Back to Search Start Over

Robust hepatitis C genotype 3a cell culture releasing adapted intergenotypic 3a/2a (S52/JFH1) viruses

Authors :
Gottwein, J.M.
Scheel, Troels Kasper Høyer
Hoegh, A.M.
Lademann, J.B.
Eugen-Olsen, J.
Lisby, G.
Bukh, J.
Gottwein, J.M.
Scheel, Troels Kasper Høyer
Hoegh, A.M.
Lademann, J.B.
Eugen-Olsen, J.
Lisby, G.
Bukh, J.
Source :
Gottwein , J M , Scheel , T K H , Hoegh , A M , Lademann , J B , Eugen-Olsen , J , Lisby , G & Bukh , J 2007 , ' Robust hepatitis C genotype 3a cell culture releasing adapted intergenotypic 3a/2a (S52/JFH1) viruses ' , Gastroenterology , vol. 133 , no. 5 , pp. 1614-26 .
Publication Year :
2007

Abstract

Udgivelsesdato: 2007-Nov<br />BACKGROUND & AIMS: Recently, full viral life cycle hepatitis C virus (HCV) cell culture systems were developed for strain JFH1 (genotype 2a) and an intragenotypic 2a/2a genome (J6/JFH). We aimed at exploiting the unique JFH1 replication characteristics to develop culture systems for genotype 3a, which has a high prevalence worldwide. METHODS: Huh7.5 cells were transfected with RNA transcripts of an intergenotypic 3a/JFH1 recombinant with core, E1, E2, p7, and NS2 of the 3a reference strain S52, and released viruses were passaged. Cultures were examined for HCV core and/or NS5A expression (immunostaining), HCV RNA titers (real-time PCR), and infectivity titers (50% tissue culture infectious dose). The role of mutations identified by sequencing of recovered S52/JFH1 viruses was analyzed by reverse genetics studies. RESULTS: S52/JFH1 and J6/JFH viruses passaged in Huh7.5 cells showed comparable growth kinetics and similar peak HCV RNA and infectivity titers. However, analysis of S52/JFH1 viruses identified 9 putative adaptive mutations in core, E2, p7, NS3, and NS5A. All 7 S52/JFH1 recombinants with an amino acid change in p7 combined with a change in NS3 or NS5A, but only 2 of 9 recombinants with individual mutations (in p7 and NS3, respectively) were fully viable without the requirement for additional mutations. The biological relevance of our system was shown by studying dependence of 3a/JFH1 infection on CD81, and its impact on distribution of intracellular lipids. CONCLUSIONS: We developed a robust intergenotypic recombinant cell culture system for HCV genotype 3a, providing a valuable tool for studies of 3a core-NS2 and related therapeutics.

Details

Database :
OAIster
Journal :
Gottwein , J M , Scheel , T K H , Hoegh , A M , Lademann , J B , Eugen-Olsen , J , Lisby , G & Bukh , J 2007 , ' Robust hepatitis C genotype 3a cell culture releasing adapted intergenotypic 3a/2a (S52/JFH1) viruses ' , Gastroenterology , vol. 133 , no. 5 , pp. 1614-26 .
Notes :
English
Publication Type :
Electronic Resource
Accession number :
edsoai.on1322548033
Document Type :
Electronic Resource