Fig. 2: PD-1 blockade in Suv39h1-KO CD8+ TILs modifies their exhaustion program and enhances their effector capacity.
From: CD8+T cell responsiveness to anti-PD-1 is epigenetically regulated by Suv39h1 in melanomas
A Representative histogram and frequency (%) of inhibitory receptors (PD-1+, TIM-3+, LAG-3+ and 2B4+) on CD8 + TILs from B16F10-OVA tumors. WT n = 6; WT + aPD-1 n = 4; KO n = 6; KO + aPD-1 n = 8. B Pie chart of co-expression (PD-1+, TIM-3+, LAG-3+, and 2B4+) on CD8+ TILs from B16F10-OVA tumors. C Representative histogram and frequency (%) of GZMb+ cells among CD8+ TILs. Cells were re-stimulated in vitro with PMA and ionomycin for 4 h. (GZMb: Granzyme b). WT n = 8; WT + aPD-1 n = 5; KO n = 7; KO + aPD-1 n = 5. D Blood cells from mice were re-stimulated ex vivo with OVA-ISIINFEKL peptide. The number of OVA-specific T cells producing IFNγ per 2.5 × 105 blood cells was determined by ELISPOT analysis. WT n = 6; WT + aPD-1 n = 6; KO n = 6; KO + aPD-1 n = 6. E Representative histogram and frequency (%) of ISG15+ among CD8 + TILs from B16F10-OVA tumors. WT n = 7; WT + aPD-1 n = 4; KO n = 5; KO + aPD-1 n = 4. F Representative dot plots and frequency (%) of OVA-specific TILS from B16F10-OVA tumors as detected using Ova-SIINFEKL/Kb tetramers coupled to two different fluorochromes. WT n = 7; WT + aPD-1 n = 6; KO n = 6; KO + aPD-1 n = 5. G Representative histogram and frequency (%) of Ki67+ among CD8+TILs from B16F10-OVA tumors. WT n = 6; WT + aPD-1 n = 4; KO n = 6; KO + aPD-1 n = 8. H Representative histogram and frequency (%) of BCL-2+ among CD8+ TILs from B16F10-OVA tumors. WT n = 6; WT + aPD-1 n = 3; KO n = 5; KO + aPD-1 n = 3. J Representative contour plots and frequency (%) of TBET+EOMES+ among CD8 + PD-1+ TILs from B16F10-OVA tumors. WT n = 6; WT + aPD-1 n = 4; KO n = 6; KO + aPD-1 n = 8. K Representative contour plots and frequency (%) of progenitor exhausted (TCF1+TIM-3−) and late exhausted (TCF1−TIM-3+) among CD8+ PD-1+ TILs from B16F10-OVA tumors. WT n = 4; WT + aPD-1 n = 4; KO n = 5; KO + aPD-1 n = 6. I Frequency (%) of Annexin V+ 7ADD− among CD8 + TILs from B16F10-OVA tumors. WT n = 6; WT + aPD-1 n = 3; KO n = 3; KO + aPD-1 n = 3. L Representative histogram and frequency (%) of transitory (CD101-) and exhausted (CD101+) among CD8+ PD-1+ TCF1-TIM-3+ TILs from B16F10-OVA tumors. WT n = 4; WT + aPD-1 n = 3; KO n = 5; KO + aPD-1 n = 6. A representative experiment out of two is shown. p values were calculated using two-tailed Mann–Whitney test. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. In all graphs, mean±s.e.m. are presented. See also Supplementary Fig. 3. Source data are provided as a Source Data file.
