Fig. 4: Investigating T/NK cells functional diversity and HBV lymphotropism in HBV-mediated HCC.

A UMAP plot of HBV and non-HBV-tumor T/NK subsets. Cluster number and cell type annotations are provided in the Figure. B Expression of marker genes correlated with T/NK subtypes (αβ T cells, γδ T cells, T memory, Tregs, CD8⁺ T, and NK cells). C T/NK subtype selected gene expression heatmap for naive, resident, inhibitory, cytokines, co-stimulatory, transcriptional factors (TF), and cell type. D Boxplots showing the ratio of KLRF1⁺, KLRD1⁺, and KLRB1⁺ NK cells in HBV(purple) and non-HBV (green) tumor patients (left panel), violin plots indicating the expression of KLRD1 in NK cells between HBV(blue) and non-HBV HCC (green) samples from the Validation cohort 2 (right panel). E UMAP colored by viral status, representing HBV-infected (green) and non-infected (red) T cells (left panel). F UMAP colored by viral status, representing HBV-infected (green) and non-infected (red) B cells (left panel); UMAP colored by viral status, representing only HBV-infected (green) B cells (right panel). G A coverage map of the HBV genome. The locations of the HBV genes, as listed in the NCBI database, are depicted at the bottom. H Representative IHC staining, showing CD3⁺HBc⁺ T cells in paired HBV and non-HBV HCC samples. Scale bar, 20 μm. I Boxplot showing the ratio of NK cells in HBV and non-HBV tumor patients. J Boxplot showing the ratio of CD4⁺, CD8⁺ T, and Th17 cells in HBV and non-HBV-HCC patients. Error bars represent mean ± standard deviation (SD). The p-values in (D, I, and J) were calculated using the unpaired Wilcoxon test. All statistical comparisons were performed on biologically independent samples (n = 5 HBV-HCC and n = 3 non-HBV-HCC) from both the discovery and validation cohort 2. See also Supplementary Figs. 8, 9 and 10.