Fig. 3.

Combined therapy with B10G5 and anti-PDL1 increases intrinsic functional potential and response to antigen-specific stimulations. B16F10-sMICB cells (4 × 10⁵ cells/mouse) were s.c. injected into syngeneic MICB/B6 host. When tumors reached a volume of 75–100 mm³, animals received i.p injection of 3 mg/kg of respective antibody every 3 days. After three injections (day 9 of therapy), animals were euthanized. Tissues were harvested for therapy-associated mechanistic studies. a, b Combined therapy significantly increases IFNγ-producing splenic CD8 T cells as assessed by ex vivo PMA/ionomycin stimulation. c, d Combined therapy significantly increased the number of splenic NKG2D⁺ CD8 T cells. e, f Combined therapy significantly increased the population of CD44⁺CD8 T cells. Data obtained at day 14 following treatment initiation. g, i Representative flow cytometry dot-plots demonstrating that combined therapy significantly increases gp100-tetramer⁺ CD8 T cell and increased IFNγ expression with gp100 peptide stimulation. Single-cell suspension of splenocytes was stained with melanoma antigen gp100-specific tetramer or evaluated for CD8 T cell IFNγ expression after stimulation with gp100 peptide overnight. h, j Summary data from the experiments presented in g and i respectively. *p < 0.05 as compared to the control group. **p < 0.05, combination therapy as compared to monotherapy