|
- 现金
- 62111 元
- 精华
- 26
- 帖子
- 30437
- 注册时间
- 2009-10-5
- 最后登录
- 2022-12-28
|
Hepatology
Original article
Proliferation of primary human hepatocytes and prevention of hepatitis B virus reinfection efficiently deplete nuclear cccDNA in vivo
Lena Allweiss1, Tassilo Volz1, Katja Giersch1, Janine Kah1, Giuseppina Raffa2, Joerg Petersen3, Ansgar W Lohse1,4, Concetta Beninati5, Teresa Pollicino2, Stephan Urban4,6, Marc Lütgehetmann1,7, Maura Dandri1,4
Author affiliations
Department of Medicine, Center for Internal Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
Department of Clinical and Experimental Medicine, University Hospital of Messina, Messina, Italy
IFI Institute for Interdisciplinary Medicine at Asklepios Clinic St. Georg, Hamburg, Germany
German Center for Infection Research (DZIF), Hamburg-Lübeck-Borstel and Heidelberg Partner Sites, Hamburg, Germany
Department of Human Pathology, University Hospital of Messina, Messina, Italy
Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany
Department of Medical Microbiology, Virology and Hygiene, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
Correspondence to Professor Dr Maura Dandri, Department of Internal Medicine, Center for Internal Medicine, University Medical Center Hamburg-Eppendorf, Martinistr. 52, Hamburg D-20246, Germany; [email protected]
Abstract
Objective The stability of the covalently closed circular DNA (cccDNA) in nuclei of non-dividing hepatocytes represents a key determinant of HBV persistence. Contrarily, studies with animal hepadnaviruses indicate that hepatocyte turnover can reduce cccDNA loads but knowledge on the proliferative capacity of HBV- infected primary human hepatocytes (PHHs) in vivo and the fate of cccDNA in dividing PHHs is still lacking. This study aimed to determine the impact of human hepatocyte division on cccDNA stability in vivo.
Methods PHH proliferation was triggered by serially transplanting hepatocytes from HBV-infected humanized mice into naïve recipients. Cell proliferation and virological changes were assessed by quantitative PCR, immunofluorescence and RNA in situ hybridisation. Viral integrations were analyzed by gel separation and deep sequencing.
Nevertheless, cell division has to cause cccDNA dilution among daughter cells and intrahepatic cccDNA loss. Nevertheless, HBV survived in sporadic non-proliferating human hepatocytes, so that This was due to reinfection of quiescent PHHs since treatment with the entry inhibitor inhibitor myrcludex-B or nucleoside analogues blocked viral spread and intrahepatic cccDNA accumulation. Viral integrations were detected both in donors and recipient mice but did not appear to contribute to antigen production.
Conclusions We demonstrate that human hepatocyte division even without involvement of cytolytic mechanisms triggers substantial cccDNA loss. This process may be fundamental to resolve self-limiting acute infection and should be considered in future therapeutic interventions along with entry inhibition strategies.
http://dx.doi.org/10.1136/gutjnl-2016-312162
|
|
发表于 2018-2-9 03:05
