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Cytotechnology

, Volume 65, Issue 3, pp 419–424 | Cite as

Impact of chicken-origin cells on adaptation of a low pathogenic influenza virus

  • Shahla Shahsavandi
  • Mohammad Majid Ebrahimi
  • Ashraf Mohammadi
  • Nima Zarrin Lebas
Original Research

Abstract

Understanding the growth dynamics of influenza viruses is an essential step in virus replication and cell-adaptation. The aim of this study was to elucidate the growth kinetic of a low pathogenic avian influenza H9N2 subtype in chicken embryo fibroblast (CEF) and chicken tracheal epithelial (CTE) cells during consecutive passages. An egg-adapted H9N2 virus was seeded into both cell culture systems. The amount of infectious virus released into the cell culture supernatants at interval times post-infection were titered and plaque assayed. The results as well as cell viability results indicate that the infectivity of the influenza virus was different among these primary cells. The egg-adapted H9N2 virus featured higher infectivity in CTE than in CEF cells. After serial passages and plaque purifications of the virus, a CTE cell-adapted strain was generated which carried amino acid substitutions within the HA stem region. The strain showed faster replication kinetics in cell culture resulting in an increase in virus titer. Overall, the present study provides the impact of cell type, multiplicity of infection, cellular protease roles in virus infectivity and finally molecular characterization during H9N2 virus adaptation procedure.

Keywords

H9N2 avian influenza virus Adaptation Chicken-origin cell Multiplicity of infection HA Cellular protease 

