Fig. 4: Intratumoral immune infiltrations within full-tumor sections following KN026 treatment.

a Representative images of segmented ROIs from two cases (OR vs non-OR) based on morphological staining of PanCK, CD3, and CD20 markers for intuitive comparison of treatment-related changes. b Heatmap of 29 individual ROIs comparing immune cells clustered in regions based on morphological segments on the x axis; red box depicts the most immune cell subtypes interacted with tumor cells. c Boxplots compared critical activation signalings within T cells and B cells in the cases of matched treated samples. P values were derived using a two-sided Wilcoxon rank-sum test. d Signature scores (y-axis) of cell types and cell states (x-axis) in responder tissues out of all T and B cell subsets in pre-treated versus post-treated tumor regions from OR. e Waterfall plots illustrating functional genes expression changes in T cells and B cells within tumor with OR following treatment. f Pairwise correlation of DEGs in T and B cells from tumor with OR following treatment. The black squares represent hierarchical clusters for modular division. g Pathway enrichment among DEGs of T cells and B cells following treatment. Left: comparison between critical up- and down-pathways within TME following effective treatment. Right: comparison of significant changed signalings in T and B cells. h Flow cytometry plots showing perforin+ and granzyme+ CD8 + T cell subset gated on all cytotoxic T cells (CD8a+TCR Vα7.2 + CD4-PD-1-). Proportion of perforin+ granzyme+ cytotoxic T cells subsets from treatment and control groups. Data points represent values from 5 individual co-cultured well and lines represent means ± SD. Statistical analysis using the Mann–Whitney test. i Cell viability of BT474 assessed by CCK8 assay in response to KN026 (100 µg/ml) treatment co-cultured with cytotoxic T cells. Triplicate measurements per group, and p values correspond to the Student’s t-test.