1887

Abstract

Serological assays with modern influenza A/H3N2 viruses have become problematic due to the progressive reduction in the ability of viruses of this subtype to bind and agglutinate red blood cells (RBCs). This is due to reduced ability of the viral haemagglutinin (HA) glycoprotein to bind to the sialic acid-containing receptors presented by these cells. Additionally, as a result of reduced HA-mediated binding in cell culture, modern A/H3N2 viruses often acquire compensatory mutations during propagation that enable binding of cellular receptors through their neuraminidase (NA) surface protein. Viruses that have acquired this NA-mediated binding agglutinate RBCs through their NA, confusing the results of serological assays designed to assess HA antigenicity. Here we confirm with a large dataset that the acquisition of mutations that confer NA binding of RBCs is a culture artefact, and demonstrate that modern A/H3N2 isolates with acquired NA-binding mutations revert to a clinical-like NA sequence after a single passage in human airway epithelial (HAE) cells.

Funding
This study was supported by the:
  • NIBSC
    • Principle Award Recipient: Jonathan Christopher Brown
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
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2019-11-08
2024-12-07
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References

  1. Matrosovich M, Matrosovich T, Carr J, Roberts NA, Klenk H-D et al. Overexpression of the alpha-2,6-sialyltransferase in MDCK cells increases influenza virus sensitivity to neuraminidase inhibitors. J Virol 2003; 77:8418–8425 [View Article][PubMed]
    [Google Scholar]
  2. Oh DY, Barr IG, Mosse JA, Laurie KL. Mdck-Siat1 cells show improved isolation rates for recent human influenza viruses compared to conventional MDCK cells; 2008; 462189–2194
  3. Lin YP, Gregory V, Collins P, Kloess J, Wharton S et al. Neuraminidase receptor binding variants of human influenza A(H3N2) viruses resulting from substitution of aspartic acid 151 in the catalytic site: a role in virus attachment?. J Virol 2010; 84:6769–6781 [View Article][PubMed]
    [Google Scholar]
  4. Zhu X, McBride R, Nycholat CM, Yu W, Paulson JC et al. Influenza virus neuraminidases with reduced enzymatic activity that avidly bind sialic acid receptors. J Virol 2012; 86:13371–13383 [View Article][PubMed]
    [Google Scholar]
  5. Xue KS, Hooper KA, Ollodart AR, Dingens AS, Bloom JD et al. Cooperation between distinct viral variants promotes growth of H3N2 influenza in cell culture. eLife 2016; 5:1–15 [View Article]
    [Google Scholar]
  6. Xue KS, Greninger AL, Pérez-Osorio A, Bloom JD. Cooperating H3N2 influenza virus variants are not detectable in primary clinical samples. mSphere 2018; 3:e00552-17 [View Article][PubMed]
    [Google Scholar]
  7. Mohr PG, Deng YM, McKimm-Breschkin JL. The neuraminidases of MDCK grown human influenza A(H3N2) viruses isolated since 1994 can demonstrate receptor binding. Virol J 2015; 12:67 [View Article][PubMed]
    [Google Scholar]
  8. Tamura D, Nguyen HT, Sleeman K, Levine M, Mishin VP et al. Cell culture-selected substitutions in influenza A(H3N2) neuraminidase affect drug susceptibility assessment. Antimicrob Agents Chemother 2013; 57:6141–6146 [View Article][PubMed]
    [Google Scholar]
  9. Mögling R, Richard MJ, Vliet Svander, Beek Rvan, Schrauwen EJA et al. Neuraminidase-mediated haemagglutination of recent human influenza A(H3N2) viruses is determined by arginine 150 flanking the neuraminidase catalytic site. J Gen Virol 2017; 98:1274–1281 [View Article][PubMed]
