Skip to main content

Functional ectodomain of the hemagglutinin-neuraminidase protein is expressed in transgenic tobacco cells as a candidate vaccine against Newcastle disease virus

Abstract

Recently, the use of plants for the production of recombinant proteins has been well demonstrated with promising outcomes. In this study, an efficient Nicotiana tabacum L. cv. Bright Yellow 2 (BY-2) cells system expressing the ectodomain of hemagglutinin-neuraminidase (eHN) protein from Newcastle disease virus (NDV) strain AF2240 was established. Transgenic tobacco BY-2 cell cultures expressing the immunogenic eHN protein were generated and the translation efficiency of eHN protein was enhanced using the 5′-untranslated region of Nicotiana tabacum alcohol dehydrogenase gene (NtADH 5′-UTR) under the control of strong cauliflower mosaic virus (CaMV 35S) promoter. Transgenic lines verified by real-time PCR showed high level of eHN mRNA transcripts and immunoblotting confirmed the presence of 66 kD eHN protein. The eHN protein was stably produced in an average of 0.2–0.4 % total soluble protein. Green fluorescent protein-tagged eHN protein was expressed and localized at the cytosol of BY-2 cell. All mice receiving purified eHN protein from transgenic tobacco BY-2 cells produced specific anti-NDV antibodies. We concluded that plant made eHN elicit immune response and can serve as candidate vaccine against NDV.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Abbreviations

eHN:

Ectodomain of hemagglutinin-neuraminidase

GFP:

Green fluorescent protein

LS:

Linsmaier and Skoog

NtADH 5′-UTR:

5′-Untranslated region of Nicotiana tabacum alcohol dehydrogenase gene

NDV:

Newcastle disease virus

References

  1. Berinstein A, Rovere CV, Asurmendi S, Gómez E, Zanetti F, Zabal O, Tozzini A, Grand DC, Toboga O, Calmante G, Barrios H, Hopp E, Carillo E (2005) Mucosal and systemic immunization elicited by Newcastle disease virus (NDV) transgenic plants as antigens. Vaccine 23:5583–5589

    PubMed  Article  CAS  Google Scholar 

  2. Faye L, Boulaflous A, Benchabane M, Gomord V, Michaud D (2005) Protein modifications in plant secretory pathway: current status and practical implications in molecular pharming. Vaccine 23:1770–1778

    PubMed  Article  CAS  Google Scholar 

  3. Girard L, Bastin M, Courtois D (2004) Expression of the human milk protein sCD14 in tobacco plant cell culture. Plant Cell Tissue Organ Cult 78:253–260

    Article  CAS  Google Scholar 

  4. Gómez E, Zoth SC, Asurmendi S, Rovere CV, Berinstein A (2009) Expression of hemagglutinin-neuraminidase glycoprotein of Newcastle disease virus in agroinfiltrated Nicotiana benthamiana plants. J Biotechnol 144:337–340

    PubMed  Article  Google Scholar 

  5. Haasen D, Kohler C, Neuhaus G, Merkle T (1999) Nuclear export of proteins in plants: AtXPO1 is the export receptor for leucine-rich nuclear export signals in Arabidopsis thaliana. Plant J 20:695–705

    PubMed  Article  CAS  Google Scholar 

  6. Hahn BS, Jeon IS, Jung YJ, Kim JB, Parl JS, Ha SH, Kim KH, Kim HM, Yang JS, Kim YH (2007) Expression of hemagglutinin-neuraminidase protein of Newcastle disease virus in transgenic tobacco. Plant Biotechnol Rep 1:85–92

    Article  Google Scholar 

  7. Ko K, Koprowski H (2005) Plant biopharming of monoclonal antibodies. Virus Res 111:93–100

    PubMed  Article  CAS  Google Scholar 

  8. Martínez CA, Giulietti AM, Rodríguez Talou J (2011) Expression of a KDEL-tagged dengue virus protein in cell suspension cultures of Nicotiana tabacum and Morinda citrifolia. Plant Cell Tissue Organ Cult 107:91–100

    Article  Google Scholar 

  9. Matsui T, Hori M, Shizawa N, Nakayama H, Shinmyo A, Yoshida K (2006) High-efficiency secretory production of peroxidase C1a using vesicular transport engineering in transgenic tobacco. J Biosci Bioeng 102:102–109

