Fig. 4

Proliferative capacity, basal apoptotic levels and tolerance-related transcriptional profiles in relation to HLA-A2. a Population doublings of A2^pos and A2^neg tumor-redirected primary CD8 T cells upon CRISPR/A2-Cas9 strategy were assessed by periodic cell counting of living cells during 10 days post-TCR sorting in the absence of cognate antigen. Data are representative of 2 independent experiments. b, c Quantification of the fraction of Ki67-positive (b) and Annexin-V-positive cells (c) in A2^pos (CRISPR/GFP) and A2^neg (CRISPR/A2) primary CD8 T cells under resting conditions. c Representative dot blots (left panel) and Annexin V quantifications (right panel) are depicted. Data are means ± SD of 4 to 5 independent experiments. * P ≤ 0.05 and *** P ≤ 0.001. d GSEA of available gene sets describing anergy [24], self-tolerance [25], and deletional tolerance [26] were found enriched in A2^pos wtc51m- versus DMβ-expressing primary CD8 T cells under steady-state culture conditions. Nominal P values and false discovery rates (FDR) are indicated for each gene set enrichment