Fig. 2

Expression of a gp100/HLA-A2-specific TCR and a MCSP-specific CAR by different T-cell populations after electroporation. ZA-activated PBMC, OKT3-activated PBMC, OKT3-stimulated MACS-isolated CD8⁺ T cells, and OKT3-stimulated MACS-isolated γ/δ T cells were expanded as described for Fig. 1. After 10–11 days, these cells were electroporated with RNA coding for the gp100/HLA-A2-specific TCR or with RNA encoding the MCSP-specific CAR. a + b TCR expression a was detected using an anti-Vbeta14 antibody and CAR expression levels b were analyzed using an anti-IgG1 antibody (black lines). TCR-transfected T cells served as negative controls (neg.; filled grey histograms) for the CAR staining and vice versa. Presented histograms are representatives out of four to seven independent experiments. c-f Short interval expression kinetics of transfected receptors on different cell populations. g-j Long interval expression kinetics of transfected receptors on different cell populations. Staining for TCR (c + e + g + i) and CAR (d + f + h + j) expression was performed at indicated time-points after electroporation using abovementioned antibodies. TCR-transfected T cells served as negative controls for CAR staining and vice versa. The percentage of positive cells is indicated. Data represent geometric means ± SEM from 4 to 7 independent experiments