Isolation and characterization of interferon lambda-resistant hepatitis C virus replicon cell lines

While hepatocytes are the major site of hepatitis C virus (HCV) infection, a number of studies have suggested that HCV can replicate in lymphocytes. However, in vitro culture systems to investigate replication of HCV in lymphocytic cells are severely limited. Robust HCV culture systems have been established using the HCV JFH-1 strain and Huh-7 cells. To gain more insights into the tissue tropism of HCV, we investigated the infection, replication, internal ribosome entry site (IRES)-dependent translation and polyprotein processing of the HCV JFH-1 strain in nine lymphocytic cell lines. HCV JFH-1 failed to infect lymphocytes and replicate, but exhibited efficient polyprotein processing and IRES-dependent translation in lymphocytes as well as in Huh-7 cells. Our results suggest that lymphocytic cells can support HCV JFH-1 translation and polyprotein processing, but may lack some host factors essential for HCV JFH-1 infection and replication.

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Isolation and characterization of hepatitis C virus resistant to a novel phenanthridinone derivative

Antiviral Chemistry and Chemotherapy ◽

10.1177/2040206616663956 ◽

2015 ◽

Vol 24 (5-6) ◽

pp. 148-154 ◽

Cited By ~ 1

Author(s):

Wataru Ito ◽

Masaaki Toyama ◽

Mika Okamoto ◽

Masanori Ikeda ◽

Koichi Watashi ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Isolation And Characterization

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Screening of Hepatitis C Virus Inhibitors Using Genotype 1a HCV Replicon Cell Lines

Current Protocols in Microbiology ◽

10.1002/9780471729259.mc1707s22 ◽

2011 ◽

Vol 22 (1) ◽

Cited By ~ 2

Author(s):

Margaret Robinson ◽

Yang Tian ◽

Nikos Pagratis ◽

William E. Delaney

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Cell Lines ◽

Replicon Cell ◽

Genotype 1A

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Ribavirin Resistance in Hepatitis C Virus Replicon-Containing Cell Lines Conferred by Changes in the Cell Line or Mutations in the Replicon RNA

Journal of Virology ◽

10.1128/jvi.79.4.2346-2355.2005 ◽

2005 ◽

Vol 79 (4) ◽

pp. 2346-2355 ◽

Cited By ~ 68

Author(s):

Julie K. Pfeiffer ◽

Karla Kirkegaard

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Cell Line ◽

Cell Lines ◽

Cell Types ◽

Resistant Cell ◽

Resistant Isolate ◽

Hcv Rna ◽

Replicon Cell ◽

Error Accumulation

ABSTRACT Ribavirin (RBV), used in combination with alpha interferon to treat hepatitis C virus (HCV) infections, is a guanosine nucleotide analog that can increase the error rate of viral RNA-dependent RNA polymerases, imbalance intracellular nucleotide pools, and cause toxicity in many cell types. To determine potential mechanisms of RBV resistance during HCV RNA replication, we passaged HCV replicon-containing cell lines in the presence of increasing concentrations of RBV. RBV-resistant, HCV replicon-containing cell lines were generated, and the majority of RBV resistance was found to be conferred by changes in the cell lines. The resistant cell lines were defective in RBV import, as measured by [3H]RBV uptake experiments. These cell lines displayed reduced RBV toxicity and reduced error accumulation during infection with poliovirus, whose replication is known to be sensitive to RBV-induced error. For one RBV-resistant isolate, two mutations in the replicon RNA contributed to the observed phenotype. Two responsible mutations resided in the C-terminal region of NS5A, G404S, and E442G and were each sufficient for low-level RBV resistance. Therefore, RBV resistance in HCV replicon cell lines can be conferred by changes in the cell line or by mutations in the HCV replicon.

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Establishment of hepatitis C virus replicon cell lines possessing interferon-resistant phenotype

Biochemical and Biophysical Research Communications ◽

10.1016/j.bbrc.2004.08.091 ◽

2004 ◽

Vol 323 (1) ◽

pp. 299-309 ◽

Cited By ~ 26

Author(s):

Katsuyuki Namba ◽

Kazuhito Naka ◽

Hiromichi Dansako ◽

Akito Nozaki ◽

Masanori Ikeda ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Cell Lines ◽

Replicon Cell ◽

Resistant Phenotype

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Tagging of NS5A expressed from a functional hepatitis C virus replicon

Journal of General Virology ◽

10.1099/vir.0.81553-0 ◽

2006 ◽

Vol 87 (3) ◽

pp. 635-640 ◽

Cited By ~ 19

Author(s):

Christopher J. McCormick ◽

Sophie Maucourant ◽

Stephen Griffin ◽

David J. Rowlands ◽

Mark Harris

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Cell Lines ◽

Magnetic Beads ◽

Fluorescent Protein ◽

Coding Region ◽

Replicon Cell ◽

Replication Complex ◽

Propionibacterium Shermanii ◽

Green Fluorescent

Knowledge of how hepatitis C virus (HCV) proteins associate with components of the host cell to form a functional replication complex is still limited. To address this issue, HCV replicon constructs were generated where either green fluorescent protein (GFP) or the Propionibacterium shermanii transcarboxylase domain (PSTCD) was introduced into the NS5A coding region. Insertion of both GFP and PSTCD was tolerated well, allowing formation of stable replicon-containing cell lines that contained viral protein and transcript levels that were comparable to those of an unmodified parental replicon. Cell lines generated from the GFP-tagged NS5A replicon allowed live-cell visualization of the location of NS5A. Cell lines generated from the PSTCD-tagged replicons allowed rapid and efficient precipitation of the PSTCD-tagged NS5A, as well as other HCV non-structural proteins, using streptavidin-coated magnetic beads. Both replicons represent useful tools that offer different but complementary ways of examining replication-complex formation in cells.

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