Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
6140108 | Virology | 2014 | 19 Pages |
Abstract
Recently, cell culture systems producing hepatitis C virus particles (HCVcc) were developed. Establishment of serum-free culture conditions is expected to facilitate development of a whole-virus inactivated HCV vaccine. We describe generation of genotype 1-6 serum-free HCVcc (sf-HCVcc) from Huh7.5 hepatoma cells cultured in adenovirus expression medium. Compared to HCVcc, sf-HCVcc showed 0.6-2.1Â log10 higher infectivity titers (4.7-6.2Â log10Â Focus Forming Units/mL), possibly due to increased release and specific infectivity of sf-HCVcc. In contrast to HCVcc, sf-HCVcc had a homogeneous single-peak density profile. Entry of sf-HCVcc depended on HCV co-receptors CD81, LDLr, and SR-BI, and clathrin-mediated endocytosis. HCVcc and sf-HCVcc were neutralized similarly by chronic-phase patient sera and by human monoclonal antibodies targeting conformational epitopes. Thus, we developed serum-free culture systems producing high-titer single-density sf-HCVcc, showing similar biological properties as HCVcc. This methodology has the potential to advance HCV vaccine development and to facilitate biophysical studies of HCV.
Keywords
IC50BmaxLDLRRCFSR-BIHRPVLDLffuHCVccAEMLVPHVR1FBSCell culture systemLow-density-lipoprotein receptorBSKBSAMOIbovine serum albuminSerum-freefocus forming unitsVaccine developmentBiophysical characterizationstandard error of the meanNeutralizationRoom temperaturefetal bovine serummedian inhibitory concentrationnonstructural proteinphycoerythrinSEMHypervariable region 1relative centrifugal forceHepatitis C virusHCVHorseradish peroxidasemultiplicity of infectionGenotypesscavenger receptor class B type I
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Authors
Christian K. Mathiesen, Tanja B. Jensen, Jannick Prentoe, Henrik Krarup, Alfredo Nicosia, Mansun Law, Jens Bukh, Judith M. Gottwein,