Journal of microbiology, epidemiology and immunobiologyJournal of microbiology, epidemiology and immunobiology0372-93112686-7613Central Research Institute for Epidemiology125410.36233/0372-9311-320Research ArticleBiological properties of domestic strain vRub-Ant of rubella virusNagievaFiraya G.<p>D. Sci. (Med.), Assoc. Prof., Head, Laboratory of hybrid cell cultures, Department of virology</p>fgn42@mail.ruhttps://orcid.org/0000-0001-8204-4899BarkovaElena P.<p>PhD (Biol.), leading researcher, Laboratory of hybrid cell cultures, Department of virology</p>e.barkova2012@yandex.ruhttps://orcid.org/0000-0002-3369-8869StroevaAlexandra D.<p>junior researcher, Laboratory of hybrid cell cultures, Department of virology</p>aleksandra.26@mail.ruhttps://orcid.org/0000-0002-4179-931XAmmurYulia I.<p>Cand. Sci. (Biol.), Head, Laboratory of experimental immunology, Department of virology</p>yulia.ammour@yahoo.frhttps://orcid.org/0000-0003-0223-5738SidorovAlexander V.<p>Cand. Sci. (Biol.), Head, Laboratory of genetics of DNA-containing viruses, Department of virology</p>sashasidorov@googlemail.comhttps://orcid.org/0000-0002-3561-8295KharchenkoOlga S.<p>researcher, Laboratory of DNA viruses, Department of virology</p>bio139@yandex.ruhttps://orcid.org/0000-0002-2169-9610BukhtoyarovGeorgy N.<p>Cand. Sci. (Biol.), researcher, Laboratory of genetics of DNA-containing viruses, Department of virology</p>Bah.234@mail.ruhttps://orcid.org/0000-0987-6885-3210SvitichOksana A.<p>D. Sci. (Med.), Corresponding Member of the Russian Academy of Sciences, Director</p>svitich_o_a@staff.sechenov.ruhttps://orcid.org/0000-0003-1757-8389ZverevVitaly V.<p>D. Sci. (Biol.), Professor, RAS Full Member, Head, Laboratory of molecular biotechnology</p>vitalyzverev@outlook.comhttps://orcid.org/0000-0001-5808-2246Mechnikov Research Institute for Vaccines and Sera0712202299550551330072022Copyright © 2022, Nagieva F.G., Barkova E.P., Stroeva A.D., Ammur Y.I., Sidorov A.V., Kharchenko O.S., Bukhtoyarov G.N., Svitich O.A., Zverev V.V.2022<p><strong>Introduction.</strong> Rubella is a mild infectious disease affecting mainly children and is caused by the rubella virus, part of the <em>Matonoviridae</em> family, genus <em>Rubivirus</em>. Rubella causes congenital rubella syndrome (CRS) and is the main cause of developmental abnormalities, especially blindness and deafness.</p>
<p>There is no specific treatment for rubella and CRS. In order to avoid possible complications from rubella infection, a live attenuated rubella vaccine based on the foreign strain of Wistar RA 27/3 rubella virus is used. However, the actual, more effective and preferred vaccine strain the rubella virus for the Russian Federation is considered to be a viral strain of rubella circulating on its territory.</p>
<p><strong>The aim</strong> of the study was to study the biological properties of the developed domestic cold-adapted strain vRub-Ant circulating in the territory of the Russian Federation.</p>
<p><strong>Materials and methods.</strong> Following cell cultures were used in the study human embryo lung diploid cell strain LECH-3, transferable cell line from embryonic kidney cells of green monkeys Vero CCL-81 and Vero ECC, human mesenchymal stem cells, human peripheral blood mononuclear cells (PBMC). Cell cultures were grown on a DMEM/F12 nutrient medium with the addition of 5% fetal bovine serum. Swabs from the pharynx and nasal passages from a child with rubella were used as clinical virus-containing material. Monoclonal anti-idiotypic antibodies m(anti-ID)Ab were used to assess the expression level of alpha/beta and gamma interferon receptors (/ and IFN-R)Ab, imitating the biological effects of alpha/beta and gamma interferons (/ and IFN) of humans. The cultural, virological, immunochemical and serological research methods were applied in the study.</p>
