title: Persistence of hepatitis C virus in a human megakaryoblastic leukaemia cell line.
abstract: Thrombocytopenia is a frequent clinical finding in patients with hepatitis C virus (HCV) infection. Platelets from patients with HCV infection have been identified as carriers of HCV RNA in our previous studies. The present study was designed to further investigate the possibility of HCV replication in megakaryoblasts from which platelets are eventually released. A megakaryoblastic cell line (MEG-01), established from a chronic myelogenous leukaemia patient 13 years ago, was used for this study. The MEG-01 cells were inoculated with fresh serum from a patient with HCV infection and renamed MEG-01-I cells. Surprisingly, both MEG-01 and MEG-01-I were positive by HCV reverse transcription-polymerase chain reaction (RT-PCR) for the existence of HCV RNA and minus-strand HCV RNA, regardless of inoculation. This was further confirmed by in situ RT-PCR. The HCV antigens, such as core, envelope, and non-structural (NS)3 and NS4, were also present in both cell lines, as identified by Western blotting and indirect immunofluorescence staining. In addition, virus-like particles were observed by electron microscopy in the MEG-01 cell line as well as in the MEG-01-I cell line. These findings indicate that the megakaryoblasts are vulnerable to HCV infection and that replication of HCV can occur in these cells. This may help us to better understand the pathogenesis of thrombocytopenia in patients with HCV infection. The MEG-01 cell line, which may have been continuously shedding HCV for years, should be a useful model for experimental research into HCV.
authors.affiliation: Center for Liver Diseases, University of Miami School of Medicine and Veterans Administration Medical Center, Miami, FL 33125, USA.
authors.forename: X
authors.initials: X
authors.lastname: Li
authors.forename: L J
authors.initials: LJ
authors.lastname: Jeffers
authors.forename: C
authors.initials: C
authors.lastname: Garon
authors.forename: E R
authors.initials: ER
authors.lastname: Fischer
authors.forename: J
authors.initials: J
authors.lastname: Scheffel
authors.forename: B
authors.initials: B
authors.lastname: Moore
authors.forename: K R
authors.initials: KR
authors.lastname: Reddy
authors.forename: M
authors.initials: M
authors.lastname: Demedina
authors.forename: E R
authors.initials: ER
authors.lastname: Schiff
mesh_terms: D005455:Fluorescent Antibody Technique; D016174:Hepacivirus / Q000502:physiology*; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive; D008533:Megakaryocytes / Q000821:virology*; D018613:Microscopy, Confocal; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured; D014779:Virus Replication
chemical_list: D012367:RNA, Viral