Figure 7. Dual laser treatment combined with the immune checkpoint blockade decreased the expression of PD-1 in TILs.

E0771 murine breast cells were injected into the flank of C57BL/6J mice. The laser treatment was performed for 5 days in a row at day 3–7. Anti-PD-1 antibody was injected at day 3 and 7. At day 16, TILs were purified from tumors and stained for surface markers and cytokines, and then analyzed by flow cytometry. (A) Percentages of FOXP3⁺ CD4⁺ T cells. (B) Representative histograms and percentages of PD-1 and (C) IFN-γ expression in CD8⁺ T cells are shown. (A-C) n= 5, 5, 4, 5, 4, 5, 5, 4 for no laser, no laser + IgG, no laser + αPD-1, dual 1064 nm 1.5 W/cm² + 1270 nm 0.25 W/cm², dual 1064 nm 1.5 W/cm² + 1270 nm 0.25 W/cm² + IgG, dual 1064 nm 1.5 W/cm² + 1270 nm 0.25 W/cm² + αPD-1, dual 1064 nm 3 W/cm² + 1270 nm 1 W/cm², dual 1064 nm 3 W/cm² + 1270 nm 1 W/cm² + αPD-1, respectively. *P < 0.05, **P < 0.01 by one-way ANOVA followed by Tukey’s multiple comparisons test.