Extended Data Fig. 3: Treatment effects on tumor-specific CD8+ T cells.
a-g, Phenotyping of CD226+ or CD226− gp70+ CD8+ T cells from dLN or tumor, as indicated, of CT26 tumor-bearing mice treated with anti-PD-L1, anti-TIGIT, or combination with or without FTY720. Frequencies of gp70+CD8+ T cells expressing Ki67 (a), naïve phenotype (b), Teff/Tem phenotype (c), Slamf6 and TCF1 co-expression (d), TCF1 and Tim3 co-expression (e), not expressing TCF1 (f) or expressing Tox (g). For a-g, data are represented as mean +/- SD with individual symbols representing individual mice (n = 5 animals per sample and treatment group) and are representative of three independent experiments. p-values are indicated where differences between two groups were determined to be statistically significant by ordinary one-way ANOVA with Tukey’s multiple comparisons test. h-m, anti-TIGIT plus anti-PD-L1 combination treatment increases frequencies of TCF1+Tim+ CD8+ T cells and decreases frequencies of Tox+ CD8+ T cells in EO771 tumor-bearing mice. Mice with established EO771 tumors were treated with isotype control Ab or anti-TIGIT combined with anti-PD-L1, then dLN and tumors were collected on day 7 post-treatment for phenotypic characterization of CD8+ T cells by flow cytometry. h, Frequencies of CD8+ T cells in dLN (left) or tumor (right) as percentage of CD45+ cells. i, Frequencies of CD8+ T cells expressing CD226. j, Frequencies of CD8+ T cells positively stained with tetramer against EO771-specific antigen p15E. k, Frequencies of p15E+CD8+ T cells expressing CD226. l, m, Frequencies of CD226+ (left) or CD226− (right) p15E+CD8+ Teff/Tem cells co-expressing TCF1 and Tim3 (l) or Tox (m). For h-m, data are represented as mean +/- SD with individual symbols represent individual mice (n = 4 animals per sample and group); data are representative of one of two independent experiments. p-values are indicated where difference were determined by unpaired t-test to be statistically significant. n-q, Frequencies of gp70+CD8+ T cells co-expressing Slamf6 and TCF1 (n), co-expressing TCF1 and Tim3 (o), Teff/Tem phenotype (p) or Tox (q) in dLN (left) or tumor (right) of CT26 tumor-bearing mice treated with anti-PD-L1 plus anti-TIGIT combination or combo with anti-CD226 Ab. For n-q, data are represented as mean +/- SD with individual symbols representing individual mice (n = 5 animals per sample and treatment group) and are representative of three independent experiments. p-values are indicated where differences between two groups were determined to be statistically significant by ordinary one-way ANOVA with Tukey’s multiple comparisons test.
