Fig. 2

Intratumoral CD4⁺FoxP3⁺ cells make up a small subset but display the highest levels of immune checkpoint protein expression. A Proportions of T cell subsets, i.e., CD8⁺, CD4⁺FoxP3⁻ and CD4⁺FoxP3⁺ in MM, NSCLC, RCC, HNSCC, EOC, and UC obtained by flow cytometry analysis of 72 freshly resected tumor specimens. B Percentage of immune checkpoint protein (ICP) positive cells in CD8⁺, CD4⁺FoxP3⁻ and CD4 + FoxP3 + T cells from 35 tumor specimens. C Mean fluorescence intensity of ICPs in CD8⁺, CD4⁺FoxP3⁻ and CD4⁺FoxP3⁺ T cells from 35 tumor specimens. Dunn’s multiple comparison test was performed independently for each ICP. D Heat map displaying the ratio of the ICP median MFI of CD4⁺FoxP3⁺ cells over CD4⁺FoxP3.⁻. Dunn’s multiple comparison test, *p value ≤ 0.05; **p value ≤ 0.01; ***p value ≤ 0.001; ****p value ≤ 0.0001. MM: Metastatic Melanoma; NSCLC: Non-Small Cell Lung Carcinoma; RCC: Renal Cell Carcinoma; HNSCC: Head and Neck Squamous Cell Carcinoma; EOC: Epithelial Ovarian Cancer; UC: Urothelial Carcinoma; ICPs: Immune Checkpoints: (CD25, CD28, CD39, 4-1BB, CTLA-4, ICOS, OX40, PD-1, PD-L1, and TIGIT)