Figure 1. T-cell subsets in PBMC, tumor samples and non-cancerous mucosa of HNSCC patients and PBMC of healthy controls.

Single cell suspensions of tumor tissue (n = 34), non-cancerous mucosa (n = 7), PBMCs of healthy controls (PBMC HC, n = 15) and PBMCs of patients with HNSCC (PBMC HNSCC, n = 34) were analyzed by flow cytometry for their expression of T-cell related antigens. (A) Scatter plots showing the number of CD45⁺ cells per μg of tumor or mucosal tissue. (B) Scatter plots comparing the percentages of CD3⁺ T cells within the CD45⁺ fraction (left), CD4⁺ (middle plot) and CD8⁺ (right) cells within the T cells fraction. (C) Depicted in bar graphs is the ratio of CD4⁺ and CD8⁺ T cells (CD3⁺ fraction) in different compartments. (D) Comparison of the rate of naïve (CD45RA⁺/CCR7⁺; left) and effector memory T cells (CD45RA⁻/CCR7⁻; right) in the T-cell fraction, shown as scatter plots. (E) Percentages of regulatory T-cell phenotypes CD4⁺/CD25⁺/CD127^low and CD4⁺/CD39⁺ within PBMC of healthy donors, HNSCC patients, HNSCC tumor tissue and normal mucosa are compared in scatter plots. (F) Bar graphs comparing the ratio of CD8⁺ cells and regulatory T-cell phenotypes CD4⁺/CD25⁺/CD127^low and CD4⁺/CD39⁺ within the CD3⁺ fraction. For statistical analysis, Mann-Whitney test was used in (A), one-way ANOVA in (B) and right plot of (D) and Kruskal-Wallis test in (C), left plot of (D), (E) and (F). Data is depicted as mean ± standard deviation. *P < 0.05; **P < 0.005; ns, not significant.