a, Experimental schematic of adoptive transfer system to generate an environment in which only CD8+ T cells lack IL-10R or IL-35R.
b, Tumor growth curve of Rag1−/− mice that were sequentially reconstituted with CD8-depleted splenocytes and CD8+ T cells from wild-type (WT) (n=11), IL-10R−/− (Il10rb−/−) (n=8), or IL-35R−/−(CD4Cre.Il6stL/L.Il12rb2 KO) (n=7) mice followed by B16 intradermal inoculation. Each measurement time point represents mean value with s.e.m. error bar. Statistical significance was determined by Two-way ANOVA with Holm-Sidak multiple comparisons (****p<0.0001).
c, Representative flow plots depicting the expression of PD-1 against the other 4 inhibitory receptors (TIM3, LAG3, TIGIT, 2B4) and SPICE plots showing the multi-inhibitory receptor expression on the donor CD8+ T cells infiltrating B16 tumor-bearing reconstituted Rag1−/− mice 18 days post-tumor inoculation as described in (a).
d, Scatter-bar graphs representing the percent distribution of donor CD8+ TILs based on the number of inhibitory receptors expressed. (WT: n=10, IL-10R−/−: n=6, IL-35R−/−: n=7). Bars represent mean values. Statistical significance was determined by One-way ANOVA with Holm-Sidak multiple comparisons (****p<0.0001 and other p-values indicated).
(a-d) Data averaged from 2 independent experiments.