Fungal protein MGL_1304 in sweat is an allergen for atopic dermatitis patients

• Published in: Journal of allergy and clinical immunology 2013-09, Vol.132 (3), p.608-615.e4
• Main Authors: Hiragun, Takaaki, MD, PhD, Ishii, Kaori, MSc, Hiragun, Makiko, MD, Suzuki, Hidenori, PhD, Kan, Takanobu, MD, Mihara, Shoji, MD, PhD, Yanase, Yuhki, PhD, Bartels, Joachim, PhD, Schröder, Jens-M., MD, PhD, Hide, Michihiro, MD, PhD
• Format: Article
• Language: English
• Subjects: Adolescent; Adult; Allergens - immunology; Allergic diseases; Allergies; Allergy and Immunology; Antigens; atopic dermatitis; Basophils - drug effects; Basophils - immunology; Biological and medical sciences; Cell Line; Cells, Cultured; Children & youth; Cloning; Deoxyribonucleic acid; Dermatitis, Atopic - immunology; DNA; DNA polymerase; Female; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Fungal Proteins - genetics; Fungal Proteins - immunology; Histamine; histamine release; Histamine Release - drug effects; Humans; IgE; Immunization; Immunoglobulin E - blood; Immunopathology; Interleukin-4 - immunology; Malassezia; Malassezia - immunology; Malassezia globosa; Male; Mass spectrometry; Mast Cells - immunology; Medical sciences; Proteins; Recombinant Proteins - pharmacology; Sarcoidosis. Granulomatous diseases of unproved etiology. Connective tissue diseases. Elastic tissue diseases. Vasculitis; Skin; Skin allergic diseases. Stinging insect allergies; Studies; Sweat; Sweat - immunology; Young Adult
• ISSN: 0091-6749; 1097-6825
• DOI: 10.1016/j.jaci.2013.03.047

Background Sweat is a major aggravating factor of atopic dermatitis (AD) and approximately 80% of patients with AD show type I hypersensitivity against sweat. Objective To identify and characterize an antigen in sweat that induces histamine release from basophils of patients with AD. Methods Basophil histamine-releasing activity in sweat was purified by a combination of chromatographies, and proteins were analyzed with mass spectrometry. Recombinant proteins of the sweat antigen were generated, and their biological characteristics were studied by immunoblots, histamine release tests, and neutralization assays. Results We identified a fungal protein, MGL_1304, derived from Malassezia globosa ( M globosa ) in the purified sweat antigen. Recombinant MGL_1304 induced histamine release from basophils of most of the patients with AD, in accordance with the semi-purified sweat antigen. Moreover, recombinant MGL_1304 abolished the binding of serum IgE of patients with AD to the semi-purified sweat antigen, or vice versa in immunoblot analysis, and attenuated the sensitization of RBL-48 mast cells expressing human FcɛRI by serum IgE. Studies of truncated mutants of MGL_1304 indicated that IgE of patients with AD recognized the conformational structure of MGL_1304 rather than short peptide sequences. Western blot analysis of the whole lysate, the culture supernatant of M globosa , and the semi-purified sweat antigen showed that MGL_1304 was produced as a minor immunological antigen of M globosa with posttranslational modification, cleaved, and secreted as a 17-kDa major histamine-releasing sweat antigen. Conclusion MGL_1304 is a major allergen in human sweat and could cause type I allergy in patients with AD.