Use of ELISpot assay to study HBs-specific B cell responses in vaccinated and HBV infected humans

Yong Liu,Chen Tian,Shan Lu,Chao Wu,Shixia Wang,Juan Xia,Rui Huang,Yuxin Chen,Xiang-An Zhao,Yang Li,Guiyang Wang

doi:10.1038/s41426-018-0034-0

Yong Liu, Chen Tian + Show 9 more

Open Access

https://doi.org/10.1038/s41426-018-0034-0

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Abstract

Hepatitis B surface antibody (HBsAb) plays a critical role in protecting against infection of hepatitis B virus (HBV) and were extensively studied in literature. At the same time, the status of hepatitis B surface antigen (HBs)-specific B cells in both vaccinated and HBV infected people received limited attention. In the current study, we adopted a highly specific B-cell Enzyme Linked ImmunoSpot (ELISpot) assay to analyze HBs-specific B cells in various clinical settings: healthy individuals with the history of HBV vaccination before and after receiving an extra HBV vaccine boost, people chronically infected with HBV (CHB) in various clinical stages, with or without a particular type anti-viral treatment, or whether receiving a dose of HBV vaccine. In all of these cases, B-cell ELISpot assay was used effectively in enumerating the frequency of HBs-specific B cells. While the focus of the current report was to establish the utility of this assay for HBV research, a number of interesting observations were made in this pilot study based on the profiles and dynamics of HBs-specific B cells in various conditions. Such information is useful to guide the future work in designing novel therapeutic strategies against CHB.

Highlights

Introduction HepatitisB virus (HBV) infection remains a major health threat in many parts of the world especially in developing countries including countries with large human populations such as China[1]
A sensitive B-cell Enzyme Linked ImmunoSpot (ELISpot) assay was used in the current study to detect and enumerate hepatitis B surface antigen (HBs)-specific memory B cells from human peripheral blood mononuclear cells (PBMCs) among healthy adult vaccinees
Among people who had been vaccinated with hepatitis B virus (HBV) vaccine in the past, HBs-specific memory B cells would be detected by the HBs-specific B cell ELISpot assay

Summary

Introduction

B virus (HBV) infection remains a major health threat in many parts of the world especially in developing countries including countries with large human populations such as China[1]. While the introduction of a subunit protein-based HBV vaccine has greatly reduced the rate of new human infections in the last several decades, the population of people who either were infected before the introduction of HBV vaccine or missed the chance of getting vaccinated is still quite large[2]. The significance of vaccine-induced antibody (HBsAb) responses against HBV surface antigen (HBsAg) in protecting against HBV infection has been well established[5,6]. How to break body’s immune tolerance to elicit protective HBsAb that can control the viral infection is a major challenge in HBV research

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