Figure 1.
Impact of IFNγ addition or depletion on H-1PV immunomodulating activity. (A) Macrophages were isolated from the peritoneal cavity of four groups (n = 3) of tumor bearing Lewis rats treated either with PBS (mock) or with an intratumoral injection of 5 × 108 pfu/rat of H-1PV (H-1PV IT) combined either with an antibody against IFNγ (H-1PV IT + αIFNγ) or recombinant IFNγ (H-1PV IT + recIFNγ). Cells were plated in 48-well plates at a density of 5 × 105 cells per well and stimulated or not with LPS. TNFα production in the supernatants was measured 24 h later by ELISA. Average values and standard deviations are shown. (B) Peritoneal macrophages (5 × 105/well) from the same groups of rats were cocultured or not with 1 × 105 HA-RPC cells at a 5:1 ratio in 48 well plates and the release of interleukins -10 and -12 was measured by ELISA. Mean cytokine ratios and standard deviations are presented. (C) Single cell suspensions of rat splenocytes were labeled with CFSE, plated in 24 well plates at 1 × 106 cells/ well and cocultured or not with 2 × 105 HA-RPC cells at a 5:1 ratio. 48 h later cells were harvested and processed for FACS analysis of proliferation. All data were median from three animals from triplicate wells. Differences were considered significant at p values below 0.05.