Human hepatocyte clonal cell lines that support persistent replication of hepatitis C virus

Masanori Ikeda, Kazuo Sugiyama, Tetsuya Mizutani, Torahiko Tanaka, Katsuaki Tanaka, Hisahiko Sekihara, Kunitada Shimotohno, Nobuyuki Kato

Research output: Contribution to journalArticle

93 Citations (Scopus)

Abstract

We previously found that a human T-cell leukemia virus type I infected T-cell line, MT-2, was susceptible to hepatitis C virus (HCV) infection, and that cloned MT-2C cells could support HCV replication more persistently than the parental MT-2 cells. Recently we found that a non-neoplastic hepatocyte line, PH5CH, showed good susceptibility to HCV infection. In this study, we cloned PH5CH cells to obtain cells that supported more persistent HCV replication, and consequently three clones (PH5CH1, PH5CH7 and PH5CH8) in which intracellular HCV RNA could be detected at least 25 days postinoculation (p.i.) were obtained. Semi-quantitative analysis of HCV RNA indicated that HCV replicated in these cloned PH5CH cells was released into the culture medium. Semi-quantitative analysis of internalized HCV RNA after treatment of cloned PH5CH cells and parental PH5CH cells with proteinase K immediately after virus inoculation revealed that PH5CH1, PH5CH7 and PH5CH8 cells contained 10-fold higher levels of HCV RNA than low susceptible cloned PH5CH or parental PH5CH cells. Furthermore, we demonstrated that HCV replication was maintained for 70-100 days in these three clonal lines when the temperature of cell culture after virus inoculation was reduced from 37 to 32°C. Moreover, we demonstrated that interferon α had antiviral effect on HCV-infected PH5CH8 cells. The three PH5CH clones obtained in this study will provide a useful tool for the study of HCV replication and proliferation, and for development of an assay system for antiviral agents. Copyright (C) 1998 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)157-167
Number of pages11
JournalVirus Research
Volume56
Issue number2
DOIs
Publication statusPublished - Aug 1998
Externally publishedYes

Fingerprint

Hepacivirus
Hepatocytes
Cell Line
Virus Replication
RNA
Virus Diseases
Antiviral Agents
Clone Cells
Viruses
Endopeptidase K
Human T-lymphotropic virus 1
Interferons
Culture Media
Cell Culture Techniques
T-Lymphocytes
Temperature

Keywords

  • HCV replication
  • Hepatitis C virus
  • Human hepatocytes
  • Persistent infection

ASJC Scopus subject areas

  • Cancer Research
  • Molecular Biology
  • Virology

Cite this

Human hepatocyte clonal cell lines that support persistent replication of hepatitis C virus. / Ikeda, Masanori; Sugiyama, Kazuo; Mizutani, Tetsuya; Tanaka, Torahiko; Tanaka, Katsuaki; Sekihara, Hisahiko; Shimotohno, Kunitada; Kato, Nobuyuki.

In: Virus Research, Vol. 56, No. 2, 08.1998, p. 157-167.

Research output: Contribution to journalArticle

Ikeda, M, Sugiyama, K, Mizutani, T, Tanaka, T, Tanaka, K, Sekihara, H, Shimotohno, K & Kato, N 1998, 'Human hepatocyte clonal cell lines that support persistent replication of hepatitis C virus', Virus Research, vol. 56, no. 2, pp. 157-167. https://doi.org/10.1016/S0168-1702(98)00063-X
Ikeda, Masanori ; Sugiyama, Kazuo ; Mizutani, Tetsuya ; Tanaka, Torahiko ; Tanaka, Katsuaki ; Sekihara, Hisahiko ; Shimotohno, Kunitada ; Kato, Nobuyuki. / Human hepatocyte clonal cell lines that support persistent replication of hepatitis C virus. In: Virus Research. 1998 ; Vol. 56, No. 2. pp. 157-167.
@article{36c0ae862fed428f94a78ffb8a41080d,
title = "Human hepatocyte clonal cell lines that support persistent replication of hepatitis C virus",
abstract = "We previously found that a human T-cell leukemia virus type I infected T-cell line, MT-2, was susceptible to hepatitis C virus (HCV) infection, and that cloned MT-2C cells could support HCV replication more persistently than the parental MT-2 cells. Recently we found that a non-neoplastic hepatocyte line, PH5CH, showed good susceptibility to HCV infection. In this study, we cloned PH5CH cells to obtain cells that supported more persistent HCV replication, and consequently three clones (PH5CH1, PH5CH7 and PH5CH8) in which intracellular HCV RNA could be detected at least 25 days postinoculation (p.i.) were obtained. Semi-quantitative analysis of HCV RNA indicated that HCV replicated in these cloned PH5CH cells was released into the culture medium. Semi-quantitative analysis of internalized HCV RNA after treatment of cloned PH5CH cells and parental PH5CH cells with proteinase K immediately after virus inoculation revealed that PH5CH1, PH5CH7 and PH5CH8 cells contained 10-fold higher levels of HCV RNA than low susceptible cloned PH5CH or parental PH5CH cells. Furthermore, we demonstrated that HCV replication was maintained for 70-100 days in these three clonal lines when the temperature of cell culture after virus inoculation was reduced from 37 to 32°C. Moreover, we demonstrated that interferon α had antiviral effect on HCV-infected PH5CH8 cells. The three PH5CH clones obtained in this study will provide a useful tool for the study of HCV replication and proliferation, and for development of an assay system for antiviral agents. Copyright (C) 1998 Elsevier Science B.V.",
keywords = "HCV replication, Hepatitis C virus, Human hepatocytes, Persistent infection",
author = "Masanori Ikeda and Kazuo Sugiyama and Tetsuya Mizutani and Torahiko Tanaka and Katsuaki Tanaka and Hisahiko Sekihara and Kunitada Shimotohno and Nobuyuki Kato",
year = "1998",
month = "8",
doi = "10.1016/S0168-1702(98)00063-X",
language = "English",
volume = "56",
pages = "157--167",
journal = "Virus Research",
issn = "0168-1702",
publisher = "Elsevier",
number = "2",

