Figure 5.

Characterization of the roles of different immune populations in TIM-3-blockade efficacy

(A) Schedule of survival experiments (NP53 cells bearing mice) performed with depleting antibodies specific for NK cells (clone K1.1), CD4⁺ T cells (clone GK1.5), and CD8⁺ T cells (clone CD8β) and anti-TIM-3 or IgG2a antibodies.

(B and C) Kaplan-Meier survival curves of mice treated with the anti-TIM-3 antibody and (B) depleting antibody specific for NK cells (n = 11), CD4 cells (n = 10), or CD8 cells (n = 11) alone or in combination (C) of NK and CD4⁺ T cells (n = 10) or NK and CD8⁺ T cells (n = 11).

(D) Kaplan-Meier survival curves of immunodeficient Rag2-γc- mice treated with anti-TIM-3 (n = 10) or IgG2a (n = 9) (log rank p = 0.01).

(E) Kaplan-Meier survival curves of mice treated with anti-TIM-3 (n = 10) or IgG2a (n = 12) or an anti-CSFR1 drug (PLX) plus the anti-TIM-3 antibody (n = 11).

(F) Flow cytometric analyses of macrophages, NK and CD8⁺ T cells per mg of tumor on day 7 after TIM-3 i.t. treatment with Vehicle+IgG2a (black), Vehicle+AbTIM-3 (blue), or PLX+AbTIM-3 (yellow).

(G) Flow cytometric analyses of microglia number of cells per mg tumor (left panel), MFI Ki67 (middle panel), and percentage of MHCII expression (right panel) on day 7.

(H) Quantification of Ki67⁺ CD4⁺ (left panel) and CD8⁺ T cells (right panel) by flow cytometry on day 7 after i.t. treatment.

(I) Flow cytometric analysis of CCL2, CCL5, CXCL10, IL-1β, and IFN-γ expression in the tumor microenvironment comparing groups on the indicated day. One-way ANOVA was performed. Bar graphs indicate the mean ± SEM (ns, p > 0.05; ^∗p < 0.05; ^∗∗p < 0.01; ^∗∗∗p < 0.001; ^∗∗∗∗p < 0.0001). See also Figures S7 and S8.