MDCK-SIAT1 Cells Show Improved Isolation Rates for Recent Human Influenza Viruses Compared to Conventional MDCK Cells

Oh, Ding Yuan; Barr, Ian G.; Mosse, Jennifer; Laurie, Karen

doi:10.1128/jcm.00398-08

Cited by 137 publications

(139 citation statements)

References 40 publications

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“…In our hands, most contemporary human influenza H1N1 and H3N2 viruses are detected earlier, and form higher virus titers in the engineered MDCK cells, than occurs in conventional MDCK cells (J. Lednicky and D. Wyatt, data to be presented elsewhere), in agreement with a recent report [72]. Apart from standard MDCK cells, we have also engineered additional cell lines that over-express SIAT1, and also, cell lines that over-express SIAT4 to catalyze the formation of -2,3 -linked sialic acid receptors (data and information to be presented elsewhere).…”

Section: Engineered Cell Lines For Virus Isolationsupporting

confidence: 81%

“…Apart from standard MDCK cells, we have also engineered additional cell lines that over-express SIAT1, and also, cell lines that over-express SIAT4 to catalyze the formation of -2,3 -linked sialic acid receptors (data and information to be presented elsewhere). With these SIAT1 and SIAT4 overexpressing cells, we have isolated influenza viruses that others were unable to, and also, as for [72], we have recovered viruses from frozen stocks that others had great difficulty with.…”

Section: Engineered Cell Lines For Virus Isolationmentioning

confidence: 99%

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The Art of Animal Cell Culture for Virus Isolation

Lednicky¹,

Wyatt²

2012

Biomedical Tissue Culture

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Section: Engineered Cell Lines For Virus Isolationsupporting

confidence: 81%

Section: Engineered Cell Lines For Virus Isolationmentioning

confidence: 99%

The Art of Animal Cell Culture for Virus Isolation

Lednicky¹,

Wyatt²

2012

Biomedical Tissue Culture

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“…Although early studies showed that the sequences of HA genes of H3N2 viruses isolated in cell culture are generally identical to those sequenced directly from nasal secretions of patients, recently circulating viruses may also undergo host selection during isolation in mammalian cells (30,49). Isolation of H3N2 viruses in MDCK cells has been progressively more difficult in recent years (49).…”

Section: Discussionmentioning

confidence: 99%

Receptor Specificity of Influenza A H3N2 Viruses Isolated in Mammalian Cells and Embryonated Chicken Eggs

Stevens

Chen

Carney

et al. 2010

J Virol

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Isolation of human subtype H3N2 influenza viruses in embryonated chicken eggs yields viruses with amino acid substitutions in the hemagglutinin (HA) that often affect binding to sialic acid receptors. We used a glycan array approach to analyze the repertoire of sialylated glycans recognized by viruses from the same clinical specimen isolated in eggs or cell cultures. The binding profiles of whole virions to 85 sialoglycans on the microarray allowed the categorization of cell isolates into two groups. Group 1 cell isolates displayed binding to a restricted set of ␣2-6 and ␣2-3 sialoglycans, whereas group 2 cell isolates revealed receptor specificity broader than that of their egg counterparts. Egg isolates from group 1 showed binding specificities similar to those of cell isolates, whereas group 2 egg isolates showed a significantly reduced binding to ␣2-6-and ␣2-3-type receptors but retained substantial binding to specific O-and N-linked ␣2-3 glycans, including ␣2-3GalNAc and fucosylated ␣2-3 glycans (including sialyl Lewis x), both of which may be important receptors for H3N2 virus replication in eggs. These results revealed an unexpected diversity in receptor binding specificities among recent H3N2 viruses, with distinct patterns of amino acid substitution in the HA occurring upon isolation and/or propagation in eggs. These findings also suggest that clinical specimens containing viruses with group 1-like receptor binding profiles would be less prone to undergoing receptor binding or antigenic changes upon isolation in eggs. Screening cell isolates for appropriate receptor binding properties might help focus efforts to isolate the most suitable viruses in eggs for production of antigenically well-matched influenza vaccines.

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“…Recent publications have reported on the expression of the human siat1 gene (ST6Gal I) in MDCK cells that was associated with an increase in ␣2,6-linked sialic acid on glycoproteins (29)(30)(31). The researchers wanted to increase the number of 6-linked sialic acids on the cell surface to enhance influenza virus sensitivity to neuraminidase inhibitor, which is the key component of antiviral drugs for influenza.…”

Section: Discussionmentioning

confidence: 99%

Conversion of MDCK cell line to suspension culture by transfecting with human siat7e gene and its application for influenza virus production

Chu

Lugovtsev

Golding

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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MDCK cells are currently being considered as an alternative to embryonated eggs for influenza virus propagation and hemagglutinin (HA) production intended for vaccine manufacturing. MDCK cells were found suitable for the virus production but their inability to grow in suspension burdens the process of scale up and hence their production capability. Anchorage-dependent MDCK cells were converted to anchorage-independent cells, capable of growing in suspension as a result of transfection with the human siat7e gene (ST6GalNac V). This gene was previously identified as having an important role in cellular adhesion when the transcriptions of genes from anchorage-dependent and anchorage-independent HeLa cells were compared. Unlike the parental MDCK cells, the siat7e-expressing cells were capable of growing in shake flasks as suspension cultures, achieving maximum concentration of 7 ؋ 10 5 cells/mL while keeping close to 100% viability throughout the growth phase. In production experiments, the siat7e-expressing cells were infected with the Influenza B/Victoria/504/2000 strain. It was determined that the cell-derived viruses retained similar antigenic properties as those obtained from egg-derived viruses and their nucleotide sequences were identical. The specific production of hemagglutinin (expressed in hemagglutination units per 10 6 cells) from the siat7e-expressing cells was approximately 20 times higher than the specific production from the parental MDCK cells. If this suspension process scales up, the production potential of HA from 10 L of siat7e-expressing cells at a concentration of 10 6 cells/mL would be equivalent to the amount of HA obtained from 10,000 embryonated eggs.anchorage-independent ͉ hemagglutinin ͉ sialyltransferase ͉ vaccine

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MDCK-SIAT1 Cells Show Improved Isolation Rates for Recent Human Influenza Viruses Compared to Conventional MDCK Cells

Cited by 137 publications

References 40 publications

The Art of Animal Cell Culture for Virus Isolation

The Art of Animal Cell Culture for Virus Isolation

Receptor Specificity of Influenza A H3N2 Viruses Isolated in Mammalian Cells and Embryonated Chicken Eggs

Conversion of MDCK cell line to suspension culture by transfecting with human siat7e gene and its application for influenza virus production

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