健壮的人和小鼠肝细胞培养模型的乙型肝炎病毒感染和复制

StephenW

J Virol. 2018 Sep 19. pii: JVI.01255-18. doi: 10.1128/JVI.01255-18. [Epub ahead of print]
Robust human and murine hepatocyte culture models of hepatitis B virus infection and replication.
Qiao L1, Sui J2, Luo G3.
Author information

1
    Department of Microbiology, University of Alabama at Birmingham School of Medicine, Birmingham, Alabama, USA.
2
    National Institute of Biological Sciences, Beijing, China.
3
    Department of Microbiology, University of Alabama at Birmingham School of Medicine, Birmingham, Alabama, USA [email protected].

Abstract

Hepatitis B virus (HBV) is a major cause of chronic liver diseases including hepatitis, cirrhosis, and hepatocellular carcinoma. HBV research has been hampered by the lack of robust cell culture and small animal models of HBV infection. The discovery of sodium taurocholate cotransporting polypeptide (NTCP) as an HBV receptor has been a landmark advance in HBV research in recent years. Ectopic expression of NTCP in nonpermissive HepG2, Huh7, and AML12 cell lines confers HBV susceptibility. However, HBV replication in these human and murine hepatocytes cell lines appeared suboptimal. In present study, we have constructed stable NTCP-expressing HepG2 and AML12 cell lines and found that HBV permissiveness is correlated with NTCP expression. More significantly, we have developed robust HBV cell culture models by treating the HBV-infected cells with dimethyl sulfoxide (DMSO) and hydrocortisone, which significantly promoted HBV replication and production. Mechanistic studies suggested that hydrocortisone significantly enhanced the transcription and expression of PGC1α and HNF4α which are known to promote HBV transcription and replication. These new human and murine hepatocyte culture systems of HBV infection and replication will accelerate the determination of molecular aspects underlying HBV infection, replication, and morphogenesis in human and murine hepatocytes. We anticipate that our HBV cell culture models will also facilitate the discovery and development of antiviral drugs towards the ultimate eradication of chronic hepatitis B.IMPORTANCEHBV research has been greatly hampered by the lack of robust cell culture and small animal models of HBV infection and propagation. The discovery of NTCP as an HBV receptor has greatly impacted the field of HBV research. Although HBV infection of the NTCP-expressing human and murine hepatocyte cell lines has been demonstrated, its replication in cell culture appeared inefficient. To further improve cell culture systems of HBV infection and replication, we have constructed NTCP-expressing HepG2 and AML-12 cell lines that are highly permissive to HBV infection. More significantly, we have found that DMSO and hydrocortisone markedly enhanced HBV transcription and replication in human and murine hepatocytes when added to cell culture medium. These new cell culture models of HBV infection and replication will facilitate HBV research and antiviral drug discovery towards ultimate elimination of chronic hepatitis B.

PMID:
    30232184
DOI:
    10.1128/JVI.01255-18

StephenW

J Virol。 2018年9月19日.pii：JVI.01255-18。 doi：10.1128 / JVI.01255-18。 [提前打印]
健壮的人和小鼠肝细胞培养模型的乙型肝炎病毒感染和复制。
乔L1，隋J2，罗G3。
作者信息

1
    阿拉巴马大学伯明翰医学院微生物学系，美国阿拉巴马州伯明翰市。
2
    国家生物科学研究所，北京，中国。
3
    阿拉巴马大学伯明翰医学院微生物学系，美国阿拉巴马州伯明翰，[email protected]。

抽象

乙型肝炎病毒（HBV）是慢性肝病的主要原因，包括肝炎，肝硬化和肝细胞癌。由于缺乏稳健的细胞培养和HBV感染的小动物模型，HBV研究受到阻碍。牛磺胆酸钠协同转运多肽（NTCP）作为HBV受体的发现是近年来HBV研究的一个里程碑式的进展。非常规HepG2，Huh7和AML12细胞系中NTCP的异位表达赋予HBV易感性。然而，这些人和小鼠肝细胞细胞系中的HBV复制似乎不是最理想的。在本研究中，我们构建了稳定的表达NTCP的HepG2和AML12细胞系，并发现HBV允许性与NTCP表达相关。更重要的是，我们通过用二甲基亚砜（DMSO）和氢化可的松处理HBV感染的细胞开发了强大的HBV细胞培养模型，这显着促进了HBV复制和生产。机理研究表明氢化可的松显着增强PGC1α和HNF4α的转录和表达，已知PGC1α和HNF4α促进HBV转录和复制。这些HBV感染和复制的新的人和小鼠肝细胞培养系统将加速确定人和小鼠肝细胞中HBV感染，复制和形态发生的分子方面。我们预计，我们的HBV细胞培养模型也将促进抗病毒药物的发现和开发，以最终根除慢性乙型肝炎。由于缺乏稳健的细胞培养和HBV感染和繁殖的小动物模型，导致医学研究受到极大阻碍。 NTCP作为HBV受体的发现极大地影响了HBV研究领域。尽管已经证实了表达NTCP的人和小鼠肝细胞系的HBV感染，但其在细胞培养物中的复制似乎是低效的。为了进一步改善HBV感染和复制的细胞培养系统，我们构建了表达NTCP的HepG2和AML-12细胞系，这些细胞系对HBV感染具有高度的许可。更重要的是，我们发现当添加到细胞培养基中时，DMSO和氢化可的松显着增强人和小鼠肝细胞中的HBV转录和复制。这些新的HBV感染和复制细胞培养模型将促进HBV研究和抗病毒药物发现，最终消除慢性乙型肝炎。

结论：
    30232184
DOI：
    10.1128 / JVI.01255-18