脏再生期间肝炎病毒持续存在，并且在体外和体内通过细胞

StephenW

Hepatitis delta virus persists during liver regeneration and is amplified through cell division both in vitro and in vivo

    Katja Giersch1, Oliver D Bhadra1, Tassilo Volz1, Lena Allweiss1, Kristoffer Riecken2, Boris Fehse2, Ansgar W Lohse1,3, Joerg Petersen4, Camille Sureau5, Stephan Urban3,6, Maura Dandri1,3, Marc Lütgehetmann7

Author affiliations

    I. Department of Internal Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
    Department of Stem Cell transplantation, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
    German Center for Infection Research (DZIF), Hamburg-Lübeck-Borstel and Heidelberg Partner sites, Germany
    IFI Institute for Interdisciplinary Medicine, Asklepios Clinic St. Georg, Hamburg, Germany
    Laboratoirede Virologie Moleculaire, INTS, Centre National de la Recherche Scientifique, Paris, France
    Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany
    Institute of Microbiology, Virology and Hygiene, University Medical Center Hamburg-Eppendorf, Hamburg, Germany

    Correspondence to Dr Maura Dandri, Department of Internal Medicine, University Medical Center Hamburg-Eppendorf, Hamburg 20246, Germany; [email protected] and Dr Marc Lütgehetmann, Institute of Microbiology, Virology and Hygiene, University Medical Center Hamburg-Eppendorf, Hamburg 20246, Germany ; [email protected]

Abstract

Objective Hepatitis delta virus (HDV) was shown to persist for weeks in the absence of HBV and for months after liver transplantation, demonstrating the ability of HDV to persevere in quiescent hepatocytes. The aim of the study was to evaluate the impact of cell proliferation on HDV persistence in vitro and in vivo.

Design Genetically labelled human sodium taurocholate cotransporting polypeptide (hNTCP)-transduced human hepatoma(HepG2) cells were infected with HBV/HDV and passaged every 7 days for 100 days in the presence of the entry inhibitor Myrcludex-B. In vivo, cell proliferation was triggered by transplanting primary human hepatocytes (PHHs) isolated from HBV/HDV-infected humanised mice into naïve recipients. Virological parameters were measured by quantitative real time polymerase chain reaction (qRT-PCR). Hepatitis delta antigen (HDAg), hepatitis B core antigen (HBcAg) and cell proliferation were determined by immunofluorescence.

Results Despite 15 in vitro cell passages and block of viral spreading by Myrcludex-B, clonal cell expansion permitted amplification of HDV infection. In vivo, expansion of PHHs isolated from HBV/HDV-infected humanised mice was confirmed 3 days, 2, 4 and 8 weeks after transplantation. While HBV markers rapidly dropped in proliferating PHHs, HDAg-positive hepatocytes were observed among dividing cells at all time points. Notably, HDAg-positive cells appeared in clusters, indicating that HDV was transmitted to daughter cells during liver regeneration even in the absence of de novo infection.

Conclusion This study demonstrates that HDV persists during liver regeneration by transmitting HDV RNA to dividing cells even in the absence of HBV coinfection. The strong persistence capacities of HDV may also explain why HDV clearance is difficult to achieve in HBV/HDV chronically infected patients.

http://dx.doi.org/10.1136/gutjnl-2017-314713

StephenW

肝脏再生期间肝炎病毒持续存在，并且在体外和体内通过细胞分裂扩增

    Katja Giersch1，Oliver D Bhadra1，Tassilo Volz1，Lena Allweiss1，Kristoffer Riecken2，Boris Fehse2，Ansgar W Lohse1,3，Joerg Petersen4，Camille Sureau5，Stephan Urban3,6，Maura Dandri1,3，MarcLütgehetmann7

作者隶属关系

    I.德国汉堡汉堡 - 埃彭多夫大学医学中心内科
    汉堡 - 埃彭多夫大学医学中心干细胞移植科，德国汉堡
    德国感染研究中心（DZIF），汉堡 - 吕贝克 - 博斯特尔和海德堡合作伙伴网站，德国
    IFI跨学科医学研究所，Asklepios Clinic St. Georg，德国汉堡
    Instratoirede Virologie Moleculaire，INTS，Centre National de la Recherche Scientifique，Paris，France
    德国海德堡海德堡大学医院传染病科，分子病毒学系
    德国汉堡汉堡 - 埃彭多夫大学医学中心微生物学，病毒学和卫生学研究所

    与德国汉堡20246汉堡 - 埃彭多夫大学医学中心内科医生Maura Dandri博士的通信; [email protected]和MarcLütgehetmann博士，德国汉堡 - 埃彭多夫大学医学中心微生物学，病毒学和卫生学研究所，德国汉堡20246; [email protected]

抽象

目的显示肝炎病毒（HDV）在没有HBV的情况下持续数周，在肝移植后持续数月，证明了HDV在静息肝细胞中持续存在的能力。该研究的目的是评估细胞增殖对体外和体内HDV持久性的影响。

设计遗传标记的人类牛磺胆酸钠协同转运多肽（hNTCP）转导的人肝癌（HepG2）细胞用HBV / HDV感染，并在进入抑制剂Myrcludex-B存在下每7天传代100天。在体内，通过将分离自HBV / HDV感染的人源化小鼠的原代人肝细胞（PHH）移植到幼稚接受者中来触发细胞增殖。通过定量实时聚合酶链反应（qRT-PCR）测量病毒学参数。通过免疫荧光测定乙型肝炎抗原（HDAg），乙型肝炎核心抗原（HBcAg）和细胞增殖。

结果尽管Myrcludex-B有15个体外细胞传代和病毒传播阻滞，克隆细胞扩增允许HDV感染的扩增。在体内，在移植后3天，2,4和8周确认从HBV / HDV感染的人源化小鼠分离的PHH的扩增。虽然HBV标志物在增殖的PHH中迅速下降，但在所有时间点在分裂细胞中观察到HDAg阳性肝细胞。值得注意的是，HDAg阳性细胞出现在簇中，表明即使没有从头感染，HDV也在肝脏再生过程中传递给子细胞。

结论本研究表明，即使在没有HBV合并感染的情况下，通过将HDV RNA传递给分裂细胞，HDV在肝再生期间仍然存在。 HDV的强持久性能力也可以解释为什么在HBV / HDV慢性感染患者中难以实现HDV清除。

http://dx.doi.org/10.1136/gutjnl-2017-314713