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Nitric oxide and its possible implication in ME/CFS (Part 2 of 2)
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Severe Restriction of Xenotropic Murine Leukemia Virus-Related Virus Replication

Discussion in 'XMRV Research and Replication Studies' started by urbantravels, Feb 16, 2011.

  1. urbantravels

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    Severe Restriction of Xenotropic Murine Leukemia Virus-Related Virus Replication and Spread in Cultured Human Peripheral Blood Mononuclear Cells

    http://jvi.asm.org/cgi/content/abstract/JVI.00046-11v1

    J. Virol. doi:10.1128/JVI.00046-11
    Copyright (c) 2011, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

    Chawaree Chaipan, Kari A. Dilley, Tobias Paprotka, Krista A. Delviks-Frankenberry, Narasimhan J. Venkatachari, Wei-Shau Hu, and Vinay K. Pathak*

    HIV Drug Resistance Program, National Cancer Institute at Frederick, Viral Mutation Section and Viral Recombination Section, Frederick, MD 21702, USA

    Abstract

    Xenotropic murine leukemia virus-related virus (XMRV) is a gammaretrovirus recently isolated from human prostate cancer and peripheral blood mononuclear cells (PBMCs) of patients with chronic fatigue syndrome (CFS). We and others have shown that host restriction factors APOBEC3G (A3G) and APOBEC3F (A3F), which are expressed in human PBMCs, inhibit XMRV in transient transfection assays involving a single cycle of viral replication. However, the recovery of infectious XMRV from human PBMCs suggested that XMRV can replicate in these cells despite the expression of APOBEC3 proteins. To determine whether XMRV can replicate and spread in cultured PBMCs even though it can be inhibited by A3G/A3F, we infected phytohemagglutinin-activated human PBMCs and A3G/A3F-positive and -negative cell lines (CEM and CEM-SS, respectively) with different amounts of XMRV and monitored virus production using quantitative real-time PCR. We found that XMRV efficiently replicated in CEM-SS cells and viral production increased by >4000-fold, but there was only a modest increase in viral production from CEM cells (<14-fold) and a decrease in activated PBMCs, indicating little or no replication and spread of XMRV. However, infectious XMRV could be recovered from the infected PBMCs by cocultivation with a canine indicator cell line, and we observed hypermutation of XMRV genomes in PBMCs. Thus, PBMCs can potentially act as a source of infectious XMRV for spread to cells that express low levels of host restriction factors. Overall, these results suggest that hypermutation of XMRV in human PBMCs constitutes one of the blocks to replication and spread of XMRV. Furthermore, hypermutation of XMRV proviruses at GG dinucleotides may be a useful and reliable indicator of human PBMC infection.

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