Extended Data Fig. 3: Host CD4 + T cells are enriched in Tc9-treated chimeric tumor.

a, Schema of experiments in Fig. 3a–e. b–g, Thy1.1⁺ Pmel-1 Tc1 or Tc9 cells were transferred into Thy1.2⁺ B6 mice bearing B16 chimeric tumor with the adjuvant treatment. Tumor-infiltrating CD45⁺ cells were isolated for CyTOF analysis. t-SNE analysis of CD3, CD4, CD8 and Thy1.2 expression (b), percentage of CD4⁺ T cells in CD45⁺ cells (c), percentage of host Thy1.2⁺ CD8⁺ T cells (d), Thy1.2⁻ transferred CD8⁺ T cells (e) in CD45⁺ cells, relative median intensity of IFNγ and GZMB in CD4⁺ T cells (f), percentage of Foxp3⁺ in CD4⁺ T cells (g). n = 2 in b-g, pooled from 6 mice. h, i, Thy1.1⁺ Pmel-1 Tc1 or Tc9 cells were transferred into Thy1.2⁺ mice bearing MC38 chimeric tumor with the adjuvant treatment, and tumors were analyzed. The number of CD4⁺ T cells, and IFNγ⁺ or GZMB⁺ CD4⁺ T cells were shown (n = 6 mice per group, one of two independent experiments). j, k, Tumors in mice treated as Fig. 3a were analyzed. MFI of IFNγ⁺ or GZMB⁺ and number of host Thy1.1⁻ CD8⁺ T cells were shown (n = 9 mice per group, pooled from two independent experiments). l, Thy1.1⁺ Pmel-1 Tc9 cells were transferred into WT or CD8^−/− mice bearing B16 chimeric tumors with the adjuvant treatment, and tumor growth curve were shown (n = 5 mice per group). m, n, Tumors in mice treated as Fig. 3a were analyzed. MFI of IFNγ⁺ or GZMB⁺ and number of transferred Thy1.1⁺ CD8⁺ T cells were shown (n = 9 mice per group, pooled from two independent experiments). o, p, Thy1.1⁺ Pmel-1 Tc1 or Tc9 cells were transferred into Thy1.2⁺ B6 mice bearing MC38 chimeric tumor with the adjuvant treatment, and tumors were analyzed. MFI of CD44, CD69 and PD-1 on CD4⁺ T cells, percentage of ICOS⁺PD-1^low CD4⁺ T cells in tumors (n = 6 mice per group). Data are presented as mean ± SEM. Two-tailed unpaired Student’s t-tests were used in h–k and m–p. Multiple t tests-one per row were used to compare tumor growth in l (day 40).

Source data