Hepatitis C virus population dynamics in human lymphocytes and hepatocytes infected in vitro.
We previously found two cell lines (MT-2 and PH5CH) that were susceptible to hepatitis C virus (HCV) infection. Analysis of the infectivity of sera from HCV-positive blood donors for MT-2 and PH5CH cells suggested the cell tropism of HCV. To investigate further the cell tropism of HCV, the dynamics of HCV populations during culture were examined using three MT-2 clones and three PH5CH clones, infected with inoculum 1B-2. To type HCV populations in these infected cells, the HCV hypervariable region 1 (HVR1) in these cloned cells was characterized by sequence analysis and HpaII digestion analysis, which could distinguish three major HVR1 types (I, II and III) derived from the inoculum 1B-2. It was found that genomes containing HVR1 type I became predominant in MT-2 clones, and genomes containing HVR1 type II became predominant in PH5CH clones during culture after inoculation. These results suggest that inoculum 1B-2 contains both lymphotropic and hepatotropic HCV species, which can be distinguished by HVR1 type. To search for cell type-specific sequences in regions other than HVR1, three HCV cDNA clones (3.4 kb of the 5' noncoding region to the nonstructural 2 region) containing HVR1 type I obtained from HCV-infected MT-2C cells, and three HCV cDNA clones containing HVR1 type II obtained from HCV-infected PH5CH7 cells were sequenced. Following a comparison of the sequences, 11 amino acids were identified as candidates for determinants of the cell tropism of HCV.
Virology Division, National Cancer Center Research Institute, Tokyo, Japan. nkato@ncc.go.jp
N
N
Kato
M
M
Ikeda
K
K
Sugiyama
T
T
Mizutani
T
T
Tanaka
K
K
Shimotohno
D000595:Amino Acid Sequence; D002461:Cell Line, Transformed; D017931:DNA Primers; D019003:Deoxyribonuclease HpaII; D016174:Hepacivirus / Q000235:genetics / Q000502:physiology*; D006526:Hepatitis C / Q000097:blood / Q000821:virology*; D006801:Humans; D008099:Liver / Q000166:cytology / Q000821:virology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D012367:RNA, Viral; D013601:T-Lymphocytes / Q000821:virology*; D014759:Viral Envelope Proteins / Q000235:genetics; D017361:Viral Nonstructural Proteins / Q000235:genetics
D017931:DNA Primers; C087035:E1 protein, Hepatitis C virus; C097010:NS2 protein, Hepatitis C virus; D012367:RNA, Viral; D014759:Viral Envelope Proteins; D017361:Viral Nonstructural Proteins; C091015:glycoprotein E2, Hepatitis C virus; D019003:Deoxyribonuclease HpaII