Fig. 4.

CCL16/CpG + anti-IL-10R mAb combination treatment induces CD8 cytotoxic T cells. a In vitro cytotoxic assay of splenocytes from tumor-free mice after triple-combination treatment. In vitro restimulation has been done for 5 days with either T1525p cells (filled circle) or SV40 Tag IV peptide (open circle) and tested for CTL activity against ⁵¹Cr-labeled syngeneic blast cells pulsed (upper panel) or not (lower panel) with SV40 Tag IV peptide. As controls, splenocytes from mice immunized three times with irradiated T1525p cells (open square) or the unrelated TRAMP tumor cell line T23 (filled square). Splenocytes from a pool of three mice per group have been used, and the experiment repeated twice. b FACS analysis of IFNγ production by CD8 T cells obtained from spleen and tumors of treated and untreated mice 5 days after the end of therapy. Ex vivo CD8 T cells are then induced in vitro to degranulate in response to a short stimulation with SV40 peptide. Upper panel representative plots showing the concomitant expression of IFNγ and CD107a; lower panel shows histograms of the percentage of IFNγ−producing CD107a+ CD8+ T cells. Mean percentage from three mice is shown; experiment has been repeated twice; naïve splenocytes have been used as controls