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In this study, the double recombinant vaccinia viruses were generated those express exogenous proteins: human granulocyte-macrophage colony-stimulating factor (GM-CSF) and the antitumor protein lactaptin in secreted and nonsecreted forms. We observed that recombinant VV-GMCSF-Lact with nonsecreted lactaptin exerted stronger cytotoxic activity than others in MDA-MB-231, BT-549 and BT-20 breast cancer cells with calculated CD50 of 0.005; 0.004 и 0.00083 PFU/cell correspondently. Strain VV-GMCSF-Lact also exhibited highest lytic activity in lung cancer cells H1299 and epidermoid carcinoma cells A-431. Normal MCF10A cells and diploid embryonic lung human cells LECH-240 were resistant to all recombinant vaccinia viruses. Strain VV-GMCSF-Lact showed the highest index of tumor selectivity in pairs normal/cancer cells: MCF10A/MDA-MB-231 (>2000) and LECH-240/H1299 (190). By flow cytometry, we demonstrated that all recombinants induced apoptosis in treated cancer cells but the rate of annexin V-positive cells was higher after treatment with VV-GMCSF-Lact than others. Thus nonsecreted lactaptin expression increased the toxicity of recombinant virus to cancer cells in the best way. It is likely that lactaptin expression inside the treated cells (without secretion outside) intensifies apoptosis and as a consequence promotes the progression of apoptotic cells to secondary necrotic cells. These results demonstrate that recombinant VV-GMCSF-Lact has good oncolytic potential and stimulate further investigation of its anticancer activity in human tumor models in vivo and to use it in the development of anticancer therapeutic agents.

Antitumor Effect of Vaccinia Virus Double Recombinant Strains Expressing Genes of Cytokine GM-CSF and Oncotoxic Peptide Lactaptin