HBV Research Protocols

There is an urgent need for centralized repositories of HBV-related materials that are readily accessible to HBV researchers globally. Critical to this will be quality assurance of the samples, and the availability of matching clinical data. This repository of HBV-related research protocols is designed to facilitate studies and the development of new drugs.

This project is designed to complement the upcoming NIAID reagents repository by making corresponding quality-controlled research protocols available freely for all researchers around the world.

When citing protocols from this database, please cite the original publications from which these protocols have been adapted. The origin of these original publications can be found within the protocols. Please also acknowledge this ICE-HBV Protocols Database. Any questions can be directed to info@ice-hbv.org

The review of these protocols has been led by Haitao Guo and the ICE-HBV working group members including Lena Allweiss, Maura Dandri, Jianming Hu, Jake Liang, Margaret Littlejohn, Peter Revill, and Barbara Testoni.

The development of the database is coordinated by Marley Easterbrook.

Hepatitis B Virus Polymerase Epsilon RNA Binding Assays

Daniel N. Clark, Scott A. Jones, Jianming Hu

Penn State University College of Medicine
  • HBV polymerase activity is assayed by measuring its ability to bind viral RNA at a secondary-structure motif, the epsilon (e) stem-loop structure.
  • HBV polymerase is expressed in cell culture to yield its binding-active form and RNA may be supplied in culture or... [Read more]

In Vitro Hepatitis B Virus Polymerase Priming Assay

Daniel N. Clark, Scott A. Jones, Jianming Hu

Penn State University College of Medicine
  • HBV polymerase activity is assayed by measuring the first step in DNA synthesis, protein priming, wherein the polymerase becomes nucleotide-labelled.
  • This in vitroassay uses a plasmid-based expression system performed in cell culture, producing a chaperone... [Read more]

HBV and HDV infection in uPA/SCID mice with humanized livers

Volz, T. 1; Lütgehetmann, M. 1,3; Dandri, M. 1,2

1. Medical Clinic, University Medic al Center Hamburg-Eppendorf, Hamburg, Germany, 2. German Centre for Infection Research (DZIF), Munich, Hamburg and Heidelberg partner sites, Germany, 3. Institute of Microbiology, Virology and Hygiene, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
  • This protocol describes the infection of liver humanized mice (here UPA/SCID mice) with hepatitis B and /or hepatitis D viruses.
  • We also address the characteristics of infection kinetics and the factors influencing viral spread in humanized mouse livers.
  • ... [Read more]

Evaluating mouse HBV-specific CD8+ T cell responses by ELISPOT assay

Safiehkhatoon Moshkani, Carolina Chiale, and Michael D. Robek

Dept. of Immunology and Microbial Disease, Albany Medical College, Albany, NY, USA
  • Immunocompetent mouse models that allow for the investigation of virus replication and the immune response to HBV include hydrodynamic injection, HBV genome transduction with adenovirus or adeno-associated virus, and HBV transgenic mice [1].
  • A number of... [Read more]

Hydrodynamic HBV transfection in mouse models

Li-Ling Wu 1,2, Hurng-Yi Wang 1, and Pei-Jer Chen 1,3,4

1. Graduate Institute of Clinical Medicine, 2. Department and Institute of Physiology, School of Medicine, National Yang-Ming University, 3. Hepatitis Research Center, and 4. Department of Medical Research, National Taiwan University College of Medicine and National Taiwan University Hospital, Taipei, Taiwan.
  • The method of hydrodynamic delivery (1, 2) was used to establish a mouse model of hepatitis B virus persistence by transfecting hepatocytes in vivo with HBV genome expressing viral antigens and replicative intermediates, resulting in production of viral particles (3).
  • ... [Read more]

Immunofluorescent Staining for the Hepatitis B Core Antigen in frozen Liver Sections

Lena Allweiss, Marc Lütgehetmann, Maura Dandri

I. Deptartment of Internal Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
  • The HBV core antigen (HBcAg) is the structural protein of the viral nucleocapsids but is also recruited to the nuclear HBV persistence form. Thus, HBcAg can be detected in the cytoplasm and nucleus of infected hepatocytes.
  • Here, we ... [Read more]

Stem Cell Derived Hepatocyte-Like Cells for Hepatitis B Virus Infection

Yuchen Xia (1), Seung Bum Park (2), T. Jake Liang (2)

1. Wuhan University, Wuhan, China 2. National Institutes of Health, Bethesda, USA
  • Human pluripotent stem cells derived hepatocyte-like cells (HLCs) are capable of expressing hepatocyte markers and host factors important for hepatic function maintenance.
  • These cells fully support HBV infection and virus-host interactions.
  • Stem cell-derived HLCs provide a new tool for antiviral drug screening and development.

Detection and characterisation of integrated Hepatitis B virus DNA using inverse nested PCR

Dr. Thomas Tu

Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany
  • The integration of HBV DNA into the host cell genome can occur during HBV infection, though it does not produce new infectious virus particles.
  • HBV DNA integration is one possible cause of HBV-induced liver cancer and could b... [Read more]