A Subclone of HuH-7 with Enhanced Intracellular Hepatitis C Virus Production and Evasion of Virus Related-cell Cycle Arrest

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2013 Jan 4

PMID 23285155

Citations 9

Authors

Asako Murayama

Nao Sugiyama

Seiko Yoshimura

Mitsuko Ishihara-Sugano

Takahiro Masaki

Sulyi Kim

Takaji Wakita

Shunji Mishiro

Takanobu Kato

Affiliations

Soon will be listed here.

Abstract

Hepatitis C virus (HCV) cell culture system with JFH-1 strain and HuH-7 cells enabled us to produce infectious HCV particles in vitro, and such system is useful to explore the anti-HCV compounds and to develop the vaccine against HCV. In the present study, we describe the derivation of a cell line that permits improved production of HCV particles. Specifically, we characterized several subclones that were isolated from the original HuH-7 cell line by limiting dilution. These HuH-7 subclones displayed a notable range of HCV production levels following transfection by full-genome JFH-1 RNA. Among these subclones, HuH-7T1 produced HCV more efficiently than other subclones and Huh-7.5.1 that is known to be highly permissive for HCV replication. Upon transfection with full-genome RNA, HCV production was increased ten-fold in HuH-7T1 compared to Huh-7.5.1. This increase in viral production correlated with increased efficiency of intracellular infectious virus production. Furthermore, HCV replication did not induce cell cycle arrest in HuH-7T1, whereas it did in Huh-7.5.1. Consequently, the use of HuH-7T1 as host cells could provide increased population of HCV-positive cells and elevated viral titer. In conclusion, we isolated a HuH-7 subclone, HuH-7T1, that supports efficient HCV production. High efficiency of intracellular infectious virus production and evasion of cell cycle arrest were important for this phenotype. We expect that the use of this cell line will facilitate analysis of the underlying mechanisms for HCV particle assembly and the cell cycle arrest caused by HCV.

Citing Articles

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IFITM3 Interacts with the HBV/HDV Receptor NTCP and Modulates Virus Entry and Infection.

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Identification of Characteristic Genomic Markers in Human Hepatoma HuH-7 and Huh7.5.1-8 Cell Lines.

Kawamoto M, Yamaji T, Saito K, Shirasago Y, Satomura K, Endo T Front Genet. 2020; 11:546106.

PMID: 33193621 PMC: 7581915. DOI: 10.3389/fgene.2020.546106.

Interferon sensitivity-determining region of hepatitis C virus influences virus production and interferon signaling.

Sugiyama R, Murayama A, Nitta S, Yamada N, Tasaka-Fujita M, Masaki T Oncotarget. 2018; 9(5):5627-5640.

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Completion of the Entire Hepatitis C Virus Life Cycle in Vero Cells Derived from Monkey Kidney.

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