Fig. 6

Activation phenotype and basal killing capacity of tumor-redirected A2^neg CD8 T cells in short-term co-cultures with NA8 target cells. a Schematic representation of the experimental design; A2^neg (CRISPR/A2) primary CD8 T cells of increased-affinity TCRs were co-cultured with A2^pos or A2^neg (CRISPR/A2) NA8 tumor cells for 3 days in the absence of cognate antigen. b Expression levels of co-activating/inhibitory receptors on A2^neg CD8 T cells after 3 days of co-culture with either A2^pos or A2^neg NA8 cells. c Co-expression of 0 to 4 co-inhibitory (PD-1, TIM-3 and 2B4) and co-activating (CD25) receptors. d Quantification of T cell population doublings (upper panel) and of NA8 cell numbers (lower panel) after 3 days of co-culture with A2^pos or A2^neg NA8 cells. e Representative images at 70 h (upper panel) and quantification (lower panel) of caspase 3/7-dependent apoptosis induced by tumor-redirected A2^neg CD8 T cells co-cultivated during 3 days with A2^pos or A2^neg NA8 cells are depicted, using IncuCyte technology. Data are expressed as means ± SD and are representative of 4 to 8 (b and c), 4 to 6 (d) and 2 (e) independent experiments. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 and **** P ≤ 0.0001