Figure 7. Antitumor efficacy of Ad-AURKA/CDK7 vaccine requires for multi-functional CD8⁺ T cells in the Renca orthotopic model. (A) Schematic diagram illustrating the design of the Renca orthotopic model and vaccination. (B) Typical image of renal orthotopic tumors in the different vaccine-treated groups at the endpoint of the experiment (n=5 mice per group). (C) The survival curve of each group in the orthotopic model (n=10 mice per group). (D) Tumor weights were calculated by the formula: Tumor mass (g)=left kidney mass − right kidney mass. (E) Tumor inhibition rate in (D). (F) Tumor-infiltrating CD8⁺ T lymphocytes were captured by immunohistochemical staining in the left kidney and the representative images were shown. The scale was 200 µm. (G, H) Statistical analysis of the proportion of CD8⁺ T cells or CD8⁺CD11c⁺ DCs in renal tumor tissues of each group. (I, J) The percentages of multifunctional CD8⁺ T lymphocytes secreting IFN-γ⁺, TNF-α⁺, IL-2⁺, TNF-α⁺IFN-γ⁺, TNF-α⁺IL-2⁺, IFN-γ⁺IL-2⁺, or TNF-α⁺IFN-γ⁺IL-2⁺ were detected by flow cytometry in spleens and tumors in each group. (K) The antigen-stimulated proliferation of CD8⁺ T cells in different groups. (L) The tumor-specific killing capability of CTL was detected by the co-culture experiment. One-way analysis of variance was used for comparisons among multiple groups. Survival analysis was performed using the log-rank (Mantel-Cox) test. The data are shown as means±SD. *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001. Ad, adenovirus; AURKA, Aurora kinase A; CDK7, cyclin-dependent kinase 7; CTL, cytotoxic T lymphocyte; DC, dendritic cell; IFN, interferon; IL, interleukin; i.m, intramuscular; s.c, subcutaneous; TNF, tumor necrosis factor.