<p><strong>Results.</strong> Attenuation of the vRub-Ant clinical isolate of rubella virus was carried out for 20 consecutive passages on LECH-3 diploid cells at a reduced temperature of 30C. The main biological markers of attenuation were determined to be ts and ca phenotypes. The avirulence of the attenuated viral strain (att-phenotype) was assessed by the level of expression of / and IFN-R. A lower level of / and IFN-R expression was found on the membranes of human PBMC induced by the vaccine strain vRub-Ant in comparison with the parent wild variant of the rubella virus. This trait,the att phenotype, is characteristic of attenuated viral strains. It has been shown that the vaccine strain vRub-Ant has lost neurotropism and was unable to bind to the membrane receptors of the brain (MRB) of guinea pig embryos, unlike its parent rubella virus strain. The high immunogenicity of the domestic cold-adapted strain vRub-Ant was confirmed by high titers of neutralizing rubella antibodies observed in guinea pigs immunized subcutaneously with one vaccination dose of the virus.</p>
<p><strong>Conclusion. </strong>A domestic attenuated vaccine strain vRub-Ant of the rubella virus that has the main biological markers of attenuation (ts-ca and att phenotypes) has been developed. The vaccine strain vRub-Ant induces a high levels of neutralizing antibodies in guinea pigs following the immunization with a single vaccination dose of the vaccine. The viral strain vRub-Ant has lost its tropism to the MRB of guinea pig embryos, unlike its parent variant.</p>rubella viruscell culturecold-adapted domestic strain of rubella virus attenuationinfectivity of the sampleimmunogenicityantigenicityneurotropicitypreparation of the membrane receptor of the brain of guinea pigsвирус краснухикультура клетокхолодоадаптированный отечественный штамм вируса краснухибиологические маркеры аттенуации вируса краснухиинфекционность образцаиммуногенностьантигенностьнейротропностьпрепарат мембранного рецептора мозга морских свинок[Gershon A.A. Rubella virus (German Meales). In: Mandell, Douglas, and Bennet´s Principles and Practice. Volume 2. Elsevier; 2015: 1875–80. https://doi.org/10.1016/B978-1-4557-4801-3.00154-5][Mawson A.R., Croft A.M. Rubella virus infection, the congenital rubella syndrome, and link to autism. Int. J. Environ. Res. Public Health. 2019; 16(19): 3543–71. https://doi.org/10.3390/ijerph16193543][Banatvala J.E., Brown D.W.G. Rubella. Lancet. 2004; 363(9415): 1127–37. https://doi.org/10.1016/S0140-6736(04)15897-2][George S.R., Viswanathan R., Sarkal G.N. Molecular aspects of the teratogenesis of rubella virus. Biol. Res. 2019; 52(1): 47. https://doi.org/10.1186/s40659-019-0254-3][Lambert N., Strebel P., Orenstein W., Icenogle J., Poland G.A. Rubella. Lancet. 2015; 385(9984): 2297–307. https://doi.org/10.1016/S0140-6736(14)60539-0][Lee J.Y., Bowden D.S. Rubella virus replication and links to teratogenicity. Clin. Microbiol. Rev. 2000; 13(4): 571–87. https://doi.org/10.1128/cmr.13.4.571-587.2000][Shishido A., Ohtawara V. Development of attenuated rubella virus vaccines in Japan. Jpn J. Med. Sci. Biol. 1976; 29(5): 227–53. https://doi.org/10.7883/yoken 952.29/227][Nagieva F.G., Barkova E.P., Stroeva A.D., Sidorov A.V., Lotte V.D., Zverev V.V., et al. Characterization of binding of Varicella zoster virus vaccine strains to preparations of mouse brain membrane receptors. Zhurnal mikrobiologii, epidemiologii i immunobiologii. 2020; 97(2): 125–33. https://doi.org/10.36233/0372-9311-2020-97-2-125-133 (in Russian)][Best L.M., Banatvala J.E. Congenital virus infections. BMJ. 1990; 300(6733): 1151–2. https://doi.org/10.1136/bmj.300.6733.1151][Plotkin S.A. The history of rubella and rubella vaccination leading to elimination. Clin. Infect. Dis. 2006; 43(Suppl. 3): S164–68. https://doi.org/10.1086/505950][Nagieva F.G., Nikulina V.G., Barkova E.P., Zubkov A.V., Kuz'mina N.S., Desyatkova R.G., et al. Monoclonal antibodies to rubella virus glycoprotein E1. Zhurnal mikrobiologii, epidemiologii i immunobiologii. 2011; 88(1): 61–7. (in Russian)][Barrett F., Ryman K., Ni H. Method of attenuating viruses. Patent US EPO922092B1; 1999.][Osokina O.V., Barkova E.P., Nagieva F.G., Zverev V.V. The vFiraVax strain for producing an attenuated live culture vaccine for the prevention of chickenpox. Patent RF № 2693440; 2019. (in Russian)][Zverev V.V., Nagieva F.G., Barkova E.P., Osokina O.V. Method of obtaining a four-component cultured live vaccine against measles, chickenpox, mumps, rubella (NT). Patent RF 2693440; 2019. (in Russian)][Nagieva F.G., Barkova E.P., Stroeva A.D., Sidorov A.V., Zverev V.V. Characterization of markers of cold-adapted candidate virus strains for live attenuated vaccines against chickenpox and shingles. Zhurnal mikrobiologii, epidemiologii i immunobiologii. 2020; 97(4): 303–11. https://doi.org/10.36233/0372-9311-2020-97-4-2 (in Russian)]