Figure 7.

Combining a modulator of infiltrating-myeloid cells and an inhibitor of PD-1/PD-L1 axis increases T cell proliferation and T cell producing IFNγ. Myeloid cells CD11b+ were isolated from tumors using BD FACSAria II (BD Biosciences). T cells were isolated from spleen of naive mice using mouse pan T cell isolation Kit (Miltenyei). CFSE-labeled T cells were stimulated with CD3/CD28 beads ratio 1:1 (Life Technologies). Stimulated T cells were cultured with CD11b+ (at a ratio CD11b:T cells of 1:1) and incubated for 72 h at 37°C. (A) Representative histograms obtained by FACS analysis showing the fluorescence intensity of CFSE-T cells gated on CD8⁺. (B) The percentage of proliferating CD8⁺ cells from the different treated groups is shown. (C) IFNγ secretion in the supernatant of the co-culture is measured 72 h following incubation at 37°C using Luminex Technology. Experiments were repeated twice (Kruskal–Wallis test, *p < 0.05).