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Whole Genome Sequencing and Phylogenetic Analysis of the Rabies Virus Strain Moscow 3253 Adapted to a Vero Cell Line

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Abstract

Molecular genetic analysis of the rabies virus strain Moscow 3253 genome adapted to Vero cells. The isolated cDNA was specifically enriched with the nucleotide sequence of the target virus genome using amplification. Whole genome sequencing was performed on the Ion PGM platform and additionally using the AB 3500xl genetic analyzer. Alignment and assembly of sequences into a complete genome and further analysis were performed using DNASTAR Lasergene 11.2 and MEGA 7. Phylogenetic analysis was carried out using 479 rabies virus genomes represented in the NCBI GenBank database. The dendrogram was constructed using the BioNumerics 7.6 software package. The genome of the rabies virus Moscow 3253 (Vero) has been deposited into NCBI GenBank (accession MF630920). The genome of the rabies virus Moscow 3253 (Vero), as compared to its original variant, has 1069 single mutations. Its phylogenetic closeness to various vaccine variants of the rabies virus genome represented in the NBCI GenBank database, which are also adapted to the Vero or BHK cell culture, has been established. A strong influence of the Vero cell culture as a reproduction medium for the rabies virus Moscow 3253 (Vero) on the acquisition of a large number of single mutations by its genome has been revealed. As has been stated in a previously published work, the physicochemical and biological properties of rabies immunoglobulin (RIG) obtained by the technology using the rabies virus strain Moscow 3253 (Vero) meet the requirements of the regulatory documentation for RIG. Thus, the use of the rabies virus adapted to the Vero cells for the production of a rabies vaccine is promising.

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REFERENCES

  1. WHO Expert Consultation on Rabies: First Report. WHO Technical Report Series 931, Geneva, 2004.

  2. Selimov, M.A., Beshenstvo (Rabies), Moscow: Meditsina, 1978.

    Google Scholar 

  3. Generalov, S.V., Abramova, E.G., Matveeva, Zh.V., Zhulidov, I.M., Lobovikova, O.A., Svintsov, R.A., et al., Culture antigen in the technology of producing rabies immunoglobulin from horse serum, Probl. Partic. Dangerous Infect., 2012, vol. 4, no. 114, pp. 65–68. https://doi.org/10.21055/0370-1069-2012-4-65-68

    Article  Google Scholar 

  4. Generalov, S.V., Abramova, E.G., Matveeva, Zh.V., Zhulidov, I.M., and Svintsov, R.A., Large scale cultivation of a fixed rabies virus strain “Moscow 3253” on Vero (B) cell line: methods and comparative analysis, Biotechnology, 2014, vol. 5, pp. 38–43.

    Google Scholar 

  5. Podchernyaeva, R.Ya., Khizhnyanova, T.M., and Mikhailova, G.R., Vero (B) cell line for the preparation of biomedical drugs, Probl. Virol., 1996, vol. 4, pp. 183–185.

    Google Scholar 

  6. Metlin, A., Paulin, L., Suomalainen, S., Neuvonen, E., Rybakov, S., Mikhalishin, V., et al., Characterization of Russian rabies virus vaccine strain RV-97, Virus Res., 2008, vol. 132, pp. 242–247. https://doi.org/10.1016/j.virusres.2007.11.016

    Article  CAS  PubMed  Google Scholar 

  7. Murphy, F.A. and Nathanson, N., The emergence of new virus diseases: an overview, Semin. Virol., 1994, vol. 5, pp. 87–102. https://doi.org/10.1006/smvy.1994.1010

    Article  Google Scholar 

  8. Yang, J., Koprowski, H., Dietzschold, B., and Fu, Z.F., Phosphorylation of rabies virus nucleoprotein regulates viral RNA transcription and replication by modulating leader RNA encapsidation, J. Virol., 1999, vol. 73, pp. 1661–1664. PMID 9882376

    Article  CAS  Google Scholar 

  9. Brzozka, K., Finke, S., and Conzelmann, K.K., Identification of the rabies virus alpha/beta interferon antagonist: Phosphoprotein P interferes with phosphorylation of interferon regulatory factor 3, J. Virol., 2005, vol. 79, pp. 7673–7681. https://doi.org/10.1128/JVI.79.12.7673-7681.2005

