Anti–4-1BB/PD-1 combination enhanced antigen-specific T-cell response. Anti–4-1BB/PD-1 combination enhanced antigen-specific T-cell response. At the end of the efficacy study as described in Fig. 1D, splenocytes were isolated and fresh frozen. Viable cells after thaw were rested for 12 hours and challenged with CD8-restricted MC38 immunodominant peptide KSPWFTTL ex vivo. IFNγ production was assessed by ELISpot. The number of spots, an indication of the number of IFNγ-producing cells, was reported. Each symbol represents an individual animal within the same treatment group. *, P < 0.05 and ****, P < 0.0001, when comparing groups as indicated by the horizontal lines. Shihao Chen et al. Cancer Immunol Res 2015;3:149-160 ©2015 by American Association for Cancer Research