Figure 2.

ESC vaccination induces effector and memory CD8⁺ T cells and antigen‐presenting cells. a,b) 2 weeks after ESCs introduction, ESC (129) + CpG + GM‐CSF‐vaccinated C57BL/6 mice showed a significant increase in cytotoxic T cells (CD8⁺granzyme‐B⁺), effector/memory CD8⁺ T cells (CD8⁺CD44⁺), reduction in percentages of MDSCs (CD11c⁺Gr‐1⁺), and increase in NK+ cells and upregulation of antigen‐presenting cells (APCs) such as DCs (CD11c⁺MHC II⁺CD86⁺) in the dLN (n = 4) and PBMCs (n = 3). c) 2 weeks after ESC (C57) injection, dLNs and PBMCs of ESC (C57)‐vaccinated C57BL/6 mice revealed an increased frequency of cytotoxic T cells (CD8⁺granzyme‐B⁺), effector/memory CD8⁺T cells (CD8⁺CD44⁺), as well as T helper cells (CD4⁺CD44⁺) (n = 3). d,e) CD8⁺ T cells are required for the antitumor effects of ESCs in a bladder cancer model. Male C57BL/6 mice (n = 5) were implanted with 5 x 10⁵ MB49 bladder cancer cells s.c. in the right flank. From day 1, 3, 6, and 9 after tumor inoculation, 10 mg kg⁻¹ of CD8⁺ T cell depletion antibody, 10 mg kg⁻¹ of CD4⁺ T cell depletion antibody, 10 mg kg⁻¹ of NK cell depletion antibody were injected intraperitoneally in mice. Tumor volume and survival time of MB49 tumor‐bearing mice were measured. Experiments were repeated three times. Data represent mean ± SEM (n = 5 per group). ANOVA with Tukey's multiple comparison test). *p < 0.05, **p < 0.001, ***p < 0.001, ****p < 0.0001.