Figure 4: TIL function is not associated with inhibitory receptor expression.

(A) Heatmap associations of relative expression of IFNγ production and inhibitory receptor frequencies (rows) for individual patient CD8⁺ TILs (columns). Red-maximum; blue-minimum, grey-no values. Data were not transformed.

(B) The percentage of CD8⁺ TILs expressing PD-1, TIGIT, CD39, TIM-3 and intracellular CTLA-4 was compared in functional (>19.3%) and hypofunctional (<19.3%) cases.

(C) The percentage of CD8⁺ TILs expressing PD-1⁺TIGIT⁺, PD-1⁺TIM-3⁺, PD-1⁺iCTLA-4⁺ combinations were compared in functional and hypofunctional cases.

(D) Representative single cell analyses showing IFNγ expression (x-axis) versus PD1 expression (y axis) for two cases. The boxes depict the percentage of PD-1⁺ IFNγ⁺ (top) or PD-1⁻ IFNγ⁺ (bottom).

(E) Representative single cell analyses showing IFNγ expression (x-axis) versus TIGIT, TIM3 and CD39 expression (y axis) for four cases. The boxes depict the percentage of TIGIT⁺ IFNγ⁺, TIM3⁺ IFNγ⁺ or CD39⁺IFNγ⁺ cells.

(F) The percentage of CD8⁺ TILs cells producing IFNγ based on their IR expression is plotted. Flow staining was performed individually for the TFLLs, and concurrently for the PBMCs, DLALs and TILS from each respective patient.

*p<0.05, ***p<0.001.