Fig. 5.

A critical role of CD40 signaling in cDC1s for the antitumor efficacy of neoantigen/TLR3/CD40 agonists vaccination and generation of antigen-specific CX3CR1⁺CD8⁺ T cells. a–d MC38-tumor bearing wild type C57BL/6 (WT) or Batf3^−/− mice (a, b) and bone marrow chimeric mice (Batf3^−/− and WT → WT or Batf3^−/− and CD40^−/− → WT) (c, d) were treated with or without Adpgk^Mut/TLR3/CD40 agonists vaccination (Vac) as described in Fig. 1a. a Tumor volume curves (left) and tumor weight (mg) in different treatment groups (n = 7 mice per group). b, c Left shows representative FACS plots showing CD8 and CX3CR1 expression gated with Adpgk^Mut Tet⁺ CD8⁺ T cells in PB. Right shows the frequency of the CX3CR1⁺ Tet⁺ CD8⁺ T cells in each group. (n = 4 (b) and 10–12 (c) mice per group). d Tumor growth and survival curves in different treatment groups (n = 10–11 mice per group). Tumors (a) and PB (b, c) were harvested 1 week after 2nd Vac. Data shown are representative of two independent experiments (a, b) and pooled from two independent experiments (c, d). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, one-way ANOVA with Tukey’s multiple comparisons (a, b and c), Mann–Whitney U test (d for tumor volume), or log-rank test (d for survival). Mean ± SEM