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Preparation and characterization of new challenge stocks of SIVmac32H J5 following rapid serial passage of virus in vivo
Authors:Ferguson D  Wade-Evans A  Elsley W  Sangster R  Silvera P  MacManus S  Davis G  Corcoran T  Berry N  Brown S  Jenkins A  Cowie J  Sethi M  Hull R  Stebbings R  Lines J  Norley S  Stott E J  Almond N
Affiliation:Division of Retrovirology, NIBSC, South Mimms, Potters Bar, Hertfordshire, UK. dferguson@nibsc.ac.uk
Abstract:BACKGROUND: A new challenge stock of the simian immunodeficiency virus SIVmacJ5 has been produced following passage in vivo. METHODS: SIVmacJ5 3/92 (J5M), was passaged serially through cynomolgus macaques (Macaca fascicularis) by intravenous inoculation of infected spleen cells isolated and prepared 14 days post-infection. Two challenge stocks, SIVmacJ5 S61MLN and SIVmacJ5 S62spl, were prepared by culture of lymphoid tissue ex vivo. RESULTS: These virus stocks appeared better adapted for replication in M. fascicularis as demonstrated by a greater persistence of recoverable live virus from the periphery and increased pathology in lymphoid tissues 20 weeks post-challenge as detected by immunohistochemistry. Sequence analysis of the envelope gene from these stocks did not identify marked diversification of sequence as a result of this procedure. CONCLUSIONS: These stocks display more robust peripheral persistence and tissue pathology in cynomolgus macaques and should prove valuable analysing recombinant vaccines based upon SIVmacJ5 transgenes.
Keywords:Cynomolgus macaque    immunohistochemistry    in situ hybridization    pathology    SIVmacJ5 S61MLN
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