Fig 1: Distinct immune microenvironment across LIHV-defined subgroups(A) Immune cell abundance comparison across subgroups using the indicated immune analysis tools across four independent iCCA cohorts. Circle color and size represent log2 fold change and p value, respectively.(B) Boxplots of CD66b+ neutrophils, CD68+ macrophages, CD3+ T cells, CD20+ B cells, and αSMA+ fibroblasts across subgroups (Mann-Whitney U test).(C) Heatmap of immune signatures and checkpoint expression summarized as mean Z scores per subgroup across four cohorts. ∗FDR < 0.05; ∗∗FDR < 0.01; ∗∗∗FDR < 0.001.(D) Heatmap of Spearman correlations between chemokine expression and neutrophil infiltration. IHC, immunohistochemistry.(E) Boxplot comparing CXCL5 expression across subgroups in the Fu-iCCA cohort (Mann-Whitney U test).(F) Dot heatmap of chemokine gene expression across major cell types in Xue’s scRNA-seq dataset.(G) Boxplots of average CXCL5 expression in tumor cells and macrophages across subgroups (Mann-Whitney U test).(H) Western blot validating CXCL5 knockdown and overexpression efficiency in RBE and HuCCT1 cells.(I) Quantification of migrated neutrophils in transwell assays co-cultured with modified RBE and HuCCT1 cells (n = 4 replicates per group; mean ± SD; Student’s t test).(J) Western blot analysis of CXCL5 overexpression in KTP cells.(K) Tumor growth curves of mice injected with control or CXCL5-overexpressing KTP cells (n = 6 per group; mean ± SEM; Student’s t test).(L) Proportion and number of infiltrated neutrophils in control and CXCL5-overexpressing KTP tumors (n = 6 per group; Student’s t test).∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.See also Figures S6 and S7 and Table S5.