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

Asako Murayama,Takanobu Kato,Takaji Wakita,Nao Sugiyama,Diane E Griffin,Peter Sarnow

doi:10.1128/mbio.00273-16

Asako Murayama, Takanobu Kato + Show 4 more

Open Access

https://doi.org/10.1128/mbio.00273-16

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Journal: mBio	Publication Date: Jun 14, 2016
Citations: 17	License type: CC BY 4.0

Affiliation: National Institute of Infectious Diseases

Abstract

ABSTRACTA hepatitis C virus (HCV) cell culture system incorporating the JFH-1 strain and the human hepatoma cell line HuH-7 enabled the production of infectious HCV particles. Several host factors were identified as essential for HCV replication. Supplementation of these factors in nonhepatic human cell lines enabled HCV replication and particle production. Vero cells established from monkey kidney are commonly used for the production of vaccines against a variety of viruses. In this study, we aimed to establish a novel Vero cell line to reconstruct the HCV life cycle. Unmodified Vero cells did not allow HCV infection or replication. The expression of microRNA 122 (miR-122), an essential factor for HCV replication, is notably low in Vero cells. Therefore, we supplemented Vero cells with miR-122 and found that HCV replication was enhanced. However, Vero cells that expressed miR-122 still did not allow HCV infection. We supplemented HCV receptor molecules and found that scavenger receptor class B type I (SRBI) was essential for HCV infection in Vero cells. The supplementation of apolipoprotein E (ApoE), a host factor important for virus production, enabled the production of infectious virus in Vero cells. Finally, we created a Vero cell line that expressed the essential factors miR-122, SRBI, and ApoE; the entire HCV life cycle, including infection, replication, and infectious virus production, was completed in these cells. In conclusion, we demonstrated that miR-122, SRBI, and ApoE were necessary and sufficient for the completion of the entire HCV life cycle in nonhuman, nonhepatic Vero cells.

Highlights

A hepatitis C virus (HCV) cell culture system incorporating the JFH-1 strain and the human hepatoma cell line HuH-7 enabled the production of infectious HCV particles
Many host factors associated with the HCV life cycle have been identified, and some of them were considered the essential factors for HCV infection, replication, and virus production in hepatocytes [14,15,16,17]
We found four polymorphisms in the CD81 open reading frame (ORF) (Fig. 2D), all of which were located in the large extracellular loop (LEL) region (Fig. 2E), which has been reported to be important for the association with HCV E2 protein [21]

Summary

Introduction

A hepatitis C virus (HCV) cell culture system incorporating the JFH-1 strain and the human hepatoma cell line HuH-7 enabled the production of infectious HCV particles. We created a Vero cell line that expressed the essential factors miR-122, SRBI, and ApoE; the entire HCV life cycle, including infection, replication, and infectious virus production, was completed in these cells. We identified the minimum essential host factors for the completion of the entire HCV life cycle in Vero cells to develop a novel HCV cell culture system. FU97 cells, derived from stomach cancer, and originally expressing essential host factors for HCV life cycle at levels comparable to those of HuH-7 cells, support HCV replication after HCV infection [18] These observations indicate the possibility that any. We identified the host factors in Vero cells essential for the completion of the entire HCV life cycle and developed a novel HCV cell culture system

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