An indispensable role of type-1 IFNs for inducing CTL-mediated complete eradication of established tumor tissue by CpG-liposome co-encapsulated with model tumor antigen

Abstract

We have evaluated the capacity of a novel, nanoparticle-based tumor vaccine to eradicate established tumors in mice. C57BL/6 mice were intradermally (i.d.) inoculated with ovalbumin (OVA)-expressing EG-7 tumor cells. When the tumor size reached 7–8 mm, the tumor-bearing mice were i.d. injected near the tumor-draining lymph node (DLN) with liposomes encapsulated with unmethylated cytosine-phosphorothioate-guanine containing oligodeoxynucleotides (CpG-ODN) (CpG-liposomes) co-encapsulated with OVA. This vaccination protocol markedly prevented the growth of the established tumor mass and ∼50% of tumor-bearing mice became completely cured. Tumor eradication correlated with the generation of OVA/H-2K^b-tetramer⁺ CTLs in the tumor DLN and at the tumor site with specific cytotoxicity toward EG-7 cells. Interestingly, tetramer⁺ CTLs failed to be induced in lymph node-deficient Aly/Aly mice. Thus, tetramer⁺ CTLs appeared to be generated in the tumor DLN and subsequently migrated into the tumor site. In vivo antibody blocking experiments revealed that CD8⁺ T cells, but not CD4⁺ T, NK or NKT cells, were the major effector cells mediating tumor eradication. CTL induction was also inhibited when vaccinated tumor-bearing mice were treated with both anti-IFN-α and anti-IFN-β mAbs but not with anti-IFN-α or anti-IFN-β mAb alone. Neither IFN-γ^−/− nor IL-12^−/− mice showed impaired induction of tetramer⁺ CTLs. Thus, these findings revealed that CpG-ODN-induced IFN-α/β, but not IL-12 or IFN-γ, is critical for the generation of tumor-specific CTLs in response to vaccination. These results highlight the potential utility of CpG-liposomes co-encapsulated with protein tumor antigens as therapeutic vaccines in cancer patients.