De novo infection and propagation of wild-type Hepatitis C virus in human T lymphocytes in vitro.
While exploring previous findings that ex vivo treatment of lymphoid cells from Hepatitis C virus (HCV)-infected individuals with T cell-stimulating mitogens augments detection of the residing virus, an in vitro HCV replication system was established, in which mitogen-induced T cell-enriched cultures served as HCV targets and the derived T cells multiplied virus during repeated serial passage. HCV replication was ascertained by detecting HCV RNA positive and negative strands, HCV NS5a and E2 proteins, release of HCV virions and nucleocapsids (confirmed by immunoelectron microscopy) and de novo infection of mitogen-induced T cells prepared from healthy donors. Further, affinity-purified normal human T lymphocytes were also susceptible to HCV infection in vitro and HCV replication was detected in pure T cells isolated from a patient with chronic hepatitis C. These results document that T cells can support propagation of HCV both in vivo and in vitro. The infection system established offers a valuable tool for in vitro studies on the entire cycle of HCV replication, virus cytopathogenicity and evaluation of antiviral agents against wild-type HCV in the natural host-cell milieu.
MacParland
Sonya A
SA
Molecular Virology and Hepatology Research, Division of Basic Medical Sciences, Faculty of Medicine, Health Sciences Centre, Memorial University, St John's, NL A1B 3V6, Canada.
Pham
Tram N Q
TNQ
Molecular Virology and Hepatology Research, Division of Basic Medical Sciences, Faculty of Medicine, Health Sciences Centre, Memorial University, St John's, NL A1B 3V6, Canada.
Gujar
Shashi A
SA
Molecular Virology and Hepatology Research, Division of Basic Medical Sciences, Faculty of Medicine, Health Sciences Centre, Memorial University, St John's, NL A1B 3V6, Canada.
Michalak
Tomasz I
TI
Discipline of Laboratory Medicine, Faculty of Medicine, Health Sciences Centre, Memorial University, St John's, NL A1B 3V6, Canada.|Molecular Virology and Hepatology Research, Division of Basic Medical Sciences, Faculty of Medicine, Health Sciences Centre, Memorial University, St John's, NL A1B 3V6, Canada.
D002478:Cells, Cultured; D016174:Hepacivirus / Q000235:genetics / Q000502:physiology*; D006801:Humans; D046529:Microscopy, Electron, Transmission; D016253:Microscopy, Immunoelectron; D012327:RNA Virus Infections; D012367:RNA, Viral / Q000032:analysis; D013601:T-Lymphocytes / Q000821:virology*; D014759:Viral Envelope Proteins / Q000032:analysis; D017361:Viral Nonstructural Proteins / Q000032:analysis; D014771:Virion / Q000648:ultrastructure; D014776:Virus Cultivation; D014779:Virus Replication; D012324:RNA-Dependent RNA Polymerase
D012367:RNA, Viral; D014759:Viral Envelope Proteins; D017361:Viral Nonstructural Proteins; C091015:glycoprotein E2, Hepatitis C virus; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase