Advantages of a single-cycle production assay to study cell culture-adaptive mutations of hepatitis C virus.

Rodney S Russell; Jean-Christophe Meunier; Shingo Takikawa; Kristina Faulk; Ronald E Engle; Jens Bukh; Robert H Purcell; Suzanne U Emerson

doi:10.1073/pnas.0800422105

Advantages of a single-cycle production assay to study cell culture-adaptive mutations of hepatitis C virus.

Rodney S Russell, Jean-Christophe Meunier, Shingo Takikawa, Kristina Faulk, Ronald E Engle, Jens Bukh, Robert H Purcell, Suzanne U Emerson

153 Citationer (Scopus)

Abstract

The JFH1 strain of hepatitis C virus (HCV) is unique among HCV isolates, in that the wild-type virus can traverse the entire replication cycle in cultured cells. However, without adaptive mutations, only low levels of infectious virus are produced. In the present study, the effects of five mutations that were selected during serial passage in Huh-7.5 cells were studied. Recombinant genomes containing all five mutations produced 3-4 logs more infectious virions than did wild type. Neither a coding mutation in NS5A nor a silent mutation in E2 was adaptive, whereas coding mutations in E2, p7, and NS2 all increased virus production. A single-cycle replication assay in CD81-deficient cells was developed to study more precisely the effect of the adaptive mutations. The E2 mutation had minimal effect on the amount of infectious virus released but probably enhanced entry into cells. In contrast, both the p7 and NS2 mutations independently increased the amount of virus released.
Udgivelsesdato: 2008-Mar-18

Originalsprog	Engelsk
Tidsskrift	Proceedings of the National Academy of Sciences of the United States of America
Vol/bind	105
Udgave nummer	11
Sider (fra-til)	4370-5
Antal sider	5
ISSN	0027-8424
DOI	https://doi.org/10.1073/pnas.0800422105
Status	Udgivet - 2008

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10.1073/pnas.0800422105

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Advantages of a single-cycle production assay to study cell culture-adaptive mutations of hepatitis C virus. / Russell, Rodney S; Meunier, Jean-Christophe; Takikawa, Shingo et al.
I: Proceedings of the National Academy of Sciences of the United States of America, Bind 105, Nr. 11, 2008, s. 4370-5.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

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abstract = "The JFH1 strain of hepatitis C virus (HCV) is unique among HCV isolates, in that the wild-type virus can traverse the entire replication cycle in cultured cells. However, without adaptive mutations, only low levels of infectious virus are produced. In the present study, the effects of five mutations that were selected during serial passage in Huh-7.5 cells were studied. Recombinant genomes containing all five mutations produced 3-4 logs more infectious virions than did wild type. Neither a coding mutation in NS5A nor a silent mutation in E2 was adaptive, whereas coding mutations in E2, p7, and NS2 all increased virus production. A single-cycle replication assay in CD81-deficient cells was developed to study more precisely the effect of the adaptive mutations. The E2 mutation had minimal effect on the amount of infectious virus released but probably enhanced entry into cells. In contrast, both the p7 and NS2 mutations independently increased the amount of virus released.",

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AU - Russell, Rodney S

AU - Meunier, Jean-Christophe

AU - Takikawa, Shingo

AU - Faulk, Kristina

AU - Engle, Ronald E

AU - Bukh, Jens

AU - Purcell, Robert H

AU - Emerson, Suzanne U

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N2 - The JFH1 strain of hepatitis C virus (HCV) is unique among HCV isolates, in that the wild-type virus can traverse the entire replication cycle in cultured cells. However, without adaptive mutations, only low levels of infectious virus are produced. In the present study, the effects of five mutations that were selected during serial passage in Huh-7.5 cells were studied. Recombinant genomes containing all five mutations produced 3-4 logs more infectious virions than did wild type. Neither a coding mutation in NS5A nor a silent mutation in E2 was adaptive, whereas coding mutations in E2, p7, and NS2 all increased virus production. A single-cycle replication assay in CD81-deficient cells was developed to study more precisely the effect of the adaptive mutations. The E2 mutation had minimal effect on the amount of infectious virus released but probably enhanced entry into cells. In contrast, both the p7 and NS2 mutations independently increased the amount of virus released.

AB - The JFH1 strain of hepatitis C virus (HCV) is unique among HCV isolates, in that the wild-type virus can traverse the entire replication cycle in cultured cells. However, without adaptive mutations, only low levels of infectious virus are produced. In the present study, the effects of five mutations that were selected during serial passage in Huh-7.5 cells were studied. Recombinant genomes containing all five mutations produced 3-4 logs more infectious virions than did wild type. Neither a coding mutation in NS5A nor a silent mutation in E2 was adaptive, whereas coding mutations in E2, p7, and NS2 all increased virus production. A single-cycle replication assay in CD81-deficient cells was developed to study more precisely the effect of the adaptive mutations. The E2 mutation had minimal effect on the amount of infectious virus released but probably enhanced entry into cells. In contrast, both the p7 and NS2 mutations independently increased the amount of virus released.

KW - Adaptation, Biological

KW - Cell Culture Techniques

KW - Cell Line, Tumor

KW - Hepacivirus

KW - Humans

KW - Mutation

KW - Virus Replication

U2 - 10.1073/pnas.0800422105

DO - 10.1073/pnas.0800422105

M3 - Journal article

C2 - 18334634

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EP - 4375

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 11

ER -

Advantages of a single-cycle production assay to study cell culture-adaptive mutations of hepatitis C virus.

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