Skip to main content
SpringerLink
Log in

Enhancement of the immune response by codelivery of hepatitis C virus recombinant DNA and proteins of the replicative complex

  • Experimental Works
  • Published:
Molecular Genetics, Microbiology and Virology Aims and scope Submit manuscript

Abstract

The plasmids encoding amino acid sequences corresponding to the hepatitis C virus (HCV) are regarded as promising candidates for anti-HCV vaccines. However, the optimal composition of these vaccines has not been determined. The goal of this work was to evaluate the immune response after immunization of mice with the DNA construct pcNS3-NS5B encoding proteins that constitute the viral replicative complex combined with the recombinant nonstructural proteins NS3 and NS5B using various adjuvants. DBA mice were immunized with the DNA construct and/or recombinant proteins three times. The antibody response was evaluated using ELISA. The cell immunity was determined using a lymphocyte proliferation test in vitro and by production and secretion of IFNγ and IL-2 measured by ELISpot and ELISA. It was found that the pcNS3-NS5B plasmid induced predominantly T-cell response, whereas the recombinant NS3 and NS5B proteins stimulated a potent humoral immune response. This made it possible for us to show for the first time statistically significant stimulation of both humoral and cell immunity after immunization with the pcNS3-NS5B plasmid in combination with the NS3 and NS5B HCV proteins. It was noteworthy that the animals developed the immune response not only to the NS3 and NS5B proteins, but also to other antigens encoded by the pcNS3-NS5B plasmid (NS4, NS5A). The most pronounced immune response was observed in mice subjected to triple immunization with the pcNS3-NS5B plasmid in combination with the gene adjuvant pcGM-CSF and recombinant proteins (NS3 and NS5B), which were injected together with IFNα. The adjuvant activity of the recombinant protein IFNα, which was shown in a model of the HCV genes and proteins for the first time, may be a prerequisite for including it in a candidate vaccine.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Horner, S.M., Activation and evasion of antiviral innate immunity by hepatitis C virus, J. Mol. Biol., 2014, vol. 426, no. 6, pp. 1198–1209.

    Article  CAS  PubMed  Google Scholar 

  2. Irshad, M., Mankotia, D.S., and Irshad, K., An insight into the diagnosis and pathogenesis of hepatitis C virus infection, World J. Gastroenterol., 2013, vol. 19, no. 44, pp. 7896–7909.

    Article  PubMed Central  PubMed  Google Scholar 

  3. Pawlotsky, J.M., Hepatitis C virus: standard-of-care treatment, Adv. Pharmacol., 2013, vol. 67, pp. 69–215.

    Google Scholar 

  4. Rupp, D. and Bartenschlager, R., Targets for antiviral therapy of hepatitis C, Semin. Liver Dis., 2014, vol. 34, no. 1, pp. 9–21.

    Article  CAS  PubMed  Google Scholar 

  5. Halliday, J., Klenerman, P., and Barnes, E., Vaccination for hepatitis C virus: closing in on an evasive target, Expert Rev. Vaccines, 2011, vol. 10, no. 5, pp. 659–672.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  6. Honegger, J.R., Zhou, Y., and Walker, C.M., Will there be a vaccine to prevent HCV infection?, Semin. Liver Dis., 2014, vol. 34, no. 1, pp. 79–88.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  7. Masalova, O.V., Lesnova, E.I., Ivanov, A.V., Pichugin, A.V., Permyakova, K.Yu., Smirnova, O.A., et al., Comparative analysis of the immune response to DNA constructions encoding hepatitis C virus nonstructural proteins, Vopr. Virusol., 2013, vol. 58, no. 2, pp. 21–28.

    CAS  PubMed  Google Scholar 

  8. Moradpour, D., Penin, F., and Rice, C.M., Replication of hepatitis C virus, Nat. Rev. Microbiol., 2007, vol. 5, pp. 453–463.

    Article  CAS  PubMed  Google Scholar 

  9. Masalova, O.V., Lesnova, E.I., Shingarova, L.N., Tunitskaia, V.L., Ulanova, T.I., Burkov, A.N., et al., The combined application of nucleotide and amino acid sequences of NS3 hepatitis C virus protein, DNA encoding granulocyte macrophage colony-stimulating factor and inhibitor of regulatory T cells induces effective immune response against hepatitis C virus, Mol. Biol., 2012, vol. 46, no. 3, pp. 525–534.

    Article  CAS  Google Scholar 

  10. Masalova, O.V., Abdulmedzhidova, A.G., Atanadze, S.N., Lakina, E.I., Semiletov, Yu.A., Burkov, A.N., et al., Characterization of a panel of monoclonal antibodies and mapping the epitopes of hepatitis C virus proteins, Dokl. Akad. Nauk, 2002, vol. 383, no. 4, pp. 545–550.

