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Adaptation of Saffold Virus-2 for High-Titer Growth in Mammalian Cells

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posted on 17.08.2012 by Shannon Hertzler, Zhiguo Liang, Treso Balint, Howard L. Lipton
Saffold viruses (SAFV) are a recently discovered group of human Cardioviruses closely related to Theiler's murine encephalomyelitis viruses (TMEV). Unlike TMEV and encephalomyocarditis virus, each of which is monotypic, SAFV are genetically diverse and include at least eight genotypes. To date, only Saffold virus 3 (SAFV-3) has been grown efficiently in mammalian cells in vitro. Here, we report the successful adaptation of SAFV-2 for efficient growth in HeLa cells after 13 passages in the alpha/beta interferon-deficient human glial cell line U118 MG. Nine amino acid changes were found in the adapted virus, with single mutations in VP2, VP3, and 2B, while 6 mutations arose in VP1. Most capsid mutations were in surface loops. Analysis of SAFV-2 revealed virus growth and cytopathic effect only in human cell lines, with large plaques forming in HeLa cells, with minimal cell association, and without using sialic acid to enter cells. Despite the limited growth of SAFV-2 in rodent cells in vitro, BALB/c mice inoculated with SAFV-2 showed antibody titers of >1:10(6), and fluorescence-activated cell sorting (FACS) analysis revealed only minimal cross-reactivity with SFV-3. Intracerebral inoculation of 6-week-old FVB/n mice produced paralysis and acute neuropathological changes, including meningeal infiltrates, encephalitis, particularly of the limbic system, and spinal cord white matter inflammation.

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This work was supported by NIH grant NS 021913, the Grant Healthcare Foundation, and the Rosztoscy Foundation fellowship for BT.

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Copyright © 2011, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved. DOI: 10.1128/JVI.00265-11

Publisher

American Society for Microbiology

Language

en_US

issn

0022-538X

Issue date

01/07/2011

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