Fig. 3

Preoperative pembrolizumab in combination with chemoradiotherapy modulates the tumor immune microenvironment. a Study design comparing PALACE cohort with control cohorts. b Stacked bar chart comparing the immune cell proportions among different groups. c UMAP plot highlighting neutrophil clusters. d Box plots comparing the proportions of RHOH⁺, ISG15⁺, and VEGFA⁺ neutrophils across different groups. e Box plots showing the proportions of CD4⁺ and CD8⁺ T cells across different groups. f UMAP plots showcasing distinct CD8⁺ T-cell clusters. g Box plot comparing the proportions of CXCL13⁺CD8⁺ T cells among different groups. h Dotplot displaying the expression of 3 signatures of previously reported tumor-reactive CD8⁺ T cells across all CD8⁺ T-cell clusters. i UMAP plots displaying distinct CD4⁺ T-cell clusters. j Box plot comparing the proportions of CXCL13⁺CD4⁺ T cells among different groups. k Dotplot showing the expression of 2 signatures of previously reported tumor-reactive CD4⁺ T cells across all CD4⁺ T-cell clusters. P values were derived from one-way ANOVA, Tukey’s test; ns: not significant, *: P < 0.05. Data are presented as mean ± SEM. BS treatment-naïve baseline, nCRT neoadjuvant chemoradiotherapy, PPCT preoperative pembrolizumab combined with chemoradiotherapy, PPCT-N non-pCR following PPCT, PPCT-P pCR following PPCT, pCR pathological complete response