Development of hepatitis C virus production reporter-assay systems using two different hepatoma cell lines

Midori Takeda, Masanori Ikeda, Yasuo Ariumi, Takaji Wakita, Nobuyuki Kato

Research output: Contribution to journalArticle

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Abstract

A hepatitis C virus (HCV) infection system was developed previously using the HCV JFH-1 strain (genotype 2a) and HuH-7 cells, and this cell culture is so far the only robust production system for HCV. In patients with chronic hepatitis C, the virological effects of pegylated interferon and ribavirin therapy differ depending on the HCV strain and the genetic background of the host. Recently, we reported the hepatoma-derived Li23 cell line, in which the JFH-1 life cycle is reproduced at a level almost equal to that in HuH-7-derived RSc cells. To monitor the HCV life cycle more easily, we here developed JFH-1 reporter-assay systems using both HuH-7- and Li23- derived cell lines. To identify any genetic mutations by long-term cell culture, HCV RNAs in HuH-7 cells were amplified 130 days after infection and subjected to sequence analysis to find adaptive mutation(s) for robust virus replication. We identified two mutations, H2505Q and V2995L, in the NS5B region. V2995L but not H2505Q enhanced JFH-1 RNA replication. However, we found that H2505Q but not V2995L enhanced HCV RNA replication of strain O (genotype 1b). We also selected highly permissive D7 cells by serial subcloning of Li23 cells. The expression levels of claudin-1 and Niemann-Pick C1-like 1 in D7 cells are higher than those in parental Li23 cells. In this study, we developed HCV JFH-1 reporter-assay systems using two distinct hepatoma cell lines, HuH-7 and Li23. The mutations in NS5B resulted in different effects on strains O and JFH-1 HCV RNA replication.

Original languageEnglish
Pages (from-to)1422-1431
Number of pages10
JournalJournal of General Virology
Volume93
Issue numberPART 7
DOIs
Publication statusPublished - Jul 2012

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Hepacivirus
Hepatocellular Carcinoma
Cell Line
Virus Replication
RNA
Mutation
Life Cycle Stages
Cell Culture Techniques
Genotype
Claudin-1
Ribavirin
Chronic Hepatitis C
Virus Diseases
Interferons
Sequence Analysis
Infection

ASJC Scopus subject areas

  • Virology

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Development of hepatitis C virus production reporter-assay systems using two different hepatoma cell lines. / Takeda, Midori; Ikeda, Masanori; Ariumi, Yasuo; Wakita, Takaji; Kato, Nobuyuki.

In: Journal of General Virology, Vol. 93, No. PART 7, 07.2012, p. 1422-1431.

Research output: Contribution to journalArticle

Takeda, Midori ; Ikeda, Masanori ; Ariumi, Yasuo ; Wakita, Takaji ; Kato, Nobuyuki. / Development of hepatitis C virus production reporter-assay systems using two different hepatoma cell lines. In: Journal of General Virology. 2012 ; Vol. 93, No. PART 7. pp. 1422-1431.
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