Cited 42 times in
Efficient long-term amplification of hepatitis B virus isolates after infection of slow proliferating HepG2-NTCP cells
DC Field | Value | Language |
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dc.contributor.author | 박준용 | - |
dc.contributor.author | 안상훈 | - |
dc.contributor.author | 한광협 | - |
dc.date.accessioned | 2019-12-18T00:29:24Z | - |
dc.date.available | 2019-12-18T00:29:24Z | - |
dc.date.issued | 2019 | - |
dc.identifier.issn | 0168-8278 | - |
dc.identifier.uri | https://ir.ymlib.yonsei.ac.kr/handle/22282913/173073 | - |
dc.description.abstract | BACKGROUND & 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.statementOfResponsibility | open | - |
dc.format | application/pdf | - |
dc.language | English | - |
dc.publisher | Elsevier | - |
dc.relation.isPartOf | JOURNAL OF HEPATOLOGY | - |
dc.rights | CC BY-NC-ND 2.0 KR | - |
dc.title | Efficient long-term amplification of hepatitis B virus isolates after infection of slow proliferating HepG2-NTCP cells | - |
dc.type | Article | - |
dc.contributor.college | College of Medicine (의과대학) | - |
dc.contributor.department | Dept. of Internal Medicine (내과학교실) | - |
dc.contributor.googleauthor | Alexander König | - |
dc.contributor.googleauthor | Jaewon Yang | - |
dc.contributor.googleauthor | Eunji Jo | - |
dc.contributor.googleauthor | Kyu Ho Paul Park | - |
dc.contributor.googleauthor | Hyun Kim | - |
dc.contributor.googleauthor | Thoa Thi Than | - |
dc.contributor.googleauthor | Xiyong Song | - |
dc.contributor.googleauthor | Xiaoxuan Qi | - |
dc.contributor.googleauthor | Xinghong Dai | - |
dc.contributor.googleauthor | Soonju Park | - |
dc.contributor.googleauthor | David Shum | - |
dc.contributor.googleauthor | Wang-Shick Ryu | - |
dc.contributor.googleauthor | Jung-Hee Kim | - |
dc.contributor.googleauthor | Seung Kew Yoon | - |
dc.contributor.googleauthor | Jun Yong Park | - |
dc.contributor.googleauthor | Sang Hoon Ahn | - |
dc.contributor.googleauthor | Kwang-Hyub Han | - |
dc.contributor.googleauthor | Wolfram Hubert Gerlich | - |
dc.contributor.googleauthor | Marc Peter Windisch | - |
dc.identifier.doi | 10.1016/j.jhep.2019.04.010 | - |
dc.contributor.localId | A01675 | - |
dc.contributor.localId | A02226 | - |
dc.contributor.localId | A04268 | - |
dc.relation.journalcode | J01441 | - |
dc.identifier.eissn | 1600-0641 | - |
dc.identifier.pmid | 31077792 | - |
dc.subject.keyword | Complete HBV life cycle | - |
dc.subject.keyword | Drug sensitivity | - |
dc.subject.keyword | HBV doubling time | - |
dc.subject.keyword | HBV spread | - |
dc.subject.keyword | HepG2-NTCP | - |
dc.subject.keyword | Kinetics of antigen | - |
dc.subject.keyword | Patient-derived HBV | - |
dc.subject.keyword | Virion secretion | - |
dc.subject.keyword | cccDNA accumulation | - |
dc.contributor.alternativeName | Park, Jun Yong | - |
dc.contributor.affiliatedAuthor | 박준용 | - |
dc.contributor.affiliatedAuthor | 안상훈 | - |
dc.contributor.affiliatedAuthor | 한광협 | - |
dc.citation.volume | 71 | - |
dc.citation.number | 2 | - |
dc.citation.startPage | 289 | - |
dc.citation.endPage | 300 | - |
dc.identifier.bibliographicCitation | JOURNAL OF HEPATOLOGY, Vol.71(2) : 289-300, 2019 | - |
dc.identifier.rimsid | 64007 | - |
dc.type.rims | ART | - |
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