Archives2019Vol. 59, No. 5pp. 503-515

Article

Designing Recombinant Plasmids Coding Hematopoietic Factors SCF, G-CSF, and FLT-3L and Study of Their Effect on Hematologic Characteristics in Mice

Vlasova O.A.1, Kravtsov I.S.2, Bagnaev I.V.1, Kovshar E.A.1, Kruglova A.A.1, Nikiforov A.S.2, Sventickaya A.M.2

1Federal State Unitary Enterprise “Scientific Center “Signal”, Moscow, Russia. 2 State Scientific-Research Testing Institute of Military Medicine, Ministry of Defense of the Russian Federation, St. Petersburg, Russia

Abstract

The primary problem of radiation biology and medicine is the search for optimal techniques effective for bone marrow acute radiation syndrome and support of oncology patients during chemotherapy. In this study, we show the perspectives of gene therapy for leukopoiesis stimulation and release of white blood cells in mouse blood flow. We have created three plasmid DNAs with the ability to translate hematopoietic factors SCF, G-CSF, and FLT-3L. We have shown expression of these factors at the levels of 5.2 ug, 2.2 ug, and 8.2 ug with secretion efficiency at 75.6, 99.1, and 88.7%, respectively. Single injection of three plasmid mix combined with electroporation leads to statistically significant increase with the maximum values on the 7th day of total leukocyte count (2.6-time increase upto 22.2 × 109/l), granular leucocyte count (4.5 time-increase upto 16.5 × 109/l) and monocyte count (2.8 time-increase upto 2.3 × 109/l), as compared to the control group in outbred white mice. We have also determined the kinetics of gene expression of three cytokines in vivo. A statistically significant increase of cytokine levels was observed from 6 hours to 3 days after a single injection of plasmid DNA mix and lasted for two weeks. Our results confirm the viability of gene therapy for hematopoiesis stimulation, which can be applied for acute radiation syndrome therapy and mobilization of hematopoietic stem cells in the patients scheduled for high-dose chemo/radiation therapy with autologous hematopoietic cell transplantation.

Keywords

plasmid DNA, hematopoiesis, gene therapy, electroporation, stem cell factor, granulocyte colony stimulating factor, FMS-like tyrosine kinase-3 ligand, bone marrow syndrome, acute radiation syndrome, leukocytes, hematopoietic cell mobilization

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