Analysis of glutathione S-transferase allergen cross-reactivity in a North American population: Relevance for molecular diagnosis Geoffrey A. Mueller,

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Presentation transcript:

Analysis of glutathione S-transferase allergen cross-reactivity in a North American population: Relevance for molecular diagnosis Geoffrey A. Mueller, PhD, Lars C. Pedersen, PhD, Jill Glesner, BS, Lori L. Edwards, BS, Josefina Zakzuk, MD, PhD, Robert E. London, PhD, L. Karla Arruda, MD, PhD, Martin D. Chapman, PhD, Luis Caraballo, MD, PhD, Anna Pomés, PhD Journal of Allergy and Clinical Immunology Volume 136, Issue 5, Pages 1369-1377 (November 2015) DOI: 10.1016/j.jaci.2015.03.015 Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 GST allergen structures. A-C (top) and D, Ribbon diagrams of the dimer. E, Superposition of monomers, and N-terminal thioredoxin-like subdomain (dashed oval). Two active site conformations of Bla g 5, and the active site loop of Der p 8 and Blo t 8 (GSH molecule in white), are shown at the bottom of A, B, and C, respectively. Arrows in 3 colors indicate how the structures in A to D are rotated with respect to E. Black arrows in A and E show the C-termini. Journal of Allergy and Clinical Immunology 2015 136, 1369-1377DOI: (10.1016/j.jaci.2015.03.015) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 IgE reactivity to GST. A, IgE reactivity to Bla g 5 and none to Der p 8 in 15 of 22 cockroach-allergic patients (from which 18 also had IgE reactivity to Dpteronyssinus by ImmunoCAP). B, IgE reactivity to Der p 8 and none to Bla g 5 in 12 of 16 mite-allergic patients. C and D, IgE from 15 Bla g 5–positive sera (Fig 2, C) and 12 Der p 8–positive sera (Fig 2, D) did not bind Asc s 13 and Blo t 8. Two positive controls are on the right. Horizontal bars represent the median of the values. Dotted lines represent a cutoff value for a positive ELISA reading as defined in the Methods section. Journal of Allergy and Clinical Immunology 2015 136, 1369-1377DOI: (10.1016/j.jaci.2015.03.015) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Inhibition of IgE reactivity to GST. A, Inhibition of IgE binding to Der p 8 by Der p 8, but not the other 3 GSTs using a pool of 3 sera from mite-allergic patients from North America. B, Inhibition of IgE binding by anti–Bla g 5 mAb, but not by the anti–Bla g 1 mAb 7C11 negative control. Serum was from a patient highly allergic to cockroach (>100 kU/L) sensitized to Bla g 1 (29.6 kU/L) and Bla g 5 (94.5 kU/L). Journal of Allergy and Clinical Immunology 2015 136, 1369-1377DOI: (10.1016/j.jaci.2015.03.015) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Inhibition of IgE reactivity to GST by Bla g 5 and Der p 8. A-E, IgE reactivity to Der p 8 was inhibited only by Der p 8 among mite-allergic patients from North America. F-H, IgE reactivity to Bla g 5 was inhibited by Bla g 5, and at approximately 100-fold less affinity by Der p 8 among cockroach-allergic patients (insets show a larger range of inhibitor concentrations). Journal of Allergy and Clinical Immunology 2015 136, 1369-1377DOI: (10.1016/j.jaci.2015.03.015) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Recognition of GST by anti–Bla g 5 mAb. Comparison of dose-response curves of 6 anti–Bla g 5 mAbs binding to (A) Bla g 5 (curves in solid lines) and to each of the other 3 GSTs: (B) Der p 8, (C) Blo t 8, and (D) Asc s 13 (curves in dashed lines). Note the differences in the x-axis: there was a 1000-fold lower concentration of mAbs required for binding to Bla g 5, compared with the other GSTs. Journal of Allergy and Clinical Immunology 2015 136, 1369-1377DOI: (10.1016/j.jaci.2015.03.015) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 Surface residue comparisons. Residue similarity of Asc s 13 (A and B), Der p 8 (C and D), and Blo t 8 (E and F) to Bla g 5. G, Der p 1 similarity to Der f 1, and epitope for cross-reactive mAb 4C1 (circle).22 H and I, Mala s 6 similarity to Cat r 1.23 Color bar represents residue similarity from low (light blue) to high (maroon) (see this article's Online Repository at www.jacionline.org). Gray represents gaps or insertions in sequence alignment. Arrows indicate how the structures are rotated with respect to each other. Journal of Allergy and Clinical Immunology 2015 136, 1369-1377DOI: (10.1016/j.jaci.2015.03.015) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 Journal of Allergy and Clinical Immunology 2015 136, 1369-1377DOI: (10.1016/j.jaci.2015.03.015) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 Journal of Allergy and Clinical Immunology 2015 136, 1369-1377DOI: (10.1016/j.jaci.2015.03.015) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E3 Journal of Allergy and Clinical Immunology 2015 136, 1369-1377DOI: (10.1016/j.jaci.2015.03.015) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E4 Journal of Allergy and Clinical Immunology 2015 136, 1369-1377DOI: (10.1016/j.jaci.2015.03.015) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions