Fig. 2.

Effective neoantigen/TLR3/CD40 stimulations correlate with generation of antigen-specific effector CX3CR1⁺ CD8⁺ T cells. a–c, MC38 tumor-bearing C57BL/6 mice were treated with PBS (NT) or Adpgk^Mut/TLR3/CD40 agonists vaccination (Vac) as described in Fig. 1a. Blood, spleens and tumors were collected 1 week after 2nd Vac. a Representative FACS plots showing CX3CR1 expression in CD8⁺ (left) or Adpgk^Mut tetramer (Tet)⁺ CD8⁺ T cells (right) in PB (upper), spleen (middle) and tumors (lower). Numbers denote percentage of CX3CR1⁺ cells. Frequency of CX3CR1⁺ cells from the CD8⁺ or Tet⁺ CD8⁺ T cells is shown in the right panels (n = 4–7 mice per group). b Scatter plot of the frequency of PB CX3CR1⁺ CD8⁺ T cells (left), CX3CR1⁺ Tet⁺ CD8⁺ T cells (middle), and Tet⁺ CD8⁺ T cells (right) against tumor volume. Correlation is shown using Pearson correlation (r) and Spearman correlation coefficients (r_s). c Representative FACS plots of CX3CR1⁻ (left) or CX3CR1⁺ (right) cells gated with Tet⁺ CD8⁺ T cells in PB. Numbers denote percentage of each marker-positive cells. Frequency of marker-positive cells from the CX3CR1⁻ or CX3CR1⁺ subset is shown in the right panels for each marker (n = 4–7 mice per group). Data shown are representative of at least two independent experiments (a–c). Two-tailed unpaired (a) and paired t-test (c). Box plots: hinges, 25th and 75th percentiles; middle line, median; whiskers, minimum to maximum value (a)