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

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Exploring the repertoire of IgE-binding self-antigens associated with atopic eczema  Sabine Zeller, MSc, Claudio Rhyner, PhD, Norbert Meyer, MD, Peter Schmid-Grendelmeier, MD, Cezmi A. Akdis, MD, Reto Crameri, PhD  Journal of Allergy and Clinical Immunology  Volume 124, Issue 2, Pages 278-285.e7 (August 2009) DOI: 10.1016/j.jaci.2009.05.015 Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 A, Recombinant proteins (2.5 μg) were separated by means of SDS-PAGE and stained with Coomassie blue. B, Specific IgE binding of the recombinant self-antigens analyzed by means of Western blotting with sera from different patients with AE. Lane 1, Actin-α; lane 2, tubulin-α; lane 3, eIF6; lane 4, HLA-DR-α; lane 5, RP1; lane 6, cyclophilin B; lane 7, thioredoxin; lane 8, molecular weight standard. Journal of Allergy and Clinical Immunology 2009 124, 278-285.e7DOI: (10.1016/j.jaci.2009.05.015) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 IgE specific for human self-antigens. Specific IgE against recombinant self-antigens determined by means of ELISA in sera from 71 patients with AE (solid diamonds, upper panel), 12 patients with psoriasis (open diamonds, middle panel), and 24 healthy control subjects (open squares, lower panel). The cutoff values for positive results are highlighted in gray and correspond to 3 times the mean of the healthy control group. Journal of Allergy and Clinical Immunology 2009 124, 278-285.e7DOI: (10.1016/j.jaci.2009.05.015) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 IgG4 specific for human eIF6. A, Specific IgG4 against recombinant eIF6 determined by means of ELISA in sera from 8 healthy control subjects (open squares, HC), 12 patients with psoriasis (open diamonds, P), and 16 patients with AE (solid diamonds). B, Correlation between eIF6-specific IgG4 and IgE analyzed by means of ELISA in sera from 16 patients with AE and 8 healthy control subjects. The Spearman rank correlation coefficient was calculated. Journal of Allergy and Clinical Immunology 2009 124, 278-285.e7DOI: (10.1016/j.jaci.2009.05.015) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Activation of basophils. Numbers of activated basophils were analyzed based on the expression of CD203c and CD63 after stimulation with recombinant self-antigens, NaCl, or anti-IgE. A, Results of 3 representative patients with AE are shown. B, Self-antigen induced activation of basophils from blood of patients with AE with self-antigen-specific IgE (upper panel), patients with AE without self-antigen–specific IgE (middle panel), and healthy control subjects (lower panel). Journal of Allergy and Clinical Immunology 2009 124, 278-285.e7DOI: (10.1016/j.jaci.2009.05.015) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Levels of IgG (A), IgG1 (C), IgG3 (E), and IgA (G) specific for recombinant eIF6 were analyzed by means of ELISA in sera from 16 patients with AE, 12 patients with psoriasis (P), and 8 healthy control subjects (HC). The correlation with eIF6-specific IgE levels is shown (B, D, F, and H). Spearman rank correlation coefficients were calculated. Journal of Allergy and Clinical Immunology 2009 124, 278-285.e7DOI: (10.1016/j.jaci.2009.05.015) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions