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Incorporation of primary patient‐derived glycoproteins into authentic infectious hepatitis C virus particles
Author(s) -
Doerrbecker Juliane,
Friesland Martina,
Riebesehl Nina,
Ginkel Corinne,
Behrendt Patrick,
Brown Richard J.P.,
Ciesek Sandra,
Wedemeyer Heiner,
Sarrazin Christoph,
Kaderali Lars,
Pietschmann Thomas,
Steinmann Eike
Publication year - 2014
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.27190
Subject(s) - virology , biology , hepatitis c virus , virus , glycoprotein , virus like particle , hepacivirus , viral replication , gene , genetics , recombinant dna
The Japanese fulminant hepatitis‐1 (JFH1)‐based hepatitis C virus (HCV) infection system has permitted analysis of the complete viral replication cycle in vitro . However, lack of robust infection systems for primary, patient‐derived isolates limits systematic functional studies of viral intrahost variation and vaccine development. Therefore, we aimed at developing cell culture models for incorporation of primary patient‐derived glycoproteins into infectious HCV particles for in‐depth mechanistic studies of envelope gene function. To this end, we first constructed a packaging cell line expressing core, p7, and NS2 based on the highly infectious Jc1 genotype (GT) 2a chimeric genome. We show that this packaging cell line can be transfected with HCV replicons encoding cognate Jc1‐derived glycoprotein genes for production of single‐round infectious particles by way of trans ‐complementation. Testing replicons expressing representative envelope protein genes from all major HCV genotypes, we observed that virus production occurred in a genotype‐ and isolate‐dependent fashion. Importantly, primary GT 2 patient‐derived glycoproteins were efficiently incorporated into infectious particles. Moreover, replacement of J6 (GT 2a) core, p7, and NS2 with GT 1a‐derived H77 proteins allowed production of infectious HCV particles with GT 1 patient‐derived glycoproteins. Notably, adaptive mutations known to enhance virus production from GT 1a‐2a chimeric genomes further increased virus release. Finally, virus particles with primary patient‐derived E1‐E2 proteins possessed biophysical properties comparable to Jc1 HCVcc particles, used CD81 for cell entry, were associated with ApoE and could be neutralized by immune sera. Conclusion : This work describes cell culture systems for production of infectious HCV particles with primary envelope protein genes from GT 1 and GT 2‐infected patients, thus opening up new opportunities to dissect envelope gene function in an individualized fashion. (H epatology 2014;60:508–520)