Hepatitis C virus molecular clones: from cDNA to infectious virus particles in cell culture

2006 ◽  
Vol 9 (4) ◽  
pp. 416-422 ◽  
Author(s):  
Ralf Bartenschlager
Keyword(s):  
Cell Culture ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Infectious Virus ◽  
Virus Particles

scholarly journals Compensatory Mutations in E1, p7, NS2, and NS3 Enhance Yields of Cell Culture-Infectious Intergenotypic Chimeric Hepatitis C Virus

2006 ◽  
Vol 81 (2) ◽  
pp. 629-638 ◽  
Author(s):  
MinKyung Yi ◽  
Yinghong Ma ◽  
Jeremy Yates ◽  
Stanley M. Lemon
Keyword(s):  
Cell Culture ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Particle Production ◽  
Infectious Virus ◽  
Cultured Cells ◽  
Virus Assembly ◽  
Infectious Hepatitis ◽  
Adaptive Mutations ◽  
Compensatory Mutations

ABSTRACT There is little understanding of mechanisms underlying the assembly and release of infectious hepatitis C virus (HCV) from cultured cells. Cells transfected with synthetic genomic RNA from a unique genotype 2a virus (JFH1) produce high titers of virus, while virus yields are much lower with a prototype genotype 1a RNA containing multiple cell culture-adaptive mutations (H77S). To characterize the basis for this difference in infectious particle production, we constructed chimeric genomes encoding the structural proteins of H77S within the background of JFH1. RNAs encoding polyproteins fused at the NS2/NS3 junction (“H-NS2/NS3-J”) and at a site of natural, intergenotypic recombination within NS2 [“H-(NS2)-J”] produced infectious virus. In contrast, no virus was produced by a chimera fused at the p7-NS2 junction. Chimera H-NS2/NS3-J virus (vH-NS2/NS3-J) recovered from transfected cultures contained compensatory mutations in E1 and NS3 that were essential for the production of infectious virus, while yields of infectious vH-(NS2)-J were enhanced by mutations within p7 and NS2. These compensatory mutations were chimera specific and did not enhance viral RNA replication or polyprotein processing; thus, they likely compensate for incompatibilities between proteins of different genotypes at sites of interactions essential for virus assembly and/or release. Mutations in p7 and NS2 acted additively and increased the specific infectivity of vH-(NS2)-J particles, while having less impact on the numbers of particles released. We conclude that interactions between NS2 and E1 and p7 as well as between NS2 and NS3 are essential for virus assembly and/or release and that each of these viral proteins plays an important role in this process.

scholarly journals Production and characterization of high-titer serum-free cell culture grown hepatitis C virus particles of genotype 1–6

Virology ◽  
2014 ◽  
Vol 458-459 ◽  
pp. 190-208 ◽  
Author(s):  
Christian K. Mathiesen ◽  
Tanja B. Jensen ◽  
Jannick Prentoe ◽  
Henrik Krarup ◽  
Alfredo Nicosia ◽  
...  
Keyword(s):  
Cell Culture ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
High Titer ◽  
Free Cell ◽  
Genotype 1 ◽  
Serum Free ◽  
Virus Particles

scholarly journals Virion-Independent Transfer of Replication-Competent Hepatitis C Virus RNA between Permissive Cells

2014 ◽  
Vol 89 (5) ◽  
pp. 2956-2961 ◽  
Author(s):  
Andrea Longatti ◽  
Bryan Boyd ◽  
Francis V. Chisari
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Infectious Virus ◽  
Immune Escape ◽  
Hcv Rna ◽  
Virus Particles ◽  
Viral Spread ◽  
Virus Rna ◽  
Huh7 Cells

In this study, we show that replication-competent subgenomic hepatitis C virus (HCV) RNA can be transferred to permissive Huh7 cells, leading to the establishment of viral RNA replication. Further, we show that these events are mediated by exosomes rather than infectious virus particles. If similar events occurin vivo, this could represent a novel, albeit inefficient, mechanism of viral spread and immune escape.

Investigations into the receptor requirements of Hepatitis C Virus (HCV) pseudoparticles and cell culture grown virions

2006 ◽  
Vol 44 (01) ◽  
Author(s):  
T von Hahn ◽  
M Flint ◽  
BD Lindenbach ◽  
A Boullier ◽  
O Quehenberger ◽  
...  
Keyword(s):  
Cell Culture ◽  
Hepatitis C Virus ◽  
Hepatitis C
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