Figure 4.

Tc17-1 cells acquire a stem-like phenotype and show long-term persistence after in vivo transfer. (A, B) GL261-Quad-bearing mice received a single IV infusion of Tc1 or Tc17-1 cells on the day after TMZ treatment, and their splenocytes were evaluated on day 7 (A) and day 50 (B) after T-cell transfer. Representative flow cytometry data of Tc1 and Tc17-1 cells before and after IV transfer, and frequency of CD44^-CD62L⁺, CD44⁺CD62L⁺ and CD44⁺CD62L^- cells in CD90.1⁺CD8⁺ T-cells (A) and CD90.1⁺ cells in CD8⁺ T-cells (B). Data are representative of 2 independent experiments and presented as mean±SD (n=4–5 per group). **P<0.001, ***p<0.0001; unpaired two-tailed Student’s t-test. (C) GL261-Quad cells were isolated from mice treated with Combo, Combo+Tc1, or Combo+Tc17-1 cells and cocultured with Pmel-1-derived CD8⁺ T-cells. IFN-γ production in the supernatant was determined by ELISA. GL261 and GL261-Quad cells were used as negative and positive controls, respectively. Data are presented as mean±SEM of 2 independent experiments performed in triplicate. *P<0.05, **p<0.001, ***p<0.0001, ****p<0.00001; one-way ANOVA with interaction followed by Tukey’s multiple comparison test. ANOVA, analysis of variance; IFN-γ, interferon-γ; IV, intravenously; TMZ, temozolomide.