VIRAL INTERLEUKIN-10 (IL-10), THE HUMAN HERPES-VIRUS-4 CELLULAR IL-10 HOMOLOG, INDUCES LOCAL ANERGY TO ALLOGENEIC AND SYNGENEIC TUMORS

After the cloning of murine cytokine synthesis inhibitory factor, it was recognized that a homologous open reading frame was encoded within the Epstein-Barr virus (human herpes virus 4). This viral protein has now been termed viral interleukin 10 (vIL-10) to reflect its protein sequence homology to ''cellular'' IL-10 (cn;-10, either murine or human IL-10). It is now widely accepted that vIL-10 shares many functions with cIL-10, principally, the ability to enhance survival of newly infected B cells and to diminish the production of IFN-gamma and IL-2 during ongoing immune reactions. The immunomodulatory effect of locally secreted vIL-10 and murine IL-IO (vIL-10) was examined in tumor models using CL8-1 (a BL6 melanoma cell line transfected with the H-2K(b) class I gene) in syngeneic animals. Although parental BL6 tumor cells grow in immunocompetent syngeneic hosts, CL8-1 are rejected. To achieve local secretion of vIL-10, we generated vIL-10 retroviral vectors. While nontransduced CL8-1 cells (1 x 10(4)) failed to grow when injected intradermally in C57BL/6 mice, CL8-1 cells (1 x 10(4)) transduced with vIL-10 formed palpable tumors and eventually killed 80% of injected animals. Suppression of tumor rejection was also noted when CL8-1 tumors with or without vIL-10 transfection were admired with syngeneic vIL-10-transfected fibroblasts and inoculated. Since the in vitro proliferation of the tumor was not altered after transduction with the vIL-10 gene and injection of vIL-10-transduced CL8-1 does not affect the rejection of nontransduced CL8-1 inoculated at a distant site, local vIL-10 secretion appears to suppress the process of immune rejection of the target cells in a dose-dependent manner. Similar results were observed for the H-2(b) MCA105 sarcoma tumor model in allogeneic BALB/c mice (H-2(d)). Although all animals that received nontransfected MCA105 rapidly rejected these tumors, MCA105 sarcomas transfected with vIL-10 remained palpable for up to 37 d. The local immunosuppressive effect of gene-delivered vIL-10 could be neutralized by anti-human IL-10 monoclonal antibody or could be reversed by the systemic administration of IL-2 or IL-12. In marked contrast, mIL-10 transfection of CL8-1 significantly suppressed tumor growth and frequently led to the rejection of tumor. Similar results were obtained for the murine tumor cell lines MC38 and MCA102. These results contrast the immunologic effects of vIL-10 and cIL-10 and argue for divergence in the function of the virally captured gene, which is presumably the result of altered interaction with one of the two derived IL-10 receptors. The present study, along with our recent study showing prolonged survival of cardiac allografts transduced with retroviral vIL-10, suggests that local vIL-10, but not mIL-10, secretion can suppress immune reactivity in both the syngeneic and allogeneic settings.