Revival, characterization, and hepatitis B virus infection of cryopreserved human fetal hepatocytes

Zhou, Ming; Huang, Yayun; Cheng, Zhikui; Zhao, Fei; Li, Jiafu; Zhi, Xiaoguang; Tian, Xin; Sun, Weijia; Hu, Kanghong

doi:10.1016/j.jviromet.2014.06.015

Cited by 8 publications

(9 citation statements)

References 39 publications

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“…For example, PHHs, the native host for HBV infection, rapidly dedifferentiate and lose susceptibility to HBV infection after being isolated and seeded (Zhou et al, 2014a). DMSO treatment can maintain PHH differentiation and susceptibility to HBV infection.…”

Section: Discussionmentioning

confidence: 99%

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Productive HBV infection of well-differentiated, hNTCP-expressing human hepatoma-derived (Huh7) cells

et al. 2017

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Feasible and effective cell models for hepatitis B virus (HBV) infection are required for investigating the complete lifecycle of this virus, including the early steps of viral entry. Resistance to dimethyl sulfoxide/polyethylene glycol (DMSO/PEG), hNTCP expression, and a differentiated state are the limiting factors for successful HBV infection models. In the present study, we used a hepatoma cell line (Huh7D hNTCP ) to overcome these limiting factors so that it exhibits excellent susceptibility to HBV infection. To achieve this goal, different hepatoma cell lines were tested with 2.5% DMSO / 4% PEG8000, and one resistant cell line (Huh7D) was used to construct a stable hNTCP-expressing cell line (Huh7D hNTCP ) using a recombinant lentivirus system. Then, the morphological characteristics and differentiation molecular markers of Huh7D hNTCP cells with or without DMSO treatment were characterized. Finally, the susceptibility of Huh7D hNTCP cells to HBV infection was assessed. Our results showed that Huh7D cells were resistant to 2.5% DMSO / 4% PEG8000, whereas the others were not. Huh7D hNTCP cells were established to express a high level of hNTCP compared to liver extracts, and Huh7D hNTCP cells rapidly transformed into a non-dividing, well-differentiated polarized phenotype under DMSO treatment. Huh7D hNTCP cells fully supported the entire lifecycle of HBV infection. This cell culture system will be useful for the analysis of host-virus interactions, which should facilitate the discovery of antiviral drugs and vaccines.

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Section: Discussionmentioning

confidence: 99%

“…Southern blot was performed as described previously www.virosin.org DECEMBER 2017 VOLUME 32 ISSUE 6 467 (Zhou et al, 2014a). Northern blot was performed as described elsewhere …”

Section: Southern Blot and Northern Blotmentioning

confidence: 99%

Productive HBV infection of well-differentiated, hNTCP-expressing human hepatoma-derived (Huh7) cells

et al. 2017

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“…Primary human hepatocytes were isolated from embryonic livers of post-mortem premature neonates (estimated gestational age, 16–24 weeks), kindly provided by Zhongnan Hospital of Wuhan University with written informed consent from participating family members. PHH were isolated using a modified two-step collagenase perfusion procedure and cultured in William’s E base medium (WEM, Gibco, Grand Island, NY) as described previously 25 26 . The ethics and protocol to use tissue samples for this study were approved by the institutional bioethics committee of Wuhan Institute of Virology (registration number: WIVH24201101) in accordance with the principles of the Declaration of Helsinki.…”

Section: Methodsmentioning

confidence: 99%

Efficient Inhibition of Hepatitis B Virus Infection by a preS1-binding Peptide

Yang

Zhou

et al. 2016

Sci Rep

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Entry inhibitors are promising novel antivirals against hepatitis B virus (HBV) infection. The existing potential entry inhibitors have targeted the cellular receptor(s). In this study, we aim to develop the first entry inhibitor that inhibits HBV infection via targeting viral particles. The preS1 segment of the large envelope glycoprotein of HBV is essential for virion attachment and infection. Previously, we obtained a preS1-binding short peptide B10 by screening a phage display peptide library using the N-terminal half of preS1 (residues 1 to 60, genotype C). We report here that by means of concatenation of B10, we identified a quadruple concatemer 4B10 that displayed a markedly increased preS1-binding activity. The main binding site of 4B10 in preS1 was mapped to the receptor binding enhancing region. 4B10 blocked HBV attachment to hepatic cells and inhibited HBV infection of primary human and tupaia hepatocytes at low nanomolar concentrations. The 4B10-mediated inhibition of HBV infection is specific as it did not inhibit the infection of vesicular stomatitis virus glycoprotein pseudotyped lentivirus or human immunodeficiency virus type 1. Moreover, 4B10 showed no binding activity to hepatic cells. In conclusion, we have identified 4B10 as a promising candidate for a novel class of HBV entry inhibitors.

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“…Considering that NTCP expression dropped sharply in ex vivo-cultured PHHs (Fig. S 8f ) 17 , 27 , 35 , the inhibitory effect of MyrB on HBV replication in PHHs was not observed (Fig. S 8 ).…”

Section: Discussionmentioning

confidence: 99%

Upregulation of HBV transcription by sodium taurocholate cotransporting polypeptide at the postentry step is inhibited by the entry inhibitor Myrcludex B

Zhao

Liu

Chen

et al. 2018

Emerging Microbes & Infections

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Sodium taurocholate cotransporting polypeptide (NTCP) is a functional receptor for hepatitis B virus (HBV) entry. However, little is known regarding whether NTCP is involved in regulating the postentry steps of the HBV life cycle. Here, we found that NTCP expression upregulated HBV transcription at the postentry step and that the NTCP-targeting entry inhibitor Myrcludex B (MyrB) effectively suppressed HBV transcription both in an HBV in vitro infection system and in mice hydrodynamically injected with an HBV expression plasmid. Mechanistically, NTCP upregulated HBV transcription via farnesoid X receptor α (FxRα)-mediated activation of the HBV EN2/core promoter at the postentry step in a manner that was dependent on the bile acid (BA)-transport function of NTCP, which was blocked by MyrB. Our findings uncover a novel role for NTCP in the HBV life cycle and provide a reference for the use of novel NTCP-targeting entry inhibitors to suppress HBV infection and replication.

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Revival, characterization, and hepatitis B virus infection of cryopreserved human fetal hepatocytes

Cited by 8 publications

References 39 publications

Productive HBV infection of well-differentiated, hNTCP-expressing human hepatoma-derived (Huh7) cells

Productive HBV infection of well-differentiated, hNTCP-expressing human hepatoma-derived (Huh7) cells

Efficient Inhibition of Hepatitis B Virus Infection by a preS1-binding Peptide

Upregulation of HBV transcription by sodium taurocholate cotransporting polypeptide at the postentry step is inhibited by the entry inhibitor Myrcludex B

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