Figure 6.

Sequential administration of anti-PD-1 and anti-Tim-3 improved the efficacy of SA-GM-CSF-anchored vaccine therapy. (A) All mice were injected subcutaneously with 1×10⁶ MB49 cells, and the tumor volume was recorded. Sequential administration of PD-1 and Tim-3 blockade with SA-GM-CSF-anchored vaccine treatment further inhibited the tumor growth compared with the other control groups (*P<0.05). (B) After the treatment was finished, splenocytes from each group were isolated and stimulated by IL-2. MB49 cells served as target cells. Supernatants were collected for a nonradioactive cytotoxicity assay (*P<0.05). (C) After the treatment was finished, splenocytes from mice treated with the sequential administration of PD-1 and Tim-3 blockade and the SA-GM-CSF-anchored vaccine were isolated and stimulated by IL-2. MB49 or RM-1 cells served as target cells. Supernatants were collected for a nonradioactive cytotoxicity assay (*P<0.05). (D) After the treatment was finished, peripheral blood was collected from each group and congealed and then the supernatants were harvested by centrifugation. The concentrations of IL-6 and progranulin (PGRN) were measured by ELISA (*P<0.05). All the experiments were replicated 3 times.