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Tissue transglutaminase generates deamidated epitopes on gluten, increasing reactivity with hydrolyzed wheat protein–sensitized IgE  Ryosuke Nakamura,

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Presentation on theme: "Tissue transglutaminase generates deamidated epitopes on gluten, increasing reactivity with hydrolyzed wheat protein–sensitized IgE  Ryosuke Nakamura,"— Presentation transcript:

1 Tissue transglutaminase generates deamidated epitopes on gluten, increasing reactivity with hydrolyzed wheat protein–sensitized IgE  Ryosuke Nakamura, PhD, Rika Nakamura, PhD, Shinobu Sakai, PhD, Reiko Adachi, PhD, Akiko Hachisuka, PhD, Atsuo Urisu, MD, PhD, Yuma Fukutomi, MD, PhD, Reiko Teshima, PhD  Journal of Allergy and Clinical Immunology  Volume 132, Issue 6, Pages e4 (December 2013) DOI: /j.jaci Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Typical EXiLE responses of serum IgE from HWP-sensitized subject (A) and patient with PedWA (B) to various treated glutens and EXiLE responses of all subjects (HWP-sensitized subjects, patients with PedWA, and healthy subjects) presented in Table E1 to gluten, gluten treated with tTG (tTG-Glu), and Glp19S (C). Values are presented as mean ± SEMs. **, ††P < .01, n-way ANOVA and the post hoc test with the Bonferroni correction. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 IgE ELISA inhibition with antigen-coated beads. Diluted sera from HWP-sensitized subjects and patients with PedWA were treated with antigen-coated beads for 2 hours, followed by centrifugation. The Glp19S-specific (A) or tTG-Glu–specific (B) IgE remaining in the supernatants was measured by using ELISA. Data are means of triplicate measurements and are shown as percentages of binding inhibition. P values of the Friedman test are shown. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig E1 Immunoblot of treated glutens with IgE from HWP-sensitized subjects. Gluten was digested with pepsin and pancreatin, followed by treatment with tTG. The glutens were resolved on SDS-PAGE and blotted with the serum of subject 2 on a polyvinylidene fluoride membrane. tTG treatment induced strong IgE binding to HMW components with a smear pattern similar to that with Glp19S, even after digestion, whereas digestion alone showed almost no effect. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig E2 Deamidation of gluten with tTG. Gluten, tTG-Glu, and Glp19S were digested with Glu-C endoproteinase. The mobility shift of tTG-Glu and Glp19S with a smear pattern can be explained by the increased negative charge of glutamic acid residues as a result of deamidation of glutamine residues. The mobility shift was completely eliminated with Glu-C treatment. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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