Fig. 1
The TME of OPSCC tumors is highly infiltrated with Th1-like T cells. Paraffin-embedded OPSCC tissue of 41 patients was analyzed by triple immunofluorescent confocal microscopy analysis with antibodies directed against CD8 (blue), Foxp3 (green) and Tbet (red). a Example of a triple immunofluorescent staining is given for a representative OPSCC sample. Triple staining results in: CD8+Foxp3–Tbet– (blue), CD8+Foxp3–Tbet+ (blue with red nucleus), CD8–Foxp3+Tbet– (CD4; green nucleus), CD8–Foxp3+Tbet+ (CD4; yellow nucleus) and CD8–Foxp3–Tbet+ (CD4; red nucleus) cells. b Scatter plot depicting (from left to right) CD8–Foxp3+Tbet+, CD8–Foxp3+Tbet–, total CD8–Foxp3+, CD8–Foxp3–Tbet+ (all CD4), CD8+Foxp3–Tbet+, CD8+Foxp3–Tbet–, total CD8+ (all CD8) and total Tbet+ (CD4 and CD8) T cell infiltrates/mm2 for each patient. Cells in the epithelium (tumor) and stroma were manually counted and represented as the number of cells per mm2 for each slide (average of five images at 250× magnification). c Scatter plots and correlation analysis between the number of cells/mm2 in the stroma (Y-axis) and the tumor (X-axis) for CD8–Foxp3+Tbet+, CD8–Foxp3+Tbet–, total CD8–Foxp3+, CD8–Foxp3–Tbet+ (all CD4; top), and CD8+Foxp3–Tbet+, CD8+Foxp3–Tbet–, total CD8+ (all CD8) and total Tbet+ (CD4 and CD8; bottom) T cell infiltrates. Pearson’s correlation with the correlation coefficient (r), the coefficient of determination (r2) and p value is depicted for each correlation analysis. * p < 0.05, ** p < 0.01, *** p < 0.001 and, **** p < 0.0001. The dotted lines represent the 95% confidence interval