Journal
INFLUENZA AND OTHER RESPIRATORY VIRUSES
Volume 11, Issue 3, Pages 263-274Publisher
WILEY
DOI: 10.1111/irv.12447
Keywords
antigenicity; influenza; MDCK cells; MDCK-SIAT1 cells; receptor binding
Categories
Funding
- Francis Crick Institute
- Cancer Research UK [FC001030]
- UK Medical Research Council [FC001030]
- Wellcome Trust [FC001030]
- MRC [MC_U117585868, MC_U117512723] Funding Source: UKRI
- Medical Research Council [MC_U117585868, MC_U117512723] Funding Source: researchfish
- The Francis Crick Institute [10030, 1783-JeGa] Funding Source: researchfish
Ask authors/readers for more resources
BackgroundTwo 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. MethodsGene 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 were examined by haemagglutination assays using red blood cells from guinea pigs, turkeys and humans. Microneutralisation assays were performed to determine how passage-acquired amino acid substitutions and polymorphisms affected virus antigenicity. ResultsViruses 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. ConclusionsCurrent 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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available