Spatial transcriptomics combined with single-cell sequencing was used to analyze the expression of immune-related cells and chemokines and receptors in the TME in human squamous cell carcinoma, such as the CXCR3 (Treg, CD8+TEM), CXCR6 (Treg, CD4+RGCC+, and CD8+ depleted cells), and CXCR4 (CD4+RGCC+, CD8+ TEMRA, and NK cells). The gene discussed is CD4; the disease is squamous cell carcinoma.