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Efficient long-term amplification of hepatitis B virus isolates after infection of slow proliferating HepG2-NTCP cells

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dc.contributor.author박준용-
dc.contributor.author안상훈-
dc.contributor.author한광협-
dc.date.accessioned2019-12-18T00:29:24Z-
dc.date.available2019-12-18T00:29:24Z-
dc.date.issued2019-
dc.identifier.issn0168-8278-
dc.identifier.urihttps://ir.ymlib.yonsei.ac.kr/handle/22282913/173073-
dc.description.abstractBACKGROUND & AIMS: As hepatitis B virus (HBV) spreads through the infected liver it is simultaneously secreted into the blood. HBV-susceptible in vitro infection models do not efficiently amplify viral progeny or support cell-to-cell spread. We sought to establish a cell culture system for the amplification of infectious HBV from clinical specimens. METHODS: An HBV-susceptible sodium-taurocholate cotransporting polypeptide-overexpressing HepG2 cell clone (HepG2-NTCPsec+) producing high titers of infectious progeny was selected. Secreted HBV progeny were characterized by native gel electrophoresis and electron microscopy. Comparative RNA-seq transcriptomics was performed to quantify the expression of host proviral and restriction factors. Viral spread routes were evaluated using HBV entry- or replication inhibitors, visualization of viral cell-to-cell spread in reporter cells, and nearest neighbor infection determination. Amplification kinetics of HBV genotypes B-D were analyzed. RESULTS: Infected HepG2-NTCPsec+ secreted high levels of large HBV surface protein-enveloped infectious HBV progeny with typical appearance under electron microscopy. RNA-seq transcriptomics revealed that HBV does not induce significant gene expression changes in HepG2-NTCPsec+, however, transcription factors favoring HBV amplification were more strongly expressed than in less permissive HepG2-NTCPsec-. Upon inoculation with HBV-containing patient sera, rates of infected cells increased from 10% initially to 70% by viral spread to adjacent cells, and viral progeny and antigens were efficiently secreted. HepG2-NTCPsec+ supported up to 1,300-fold net amplification of HBV genomes depending on the source of virus. Viral spread and amplification were abolished by entry and replication inhibitors; viral rebound was observed after inhibitor discontinuation. CONCLUSIONS: The novel HepG2-NTCPsec+ cells efficiently support the complete HBV life cycle, long-term viral spread and amplification of HBV derived from patients or cell culture, resembling relevant features of HBV-infected patients. LAY SUMMARY: Currently available laboratory systems are unable to reproduce the dynamics of hepatitis B virus (HBV) spread through the infected liver and release into the blood. We developed a slowly dividing liver-derived cell line which multiplies infectious viral particles upon inoculation with patient- or cell culture-derived HBV. This new infection model can improve therapy by measuring, in advance, the sensitivity of a patient's HBV strain to specific antiviral drugs.-
dc.description.statementOfResponsibilityopen-
dc.formatapplication/pdf-
dc.languageEnglish-
dc.publisherElsevier-
dc.relation.isPartOfJOURNAL OF HEPATOLOGY-
dc.rightsCC BY-NC-ND 2.0 KR-
dc.titleEfficient long-term amplification of hepatitis B virus isolates after infection of slow proliferating HepG2-NTCP cells-
dc.typeArticle-
dc.contributor.collegeCollege of Medicine (의과대학)-
dc.contributor.departmentDept. of Internal Medicine (내과학교실)-
dc.contributor.googleauthorAlexander König-
dc.contributor.googleauthorJaewon Yang-
dc.contributor.googleauthorEunji Jo-
dc.contributor.googleauthorKyu Ho Paul Park-
dc.contributor.googleauthorHyun Kim-
dc.contributor.googleauthorThoa Thi Than-
dc.contributor.googleauthorXiyong Song-
dc.contributor.googleauthorXiaoxuan Qi-
dc.contributor.googleauthorXinghong Dai-
dc.contributor.googleauthorSoonju Park-
dc.contributor.googleauthorDavid Shum-
dc.contributor.googleauthorWang-Shick Ryu-
dc.contributor.googleauthorJung-Hee Kim-
dc.contributor.googleauthorSeung Kew Yoon-
dc.contributor.googleauthorJun Yong Park-
dc.contributor.googleauthorSang Hoon Ahn-
dc.contributor.googleauthorKwang-Hyub Han-
dc.contributor.googleauthorWolfram Hubert Gerlich-
dc.contributor.googleauthorMarc Peter Windisch-
dc.identifier.doi10.1016/j.jhep.2019.04.010-
dc.contributor.localIdA01675-
dc.contributor.localIdA02226-
dc.contributor.localIdA04268-
dc.relation.journalcodeJ01441-
dc.identifier.eissn1600-0641-
dc.identifier.pmid31077792-
dc.subject.keywordComplete HBV life cycle-
dc.subject.keywordDrug sensitivity-
dc.subject.keywordHBV doubling time-
dc.subject.keywordHBV spread-
dc.subject.keywordHepG2-NTCP-
dc.subject.keywordKinetics of antigen-
dc.subject.keywordPatient-derived HBV-
dc.subject.keywordVirion secretion-
dc.subject.keywordcccDNA accumulation-
dc.contributor.alternativeNamePark, Jun Yong-
dc.contributor.affiliatedAuthor박준용-
dc.contributor.affiliatedAuthor안상훈-
dc.contributor.affiliatedAuthor한광협-
dc.citation.volume71-
dc.citation.number2-
dc.citation.startPage289-
dc.citation.endPage300-
dc.identifier.bibliographicCitationJOURNAL OF HEPATOLOGY, Vol.71(2) : 289-300, 2019-
dc.identifier.rimsid64007-
dc.type.rimsART-
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Internal Medicine (내과학교실) > 1. Journal Papers

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