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.
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British Columbia Ctr Dis Control, Vancouver, BC, Canada
Univ British Columbia, Vancouver, BC V5Z 1M9, CanadaBritish Columbia Ctr Dis Control, Vancouver, BC, Canada
Skowronski, D. M.
;
Sabaiduc, S.
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British Columbia Ctr Dis Control, Vancouver, BC, CanadaBritish Columbia Ctr Dis Control, Vancouver, BC, Canada
Sabaiduc, S.
;
Chambers, C.
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British Columbia Ctr Dis Control, Vancouver, BC, CanadaBritish Columbia Ctr Dis Control, Vancouver, BC, Canada
Chambers, C.
;
Eshaghi, A.
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Publ Hlth Ontario, Toronto, ON, CanadaBritish Columbia Ctr Dis Control, Vancouver, BC, Canada
Eshaghi, A.
;
Gubbay, J. B.
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Publ Hlth Ontario, Toronto, ON, Canada
Univ Toronto, Toronto, ON, CanadaBritish Columbia Ctr Dis Control, Vancouver, BC, Canada
Gubbay, J. B.
;
Krajden, M.
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British Columbia Ctr Dis Control, Vancouver, BC, Canada
Univ British Columbia, Vancouver, BC V5Z 1M9, CanadaBritish Columbia Ctr Dis Control, Vancouver, BC, Canada
Krajden, M.
;
Drews, S. J.
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Univ Alberta, Edmonton, AB, Canada
Alberta Prov Lab, Edmonton, AB, CanadaBritish Columbia Ctr Dis Control, Vancouver, BC, Canada
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New York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA
SUNY Albany, Sch Publ Hlth, Dept Biomed Sci, Albany, NY 12201 USANew York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA
Zhou, Bin
;
Donnelly, Matthew E.
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New York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USANew York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA
Donnelly, Matthew E.
;
Scholes, Derek T.
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New York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USANew York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA
Scholes, Derek T.
;
George, Kirsten St.
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New York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USANew York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA
George, Kirsten St.
;
Hatta, Masato
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Univ Wisconsin, Sch Vet Med, Dept Pathobiol Sci, Madison, WI 53706 USANew York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA
Hatta, Masato
;
Kawaoka, Yoshihiro
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Univ Wisconsin, Sch Vet Med, Dept Pathobiol Sci, Madison, WI 53706 USA
Univ Tokyo, Inst Med Sci, Dept Microbiol & Immunol, Div Virol, Tokyo 1088639, JapanNew York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA
Kawaoka, Yoshihiro
;
Wentworth, David E.
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h-index: 0
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New York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA
SUNY Albany, Sch Publ Hlth, Dept Biomed Sci, Albany, NY 12201 USANew York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA
机构:
British Columbia Ctr Dis Control, Vancouver, BC, Canada
Univ British Columbia, Vancouver, BC V5Z 1M9, CanadaBritish Columbia Ctr Dis Control, Vancouver, BC, Canada
Skowronski, D. M.
;
Sabaiduc, S.
论文数: 0引用数: 0
h-index: 0
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British Columbia Ctr Dis Control, Vancouver, BC, CanadaBritish Columbia Ctr Dis Control, Vancouver, BC, Canada
Sabaiduc, S.
;
Chambers, C.
论文数: 0引用数: 0
h-index: 0
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British Columbia Ctr Dis Control, Vancouver, BC, CanadaBritish Columbia Ctr Dis Control, Vancouver, BC, Canada
Chambers, C.
;
Eshaghi, A.
论文数: 0引用数: 0
h-index: 0
机构:
Publ Hlth Ontario, Toronto, ON, CanadaBritish Columbia Ctr Dis Control, Vancouver, BC, Canada
Eshaghi, A.
;
Gubbay, J. B.
论文数: 0引用数: 0
h-index: 0
机构:
Publ Hlth Ontario, Toronto, ON, Canada
Univ Toronto, Toronto, ON, CanadaBritish Columbia Ctr Dis Control, Vancouver, BC, Canada
Gubbay, J. B.
;
Krajden, M.
论文数: 0引用数: 0
h-index: 0
机构:
British Columbia Ctr Dis Control, Vancouver, BC, Canada
Univ British Columbia, Vancouver, BC V5Z 1M9, CanadaBritish Columbia Ctr Dis Control, Vancouver, BC, Canada
Krajden, M.
;
Drews, S. J.
论文数: 0引用数: 0
h-index: 0
机构:
Univ Alberta, Edmonton, AB, Canada
Alberta Prov Lab, Edmonton, AB, CanadaBritish Columbia Ctr Dis Control, Vancouver, BC, Canada
机构:
New York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA
SUNY Albany, Sch Publ Hlth, Dept Biomed Sci, Albany, NY 12201 USANew York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA
Zhou, Bin
;
Donnelly, Matthew E.
论文数: 0引用数: 0
h-index: 0
机构:
New York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USANew York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA
Donnelly, Matthew E.
;
Scholes, Derek T.
论文数: 0引用数: 0
h-index: 0
机构:
New York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USANew York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA
Scholes, Derek T.
;
George, Kirsten St.
论文数: 0引用数: 0
h-index: 0
机构:
New York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USANew York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA
George, Kirsten St.
;
Hatta, Masato
论文数: 0引用数: 0
h-index: 0
机构:
Univ Wisconsin, Sch Vet Med, Dept Pathobiol Sci, Madison, WI 53706 USANew York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA
Hatta, Masato
;
Kawaoka, Yoshihiro
论文数: 0引用数: 0
h-index: 0
机构:
Univ Wisconsin, Sch Vet Med, Dept Pathobiol Sci, Madison, WI 53706 USA
Univ Tokyo, Inst Med Sci, Dept Microbiol & Immunol, Div Virol, Tokyo 1088639, JapanNew York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA
Kawaoka, Yoshihiro
;
Wentworth, David E.
论文数: 0引用数: 0
h-index: 0
机构:
New York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA
SUNY Albany, Sch Publ Hlth, Dept Biomed Sci, Albany, NY 12201 USANew York State Dept Hlth, Wadsworth Ctr, Albany, NY 12201 USA