Production and characterization of high-titer serum-free cell culture grown hepatitis C virus particles of genotype 1-6
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Production and characterization of high-titer serum-free cell culture grown hepatitis C virus particles of genotype 1-6. / Mathiesen, Christian K; Jensen, Tanja B; Prentoe, Jannick; Krarup, Henrik; Nicosia, Alfredo; Law, Mansun; Bukh, Jens; Gottwein, Judith M.
In: Virology, Vol. 458-459, 06.2014, p. 190-208.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Production and characterization of high-titer serum-free cell culture grown hepatitis C virus particles of genotype 1-6
AU - Mathiesen, Christian K
AU - Jensen, Tanja B
AU - Prentoe, Jannick
AU - Krarup, Henrik
AU - Nicosia, Alfredo
AU - Law, Mansun
AU - Bukh, Jens
AU - Gottwein, Judith M
N1 - Copyright © 2014 Elsevier Inc. All rights reserved.
PY - 2014/6
Y1 - 2014/6
N2 - 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 0.6-2.1 log10 higher infectivity titers (4.7-6.2 log10 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.
AB - 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 0.6-2.1 log10 higher infectivity titers (4.7-6.2 log10 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.
KW - Cell Line, Tumor
KW - Cell Survival
KW - Culture Media, Serum-Free
KW - Gene Expression Regulation, Viral
KW - Genotype
KW - Hepacivirus
KW - Humans
KW - Virus Cultivation
U2 - 10.1016/j.virol.2014.03.021
DO - 10.1016/j.virol.2014.03.021
M3 - Journal article
C2 - 24928051
VL - 458-459
SP - 190
EP - 208
JO - Virology
JF - Virology
SN - 0042-6822
ER -
ID: 131071143