}

TY - JOUR

T1 - Human hepatocyte clonal cell lines that support persistent replication of hepatitis C virus

AU - Ikeda, Masanori

AU - Sugiyama, Kazuo

AU - Mizutani, Tetsuya

AU - Tanaka, Torahiko

AU - Tanaka, Katsuaki

AU - Sekihara, Hisahiko

AU - Shimotohno, Kunitada

AU - Kato, Nobuyuki

PY - 1998/8

Y1 - 1998/8

N2 - We previously found that a human T-cell leukemia virus type I infected T-cell line, MT-2, was susceptible to hepatitis C virus (HCV) infection, and that cloned MT-2C cells could support HCV replication more persistently than the parental MT-2 cells. Recently we found that a non-neoplastic hepatocyte line, PH5CH, showed good susceptibility to HCV infection. In this study, we cloned PH5CH cells to obtain cells that supported more persistent HCV replication, and consequently three clones (PH5CH1, PH5CH7 and PH5CH8) in which intracellular HCV RNA could be detected at least 25 days postinoculation (p.i.) were obtained. Semi-quantitative analysis of HCV RNA indicated that HCV replicated in these cloned PH5CH cells was released into the culture medium. Semi-quantitative analysis of internalized HCV RNA after treatment of cloned PH5CH cells and parental PH5CH cells with proteinase K immediately after virus inoculation revealed that PH5CH1, PH5CH7 and PH5CH8 cells contained 10-fold higher levels of HCV RNA than low susceptible cloned PH5CH or parental PH5CH cells. Furthermore, we demonstrated that HCV replication was maintained for 70-100 days in these three clonal lines when the temperature of cell culture after virus inoculation was reduced from 37 to 32°C. Moreover, we demonstrated that interferon α had antiviral effect on HCV-infected PH5CH8 cells. The three PH5CH clones obtained in this study will provide a useful tool for the study of HCV replication and proliferation, and for development of an assay system for antiviral agents. Copyright (C) 1998 Elsevier Science B.V.

AB - We previously found that a human T-cell leukemia virus type I infected T-cell line, MT-2, was susceptible to hepatitis C virus (HCV) infection, and that cloned MT-2C cells could support HCV replication more persistently than the parental MT-2 cells. Recently we found that a non-neoplastic hepatocyte line, PH5CH, showed good susceptibility to HCV infection. In this study, we cloned PH5CH cells to obtain cells that supported more persistent HCV replication, and consequently three clones (PH5CH1, PH5CH7 and PH5CH8) in which intracellular HCV RNA could be detected at least 25 days postinoculation (p.i.) were obtained. Semi-quantitative analysis of HCV RNA indicated that HCV replicated in these cloned PH5CH cells was released into the culture medium. Semi-quantitative analysis of internalized HCV RNA after treatment of cloned PH5CH cells and parental PH5CH cells with proteinase K immediately after virus inoculation revealed that PH5CH1, PH5CH7 and PH5CH8 cells contained 10-fold higher levels of HCV RNA than low susceptible cloned PH5CH or parental PH5CH cells. Furthermore, we demonstrated that HCV replication was maintained for 70-100 days in these three clonal lines when the temperature of cell culture after virus inoculation was reduced from 37 to 32°C. Moreover, we demonstrated that interferon α had antiviral effect on HCV-infected PH5CH8 cells. The three PH5CH clones obtained in this study will provide a useful tool for the study of HCV replication and proliferation, and for development of an assay system for antiviral agents. Copyright (C) 1998 Elsevier Science B.V.

KW - HCV replication

KW - Hepatitis C virus

KW - Human hepatocytes

KW - Persistent infection

UR - http://www.scopus.com/inward/record.url?scp=0032143649&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032143649&partnerID=8YFLogxK

U2 - 10.1016/S0168-1702(98)00063-X

DO - 10.1016/S0168-1702(98)00063-X

M3 - Article

C2 - 9783464

AN - SCOPUS:0032143649

VL - 56

SP - 157

EP - 167

JO - Virus Research

JF - Virus Research

SN - 0168-1702

IS - 2

ER -