Figure 4: TCE-IL2_REH combination therapy promotes anti-tumor TIL responses and differentially affects CD4⁺ and CD8⁺ TILs.

(A) Schematic overview of experimental workflow. Tumors were removed at day 6 after treatment prior to MULTI-seq barcoding, T cell enrichment, and paired scRNA-seq and scTCR-seq.

(B) Uniform manifold approximation and projection (UMAP) visualization of CD4⁺ TIL (left) and CD8⁺ TIL (right) gene expression space colored by subtype annotation.

(C) UMAP visualization of CD4⁺ TIL gene expression space colored by treatment groups (left) with bar charts of CD4⁺ TIL subtype proportions in each treatment group. * p < 0.05 and proportion ratio ± 0.5, propeller test.

(D) Bar charts of CD8⁺ TIL subtype proportions in each treatment group. * p < 0.05 and proportion ratio ± 0.5, propeller test.

(E) Z-score heatmap of the average expression of treatment-specific Tex marker genes in each treatment group. Genes grouped by hierarchical clustering of the z-score matrix.

(F) Violin plots of a subset of treatment-specific Tex marker genes in each treatment group. * p < 0.05 Bonferroni-corrected Wilcoxon rank-sum test.

(G) Z-score heatmap of the average expression of treatment-specific effector CD8⁺ TIL marker genes in each treatment group. Genes grouped by hierarchical clustering of the z-score matrix.

(H) Violin plots of a subset of treatment-specific effector CD8⁺ TIL marker genes in each treatment group. * p < 0.05 Bonferroni-corrected Wilcoxon rank-sum test.

See also Supplemental Figure S5.