Target cell lysis by ZnT8186–194-reactive CD8+ T cell clones.

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Target cell lysis by ZnT8186–194-reactive CD8+ T cell clones. Target cell lysis by ZnT8186–194-reactive CD8+ T cell clones. (A to C) Lysis of K562-A2 cells transfected (open triangles) or not (open circles) with a full-length ZnT8 plasmid and cultured for 24 hours with clones D222D 2 (A), H017N A1 (B), or H314C 6C4 (C). Filled symbols indicate ZnT8186–194-pulsed target cells (10 μM). Results are presented as mean ± SEM of triplicate wells from two separate experiments. (D to G) Real-time cytotoxicity for the indicated clones versus HLA-A2+ ECN90 (white triangles) or control HLA-A2− EndoC-βH2 β cell targets (white circles) (E/T, 2:1). Black and gray symbols indicate the corresponding targets pulsed with 10 μM ZnT8186–194 or GAD114–122 peptide, respectively [ZnT8186–194 or MelanA26–35 for the H004N clone MelanA in panel (G)]. Means ± SEM of triplicate measurements are shown at each time point. Results are representative of at least two separate experiments. (H) Percent maximal HLA-A2+ ECN90 and HLA-A2− EndoC-βH2 β cell lysis by the indicated clones (T1D, gray symbols; healthy, white symbols; control H004N clone MelanA, horizontal dotted line) in the absence or presence of the ZnT8186–194 peptide. Bars indicate median values. Lysis was calculated from the cytotoxicity profiles as in (D) to (G). Slobodan Culina et al. Sci. Immunol. 2018;3:eaao4013 Copyright © 2018 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works