Figure 9. αCD40-induced monocyte/macrophage priming generates atypical tumor-specific T cells that lack antitumor activity in Batf3^–/– mice.

(A) Batf3^–/– spleen weight after various cell depletions + CD40 agonist. (B) Batf3^–/– tumor weight after various cell depletions + CD40 agonist. Each dot is an independent animal. Data are representative of n = 3–8 mice per group and are pooled from 3 independent experiments. (C) Percentage change in tumor mass in Batf3^–/– mice treated with CD40 agonist and the indicated cell depletion strategies as compared with Batf3^–/– mice treated with CD40 agonist only. (D) Representative Klrg1 and Lag3 staining of CD8⁺tetramer⁺ T cells from spleens and tumors from untreated Batf3^+/+ mice, and Batf3^+/+ and Batf3^–/– mice treated with αCD40 on day 14 after tumor. (E) Quantification of D, gated on CD8⁺tetramer⁺ T cells. (F) Representative Foxp3 and Helios staining of CD8⁺tetramer⁺ T cells in spleen and tumor from untreated Batf3^+/+ mice and Batf3^+/+ and Batf3^–/– mice treated with αCD40 on day 14 posttumor. (G) Quantification of F, gated on CD8⁺tetramer⁺ T cells. (H) Representative Klrg1 and Lag3 staining of CD8⁺tetramer⁺Foxp3⁺ (red) and CD8⁺tetramer⁺ Foxp3^– (gray) T cells from spleens of tumor-bearing Batf3^–/– mice treated with αCD40. (I) Phenotype of splenic CD8⁺tetramer⁺Foxp3⁺ T cells from tumor-bearing Batf3^–/– mice treated with αCD40 on day 14 posttumor (day 7 after αCD40). (J) Functional analysis of Gitr⁺ and Gitr^– CD8⁺ T cells following a 5-hour restimulation with CB_101-109 peptide. n = 3–9 mice per group. Data are mean ± SEM and pooled from at least 2 independent experiments. Each dot is an independent animal. One-way ANOVA with a Tukey’s posttest (B, C, E, and G). Paired Student’s t test (J).