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

ABSTRACT 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. Several host factors were identified as essential for HCV replication. Supplementation of these factors in nonhepatic human...

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Autores principales:	Asako Murayama, Nao Sugiyama, Takaji Wakita, Takanobu Kato
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2016
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/c691eb3b6159465b98ee994defa015ab
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spelling	oai:doaj.org-article:c691eb3b6159465b98ee994defa015ab2021-11-15T15:50:16ZCompletion of the Entire Hepatitis C Virus Life Cycle in Vero Cells Derived from Monkey Kidney10.1128/mBio.00273-162150-7511https://doaj.org/article/c691eb3b6159465b98ee994defa015ab2016-07-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00273-16https://doaj.org/toc/2150-7511ABSTRACT 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. 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. IMPORTANCE HCV is a major cause of chronic liver diseases worldwide, and an effective prophylactic HCV vaccine is needed. For safety reasons, the current HCV cell culture system using HuH-7 cells, which was established from a hepatocellular carcinoma, is not suitable for the production of a vaccine against HCV. A robust HCV production system using non-cancer-derived cells is indispensable for this purpose. In this study, we wanted to establish a novel HCV cell culture system using Vero cells, which are widely used in the production of vaccines against different viruses. 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. A cell culture system that uses Vero cells will be useful not only for HCV vaccine production but also for the further elucidation of the mechanisms of various HCV-host interactions.Asako MurayamaNao SugiyamaTakaji WakitaTakanobu KatoAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 7, Iss 3 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Asako Murayama Nao Sugiyama Takaji Wakita Takanobu Kato Completion of the Entire Hepatitis C Virus Life Cycle in Vero Cells Derived from Monkey Kidney
description	ABSTRACT 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. 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. IMPORTANCE HCV is a major cause of chronic liver diseases worldwide, and an effective prophylactic HCV vaccine is needed. For safety reasons, the current HCV cell culture system using HuH-7 cells, which was established from a hepatocellular carcinoma, is not suitable for the production of a vaccine against HCV. A robust HCV production system using non-cancer-derived cells is indispensable for this purpose. In this study, we wanted to establish a novel HCV cell culture system using Vero cells, which are widely used in the production of vaccines against different viruses. 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. A cell culture system that uses Vero cells will be useful not only for HCV vaccine production but also for the further elucidation of the mechanisms of various HCV-host interactions.
format	article
author	Asako Murayama Nao Sugiyama Takaji Wakita Takanobu Kato
author_facet	Asako Murayama Nao Sugiyama Takaji Wakita Takanobu Kato
author_sort	Asako Murayama
title	Completion of the Entire Hepatitis C Virus Life Cycle in Vero Cells Derived from Monkey Kidney
title_short	Completion of the Entire Hepatitis C Virus Life Cycle in Vero Cells Derived from Monkey Kidney
title_full	Completion of the Entire Hepatitis C Virus Life Cycle in Vero Cells Derived from Monkey Kidney
title_fullStr	Completion of the Entire Hepatitis C Virus Life Cycle in Vero Cells Derived from Monkey Kidney
title_full_unstemmed	Completion of the Entire Hepatitis C Virus Life Cycle in Vero Cells Derived from Monkey Kidney
title_sort	completion of the entire hepatitis c virus life cycle in vero cells derived from monkey kidney
publisher	American Society for Microbiology
publishDate	2016
url	https://doaj.org/article/c691eb3b6159465b98ee994defa015ab
work_keys_str_mv	AT asakomurayama completionoftheentirehepatitiscviruslifecycleinverocellsderivedfrommonkeykidney AT naosugiyama completionoftheentirehepatitiscviruslifecycleinverocellsderivedfrommonkeykidney AT takajiwakita completionoftheentirehepatitiscviruslifecycleinverocellsderivedfrommonkeykidney AT takanobukato completionoftheentirehepatitiscviruslifecycleinverocellsderivedfrommonkeykidney
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Completion of the Entire Hepatitis C Virus Life Cycle in Vero Cells Derived from Monkey Kidney

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