    PubMed  Article  CAS  Google Scholar 

  10. Matsui T, Asao H, Ki M, Sawada K, Kato K (2009) Transgenic lettuce producing a candidate protein for vaccine against edema disease. Biosci Biotechnol Biochem 73:1628–1634

    PubMed  Article  CAS  Google Scholar 

  11. Nakayama H, Yoshida K, Ono H, Murooka Y, Shinmyo A (2000) Ectoine, the compatible solute of Halomonas elongata, confers hyperosmotic tolerance in cultured tobacco cells. Plant Physiol 122:1239–1246

    PubMed  Article  CAS  Google Scholar 

  12. Obembe OO, Popoola JO, Leelavathi S, Reddy SV (2010) Advances in plant molecular farming. Biotechnol Adv 29:210–222

    PubMed  Article  Google Scholar 

  13. Ong HKA, Ali AM, Omar AR, Yusoff K (2000) Cloning and expression of the HN gene from velogenic viscerotropic Newcastle disease virus strain AF2240 in Sf9 insect cells. Cytotechnology 32:243–251

    PubMed  Article  CAS  Google Scholar 

  14. Pires AS, Rosa S, Castanheira S, Fevereiro P, Abranches R (2012) Expression of a recombinant human erythropoietin in suspension cultures of Arabidopsis, tobacco and Medicago. Plant Cell Tissue Organ Cult. doi:10.1007/s11240-012-0141-x

    Google Scholar 

  15. Satoh J, Kato K, Shinmyo A (2004) The 5′-untranslated region of the tobacco alcohol dehydrogenase gene functions as an effective translational enhancer in plant. J Biosci Bioeng 98:1–8

    PubMed  CAS  Google Scholar 

  16. Soria-Guerra RE, Moreno-Fierros L, Roslaes-Mendoza S (2011) Two decades of plant-based candidates vaccine: a review of the chimeric protein approaches. Plant Cell Rep 30:1367–1382

    PubMed  Article  CAS  Google Scholar 

  17. Sorrentino A, Schillberg S, Fisher R, Rao R, Porta R, Mariniello L (2005) Recombinant human tissue transglutaminase produced into tobacco suspension cell cultures is active and recognizes autoantibodies in the serum of coeliac patients. Int J Biochem Cell Biol 37:842–851

    PubMed  Article  CAS  Google Scholar 

  18. Sun QY, Ding LW, Lomomossoff GP, Sun YB, Luo M, Li CQ, Jiang L, Xu ZF (2011) Improved expression and purification of recombinant human serum albumin from transgenic tobacco suspension culture. J Biotechnol 155:164–172

    PubMed  Article  CAS  Google Scholar 

  19. Urreta I, Oyanguren I, Castanón S (2011) Tobacco as biofactory for biologically active hPL production: a human hormone with the potential applications in type-1 diabetes. Transgenic Res 20:721–733

    PubMed  Article  CAS  Google Scholar 

  20. Wong SK, Tan WS, Omar AR, Tan CS, Yusoff K (2009) Immunogenic properties of recombinant ectodomian of Newcastle disease virus hemagglutinin-neuraminidase protein expressed in Escherichia coli. Act Virol 53:35–41

    Article  CAS  Google Scholar 

  21. Yusoff K, Tan WS (2001) Newcastle disease virus: macromolecules and opportunities. Avian Pathol 30:439–455

    PubMed  Article  CAS  Google Scholar 

Download references

Acknowledgments

Authors are thankful to the members of plant biotechnology and virology laboratories in UPM for their technical support.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Maziah Mahmood.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Lai, K.S., Yusoff, K. & Mahmood, M. Functional ectodomain of the hemagglutinin-neuraminidase protein is expressed in transgenic tobacco cells as a candidate vaccine against Newcastle disease virus. Plant Cell Tiss Organ Cult 112, 117–121 (2013). https://doi.org/10.1007/s11240-012-0214-x

Download citation

Keywords

  • Hemagglutinin-neuraminidase
  • Newcastle disease virus
  • NtADH 5′-UTR
  • Plant-based vaccine