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Cassette Construction and Co-Expression of NA and F Proteins of H1N1 Influenza Virus and Newcastle Disease Virus in Saccharomyces cerevisiae

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Abstract

The aim of this study is to optimize the design, construction, and expression of recombinant protein, NA-F, derived from pathogenic influenza and Newcastle viruses within the eukaryotic host Saccharomyces cerevisiae. The chimeric gene was constructed using a homologous recombination method, and the target genes were cloned onto the T&A vector using PCR cloning techniques. Subsequently, the chimeric gene was expressed in S. cerevisiae, and its presence was confirmed through extracellular expression validation, Western blotting, and specific antibody detection. The recombinant protein displayed significant biological activity, demonstrating its functional characteristics and potential for use in immunization. The chimeric gene structure produced in this study holds promise as a candidate to replace existing vaccines worldwide, considering the high prevalence of influenza and Newcastle viruses in poultry and the need for a universal subunit vaccine. However, further research in animal and human models is essential to ensure safety before widespread application.

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REFERENCES

  1. Kim, S.H. and Samal, S.K., Innovation in Newcastle disease virus vectored avian influenza vaccines, Viruses, 2019, vol. 11, no. 3, p. 300.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Bouvier, N.M., and Palese, P., The biology of influenza viruses, Vaccine, 2008, vol. 26, no. 4, Suppl., pp. D49–D53.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Bai, Y., Jones, J.C., Wong, S.-S., and Zanin, M., Antivirals targeting the surface glycoproteins of influenza virus: Mechanisms of action and resistance, Viruses, 2021, vol. 13, p. 624.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Allen, V., Longley, N., Galloway, J.B., and Bechman, K., in Handbook of Systemic Autoimmune Diseases, Atzeni, F., Galloway, J.B., Gomez-Reino, J.J., and Galli, M., Eds., Elsevier, 2020, pp. 109–142.

    Google Scholar 

  5. Rehman, Z.U., Ren, S., Butt, S.L., Manzoor, Z., Iqbal, J., Anwar, M.N., Sun, Y., Qiu, X., Tan, L., Liao, Y., Song, C., Liu, W., Meng, C., and Ding, C., Newcastle disease virus induced pathologies severely affect the exocrine and endocrine functions of the pancreas in chickens, Genes (Basel), 2021, vol. 12, no. 4, p. 495.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Brown, V.R. and Bevins, S.N., A review of virulent Newcastle disease viruses in the United States and the role of wild birds in viral persistence and spread, Vet. Res., 2017, vol. 48, no. 1, p. 68.

    Article  PubMed  PubMed Central  Google Scholar 

  7. Ganar, K., Das, M., Sinha, S., and Kumar, S., Newcastle disease virus: current status and our understanding, Virus Res., 2014, vol. 184, pp. 71–81.

    Article  CAS  PubMed  Google Scholar 

  8. Hosseini, S.S., Kolyani, K.A., Tabatabaei, R.R., Goudarzi, H., Sepahi, A.A., and Salemi, M., In silico prediction of B and T cell epitopes based on NDV fusion protein for vaccine development against Newcastle disease virus, Vet. Res. Forum, 2021, vol. 12, no. 2, p. 157.

    PubMed  PubMed Central  Google Scholar 

  9. Liu, M.-M., Cheng, J.-L., Yu, X.-H., Qin, Z.-M., Tian, F.-L., and Zhang, G.-Z., Generation by reverse genetics of an effective attenuated Newcastle disease virus vaccine based on a prevalent highly virulent Chinese strain, Biotechnol. Lett., 2015, vol. 37, pp. 1287–1296.

    Article  CAS  PubMed  Google Scholar 

  10. Takrim, S., Motamedi, M., Jafari, M., Amani, J., and Salmanian, A., Expression, purification and immunogenicity evaluation of recombinant fusion protein (F) from Newcastle virus in animal model, Modares J. Biotechnol., 2019, vol. 10, pp. 15–21.

    Google Scholar 

  11. Berinstein, A., Vazquez-Rovere, C., Asurmendi, S., Gómez, E., Zanetti, F., Zabal, O., Tozzini, A., Grand, D.C., Taboga, O., Calamante, G., Barrios, H., Hopp, E., and Carrillo, E., Mucosal and systemic immunization elicited by Newcastle disease virus (NDV) transgenic plants as antigens, Vaccine, 2005, vol. 23, pp. 5583–5589.

    Article  CAS  PubMed  Google Scholar 

  12. Bello, M.B., Yusoff, K., Ideris, A., Hair-Bejo, M., Peeters, B.P., and Omar, A.R., Diagnostic and vaccination approaches for Newcastle disease virus in poultry: The current and emerging perspectives, BioMed Res. Int., 2018, vol. 2018, p. 7278459.

    Article  PubMed  PubMed Central  Google Scholar 

  13. Garg, R., Brownlie, R., Latimer, L., Gerdts, V., Potter, A., and van Drunen Littel-van den Hurk, S., A chimeric glycoprotein formulated with a combination adjuvant induces protective immunity against both human respiratory syncytial virus and parainfluenza virus type 3, Antiviral Res., 2018, vol. 158, pp. 78–87.

    Article  CAS  PubMed  Google Scholar 

  14. Ogonczyk Makowska, D., Hamelin, M., and Boivin, G., Engineering of live chimeric vaccines against human metapneumovirus, Pathogens (Basel, Switz.), 2020, vol. 9, no. 2, p. 135.

  15. Xu, X., Qian, J., Qin, L., Li, J., Xue, C., Ding, J., Wang, W., Ding, W., Yin, R., Jin, N., and Ding, Z., Chimeric Newcastle disease virus-like particles containing DC-binding peptide-fused haemagglutinin protect chickens from virulent Newcastle disease virus and H9N2 avian influenza virus challenge, Virol. Sin., 2020, vol. 35, pp. 455–467.