    [Google Scholar]
  10. Chambers BS, Li Y, Hodinka RL, Hensley SE. Recent H3N2 influenza virus clinical isolates rapidly acquire hemagglutinin or neuraminidase mutations when propagated for antigenic analyses. J Virol 2014; 88:10986–10989 [View Article][PubMed]
    [Google Scholar]
  11. Skowronski DM, Sabaiduc S, Chambers C, Eshaghi A, Gubbay JB et al. Mutations acquired during cell culture isolation may affect antigenic characterisation of influenza A(H3N2) clade 3C.2a viruses. Eurosurveillance 2016; 21:1–7 [View Article]
    [Google Scholar]
  12. Lin Y, Wharton SA, Whittaker L, Dai M, Ermetal B et al. The characteristics and antigenic properties of recently emerged subclade 3C.3a and 3C.2a human influenza A(H3N2) viruses passaged in MDCK cells. Influenza Other Respi Viruses 2017; 11:263–274 [View Article]
    [Google Scholar]
  13. Lin YP, Xiong X, Wharton SA, Martin SR, Coombs PJ et al. Evolution of the receptor binding properties of the influenza A(H3N2) hemagglutinin. Proc Natl Acad Sci USA 2012; 109:21474–21479 [View Article][PubMed]
    [Google Scholar]
  14. Medeiros R, Escriou N, Naffakh N, Manuguerra JC, van der Werf S et al. Hemagglutinin residues of recent human A(H3N2) influenza viruses that contribute to the inability to agglutinate chicken erythrocytes. Virology 2001; 289:74–85 [View Article][PubMed]
    [Google Scholar]
  15. Bloom JD, Gong LI, Baltimore D. Permissive secondary mutations enable the evolution of influenza oseltamivir resistance. Science 2010; 328:1272–1275 [View Article][PubMed]
    [Google Scholar]
  16. Byrd-Leotis L, Cummings RD, Steinhauer DA. The interplay between the host receptor and influenza virus hemagglutinin and neuraminidase. Int J Mol Sci 2017; 18:1541 [View Article]
    [Google Scholar]
  17. Kaverin NV, Gambaryan AS, Bovin NV, Rudneva IA, Shilov AA et al. Postreassortment changes in influenza A virus hemagglutinin restoring HA-NA functional match. Virology 1998; 244:315–321 [View Article][PubMed]
    [Google Scholar]
  18. Mitnaul LJ, Matrosovich MN, Castrucci MR, Tuzikov AB, Bovin NV et al. Balanced hemagglutinin and neuraminidase activities are critical for efficient replication of influenza A virus. J Virol 2000; 74:6015–6020 [View Article][PubMed]
    [Google Scholar]
  19. Park S, Il Kim J, Lee I, Bae JY, Yoo K et al. Adaptive mutations of neuraminidase stalk truncation and deglycosylation confer enhanced pathogenicity of influenza A viruses. Sci Rep 2017; 7:10928 [View Article][PubMed]
    [Google Scholar]
  20. Wagner R, Matrosovich M, Klenk H-D. Functional balance between haemagglutinin and neuraminidase in influenza virus infections. Rev Med Virol 2002; 12:159–166 [View Article][PubMed]
    [Google Scholar]
  21. Takada K, Kawakami C, Fan S, Chiba S, Zhong G et al. A humanized MDCK cell line for the efficient isolation and propagation of human influenza viruses. Nat Microbiol 2019; 4:1268–1273 [View Article][PubMed]
    [Google Scholar]
  22. Nicolson C, Major D, Wood JM, Robertson JS. Generation of influenza vaccine viruses on Vero cells by reverse genetics: an H5N1 candidate vaccine strain produced under a quality system. Vaccine 2005; 23:2943–2952 [View Article][PubMed]
    [Google Scholar]
  23. Zhou B, Donnelly ME, Scholes DT, St George K, Hatta M et al. Single-reaction genomic amplification accelerates sequencing and vaccine production for classical and swine origin human influenza A viruses. J Virol 2009; 83:10309–10313 [View Article][PubMed]
    [Google Scholar]
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