References

  1. Alexander DJ (2003) Report on avian influenza in the Eastern Hemisphere during 1997–2002. Avian Dis 47:792–797CrossRefGoogle Scholar
  2. Böttcher E, Matrosovich T, Beyerle M, Klenk HD, Garten W, Matrosovich M (2006) Proteolytic activation of influenza viruses by serine proteases TMPRSS2 and HAT from human airway epithelium. J Virol 80:9896–9898CrossRefGoogle Scholar
  3. Brown IH, Banks J, Manvell RJ, Essen SC, Shell W, Slomka M, Londt B, Alexander DJ (2006) Recent epidemiology and ecology of influenza A viruses in avian species in Europe and the Middle East. Dev Biol (Basel) 124:45–50Google Scholar
  4. Bugge TH, Antalis TM, Wu Q (2009) Type II transmembrane serine proteases. J Biol Chem 284:23177–23181CrossRefGoogle Scholar
  5. Butt KM, Smith GJD, Chen H, Zhang LJ, Connie Leung YH, Xu KM, Lim W, Webster RG, Yuen KY, Malik Peiris JS, Guan Y (2005) Human infection with an avian H9N2 influenza A virus in Hong Kong in 2003. J Clin Microbiol 43:5760–5767CrossRefGoogle Scholar
  6. Chaipan C, Kobasa D, Bertram S, Glowacka I, Steffen I, Tsegaye TS, Takeda M, Bugge TH, Kim S, Park Y, Marzi A, Pohlmann S (2009) Proteolytic activation of the 1918 influenza virus hemagglutinin. J Virol 83:3200–3211CrossRefGoogle Scholar
  7. Klenk HD, Matrosovich M, Stech J (2007) Avian influenza: molecular mechanisms of pathogenesis and host range. In: Mettenleiter TC, Sobrino F (eds) Animal viruses: molecular biology. Caister Academic, Norfolk, pp 253–303Google Scholar
  8. Lee CW, Jung K, Jadhao SJ, Suarez DL (2008) Evaluation of chicken-origin (DF-1) and quail-origin (QT-6) fibroblast cell lines for replication of avian influenza viruses. J Virol Methods 153:22–28CrossRefGoogle Scholar
  9. Liu X, Sun L, Yu M, Wang Z, Xu C, Xue Q, Zhang K, Ye X, Kitamura Y, Liu W (2009) Cyclophilin A interacts with influenza A virus M1 protein and impairs the early stage of the viral replication. Cell Microbiol 11:730–741CrossRefGoogle Scholar
  10. Matrosovich M, Klenk HD (2003) Natural and synthetic sialic acid-containing inhibitors of influenza virus receptor binding. Rev Med Virol 13:85–97CrossRefGoogle Scholar
  11. Matrosovich MN, Krauss S, Webster RG (2001) H9N2 influenza A viruses from poultry in Asia have human virus-like receptor specificity. Virology 281:156–162CrossRefGoogle Scholar
  12. Matrosovich MN, Matrosovich TY, Gray T, Roberts NA, Klenk HD (2004) Human and avian influenza viruses target different cell types in cultures of human airway epithelium. Proc Natl Acad Sci USA 101:4620–4624CrossRefGoogle Scholar
  13. Moresco KA, Stallknecht DE, Swayne DE (2010) Evaluation and attempted optimization of avian embryos and cell culture methods for efficient isolation and propagation of low pathogenicity avian influenza viruses. Avian Dis 54:622–626CrossRefGoogle Scholar
  14. Office International des Epizooties (OIE) (2008) Avian influenza. In: Manual of diagnostic tests and vaccines for terrestrial animals, 5th edn. Office International des Epizooties, Paris, France (Chapter 2.7.12)Google Scholar
  15. Okumura Y, Takahashi E, Yano M, Ohuchi M, Daidoji T, Nakaya T, Böttcher E, Garten W, Klenk HD, Kido H (2010) Novel type II transmembrane serine proteases, MSPL and TMPRSS13, Proteolytically activate membrane fusion activity of the hemagglutinin of highly pathogenic avian influenza viruses and induce their multicycle replication. J Virol 84:5089–5096CrossRefGoogle Scholar
  16. Saito T, Lim W, Suzuki T, Suzuki Y, Kida H, Nishimura SI, Tashiro M (2001) Characterization of a human H9N2 influenza virus isolated in Hong Kong. Vaccine 20:125–133CrossRefGoogle Scholar
  17. Scull MA, Gillim-Ross L, Santos C, Roberts KL, Bordonali E, Subbarao K, Barclay WS, Pickles RJ (2009) Avian influenza virus glycoproteins restrict virus replication and spread through human airway epithelium at temperatures of the proximal airways. PLoS Pathog 5:e1000424. doi: 10.1371/journal.ppat.1000424 CrossRefGoogle Scholar
  18. Shahsavandi S, Salmanian AH, Ghorashi SA, Masoudi S, Ebrahimi MM (2012) Evolutionary characterization of hemagglutinin gene of H9N2 influenza viruses isolated from Asia. Res Vet Sci 93:234–239CrossRefGoogle Scholar
  19. Shen C-I, Wang C-H, Shen S-C, Lee H-C, Liao J-W, Lia J-W, Su H-L (2011) The infection of chicken tracheal epithelial cells with a H6N1 avian influenza virus. PLoS ONE 6:e18894. doi: 10.1371/journal.pone.0018894 CrossRefGoogle Scholar
  20. Steinhauer DA (1999) Role of hemagglutinin cleavage for the pathogenicity of influenza virus. Virology 25:1–20CrossRefGoogle Scholar
  21. Swayne DE, Senne DA, Beard CW (2008) Avian influenza. In: A laboratory manual for the isolation and identification of avian pathogens, 5th edn. American Association of Avian Pathologists, Jacksonville, pp 128–134Google Scholar
  22. Thompson KAS, Yin J (2010) Population dynamics of an RNA virus and its defective interfering particles in passage cultures. Virol J 7:257CrossRefGoogle Scholar
  23. Thompson CI, Barclay WS, Zambon MC, Pickles RJ (2006) Infection of human airway epithelium by human and avian strains of influenza A virus. J Virol 80:8060–8068CrossRefGoogle Scholar
  24. Webster RG, Bean WJ, Gorman OT, Chambers TM, Kawaoka Y (1992) Evolution and ecology of influenza A viruses. Microbiol Rev 56:152–179Google Scholar
  25. Xu C, Meng S, Liu X, Sun L, Liu W (2010) Chicken cyclophilin A is an inhibitory factor to influenza virus replication. Virol J 7:372CrossRefGoogle Scholar
  26. Zaffuto KM, Estevez CN, Afonso CL (2008) Primary chicken tracheal cell culture system for the study of infection with avian respiratory viruses. Avian Pathol 37:25–31CrossRefGoogle Scholar
  27. Zhang H (2009) Tissue and host tropism of influenza viruses: Importance of quantitative analysis. Sci China Ser C-Life Sci 52:1101–1110CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Shahla Shahsavandi
    • 1
  • Mohammad Majid Ebrahimi
    • 1
  • Ashraf Mohammadi
    • 1
  • Nima Zarrin Lebas
    • 1
  1. 1.Razi Vaccine & Serum Research InstituteKarajIran

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