Extended Data Fig. 5: Host CD4+ T cell is required for Tc9-mediated tumor restraint.
a, Cytotoxicity of Pmel-1 Thy1.1+ Tc9 cells against target cells was determined by a 6-hour cytotoxicity assay at 10:1 E:T ratio. Target cells were WT B16 cells, relapsed B16 cells from tumors on day 40, B16gp100-kd cells and B16gp100-ko cells (n = 6 biologically independent samples per group, pooled from three independent experiments). b, Schema of experiments in Fig. 4i. c, Schema of experiments in Extended Data Fig. 5d. d, Thy1.1+ Pmel-1 Tc1 cells were transferred into Thy1.2+ B6 mice bearing chimeric (B16 and B16gp100-kd) tumors with the adjuvant treatment, and IgG isotype or αCD4 antibodies were injected on day 20 after tumor cell inoculation. Tumor growth curves of treated mice were shown (n = 5 mice per group). e, Representative plots of CD4+ and CD8+ cell percentages in peripheral blood of WT and Cd4−/− mice. f, g, on day 0, mice (n = 6 for each group) were inoculated s.c. in the right back flank with 0.4 million WT B16 and 0.2 million B16gp100-kd chimeric tumor cells. On day 8, one dose of CTX was given intraperitoneally at 200 mg/kg body weight. On day 9 after tumor inoculation, mice were treated with intravenous injection of 2 million Tc1 or Tc9 cells, followed by i.v. injection of 0.5 million peptide-pulsed bone marrow-derived DCs and 4 doses of rhIL-2 (6 × 105 U). On day 20, mice were injected intraperitoneal every three days with 200μg of InVivoMAb anti mouse NK1.1 or IgG. Schema of experiments in f and tumor growth curve in g. Data are presented as mean ± SEM. One-way ANOVA was used in a; Multiple t tests-one per row were used to compare tumor growth in d (day 40).