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Chenik, M., Chebli, K., Gaudin, Y., and Blondel, D., In vivo interaction of rabies virus Phosphoprotein (P) and nucleoprotein (N), existence of two N binding sites on P protein, J. Gen. Virol., 1994, vol. 75, pp. 2889–2896. https://doi.org/10.1099/0022-1317-75-11-2889

    Article  CAS  PubMed  Google Scholar 

  11. Fu, Z.F., Zheng, Y., Wunner, W.H., Koprowski, H., and Dietzschold, B., Both the N- and the C-terminal domains of the nominal phosphoprotein of rabies virus are involved in binding to the nucleoprotein, J. Virol., 1994, vol. 200, pp. 590–597. https://doi.org/10.1006/viro.1994.1222

    Article  CAS  Google Scholar 

  12. Sissoeff, L., Mousli, M., England, P., and Tuffereau, C., Stable trimerization of recombinant rabies virus glycoprotein ectodomain is required for interaction with the p75NTR receptor, J. Gen. Virol., 2005, vol. 86, pp. 2543–2552. https://doi.org/10.1099/vir.0.81063-0

    Article  CAS  PubMed  Google Scholar 

  13. Nakahara, T., Toriumi, H., Irie, T., Takahashi, T., Ameyama, S., Mizukoshi, M., et al., Characterization of a slow-migrating component of the rabies virus matrix protein strongly associated with the viral glycoprotein, Microbiol. Immunol., 2003, vol. 47, pp. 977–988. https://doi.org/10.1111/j.1348-0421.2003.tb03458.x

    Article  CAS  PubMed  Google Scholar 

  14. Evans, J.S., Selden, D., Wu, G., Wright, E., Horton, D.L., Fooks, A.R., et al., Antigenic site changes in the rabies virus glycoprotein dictates functionality and neutralizing capability against divergent lyssaviruses, J. Gen. Virol., 2018, vol. 99, no. 2, pp. 169–180. https://doi.org/10.1099/jgv.0.000998

    Article  CAS  PubMed  Google Scholar 

  15. Tuchkov, I.V., Krasnov, Ya.M., Goryaev, A.A., Matveeva, Zh.V., Stepanov, A.V., Maiorov, N.V., et al., Nucleotide sequence and phylogenetic analysis of G glycoprotein of the rabies virus strain “Moscow 3253” from Russia, Probl. Partic. Dangerous Infect., 2013, no. 4, pp. 73–75. https://doi.org/10.21055/0370-1069-2013-4-73-75

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Authors and Affiliations

  1. Russian Research Anti-Plague Institute Microbe, 410071, Saratov, Russia

    Ya. M. Krasnov, Zh. V. Alkhova, S. V. Generalov, I. V. Tuchkov, E. A. Naryshkina, N. A. Sharapova, E. G. Abramova & A. K. Nikiforov

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  1. Ya. M. Krasnov
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  2. Zh. V. Alkhova
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  3. S. V. Generalov
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  4. I. V. Tuchkov
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  5. E. A. Naryshkina
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  6. N. A. Sharapova
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  7. E. G. Abramova
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  8. A. K. Nikiforov
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Correspondence to Ya. M. Krasnov.

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The authors declare that they have no conflict of interest. This article does not contain any studies involving animals or human participants performed by any of the authors.

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Translated by M. Novikova

ADDITIONAL INFORMATION

Krasnov Ya.M.—https://orcid.org/0000-0002-4909-2394; e-mail: rusrapi@microbe.ru

Alkhova Zh.V.—https://orcid.org/0000-0001-7681-5773

Generalov S.V.—https://orcid.org/0000-0003-1461-5383

Tuchkov I.V.—https://orcid.org/0000-0003-0712-8659

Naryshkina E.A.—https://orcid.org/0000-0002-9190-099X

Sharapova N.A.—https://orcid.org/0000-0002-5289-7783

Abramova E.G.—https://orcid.org/0000-0002-8798-1547

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Krasnov, Y.M., Alkhova, Z.V., Generalov, S.V. et al. Whole Genome Sequencing and Phylogenetic Analysis of the Rabies Virus Strain Moscow 3253 Adapted to a Vero Cell Line. Mol. Genet. Microbiol. Virol. 35, 237–242 (2020). https://doi.org/10.3103/S0891416820040060

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  • DOI: https://doi.org/10.3103/S0891416820040060

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