BackgroundThe CD274 (PD-L1)/PDCD1 (PD-1) immune checkpoint interaction may promote cancer progression, but the expression patterns and prognostic significance of PD-L1 and PD-1 in the colorectal cancer microenvironment are inadequately characterised.MethodsWe used a custom 9-plex immunohistochemistry assay to quantify the expression patterns of PD-L1 and PD-1 in macrophages, T cells, and tumour cells in 910 colorectal cancer patients. We evaluated cancer-specific mortality according to immune cell subset densities using multivariable Cox regression models.ResultsCompared to PD-L1–macrophages, PD-L1+macrophages were more likely M1-polarised than M2-polarised and located closer to tumour cells. PD-L1+macrophage density in the invasive margin associated with longer cancer-specific survival [Ptrend= 0.0004, HR for the highest vs. lowest quartile, 0.52; 95% CI: 0.34–0.78]. T cell densities associated with longer cancer-specific survival regardless of PD-1 expression (Ptrend< 0.005 for both PD-1+and PD-1–subsets). Higher densities of PD-1+T cell/PD-L1+macrophage clusters associated with longer cancer-specific survival (Ptrend< 0.005).ConclusionsPD-L1+macrophages show distinct polarisation profiles (more M1-like), spatial features (greater co-localisation with tumour cells and PD-1+T cells), and associations with favourable clinical outcome. Our comprehensive multimarker assessment could enhance the understanding of immune checkpoints in the tumour microenvironment and promote the development of improved immunotherapies.