Construction of recombinant vaccinia virus expressing GM-CSF and its use as tumor vaccine.

We made several generic plasmids for construction of recombinant vaccinia virus (rvv) expressing foreign proteins in high yield. Rvvs expressing biologically active Escherichia coli beta-galactosidase (rvv-lacZ) and the cytokine murine GM-CSF (rvv-mGM-CSF) were constructed by using these plasmids. To obtain attenuated rvv, cDNA for these proteins was inserted in the thymidine kinase gene of vaccinia virus. Their expression was controlled by vaccinia early/late promoter, 7.5 K so that these proteins could be expressed in the infected cells throughout the life cycle of the virus. Female C57BL/6 mice were immunized subcutaneously with B16-F10 melanoma cells infected with rvv, and 2 weeks later challenged with viable B16 cells. Mice immunized with rvv-mGM-CSF showed delay in tumor development, smaller tumor volumes and longer survival time compared with unimmunized mice, as well as mice immunized with rvv-lacZ. Mice immunized with rvv-mGM-CSF followed by a booster injection after 1 week responded slightly better than those immunized once, but this difference was not statistically significant. These results suggested that rvv-mGM-CSF could be a promising vaccine for cancer therapy.