Fig. 5: Proliferative T cell exhibits robust antitumor immunity under electrical stimulation.

a Graphic illustration of the study design for the investigation the role of proliferative T cells in antitumor immunity. OT-I cells were cultured on HC nanocomposite membranes. OVA peptides (2 μg/mL) and IL-2 (10 U/mL) were used to stimulate T cell activation and ensuing proliferation. CFSE^lo and CFSE^hi cells were isolated by BD FACS Aria II flow cytometer and subjected to RNA-sequencing. b Volcano plot analysis of gene expression in CFSE^lo and CFSE^hi T cells. Red, genes upregulated in CFSE^lo T cells. Blue, genes downregulated in CFSE^lo T cells. Statistical significance was assessed by the Wald test. c Genes that were significantly upregulated in CFSE^lo T cells were analyzed using GO database. Statistical significance was assessed by oa ne-sided hypergeometric test. d Macroscopic evaluation of tumors from mice intravenously injected with CFSE^lo and CFSE^hi T cells (n = 5 mice). e Tumor volume of mice intravenously injected with CFSE^lo and CFSE^hi T cells cultured on HC nanocomposite membrane was monitored over time (n = 5 mice, mean ± sem, ***P = 0.0005 (Day 13), ***P = 0.0008 (Day 15), ***P = 0.0003 (Day 19), ****P < 0.0001, two-tailed unpaired Student’s t test). f Quantity of tumor-infiltrating immune cells in mice intravenously injected with CFSE^lo and CFSE^hi T cells (n = 5 mice, mean ± sem, **P = 0.0055, two-tailed unpaired Student’s t test). g Flow cytometric analysis of the frequency of CD8⁺ T cells in tumors from mice intravenously injected with CFSE^lo and CFSE^hi T cells (n = 5 mice, mean ± sem, ****P < 0.0001, two-tailed unpaired Student’s t test). h Flow cytometric analysis of the expression of IFNγ in tumor-infiltrated OT-I cells (n = 5 mice, mean ± sem, **P = 0.0042, two-tailed unpaired Student’s t test). i Flow cytometric analysis of the expression of TNF in tumor-infiltrated OT-I cells (n = 5 mice, mean ± sem, **P = 0.0013, two-tailed unpaired Student’s t test). j Flow cytometric analysis of the expression of GZMB in tumor-infiltrated OT-I cells (n = 5 mice, mean ± sem, ***P = 0.0001, two-tailed unpaired Student’s t test). k Flow cytometric analysis of the expression of KLRG1 in tumor-infiltrated OT-I cells (n = 5 mice, mean ± sem, ****P < 0.0001, two-tailed unpaired Student’s t-test). Source data are provided as a Source Data file.