Figure 5. Multifunctional CD8⁺ T cells exerted an indispensable role in the antitumor effects of Ad-AURKA/CDK7 vaccine. (A–D) The percentages of multifunctional CD8⁺ T cells secreting TNF-α⁺IFN-γ⁺, TNF-α⁺IL-2⁺, IFN-γ⁺IL-2⁺, or TNF-α⁺IFN-γ⁺IL-2⁺ in spleens of each group were detected by flow cytometry after continuous stimulation with AURKA/CDK7 antigens for 96 hours. (E–H) The proportions of tumor-infiltrating multifunctional CD8⁺ T lymphocytes secreting TNF-α⁺IFN-γ⁺, TNF-α⁺IL-2⁺, IFN-γ⁺IL-2⁺ or TNF-α⁺IFN-γ⁺IL-2⁺ in tumor tissues of each group were analyzed on day 35 after Renca tumor inoculation. (I) The tumor weights of mice in each group were measured at the end of CD8 depletion. (J) Tumor inhibition rates in (I). (K, L) The percentages of CD8⁺ T cells or CD8⁺CD11c⁺ DCs were detected in spleens and tumor tissues of each group in the CD8⁺ T-cell depletion assay. The two-tailed independent Student’s t-test was used to analyze two-group comparisons. One-way analysis of variance was used for comparisons among multiple groups. The data are shown as means±SD, with n=5 mice per group. *p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001. Ad, adenovirus; AURKA, Aurora kinase A; CDK7, cyclin-dependent kinase 7; IFN, interferon; IL, interleukin; TIL, tumor-infiltrating leukocytes; TNF, tumor necrosis factor.