In previous reports, systemic administration of a stimulatory
monoclonal antibody directed against the
4-1BB receptor had no effect on survival or
tumor burden in mice inoculated with the poorly immunogenic B16-F10
melanoma. We combined
IL-12 gene transfer with 4-1BB costimulation to explore a previously noted cooperative anti-
tumor effect against this model
tumor. We hypothesize that the innate immune response mediated by IL-12-activated natural killer (NK) cells initiates the activation of the immune system, leading to the priming of T cells, whereas 4-1BB costimulation enhances the function of primed
tumor-specific T cells. The effect of the combination
therapy on the growth of subcutaneous (s.c.)
tumors and pulmonary
metastasis was examined. The combination
therapy significantly retarded the growth of subcutaneously-inoculated
tumors, and 50% of
tumor-bearing mice survived with complete
tumor regression. In contrast, neither
IL-12 gene transfer nor anti-4-1BB antibody administration alone was as effective. Enhanced CTL activity against both B16-F10
tumor cells and TRP-2-pulsed EL4 syngeneic
tumor cells was observed in
tumor-bearing animals treated with the combination
therapy 2 weeks
after treatment and, in long-term survivors from this combination
therapy, at >120 days. In a pulmonary metastatic model, only the combination
therapy generated significant protection against
metastasis. In vivo depletion of NK or CD8(+) but not CD4(+) subsets eliminated the protective immunity. Furthermore, NK cell depletion significantly reduced both
tumor-specific CTL activity and the number of
tumor-specific IFN-gamma-producing cells, suggesting that this synergistic effect requires the participation of both NK and CD8(+) T cells.