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.

An Optimized Ex Vivo Flurospot assay to identify multi-functional HBV-specific T cells in Chronic Hepatitis B patients

Conan Chua, Aman Mehrotra, Dr. Adam Gehring

Toronto Centre for Liver Disease, Toronto, Ontario, Canada
During a traditional FluroSpot assay, low HBV Specific T cell frequencies have hindered effective ex vivo analysis. We overcame this obstacle to measure ex vivo T cell responses in CHB patients, by modifying the key variables of cell number and the peptide pulsing method to improve ex vivo detection of HBV-specific T cells.

Fluorescent-bait labelling for the ex vivo detection of HBV antigen-specific B cells

Nina Le Bert (1), Loghman Salimzadeh (1), Alice R. Burton (2), Mala K. Maini (2) Antonio Bertoletti (1)

(1) Emerging Infectious Diseases Program, Duke-NUS Medical School, Singapore (2) Division of Infection and Immunity, UCL, London, UK
Fluorescently conjugated antigen-bait systems have been extensively used to identify antigen-specific B cells and probe humoral immunity across different settings. Using this principle, HBV antigens are used to bind the B cell receptor (BCR), permitting an... [Read more]

Tetramer enrichment for HBV-specific CD8+ T cells

Kathrin Heim, Nina Hensel, Maike Hofmann, Rober Thimme

Department of Medicine II, University Hospital Freiburg, Freiburg, Germany
The frequency of HBV-specific CD8+ T cells can be very low, depending on the specificity. Therefore, they cannot always be detected by conventional ex vivo tetramer staining using 106 PBMCs. Tetramer-based magnetic enrichment enables the detection of these rare virus-spec... [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]