Several in silico immune deconvolution published methods20–25 were benchmarked against molecular protein staining of CD4+ and CD8+ cells, tumour cellularity and copy number profiles (Methods, Supplementary Fig. 7), and the best-performing method, the Danaher et al.22 signature was used to investigate the tumour-infiltrating immune cell populations within the 120 ESCC tumours. This evidence concerns the gene CD4 and esophageal squamous cell carcinoma.