Skip to main content
Log in

Expression in Plants of a Recombinant Protein Based on Flagellin Linked to Conservative Fragments of M2 Protein and Hemagglutintin of Influenza Virus

  • Published:
Applied Biochemistry and Microbiology Aims and scope Submit manuscript

Abstract

The composition of traditional influenza vaccines require nearly annual updates due to the high variability of the influenza virus. The use of conservative viral antigens, the extracellular domain of the transmembrane protein M2, and fragments of the second subunit of hemagglutinin provides the opportunity to create recombinant broad-spectrum vaccines. Bacterial flagellin was used as a mucosal adjuvant to increase the immunogenicity of these conservative antigens. Recombinant proteins based on flagellin simultaneously containing M2e and a fragment of the hemagglutinin stem region were expressed in Nicotiana benthamiana plants using a self-replicating vector based on the potato virus X genome. Methods for their isolation from plants and purification have been developed. The developed expression system can be used to produce a new candidate influenza vaccine in plants.

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

Access this article

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. Webster, R.G., Bean, W.J., Gorman, O.T., Chambers, T.M., and Kawaoka, Y., Microbiol. Rev., 1992, vol. 56, pp. 152–179.

    PubMed  PubMed Central  CAS  Google Scholar 

  2. Neirynck, S., Deroo, T., Saelens, X., Vanlandschoot, P., Jou, W.M., and Fiers, W., Nat. Med., 1999, vol. 5, no. 10, pp. 1157–1163.

    Article  PubMed  CAS  Google Scholar 

  3. Fiers, W., De Filette, M., El Bakkouri, K., Schepens, B., Roose, K., Schotsaert, M., Birkett, A., and Saelens, X., Vaccine, 2009, vol. 27, no. 45, pp. 6280–6283.

    Article  PubMed  CAS  Google Scholar 

  4. Ito, T., Gorman, O.T., Kawaoka, Y., Bean, W.J., and Webster, R.G., J. Virol., 1991, vol. 65, no. 10, pp. 5491–5498.

    PubMed  PubMed Central  CAS  Google Scholar 

  5. Feng, J., Zhang, M., Mozdzanowska, K., Zharikova, D., Hoff, H., Wunner, W., Couch, R.B., and Gerhard, W., Virol. J., 2006, vol. 3, p.102.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  6. Iwasaki, A. and Medzhitov, R., Nat. Immunol., 2004, vol. 5, no. 10, pp. 987–995.

    Article  PubMed  CAS  Google Scholar 

  7. McDonald, W.F., Huleatt, J.W., Foellmer, H.G., Hewitt, D., Tang, J., Desai, P., Price, A., Jacobs, A., Takahashi, V.N., Huang, Y., Nakaar, V., Alexopoulou, L., Fikrig, E., and Powell, T.J., J. Infect. Dis., 2007, vol. 195, no. 11, pp. 1607–1617.

    Article  PubMed  CAS  Google Scholar 

  8. Huleatt, J.W., Nakaar, V., Desai, P., Huang, Y., Hewitt, D., Jacobs, A., Tang, J., McDonald, W., Song, L., Evans, R.K., Umlauf, S., Tussey, L., and Powell, T.J., Vaccine, 2008, vol. 26, no. 2, pp. 201–214.

    Article  PubMed  CAS  Google Scholar 

  9. Stepanova, L., Kotlyarov, R., Kovaleva, A., Potapchuk, M., Korotkov, A., Sergeeva, M., Kasianenko, M.A., Kuprianov, V.V., Ravin, N.V., Tsybalova, L.M., Skryabin, K.G., and Kiselev, O.I., PLoS One, 2015, vol. 10, no. 3. e0119520.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  10. Kusnadi, A.R., Nikolov, Z.L., and Howard, J.A., Biotechnol. Bioeng., 1997, vol. 56, no. 5, pp. 473–484.

    Article  PubMed  CAS  Google Scholar 

  11. Gleba, Y., Klimyuk, V., and Marillonnet, S., Curr. Opin. Biotechnol., 2007, vol. 18, no. 2, pp. 134–141.

    Article  PubMed  CAS  Google Scholar 

  12. Kapila, J., De Rycke, R., van Montagu, M., and Angenon, G., Plant Sci., 1997, vol. 122, no. 1, pp. 101–108.

    Article  CAS  Google Scholar 

  13. Marillonnet, S., Thoeringer, C., Kandzia, R., Klimyuk, V., and Gleba, Y., Nat. Biotechnol., 2005, vol. 23, no. 6, pp. 718–723.

    Article  PubMed  CAS  Google Scholar 

  14. Thuenemann, E.C., Lenzi, P., Love, A.J., Taliansky, M., Becares, M., Zuniga, S., Enjuanes, L., Zahmanova, G.G., Minkov, I.N., Matic, S., Noris, E., Meyers, A., Hattingh, A., Rybicki, E.P., Kiselev, O.I., Ravin, N.V., Eldarov, M.A., Skryabin, K.G., and Lomonossoff, G.P., Curr. Pharm. Des., 2013, vol. 19, no. 31, pp. 5564–5573.

    Article  PubMed  CAS  Google Scholar 

  15. Lomonossoff, G.P. and D’Aoust, M.A., Science, 2016, vol. 353, no. 6305, pp. 1237–1240.

    Article  PubMed  CAS  Google Scholar 

  16. D'Aoust, M.A., Lavoie, P.O., Couture, M.M., Trepanier, S., Guay, J.M., Dargis, M., Mongrand, S., Landry, N., Ward, B.J., and Vezina, L.P., Plant Biotechnol. J., 2008, vol. 6, no. 9, pp. 930–940.

