Figure 4.

Co-regulatory molecules show specific clustering patterns on mature Vδ1 differentiation stages and are co-expressed in OvCA. The surface expression of TIGIT, PD-1, and CD39 on Vδ1 T cells was compared between differentiation stages based on CD27 and CD45RA expression in OvCA PBL (triangles, n = 17), MAL (squares, n = 16), and TIL (diamonds, n = 8). CD27⁺CD45RA⁺: naïve (NA); CD27⁺CD45RA⁻: central memory (CM); CD27⁻CD45RA⁻: effector memory (EM); CD27⁻CD45RA⁺: terminally differentiated effector memory (TEMRA); CD27⁻CD45RA^high: TEMRA subpopulation with high CD45RA expression. (A) Summary data show the frequency of CRR⁺ cells in the respective differentiation stage vs. all other stages taken together. Upper row: TIGIT⁺ cells in all TEMRA (light blue, left panel) and TEMRA^high Vδ1 T cells (dark blue, right panel) vs. other (black; for TEMRA: NA, CM, and EM; for TEMRA^high: NA, CM, EM, and TEMRA^low/int). Lower row: PD-1⁺ cells in CM (green, left panel) and CD39⁺ cells in EM Vδ1 T cells (red, right panel) vs. other (black; for CM: NA, EM, and TEMRA; for EM: NA, CM, and TEMRA). p-values were obtained by the Wilcoxon matched-pairs signed-rank test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. (B) Representative flow cytometry data illustrate the expression of TIGIT (left column), PD-1 (middle column), and CD39 (right column) in Vδ1 T cells based on the expression of CD27 and CD45RA in OvCA PBL (first row), MAL (second row), and TIL (third row), respectively. (C) Summary data depict the expression of PD-1, TIM-3, and CD39 on TIGIT⁺ (orange) vs. TIGIT⁻ (black) cells on Vδ1 T cells. p-values were obtained by the Wilcoxon matched-pairs signed-rank test. * p < 0.05, ** p < 0.01, *** p < 0.001. (D) Representative flow cytometry data illustrate the (co-)expression of TIGIT and PD-1 (first row), TIM-3 (second row), and CD39 (third row) in Vδ1 T cells in OvCA PBL (left column), MAL (middle column), and TIL (right column), respectively.