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

TY - JOUR

T1 - Advantages of a single-cycle production assay to study cell culture-adaptive mutations of hepatitis C virus

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

PY - 2008/3/18

Y1 - 2008/3/18

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

SN - 0027-8424

VL - 105

SP - 4370

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 -