    Article  PubMed  PubMed Central  Google Scholar 

  16. Wei, C.-J., Crank, M.C., Shiver, J., Graham, B.S., Mascola, J.R., and Nabel, G.J., Next-generation influenza vaccines: Opportunities and challenges, Nat. Rev. Drug Discovery, 2020, vol. 19, pp. 239–252.

    Article  CAS  PubMed  Google Scholar 

  17. Ravikumar, R., Chan, J., and Prabakaran, M., Vaccines against major poultry viral diseases: Strategies to improve the breadth and protective efficacy, Viruses, 2022, vol. 14, p. 1195.

  18. Hoffmann, E., Stech, J., Guan, Y., Webster, R., and Perez, D., Universal primer set for the full-length amplification of all influenza A viruses, Arch. Virol., 2001, vol. 146, pp. 2275–2289.

    Article  CAS  PubMed  Google Scholar 

  19. Selim, K.M., Selim, A., Arafa, A., Hussein, H.A., and Elsanousi, A.A., Molecular characterization of full fusion protein (F) of Newcastle disease virus genotype VIId isolated from Egypt during 2012–2016, Vet. World, 2018, vol. 11, pp. 930–938.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Karlsson, E.A., Horm, S.V., Tok, S., Tum, S., Kalpravidh, W., Claes, F., Osbjer, K., and Dussart, P., Avian influenza virus detection, temporality and co-infection in poultry in Cambodian border provinces, 2017–2018, Emerging Microbes Infect., 2019, vol. 8, pp. 637–639.

    Article  Google Scholar 

  21. Ingrao, F., Duchatel, V., Rodil, I.F., Steensels, M., Verleysen, E., Mast, J., and Lambrecht, B., The expression of hemagglutinin by a recombinant Newcastle disease virus causes structural changes and alters innate immune sensing, Vaccines (Basel), 2021, vol. 9, no. 7, p. 758.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Gebauer, M., Hürlimann, H.C., Behrens, M., Wolff, T., and Behrens, S.-E., Subunit vaccines based on recombinant yeast protect against influenza A virus in a one-shot vaccination scheme, Vaccine, 2019, vol. 37, pp. 5578–5587.

    Article  CAS  PubMed  Google Scholar 

  23. Bourret, V., Avian influenza viruses in pigs: An overview, Vet. J., 2018, vol. 239, pp. 7–14.

    Article  CAS  PubMed  Google Scholar 

  24. Hatami, S., Fotouhi, F., Hashemi, M., Salehi, M., Ghaderi, E., Shokohi, H., and Farahmand, B., Prokaryotic expression of chimeric trimer protein (3M2e-HA2-NP) derived from conserved domains of influenza virus A/H1N1 as a promising universal subunit vaccine, Sci. J. Kurdistan Univ. Med. Sci., 2017, vol. 22, pp. 111–120.

    Google Scholar 

  25. Bouvier, N.M. and Palese, P., The biology of influenza viruses, Vaccine, 2008, vol. 26, pp. D49–D53.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Ha, Y., Stevens, D.J., Skehel, J.J., and Wiley, D.C., H5 avian and H9 swine influenza virus haemagglutinin structures: Possible origin of influenza subtypes, EMBO J., 2002, vol. 21, pp. 865–875.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Skehel, J.J. and Wiley, D.C., Influenza haemagglutinin, Vaccine, 2002, vol. 20, pp. S51–S54.

    Article  CAS  PubMed  Google Scholar 

  28. Orabi, A., Hussein, A., Saleh, A.A., El-Magd, M.A., and Munir, M., Evolutionary insights into the fusion protein of Newcastle disease virus isolated from vaccinated chickens in 2016 in Egypt, Arch. Virol., 2017, vol. 162, pp. 3069–3079.

    Article  CAS  PubMed  Google Scholar 

  29. Kingsman, S.M., Kingsman, A.J., Dobson, M.J., Mellor, J., and Roberts, N.A., Heterologous gene expression in Saccharomyces cerevisiae, Biotechnol. Genet. Eng. Rev., 1985, vol. 3, pp. 377–416.

    Article  CAS  PubMed  Google Scholar 

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ACKNOWLEDGMENTS

We greatly appreciate the dedicated efforts of the laboratory staff at the Department of Molecular Biology and Genetic, Tehran Central Branch, Islamic Azad University, and the personnel of the Molecular Department at Baghiyatollah al-Azam Hospital as well as Dr. Mohammadreza Shafaati from the Department of Cellular and Molecular Biology, Faculty of Basic Sciences, Hamedan Branch, Islamic Azad University, Hamadan.

Funding

This work was supported by ongoing institutional funding. No additional grants to carry out or direct this particular research were obtained.

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  1. Department of Biology, Islamic Azad University Central Tehran Branch, Tehran, Iran

    A. Seyed Hamzehzadeh & A. Hesampour

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  1. A. Seyed Hamzehzadeh
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Contributions

Ardeshir Hesampour conceptualized and designed the experiments. Atefeh Seyed Hamzehzadeh executed the experiments, analyzed the data, and drafted the manuscript. Ardeshir Hesampour reviewed and edited the paper. All authors have reviewed and approved the final version of the manuscript for publication.

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Correspondence to A. Hesampour.

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Hamzehzadeh, A.S., Hesampour, A. Cassette Construction and Co-Expression of NA and F Proteins of H1N1 Influenza Virus and Newcastle Disease Virus in Saccharomyces cerevisiae. Mol. Genet. Microbiol. Virol. 38, 293–303 (2023). https://doi.org/10.3103/S0891416823040134

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  • DOI: https://doi.org/10.3103/S0891416823040134

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