Fig. 6: The mechanism of tumor-driven CD8⁺ T_RM cell activation in type 3 TETs.

(a) Same t-SNE plots as (Fig. 2c), colored by CD8⁺ T_RM cells and other cells. (b) Bar plot showing the frequencies of CD8⁺ T_RM cells among the three groups (n = 3, 16 and 6 for N, T1 + T2 and T3, respectively. Data are presented as the mean ± s.e.m. P values in the figure were determined by an unpaired two-tailed Student’s t test). (c) Heatmap showing the number of interactions between CD8⁺ T_RM cells and epithelial cell subpopulations. (d) Heatmap of chemokine interactions among T cells and epithelial cells, normalized per column. (e–f) Flow cytometric analysis of CXCR3 expression among CD4⁺ T cells, CD8⁺ non-T_RM cells and CD8⁺T_RM cells in TETs. (g) Heatmap showing the expression of activation-associated proteins in CD8⁺ T_RM cells in each group by CyTOF and normalized total matrix. (h) Dot plot of the expression of cell activation- and TCR activation-associated genes within CD8⁺ T_RM cells and other T-cell subsets. (i-j) Flow cytometric analysis of IFN-γ and CD39 expression among CD4⁺ T cells, CD8⁺ non-T_RM cells and CD8⁺T_RM cells in TETs. (k) KEGG pathway analysis of genes that were significantly (P  <  0.01) upregulated in CD8⁺ T_RM cells compared to other T cells. (l) Heatmap of chemokine interactions among CD8⁺ T_RM cells and other cell types, normalized per column. (m–n) Flow cytometric analysis of CXCL13 expression among CD4⁺ T cells, CD8⁺ non-T_RM cells and CD8⁺ T_RM cells in TETs. (o–p) Representative IF staining images showing CD103 (red), EPCAM (green), CD11c (green), CD20 (pale blue) and DAPI (nuclei, blue) in type 3 TET samples. The interactions between epithelial cells and T_RM cells are indicated by white arrows (o). The interactions between T_RM cells and DCs are indicated by yellow arrows, and those between T_RM cells and B cells are indicated by white arrows (p). Scale bar: 20 μm. Experiment was performed in three independent samples with similar results. (f, j, n) n = 8(f), 8 (j-left), 6 (j-right),6(n), respectively. Data are presented as the mean ± s.e.m. P values in the figure were determined by a two-tailed paired Student’s t test. Source data are provided as a Source Data file.