Extended Data Fig. 8: Immunosuppressive CXCR6+ DP8 cells infiltrate AT3 tumor.
(a) CD103+CD8+ T cells decrease in TILs of A-AT3 mice. (b) Tumor weight (g, at day 35 post tumor challenge) in A-At3 mice treated with Abs to deplete CD4+ or CD8+ T cells or control IgG; (c) CXCL16 shRNA knockdown in the AT3 cells decrease tumor growth (left panel) and production of CXCL16 (ELISA quantification of CM of indicated cells, right panel); (d) Representative dot plot showing presence of CD4+ and CD8+ T cells in TILs of A-AT3 mice; (e and f) CD3+ T cells (eFluor450 labelled and FACS-sorted from spleen of young mice) were stimulated with anti-CD3/28 beads in the presence or absence of sorted splenic DP8 and CD73+CD39− CD8+ T cells from Y-WT and A-WT (e) or spleen of Y-AT3 and A-WT (f) mice. Shown is frequency of proliferated CD4+ T cells after 5 days co-culture. (g and h) DP8 cells suppress target CD4+ T cells using CD39. The same suppression assay as in e was performed in the presence or absence of 10 µM POM1 (inhibitor of CD39 activity; (h) POM1 does not reduce viability of target CD3+ T cells. Shown is frequency ± SEM (a, b,c, e-i); (i) While adoptively transferred CD8+ T cells from Y-WT reduces AT3 tumor in A-Rag KO mice, A-WT CD8+ T cells failed to do so. Injection diagrams were created in BioRender.com and numbers represent the days of injection. Each symbol is for a single mouse, n = 7-8 (a), n = 3-7 (b), n = 6-9 (c), n = 3-6 (e–g) and n = 6-7 (i) mice per group. Experiments were reproduced three times. * P < 0.05; ** P < 0.01; *** P < 0.001 in unpaired Mann-Whitney test and Kruskal-Wallis test.
