Infection courses, virological features and IFN-α responses of HBV genotypes in cell culture and animal models

Min Zhang; Zhensheng Zhang; Michio Imamura; Mitsutaka Osawa; Yuji Teraoka; Jason Piotrowski; Yuji Ishida; Vitina Sozzi; Peter A Revill; Takeshi Saito; Kazuaki Chayama; T Jake Liang

doi:10.1016/j.jhep.2021.07.030

Infection courses, virological features and IFN-α responses of HBV genotypes in cell culture and animal models

J Hepatol. 2021 Dec;75(6):1335-1345. doi: 10.1016/j.jhep.2021.07.030. Epub 2021 Aug 4.

Authors

Affiliations

¹ Liver Diseases Branch, NIDDK, NIH, Bethesda, MD, USA.
² Department of Gastroenterology and Metabolism, Graduate School of Biomedical & Health Science, Hiroshima University, Hiroshima, Japan.
³ Division of Gastrointestinal and Liver Diseases, Department of Medicine, University of Southern California, Keck School of Medicine, Los Angeles, CA, USA; PhoenixBio Co., Ltd., Higashi-Hiroshima, Hiroshima, Japan.
⁴ Victorian Infectious Diseases Reference Laboratory, Royal Melbourne Hospital at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia.
⁵ Division of Gastrointestinal and Liver Diseases, Department of Medicine, University of Southern California, Keck School of Medicine, Los Angeles, CA, USA.
⁶ Collaborative Research Laboratory of Medical Innovation, Hiroshima University, Hiroshima, Japan.
⁷ Liver Diseases Branch, NIDDK, NIH, Bethesda, MD, USA. Electronic address: jliang@nih.gov.

Abstract

Background & aims: HBV consists of 9 major genotypes (A to I), 1 minor strain (designated J) and multiple subtypes, which may be associated with different clinical characteristics. As only cell lines expressing genotype D3 have been established, herein, we aimed to establish stable cell lines producing high-titer cell culture-generated HBV (HBVcc) of different genotypes and to explore their infectivity, virological features and responses to treatment.

Methods: Stable cell lines producing high titers of HBV genotype A2, B2, C1, E, F1b and H were generated by transfecting plasmids containing a replication-competent 1.3x length HBV genome and an antibiotic marker into HepG2 cells that can support HBV replication. Clones with the highest levels of HBV DNA and/or HBeAg were selected and expanded for large-scale purification of HBVcc. HBVcc of different genotypes were tested in cells and a humanized chimeric mouse model.

Results: HBVcc genotypes were infectious in mouse-passaged primary human hepatocytes (PXB cells) and responded differently to human interferon (IFN)-α with variable kinetics of reduction in HBV DNA, HBeAg and HBsAg. HBVcc of all genotypes were infectious in humanized chimeric mice but with variable kinetics of viremia and viral antigen production. Treatment of infected mice with human IFN-α resulted in modest and variable reductions of viremia and viral antigenemia. HBVcc passaged in humanized chimeric mice (HBVmp) infected PXB cells much more efficiently than that of the original HBVcc viral stock.

Conclusions: Herein, we generated stable cell lines producing HBV of various genotypes that are infectious in vitro and in vivo. We observe genotype-associated variations in viral antigen production, infection kinetics and responses to human IFN-α treatment in these models.

Lay summary: Stable cell lines producing high-titer cell culture-generated hepatitis B virus (HBV) of various genotypes were established. HBV genotypes showed stable infectivity in both in vitro and in vivo models, which are valuable tools for antiviral development.

Keywords: Virus genotypes; infection model; liver disease; stable cell lines; treatment.

Published by Elsevier B.V.

Publication types

Research Support, N.I.H., Intramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Culture Techniques / methods
Cell Culture Techniques / statistics & numerical data
Disease Models, Animal
Genotype*
Hepatitis B / complications*
Hepatitis B / genetics
Hepatitis B virus / drug effects
Hepatitis B virus / pathogenicity
Mice

Abstract

Publication types

MeSH terms

Grants and funding