1887

Abstract

Influenza A virus mutates rapidly, allowing it to escape natural and vaccine-induced immunity. Neuraminidase (NA) is a surface protein capable of cleaving the glycosidic linkages of neuraminic acids to release newly formed virions from infected cells. Genetic variants within a viral population can influence the emergence of pandemic viruses as well as drug susceptibility and vaccine effectiveness. In the present study, 55 clinical specimens from patients infected with the 2009 pandemic influenza A/H1N1 virus, abbreviated as A(H1N1)pdm09, during the 2015–2016 outbreak season in Taiwan were collected. Whole genomes were obtained through next-generation sequencing. Based on the published sequences from A(H1N1)pdm09 strains worldwide, a mixed population of two distinct variants at NA position 151 was revealed. We initially reasoned that such a mixed population may have emerged during cell culture. However, additional investigations confirmed that these mixed variants were detectable in the specimens of patients. To further investigate the role of the two NA-151 variants in a dynamic population, a reverse genetics system was employed to generate recombinant A(H1N1)pdm09 viruses. It was observed that the mixture of the two distinct variants was characterized by a higher replication rate compared to the recombinant viruses harbouring a single variant. Moreover, an NA inhibition assay revealed that a high frequency of the minor NA-151 variant in A(H1N1)pdm09 was associated with a reduced susceptibility to NA inhibitors. We conclude that two distinct NA-151 variants can be identified in patient specimens and that such variants may increase viral replication and NA activity.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.001258
2019-04-17
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/jgv/100/5/752.html?itemId=/content/journal/jgv/10.1099/jgv.0.001258&mimeType=html&fmt=ahah

