Fig. 1.

Prognostic value of CD8⁺T cells and CD68⁺CD163⁺macrophages revealed by comprehensive characterization of immune cell subsets in 286 therapy-naïve colon cancers (See also Figures S1–S4). (a) Immune marker combinations in IHC panels define classes and subclasses of immune cells (See also Supplementary Table S1). (b) Forest plot of univariable associations of tissue immune cell densities translated into three-level categorised values, with OS in patients of stage I-III. Filled squares, hazard ratios (HR); whiskers, 95% confidence intervals (CI), ∗p < 0.050 (Cox regression). (c) Representative multiplex macrophage marker staining of colon cancer tissue. Expression of two markers, CD68 (red) and CD163 (green) with nuclear DAPI staining (white), visualised in pseudocolours, identified three cell types (insets), M1-like macrophages, M2-like macrophages and CD68⁻CD163⁺ cells. (d) Venn diagram of the counts of cells in the entire cohort expressing CD68 only (red, n = 9.0 × 10⁵), CD163 only (green, n = 1.9 × 10⁵) or both markers (gold, n = 4.4 × 10⁴). (e) Density of three macrophage subsets in patient tumours. Boxes, median and interquartile range (IQR) of the ratios; whiskers, 1.5 IQR. (f) Signature of immune activation (SIA), defined as the ratio of CD8⁺ cell density to the sum of CD8⁺ and M2-like cell densities.