In current study, the infiltration of four immune cell types (CD8+ /CD4+/Foxp3+/CD33+ cells), the expression of both inhibitory (PD-1/PD-L1/Tim-3/LAG-3) and stimulatory checkpoints (OX-40/ICOS), and IDO1 were evaluated in two independent cohorts with ESCC (95 patients in the primary cohort and 55 patients in the validation cohort). This evidence concerns the gene CD4 and esophageal squamous cell carcinoma.