References

  1. Dawood FS, Jain S, Finelli L, Shaw MW, Lindstrom S et al. Emergence of a novel swine-origin influenza A (H1N1) virus in humans. N Engl J Med 2009; 360:2605–2615 [View Article][PubMed]
    [Google Scholar]
  2. Xue KS, Hooper KA, Ollodart AR, Dingens AS, Bloom JD. Cooperation between distinct viral variants promotes growth of H3N2 influenza in cell culture. Elife 2016; 5:e13974 [View Article][PubMed]
    [Google Scholar]
  3. Xue KS, Greninger AL, Pérez-Osorio A, Bloom JD. Cooperating H3N2 Influenza Virus Variants Are Not Detectable in Primary Clinical Samples. mSphere 2018; 3: [View Article][PubMed]
    [Google Scholar]
  4. Mishin VP, Sleeman K, Levine M, Carney PJ, Stevens J et al. The effect of the MDCK cell selected neuraminidase D151G mutation on the drug susceptibility assessment of influenza A(H3N2) viruses. Antiviral Res 2014; 101:93–96 [View Article][PubMed]
    [Google Scholar]
  5. Meijer A, Rebelo-de-Andrade H, Correia V, Besselaar T, Drager-Dayal R et al. Global update on the susceptibility of human influenza viruses to neuraminidase inhibitors, 2012-2013. Antiviral Res 2014; 110:31–41 [View Article][PubMed]
    [Google Scholar]
  6. Gubareva LV, Besselaar TG, Daniels RS, Fry A, Gregory V et al. Global update on the susceptibility of human influenza viruses to neuraminidase inhibitors, 2015-2016. Antiviral Res 2017; 146:12–20 [View Article][PubMed]
    [Google Scholar]
  7. Okomo-Adhiambo M, Nguyen HT, Sleeman K, Sheu TG, Deyde VM et al. Host cell selection of influenza neuraminidase variants: implications for drug resistance monitoring in A(H1N1) viruses. Antiviral Res 2010; 85:381–388 [View Article][PubMed]
    [Google Scholar]
  8. Kilander A, Rykkvin R, Dudman SG, Hungnes O. Observed association between the HA1 mutation D222G in the 2009 pandemic influenza A(H1N1) virus and severe clinical outcome, Norway 2009-2010. Euro Surveill 2010; 15: [View Article]
    [Google Scholar]
  9. Mak GC, Au KW, Tai LS, Chuang KC, Cheng KC et al. Association of D222G substitution in haemagglutinin of 2009 pandemic influenza A (H1N1) with severe disease. Euro Surveill 2010; 15:[PubMed]
    [Google Scholar]
  10. Chen H, Wen X, To KK, Wang P, Tse H et al. Quasispecies of the D225G substitution in the hemagglutinin of pandemic influenza A(H1N1) 2009 virus from patients with severe disease in Hong Kong, China. J Infect Dis 2010; 201:1517–1521 [View Article][PubMed]
    [Google Scholar]
  11. Manchanda H, Seidel N, Blaess MF, Claus RA, Linde J et al. Differential Biphasic Transcriptional Host Response Associated with Coevolution of Hemagglutinin Quasispecies of Influenza A Virus. Front Microbiol 2016; 7:1167 [View Article][PubMed]
    [Google Scholar]
  12. Newitt S, Mironenko A, Holubka O, Zaika O, Gubar O et al. Rapid risk assessment during the early weeks of the 2015-2016 influenza season in Ukraine. Influenza Other Respir Viruses 2018; 12:241–249 [View Article][PubMed]
    [Google Scholar]
  13. Gong YN, Kuo RL, Chen GW, Shih SR. Centennial review of influenza in Taiwan. Biomed J 2018; 41:234–241 [View Article][PubMed]
    [Google Scholar]
  14. Hsieh YC, Tsao KC, Huang CT, Chang KY, Huang YC et al. Clinical characteristics of patients with laboratory-confirmed influenza A(H1N1)pdm09 during the 2013/2014 and 2015/2016 clade 6B/6B.1/6B.2-predominant outbreaks. Sci Rep 2018; 8:15636 [View Article][PubMed]
    [Google Scholar]
  15. Samson M, Pizzorno A, Abed Y, Boivin G. Influenza virus resistance to neuraminidase inhibitors. Antiviral Res 2013; 98:174–185 [View Article][PubMed]
    [Google Scholar]
  16. Jonges M, Welkers MR, Jeeninga RE, Meijer A, Schneeberger P et al. Emergence of the virulence-associated PB2 E627K substitution in a fatal human case of highly pathogenic avian influenza virus A(H7N7) infection as determined by Illumina ultra-deep sequencing. J Virol 2014; 88:1694–1702 [View Article][PubMed]
    [Google Scholar]
  17. 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]
  18. Okomo-Adhiambo M, Sleeman K, Ballenger K, Nguyen HT, Mishin VP et al. Neuraminidase inhibitor susceptibility testing in human influenza viruses: a laboratory surveillance perspective. Viruses 2010; 2:2269–2289 [View Article][PubMed]
    [Google Scholar]
  19. Mori K, Murano K, Ohniwa RL, Kawaguchi A, Nagata K. Oseltamivir expands quasispecies of influenza virus through cell-to-cell transmission. Sci Rep 2015; 5:9163 [View Article][PubMed]
    [Google Scholar]
  20. Gulati U, Wu W, Gulati S, Kumari K, Waner JL et al. Mismatched hemagglutinin and neuraminidase specificities in recent human H3N2 influenza viruses. Virology 2005; 339:12–20 [View Article][PubMed]
    [Google Scholar]
  21. Neverov AD, Kryazhimskiy S, Plotkin JB, Bazykin GA. Coordinated Evolution of Influenza A Surface Proteins. PLoS Genet 2015; 11:e1005404 [View Article][PubMed]
    [Google Scholar]
  22. Mitrasinovic PM. On the structure-based design of novel inhibitors of H5N1 influenza A virus neuraminidase (NA). Biophys Chem 2009; 140:35–38 [View Article][PubMed]
    [Google Scholar]
  23. Sievers F, Wilm A, Dineen D, Gibson TJ, Karplus K et al. Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Mol Syst Biol 2011; 7:539 [View Article][PubMed]
    [Google Scholar]
  24. Gong YN, Chen GW, Yang SL, Lee CJ, Shih SR et al. A Next-Generation Sequencing Data Analysis Pipeline for Detecting Unknown Pathogens from Mixed Clinical Samples and Revealing Their Genetic Diversity. PLoS One 2016; 11:e0151495 [View Article][PubMed]
    [Google Scholar]
  25. Hsieh EF, Lin SJ, Mok CK, Chen GW, Huang CH et al. Altered pathogenicity for seasonal influenza virus by single reassortment of the RNP genes derived from the 2009 pandemic influenza virus. J Infect Dis 2011; 204:864–872 [View Article][PubMed]
    [Google Scholar]
  26. Hoffmann E, Neumann G, Kawaoka Y, Hobom G, Webster RG. A DNA transfection system for generation of influenza A virus from eight plasmids. Proc Natl Acad Sci USA 2000; 97:6108–6113 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.001258
Loading
/content/journal/jgv/10.1099/jgv.0.001258
Loading

Data & Media loading...

Supplements

Supplementary File 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error