Highly effective production of biologically active, secreted, human granulocyte-macrophage colony-stimulating factor by recombinant vaccinia virus

• Published in: Applied biochemistry and microbiology 2016-12, Vol.52 (7), p.685-691
• Main Authors: Grazhdantseva, A. A., Sivolobova, G. F., Tkacheva, A. V., Gileva, I. P., Kuligina, E. V., Rikhter, V. A., Kochneva, G. V.
• Format: Article
• Language: English
• Subjects: Biochemistry; Bioengineering; Biological activity; Biological effects; Biology; Biomedical and Life Sciences; Cell culture; Cell growth; Cell proliferation; Colonies; Colony-stimulating factor; Cytokines; Erythroleukemia; Gene Engineering; Gene expression; Granulocyte-macrophage colony stimulating factor; Granulocytes; Insertion; Kinases; Life Sciences; Mammalian cells; Medical Microbiology; Microbiology; mRNA; Oncolysis; Producers; Selection; Thymidine; Thymidine kinase; Vaccines; Viruses
• ISSN: 0003-6838; 1608-3024
• DOI: 10.1134/S0003683816070036

The recombinant vaccinia virus strain VV-GMCSF-S1/3, which contains an insertion of full-length DNA copy of messenger RNA of human granulocyte-macrophage colony-stimulating factor (GM-CSF) in the structural part of the viral thymidine kinase gene, was obtained. The expression of the GM-CSF gene as a part of the recombinant virus is under the control of the native vaccinia virus promoter р7.5K; this results in the production of a mature form of the secreted protein with a molecular mass of 32 kDa. The biological activity of GM-CSF was evaluated by stimulation of the proliferation of cytokine-dependent human TF-1 erythroleukemia cells. The secretion level of biologically active human GM-CSF in the system of recombinant vaccinia virus/mammalian cells was 1–40 μg/mL of culture medium. The recombinant strain VV-GMCSF-S1/3 can be used as a producer of the glycosylated mature form of human GM-CSF, as well as a vector for the construction of oncolytic viruses and multivalent vaccine preparations.