Hsp110 over-expression increases the immunogenicity of the murine CT26 colon tumor

Abstract.

Several studies have suggested a positive correlation between heat shock protein (hsp) expression and tumor immunogenicity. Independently, many studies have shown that hsp purified from tumors can be used as a tumor-specific vaccine. In this study, we have explored the connection between hsp expression and anti-tumor immunity by transducing murine CT26 colon carcinoma cells with the cDNA of a major hsp, i.e. hsp110. We have shown that over-expression of hsp110 has no effect on CT26 tumor cell growth in vitro, and does not inhibit their anchorage-independent growth capacity. However, in situ, hsp110 over-expressing CT26 tumor (CT26-hsp110) grew at a significantly reduced rate as compared to the wild-type CT26 tumor in immunocompetent mice. Moreover, immunization of mice with inactivated CT26-hsp110 cells significantly inhibited the growth of wild-type CT26 tumor. This immunity was associated with an increased frequency of tumor-specific T cells after vaccination. An in vivo antibody depletion assay demonstrated that inactivated CT26-hsp110 cells elicited anti-tumor responses involving CD8⁺ T cells and natural killer (NK) cells, but not CD4⁺ T cells. Lastly, the effect of the addition of granulocyte–macrophage colony stimulating factor (GM–CSF) to these vaccine formulations was determined. Mice immunized with irradiated CT26-hsp110 cells combined with GM–CSF-producing bystander cells revealed a complete inhibition of CT26 tumor growth, indicating a synergy between inactivated CT26-hsp110 vaccine activity and GM–CSF. These observations demonstrate that manipulation of hsp110 expression in tumors, specifically when combined with GM–CSF, represents a potentially powerful approach to cancer vaccine formulation.