De novo infection and propagation of wild-type Hepatitis C virus in human T lymphocytes in vitro

1 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 2 Discipline of Laboratory Medicine, Faculty of Medicine, Health Sciences Centre, Memorial University, St John's...

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Published in:Journal of general virology 2006-12, Vol.87 (12), p.3577-3586
Main Authors: MacParland, Sonya A, Pham, Tram N. Q, Gujar, Shashi A, Michalak, Tomasz I
Format: Article
Language:English
Subjects:
Biological and medical sciences
Cells, Cultured
Fundamental and applied biological sciences. Psychology
Hepacivirus - genetics
Hepacivirus - physiology
Hepatitis C virus
Humans
Microbiology
Microscopy, Electron, Transmission
Microscopy, Immunoelectron
Miscellaneous
RNA Virus Infections
RNA, Viral - analysis
T-Lymphocytes - virology
Viral Envelope Proteins - analysis
Viral Nonstructural Proteins - analysis
Virion - ultrastructure
Virology
Virus Cultivation
Virus Replication
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Summary:1 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 2 Discipline of Laboratory Medicine, Faculty of Medicine, Health Sciences Centre, Memorial University, St John's, NL A1B 3V6, Canada Correspondence Tomasz I. Michalak timich{at}mun.ca 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. A supplementary figure is available in JGV Online.
ISSN:0022-1317
1465-2099
DOI:10.1099/vir.0.81868-0