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Influenza Other Respir Viruses. 2017 Feb 5. doi: 10.1111/irv.12447. [Epub ahead of print]

The characteristics and antigenic properties of recently emerged subclade 3C.3a and 3C.2a human influenza A(H3N2) viruses passaged in MDCK cells.

Author information

  • 1The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK.
  • 2Centre for Health Protection, Department of Health, 382 Nam Cheong Street, Hong Kong Special Administrative Region, China.
  • 3Instituto Nacional de Enfermedades Infecciosas-ANLIS "Dr.Carlos G.Malbran", Av. Velez Sarsfield 563, C1282AFF, Buenos Aires, Argentina.

Abstract

BACKGROUND:

Two new subclades of influenza A(H3N2) viruses became prominent during the 2014-2015 Northern Hemisphere influenza season. The HA glycoproteins of these viruses showed sequence changes previously associated with alterations in receptor-binding properties. To address how these changes influence virus propagation, viruses were isolated and propagated in conventional MDCK cells and MDCK-SIAT1 cells, cells with enhanced expression of the human receptor for the virus, and analysed at each passage.

METHODS:

Gene sequence analysis was undertaken as virus was passaged in conventional MDCK cells and MDCK-SIAT1 cells. Alterations in receptor recognition associated with passage of virus was examined by haemagglutination assays using red blood cells from guinea pigs, turkeys and humans. Micro-neutralisation assays were performed to determine how passage-acquired amino acid substitutions and polymorphisms affected virus antigenicity.

RESULTS:

Viruses were able to infect MDCK-SIAT1 cells more efficiently than conventional MDCK cells. Viruses of both the 3C.2a and 3C.3a subclades showed greater sequence change on passage in conventional MDCK cells than in MDCK-SIAT1 cells, with amino acid substitutions being seen in both HA and NA glycoproteins. However, virus passage in MDCK-SIAT1 cells at low inoculum dilutions showed reducing infectivity on continued passage.

CONCLUSIONS:

Current H3N2 viruses should be cultured in the MDCK-SIAT1 cell-line to maintain faithful replication of the virus, and at an appropriate multiplicity of infection to retain infectivity. This article is protected by copyright. All rights reserved.

KEYWORDS:

Antigenicity; Influenza; MDCK cells; MDCK-SIAT1 cells; Receptor binding

PMID:
28164446
DOI:
10.1111/irv.12447
[PubMed - as supplied by publisher]
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