    Google Scholar 

  11. Mukovnya, A.V., Tunitskaya, V.L., Khandazhinskaya, A.L., Golubeva, N.A., Zakirova, N.F., Ivanov, A.V., et al., Hepatitis C virus helicase/NTPase: an efficient expression system and new inhibitors, Biokhimiya, 2008, vol. 73, no. 6, pp. 660–669.

    Article  CAS  Google Scholar 

  12. Ulanova, T.I., Puzyrev, V.F., Burkov, A.N., and Obyadina, A.P., Impact of the heterogenicity of amino acid sequence on the immunoreactivity of an antigenic epitopic complex localized within amino acids 1192–1456 of protein NS3 protein of hepatitis C virus, Vopr. Virusol., 2006, vol. 51, no. 1, pp. 28–30.

    CAS  PubMed  Google Scholar 

  13. Ivanov, A.V., Korovina, A.N., Tunitskaya, V.L., Kostyuk, D.A., Rechinsky, V.O., Kukhanova, M.K., et al., Development of the system ensuring a high-level expression of hepatitis C virus nonstructural NS5B and NS5A proteins, Protein Expr. Purif., 2006, vol. 48, no. 1, pp. 14–23.

    Article  CAS  PubMed  Google Scholar 

  14. Masalova, O.V., Lesnova, E.I., Grabovetskiy, V.V., Smirnova, O.A., Ulanova, T.I., Burkov, A.N., et al., DNA immunization with a plasmid carrying the gene of hepatitis C virus protein 5A (NS5A) induces an effective cellular immune response, Mol. Biol., 2010, vol. 44, no. 2, pp. 245–253.

    Article  CAS  Google Scholar 

  15. Masalova, O.V. and Kushch, A.A., Monoclonal antibodies to hepatitis C virus proteins as a tool for antigenic determinant mapping, hepatitis C diagnostics and investigation of viral pathogenesis, Ross. Bioterap. Zh., 2003, vol. 2, no. 3, pp. 7–24.

    Google Scholar 

  16. Thimme, R., Neumann-Haefelin, C., Boettler, T., and Blum, H., Adaptive immune responses to hepatitis C virus: from viral immunobiology to a vaccine, J. Biol. Chem., 2008, vol. 389, no. 5, pp. 457–467.

    CAS  Google Scholar 

  17. Masalova, O.V., Lesnova, E.I., Pichugin, A.V., Melnikova, T.M., Grabovetsky, V.V., Petrakova, N.V., et al., The successful immune response against hepatitis C nonstructural protein 5A (NS5A) requires heterologous DNA/protein immunization, Vaccine, 2010, vol. 28, no. 8, pp. 1987–1996.

    Article  CAS  PubMed  Google Scholar 

  18. Alvarez-Lajonchere, L., Shoukry, N.H., Grá, B., Amador-Cañizares, Y., Helle, F., Bédard, N., et al., Immunogenicity of CIGB-230, a therapeutic DNA vaccine preparation, in HCV-chronically infected individuals in a phase I clinical trial, J. Viral Hepatitis, 2009, vol. 16, no. 3, pp. 156–167.

    Article  CAS  Google Scholar 

  19. Ghorbani, M., Nass, T., Azizi, A., Soare, C., Aucoin, S., Giulivi, A., et al., Comparison of antibody- and cell-mediated immune responses after intramuscular hepatitis C immunizations of BALB/c mice, Viral Immunol., 2005, vol. 18, no. 4, pp. 637–648.

    Article  CAS  PubMed  Google Scholar 

  20. Aricò, E. and Belardelli, F., Interferon-α as antiviral and antitumor vaccine adjuvants: mechanisms of action and response signature, J. Interferon Cytokine Res., 2012, vol. 32, no. 6, pp. 235–247.

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Ivanovsky Institute of Virology, ul. Gamalei 16, Moscow, 123098, Russia

    O. V. Masalova, E. I. Lesnova, K. Yu. Permyakova & A. A. Kushch

  2. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, ul. Vavilova 32, Moscow, 119991, Russia

    A. V. Ivanov & V. L. Tunitskaya

Authors
  1. O. V. Masalova
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E. I. Lesnova
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. K. Yu. Permyakova
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A. V. Ivanov
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. V. L. Tunitskaya
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. A. A. Kushch
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to O. V. Masalova.

Additional information

Original Russian Text © O.V. Masalova, E.I. Lesnova, K.Yu. Permyakova, A.V. Ivanov, V.L. Tunitskaya, A.A. Kushch, 2015, published in Molekulyarnaya Genetika, Mikrobiologiya i Virusologiya, 2015, No. 1, pp. 36–41.

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Masalova, O.V., Lesnova, E.I., Permyakova, K.Y. et al. Enhancement of the immune response by codelivery of hepatitis C virus recombinant DNA and proteins of the replicative complex. Mol. Genet. Microbiol. Virol. 30, 39–47 (2015). https://doi.org/10.3103/S0891416815010061

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.3103/S0891416815010061

Keywords

Search

Navigation