Figure 4. Batf3 is required for tumor-specific T cell priming to a pancreas tumor antigen.

(A) Tumor radiance at 7 days posttumor in Batf3^+/+ and Batf3^–/– mice. (B) Survival of Batf3^+/+ or Batf3^–/– mice bearing nAg⁺ tumors. Log-rank (Mantel-Cox) test, n = 4–6 mice per group. (C) Spleen and orthotopic tumor weights from Batf3^+/+ and Batf3^–/– mice at day 14 posttumor. Student’s t tests. (D) Frequency of immune cells in spleen gated on CD45⁺ cells (left) and cell number normalized to spleen gram (right) at day 14. Student’s t test. Dotted red line indicates limit of detection. (E) Frequency of intratumoral immune cells gated on CD45⁺ cells (left) and cell number normalized to tumor gram (right) at day 14. Student’s t test. Dotted red line indicates limit of detection. (F) Representative CB_101-109:H-2D^b tetramer staining gated on CD8⁺ T cells at day 14. (G) Frequency of CB_101-109:H-2D^b-specific T cells at day 14. Student’s t test. (H) Number of CB_101-109:H-2D^b-specific T cells isolated from spleen (Spl) or tumors (PDA) and normalized to tissue gram on day 14. Student’s t test. (I) Representative plots of CB_101-109:H-2D^b-specific T cell tetramer and PD-1 gated on CD8⁺ T cells of spleens of naive mice or mice vaccinated with CB_101-109 peptide, CD40 agonist, and Poly:(IC) on day 7. Data are graphed below. One-way ANOVA with Tukey’s posttest. (J) Foxp3 and CD44 staining of live CD45⁺CD4⁺ T cells isolated from spleens and tumors at day 14. (K) Frequency of Foxp3⁺CD4⁺ T cells gated on total CD4⁺ T cells in spleen (Spl) and tumor (PDA). Student’s t test. (L) Effector CD8⁺ T cell/Treg ratio in tumors from Batf3^+/+ and Batf3^–/– mice. (M) Frequency of CD4⁺Foxp3^– T cells that coexpress Fr4 and CD73. (C–E, G, H, and K–M) Each dot is an independent mouse. Data are mean ± SEM. n = 3–6 mice per group and 2–3 independent experiments. *P < 0.05, **P < 0.005, and ***P < 0.0005.