    Article  PubMed  CAS  Google Scholar 

  17. Shoji, Y., Jones, R.M., Mett, V., Chichester, J.A., Musiychuk, K., Sun, X., Tumpey, T.M., Green, B.J., Shamloul, M., Norikane, J., Bi, H., Hartman, C.E., Bottone, C., Stewart, M., Streatfield, S.J., and Yusibov, V., Hum. Vaccin. Immunother., 2013, vol. 9, no. 3, pp. 533–560.

    Article  Google Scholar 

  18. Ravin, N.V., Kotlyarov, R.Yu., Mardanova, E.S., Kuprianov, V.V., Migunov, A.I., Stepanova, L.A., Tsybalova, L.M., Kiselev, O.I., and Skryabin, K.G., Biokhimiya, 2012, vol. 77, no. 1, pp. 43–52.

    Google Scholar 

  19. Petukhova, N.V., Gasanova, T.V., Stepanova, L.A., Rusova, O.A., Potapchuk, M.V., Korotkov, A.V., Skurat, E.V., Tsybalova, L.M., Kiselev, O.I., Ivanov, P.A., and Atabekov, J.G., Curr. Pharm. Des, 2013, vol. 19, no. 31, pp. 5587–5600.

    Article  PubMed  CAS  Google Scholar 

  20. Mardanova, E.S., Kotlyarov, R.Y., Kuprianov, V.V., Stepanova, L.A., Tsybalova, L.M., Lomonosoff, G.P., and Ravin, N.V., BMC Biotechnology, 2015, vol. 15, p.42.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  21. Wang, T.T., Tan, G.S., Hai, R., Pica, N., Ngai, L., Ekiert, D.C., Wilson, I.A., García-Sastre, A., Moran, T.M., and Palese, P., Proc. Natl. Acad. Sci. U. S. A., 2010, vol. 107, no. 44, pp. 18979–18984.

    Article  PubMed  PubMed Central  Google Scholar 

  22. Gamblin, S.J. and Skehel, J.J., J. Biol. Chem., 2010, vol. 285, no. 37, pp. 28403–28409.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  23. Nachbagauer, R. and Krammer, F., Clin. Microbiol. Infect., 2017, vol. 23, no. 4, pp. 222–228.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  24. Zhang, H., Wang, L., Compans, R.W., and Wang, B.Z., Viruses, 2014, vol. 6, no. 5, pp. 1974–1991.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  25. Stepanova, L.A., Sergeeva, M.V., Shuklina, M.A., Shaldzhyan, A.A., Potapchuk, M.V., Korotkov, A.V., and Tsybalova, L.M., Acta Naturae, 2016, vol. 8, no. 2, pp. 129–140.

    Google Scholar 

  26. Guo, Y., He, L., Song, N., Li, P., Sun, S., Zhao, G., Tai, W., Jiang, S., Du, L., and Zhou, Y., Microbes Infect., 2017, vol. 19, no. 12, pp. 641–647.

    Article  PubMed  CAS  Google Scholar 

  27. Mardanova, E.S., Blokhina, E.A., Tsybalova, L.M., Peyret, H., Lomonossoff, G.P., and Ravin, N.V., Front. Plant Sci., 2017, vol. 8, p.247.

    Article  PubMed  PubMed Central  Google Scholar 

  28. De Filette, M., Fiers, W., Martens, W., Birkett, A., Ramne, A., Löwenadler, B., Lycke, N., Jou, W.M., and Saelens, X., Vaccine, 2006, vol. 24, nos. 44–46, pp. 6597–6601.

    Article  PubMed  CAS  Google Scholar 

  29. Kotlyarov, R.Yu., Kupriyanov, V.V., Migunov, A.I., Stepanova, L.A., Tsybalova, L.M., Kiselev, O.I., Ravin, N.V., and Skryabin, K.G., Acta Naturae, 2010, vol. 2, no. 2, pp. 75–80.

    Google Scholar 

  30. De Filette, M., Min Jou, W., Birkett, A., Lyons, K., Schultz, B., Tonkyro, A., Resch, S., and Fiers, W., Virology, 2005, vol. 337, no. 1, pp. 149–161.

    Article  PubMed  CAS  Google Scholar 

  31. Ravin, N.V., Blokhina, E.A., Kuprianov, V.V., Stepanova, L.A., Shaldjan, A.A., Kovaleva, A.A., Tsybalova, L.M., and Skryabin, K.G., Vaccine, 2015, vol. 33, no. 29, pp. 3392–3397.

    Article  PubMed  CAS  Google Scholar 

  32. Nemchinov, L.G. and Natilla, A., Protein Expr. Purif., 2007, vol. 56, no. 2, pp. 153–159.

    Article  PubMed  CAS  Google Scholar 

  33. Matić, S., Rinaldi, R., Masenga, V., and Noris, E., BMC Biotechnol., 2011, vol. 11, p. 106.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to N. V. Ravin.

Additional information

Original Russian Text © E.A. Blokhina, E.S. Mardanova, L.M. Tsybalova, N.V. Ravin, 2018, published in Prikladnaya Biokhimiya i Mikrobiologiya, 2018, Vol. 54, No. 4.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Blokhina, E.A., Mardanova, E.S., Tsybalova, L.M. et al. Expression in Plants of a Recombinant Protein Based on Flagellin Linked to Conservative Fragments of M2 Protein and Hemagglutintin of Influenza Virus. Appl Biochem Microbiol 54, 404–409 (2018). https://doi.org/10.1134/S0003683818040026

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S0003683818040026

Keywords

Navigation