Identification of drug- and drug-metabolite immune responses originating from both naive and memory T cells Andrew Gibson, MRes, Lee Faulkner, PhD, Sally.

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Presentation transcript:

Identification of drug- and drug-metabolite immune responses originating from both naive and memory T cells Andrew Gibson, MRes, Lee Faulkner, PhD, Sally Wood, BSc, B. Kevin Park, PhD, Dean J. Naisbitt, PhD Journal of Allergy and Clinical Immunology Volume 140, Issue 2, Pages 578-581.e5 (August 2017) DOI: 10.1016/j.jaci.2016.11.032 Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Naive and memory T cells were exposed to SMX-NO and SMX for 8 days and then recall responses tested. Proliferative responses to SMX-NO (A) and SMX (B) were measured by [3H]-thymidine incorporation (n = 3). Error bars indicate SD of triplicate wells (*P ≤ .05, **P ≤ .005, and ***P < .001). IL-13 secretion to SMX-NO (C) and SMX (D) was measured by ELISpot (n = 3). Journal of Allergy and Clinical Immunology 2017 140, 578-581.e5DOI: (10.1016/j.jaci.2016.11.032) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 T-cell clones were isolated from 2 donors following exposure of naive and memory T cells to SMX-NO and SMX. A, Generation and phenotype of T-cell clones. B and C, Proliferation (SMX, 2 from naive and 5 from memory precursors; SMX-NO, 5 from naive and 2 from memory precursors) and (D and E) secretion of cytokines/cytolytic molecules by T-cell clones isolated following SMX and SMX-NO exposure. Error bars indicate SD of triplicate wells (*P ≤ .05, **P ≤ .005, and ***P < .001). Journal of Allergy and Clinical Immunology 2017 140, 578-581.e5DOI: (10.1016/j.jaci.2016.11.032) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 Overview of study design and results. Delineation of naive and memory T-cell responses to SMX-derived antigens in drug-naive donors. Journal of Allergy and Clinical Immunology 2017 140, 578-581.e5DOI: (10.1016/j.jaci.2016.11.032) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 Comparative ability of DCs and B cells to induce naive and memory T-cell responses to drug-antigen in vitro. Naive and memory T cells from drug-naive donors were exposed to SMX-NO and SMX for 8 days in the presence of EBV-transformed B cells (A) or DCs (B), and then recall responses tested by [3H]-thymidine incorporation. Error bars indicate SD of triplicate wells. Journal of Allergy and Clinical Immunology 2017 140, 578-581.e5DOI: (10.1016/j.jaci.2016.11.032) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E3 Antigen cross-reactivity of T-cell clones derived from priming in drug-naive donors and hypersensitive patients. Naive and memory T cells from drug-naive donors were exposed to SMX and SMX-NO for 8 days and then recall responses tested. Cross-reactivity to SMX-NO and SMX was measured by [3H]-thymidine incorporation and/or ELISpot after exposure of naive and memory T cells to SMX-NO (A, n = 3) and SMX (B, n = 3), or exposure of representative SMX-NO– (C) and SMX-responsive (D) T-cell clones from hypersensitive patients. Error bars indicate SD of triplicate wells (*P ≤ .05, **P ≤ .005, and ***P < .001). Journal of Allergy and Clinical Immunology 2017 140, 578-581.e5DOI: (10.1016/j.jaci.2016.11.032) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E4 MHC I and II restriction of SMX-responsive T-cell clones derived from drug-naive donors. Naive or memory-derived SMX-responsive T-cell clones from drug-naive donors were cultured with autologous EBV-transformed B cells and SMX, with or without MHC I– or II–blocking antibodies for 48 hours. Proliferative responses were measured by [3H]-thymidine incorporation for a further 16 hours. Error bars indicate SD of triplicate wells (*P ≤ .05, **P ≤ .005, and ***P < .001). Journal of Allergy and Clinical Immunology 2017 140, 578-581.e5DOI: (10.1016/j.jaci.2016.11.032) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions