DENDRITIC CELLS ELECTROPORATION WITH DNA-PLASMID ENCODING IL-10

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Abstract

Dendritic cells (DC) are widely used to produce vaccines for the treatment of various types of diseases, such as malignant neoplasms, autoimmune disorders, rejection and GVHD. To control the functional properties of DCs, they are transfected with various immunoregulatory proteins, including IL-10. In this paper, the optimization of the method of electrical transfection (electroporation) of DCs with a DNA plasmid encoding IL-10 was performed. The effi ciency of DC electroporation (EP) reaches 45‒72% with cell death below 7%. The active expression of IL-10 in transfected DC was shown within 3 days after EP. According to the results of the work, EP was recognized as an eff ective method for obtaining transfected DCs for the production of cellular vaccines.

About the authors

V. P. Tereshchenko

FSBSI Research institute of fundamental and clinical immunology

Author for correspondence.
Email: tervp@ngs.ru

Ph.D studentin laboratory of molecular immunology,

Novosibirsk

Russian Federation

J. N. Khantakova

FSBSI Research institute of fundamental and clinical immunology

Email: fake@neicon.ru

Ph.D, researcherin laboratory of molecular immunology,

Novosibirsk

Russian Federation

V. V. Kurilin

FSBSI Research institute of fundamental and clinical immunology

Email: fake@neicon.ru

Ph.D, researcherin laboratory of molecular immunology,

Novosibirsk

Russian Federation

A. N. Silkov

FSBSI Research institute of fundamental and clinical immunology

Email: fake@neicon.ru

Ph.D, researcherin laboratory of molecular immunology, head of RIFCI,

Novosibirsk

Russian Federation

R. A. Maksyutov

SRC of Virology and Biotechnology “Vector”

Email: fake@neicon.ru

Ph.D, researcher,

Koltsovo

Russian Federation

References

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  2. Akbari O., DeKruyff R. H. &Umetsu D. T. Pulmonary dendritic cells producing IL-10 mediate tolerance induced by respiratory exposure to antigen. (2001) Nature Immunol 2: 725–731.
  3. Fiorentino D. F., Zlotnik A., Mosmann T. R., Howard M. & O’Garra A. IL-10 inhibits cytokine production by activated macrophages. J. Immunol. 147, 3815–3822 (1991).
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  5. Yin H., Kanasty R. L., Eltoukhy A. A., Vegas A. J., Dorkin J. R., Anderson D. G. Non-viral vectors for genebased therapy. Nat Rev Genet. 2014 Aug;15(8):541–55.

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Copyright (c) 2019 Tereshchenko V.P., Khantakova J.N., Kurilin V.V., Silkov A.N., Maksyutov R.A.

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