Full-Length GB Virus C (Hepatitis G Virus) RNA Transcripts Are Infectious in Primary CD4-Positive T Cells

Abstract

GB virus C (GBV-C or hepatitis G virus) is a recently described flavivirus which frequently leads to chronic viremia in humans. Although GBV-C is associated with acute posttransfusion hepatitis, it is not clear if the virus is pathogenic for humans. We constructed a full-length cDNA from the plasma of a person with chronic GBV-C viremia. Peripheral blood mononuclear cells (PBMCs) transfected with full-length RNA transcripts from this GBV-C clone resulted in viral replication. This was demonstrated by serial passage of virus from cell culture supernatants, detection of increasing concentrations of positive-and negative-sense GBV-C RNA over time, and the detection of the GBV-C E2 antigen by confocal microscopy. In addition, two types of GBV-C particles were identified in cell lysates; these particles had buoyant densities of 1.06 and 1.12 to 1.17 g/ml in sucrose gradients. PBMCs sorted for expression of CD4 contained 100-fold-more GBV-C RNA than CD4-negative cells. Taken together, these data demonstrate that RNA transcripts from GBV-C full-length cDNA are infectious in primary CD4-positive T cells. In contrast, RNA transcripts from an infectious hepatitis C virus clone did not replicate in the same cell culture system. Infectious RNA transcripts from GBV-C cDNA should prove useful for studying viral replication and may allow identification of differences between GBV-C and hepatitis C virus cultivation in vitro.

Tropism of human pegivirus (formerly known as GB virus C/hepatitis G virus) and host immunomodulation: insights into a highly successful viral infection

“…As noted above, early studies proposed that the virus was hepatotropic; however, several lines of evidence suggest that the virus is lymphotropic (Fogeda et al, 1999;George et al, 2006;Xiang et al, 2000Xiang et al, , 2001. Specifically, PBMCs from HPgVinfected individuals maintained in culture ex vivo release virus into culture media and virus production can persist for at least 35 days in culture (Fogeda et al, 1999;George et al, 2003;Rydze et al, 2012).…”

“…Specifically, PBMCs from HPgVinfected individuals maintained in culture ex vivo release virus into culture media and virus production can persist for at least 35 days in culture (Fogeda et al, 1999;George et al, 2003;Rydze et al, 2012). Furthermore, serumderived virus from infected individuals is capable of infecting PBMCs in vitro (Chivero et al, 2014;Xiang et al, 2000).…”

“…Specifically, PBMCs from HPgVinfected individuals maintained in culture ex vivo release virus into culture media and virus production can persist for at least 35 days in culture (Fogeda et al, 1999;George et al, 2003;Rydze et al, 2012). Furthermore, serumderived virus from infected individuals is capable of infecting PBMCs in vitro (Chivero et al, 2014;Xiang et al, 2000).In cell culture, HPgV replication is at best intermittent and produces low titres of intracellular RNA or virus released into cell culture media. The best-characterized in vitro methods described to date for HPgV replication utilize primary PBMCs (Fogeda et al, 1999;George et al, 2003George et al, , 2006Rydze et al, 2012;Xiang et al, 2000).…”

“…Furthermore, serumderived virus from infected individuals is capable of infecting PBMCs in vitro (Chivero et al, 2014;Xiang et al, 2000).…”

Primary screening of blood donors by nat testing for HCV-RNA: development of an "in-house" method and results

“…However, if evaluating rejection due to the initially reactive (pools)/ repeatedly reactive (individual sample) analysis, the specificity is > 99.9% since only five samples displayed this pattern and were further non reactive in the confirmatory assay. These five NAT false-positives amplicons were sequenced, revealing the presence of an HGV "Iowan" strain 38 in four of them. Those four were also reactive for two distinct HGV-specific molecular assays, confirming their status of HGV carriers.…”