Fig. 3. Target cell lysis by ZnT8_186–194-reactive CD8⁺ T-cell clones.

(A–C) Lysis of K562-A2 cells transfected (open triangles) or not (open circles) with a full-length ZnT8 plasmid and cultured for 24 h with clones D222D 2 (A), H017N A1 (B) or H314C 6C4 (C). Filled symbols indicate ZnT8_186–194-pulsed target cells (10 µM). Results are presented as mean±SEM of triplicate wells from two separate experiments. (D–G) Real-time cytotoxicity for the indicated clones vs. HLA-A2⁺ ECN90 (white triangles) or control HLA-A2⁻ EndoC-βH2 β-cell targets (white circles) (E/T 2/1). Black and grey symbols indicate the corresponding targets pulsed with 10 µM ZnT8_186–194 or GAD_114–122 peptide, respectively. Mean±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, grey symbols; healthy, white symbols; control, horizontal dotted line) in the absence or presence of ZnT8_186–194 peptide. Bars indicate median values. Lysis was calculated from the cytotoxicity profiles as in panels D–G.