Fig. 4.

CD40 signaling is required for the antitumor efficacy of neoantigen/TLR3/CD40 agonists and generation of antigen-specific CX3CR1⁺ CD8⁺ T cells. a Left shows tumor volume curves in MC38 tumor-bearing wild type C57BL/6 mice (WT) or CD40^−/− mice treated with or without Adpgk^Mut/TLR3/CD40 agonists vaccination (Vac) as described in Fig. 1a (n = 7 mice per group). Right shows tumor weight. Tumors were harvested 1 week after 2nd vaccination (Vac). b, c Left shows representative FACS plots showing CD8 and CX3CR1 expression gated with Adpgk^Mut Tet⁺ CD8⁺ T cells in PB of MC38-tumor bearing WT or CD40^−/− mice (b) and bone marrow chimeric mice (WT → WT or CD40^−/− → WT) (c) treated with or without Vac. Numbers denote percentage of the CX3CR1⁺ subset. Right shows the frequency of the CX3CR1⁺ subset in each group (n = 4–8 mice per group). PB was harvested 1 week after 2nd Vac. Data shown are representative of two independent experiments (a). NS: not significant, ***P < 0.001, ****P < 0.0001, one-way ANOVA with Tukey’s multiple comparisons. Mean ± SEM