrier-bound, nonallergenic Ole e 1 peptides for vaccination against olive pollen allergy Teresa E. Twaroch, MSc, Margit Focke, PhD, Vera Civaj, Milena.

The effects of gastric digestion on codfish allergenicity

Allergy multivaccines created by DNA shuffling of tree pollen allergens Michael Wallner, PhD, Angelika Stöcklinger, MSc, Theresa Thalhamer, MSc, Barbara.

CCL17/thymus and activation-regulated chemokine induces calcitonin gene–related peptide in human airway epithelial cells through CCR4 Kandace Bonner,

Presentation transcript:

Fungal protein MGL_1304 in sweat is an allergen for atopic dermatitis patients Takaaki Hiragun, MD, PhD, Kaori Ishii, MSc, Makiko Hiragun, MD, Hidenori Suzuki, PhD, Takanobu Kan, MD, Shoji Mihara, MD, PhD, Yuhki Yanase, PhD, Joachim Bartels, PhD, Jens-M. Schröder, MD, PhD, Michihiro Hide, MD, PhD Journal of Allergy and Clinical Immunology Volume 132, Issue 3, Pages 608-615.e4 (September 2013) DOI: 10.1016/j.jaci.2013.03.047 Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Identification of MGL_1304 in human sweat. QRX was purified with an Aqua 5μ-C18-200Å column (A) and with a Jupiter 5μ-C18-300Å HPLC column (B) based on histamine-releasing activity (black bars in lower panels). The fraction indicated by arrows in Fig 1, A was further purified in Fig 1, B. The peptide sequence identified by mass spectrometry is shown in Fig 1, B. Journal of Allergy and Clinical Immunology 2013 132, 608-615.e4DOI: (10.1016/j.jaci.2013.03.047) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 rMGL_1304 induces histamine release from basophils of patients with AD. A, TF, TF-rMGL_1304, and rMGL_1304 were stained by CBB (left), or blotted with anti-His (center) and AD serum (right). B, Histamine release in response to rMGL_1304 from basophils of patients with AD and of healthy controls (HC). C, Supernatants of COS7 cells transfected with mock, MGL_1304, or Der f 1 were blotted with anti-Myc. D, Plot of histamine release from basophils of each patient with AD induced by QR and rMGL_1304. CBB, Coomassie brilliant blue. Journal of Allergy and Clinical Immunology 2013 132, 608-615.e4DOI: (10.1016/j.jaci.2013.03.047) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Correlation between histamine release against MGL_1304 and specific IgE against Malassezia and Candida. Plot of histamine release from basophils of each patient with AD induced by rMGL_1304 and specific IgE against Malassezia (A) and Candida (B) quantified by ImmunoCAP. Journal of Allergy and Clinical Immunology 2013 132, 608-615.e4DOI: (10.1016/j.jaci.2013.03.047) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 rMGL_1304 captures immunoreactive IgE against the semi-purified sweat antigen from sera of patients with AD. A, Antigens (rDer f 1, QRX, and rMGL_1304) were blotted with sera of patients with AD, sera pretreated with rMGL_1304, or sera pretreated with QRX. B, RBL-48 cells were sensitized by sera treated with TF or TF-MGL_1304, and then stimulated with anti-IgE, QR, or the mite extract (Mite-Df). Data were expressed as mean ± SEM of net release of β-hexosaminidase (%) in 3 independent experiments done in duplicate. Journal of Allergy and Clinical Immunology 2013 132, 608-615.e4DOI: (10.1016/j.jaci.2013.03.047) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 IgE of patients with AD recognizes the conformational structure of MGL_1304. A, A schematic figure of MGL_1304 mutants. The density of dots of bars indicated the histamine-releasing activity and IgE-binding ability. The bars highlighted by double lines represent immunoreactivity to Smith-2. B and E, The truncated proteins were blotted with anti-His, sera from patients with AD, or Smith-2. C and D, Histamine release from basophils with rMGL_1304 mutants that lack amino acids in the N-terminus (Fig 5, C) or the C-terminus (Fig 5, D). Journal of Allergy and Clinical Immunology 2013 132, 608-615.e4DOI: (10.1016/j.jaci.2013.03.047) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 MGL_1304 is cleaved and secreted into sweat from M globosa. A, Histamine-releasing activity was purified from sweat (QRX), culture supernatants (M. sup. [purified]), and lysates (M. lys. [purified]) of M globosa. The purified proteins were blotted with Smith-2. B, The whole lysate (M. lys. [whole]) and whole culture supernatant (M. sup. [whole]) of M globosa, and QRX were blotted by sera of patients with AD or sera pretreated with rMGL_1304. Journal of Allergy and Clinical Immunology 2013 132, 608-615.e4DOI: (10.1016/j.jaci.2013.03.047) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 7 MGL_1304 induces degranulation of LAD-2 cells and cytokine production of human basophils. A, LAD-2 cells were sensitized with sera of HC and patients with AD, and then stimulated with MGL_1304 purified from sweat. The release percent of β-hexosaminidase was calculated from 3 independent experiments (mean ± SEM). B, Purified human basophils from AD were stimulated with MGL_1304 from different sources. The released IL-4 level was measured in triplicate (mean ± SEM). Similar results were obtained in 2 independent experiments with basophils of different donors. HC, Healthy controls. Journal of Allergy and Clinical Immunology 2013 132, 608-615.e4DOI: (10.1016/j.jaci.2013.03.047) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 Summary of mass spectrometric analysis. Mass spectrometric amino acid sequence analysis of the protein corresponding to the histamine-release activity containing UV-absorbing peak (Fig 1) revealed a partial peptide sequence, HKMSVGDSESPGNMRSFCTKPYSSK, which by Blast search was revealed to be identical to 51-75 of amino acid sequence of MGL_1304 from Malassezia globosa. Journal of Allergy and Clinical Immunology 2013 132, 608-615.e4DOI: (10.1016/j.jaci.2013.03.047) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 HRTs by the use of rMGL_1304 mutants. Histamine release with mutant rMGL_1304 proteins that lack amino acids in the N-terminus (A and B) or the C-terminus (C and D). Each panel shows independent data for a patient with AD performed in duplicate. Journal of Allergy and Clinical Immunology 2013 132, 608-615.e4DOI: (10.1016/j.jaci.2013.03.047) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E3 Immunoreactive IgE against the semi-purified sweat antigen does not cross-react with hMnSOD. RBL-48 cells were sensitized with serum of a patient with AD that was pretreated with TF, TF-rMGL_1304, or TF-rhMnSOD, and then stimulated with anti-IgE, QR, or the mite extract (Mite-Df), respectively. Data were expressed as mean ± SEM of net release of β-hexosaminidase (%) performed in triplicate. Journal of Allergy and Clinical Immunology 2013 132, 608-615.e4DOI: (10.1016/j.jaci.2013.03.047) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions