Figure 5
Mice injected with anti-TIM-3/CD137 mAb developed a tumor antigen-specific CTL response. A, mice injected i.p. with 1 × 106 ID8 cells 10 day earlier were injected thrice at 4 days interval with 250 μg of anti-TIM-3/CD137 mAb. Seven days after the last mAb injection, splenocytes from treated mice were cultured in the presence or absence of H-2Db-restricted mesothelin-derived peptides or control HPV-E7-derived peptide for 3 days and IFN-γ production in the supernatants were assayed by ELISA. B, mice were treated as described in (A). Seven days after the last mAb injection, splenocytes (5 × 106) from 3 mice were incubated with 5 × 105 UV-irradiated ID8 cells for 4 days prior to subject to analysis of antigen-specific CTL activity by CytoTox 96 Non-radioactive cytotoxicity assay using EL4 cells pulsed with H-2Db-restricted mesothelin or HPV-E7 peptide as target cells. The killing activity was also evaluated in the presence of anti-CD4, anti-CD8 or control antibody (C). Data were expressed as M ± SEM of triplicate wells.