Immunization With Fc-Based Recombinant Epstein-Barr Virus gp350 Elicits Potent Neutralizing Humoral Immune Response in a BALB/c Mice Model.

Bingchun Zhao,Musheng Zeng,Qisheng Feng,Haojiong Zhang,Xiao Zhang,Claude Krummenacher,Lin Feng,Yixin Zeng,Shuo Song,Miao Xu,Ling Gao,Yuting Xu

doi:10.3389/fimmu.2018.00932

Abstract

Epstein–Barr virus (EBV) was the first human virus proved to be closely associated with tumor development, such as lymphoma, nasopharyngeal carcinoma, and EBV-associated gastric carcinoma. Despite many efforts to develop prophylactic vaccines against EBV infection and diseases, no candidates have succeeded in effectively blocking EBV infection in clinical trials. Previous investigations showed that EBV gp350 plays a pivotal role in the infection of B-lymphocytes. Nevertheless, using monomeric gp350 proteins as antigens has not been effective in preventing infection. Multimeric forms of the antigen are more potently immunogenic than monomers; however, the multimerization elements used in previous constructs are not approved for human clinical trials. To prepare a much-needed EBV prophylactic vaccine that is potent, safe, and applicable, we constructed an Fc-based form of gp350 to serve as a dimeric antigen. Here, we show that the Fc-based gp350 antigen exhibits dramatically enhanced immunogenicity compared with wild-type gp350 protein. The complete or partial gp350 ectodomain was fused with the mouse IgG2a Fc domain. Fusion with the Fc domain did not impair gp350 folding, binding to a conformation-dependent neutralizing antibody (nAb) and binding to its receptor by enzyme-linked immunosorbent assay and surface plasmon resonance. Specific antibody titers against gp350 were notably enhanced by immunization with gp350-Fc dimers compared with gp350 monomers. Furthermore, immunization with gp350-Fc fusion proteins elicited potent nAbs against EBV. Our data strongly suggest that an EBV gp350 vaccine based on Fc fusion proteins may be an efficient candidate to prevent EBV infection in clinical applications.

Highlights

Epstein–Barr virus (EBV), known as human herpesvirus 4, is a highly prevalent virus from the Herpesviridae γ subfamily (1)
For purification of soluble gp350-ECD123, a 6× His tag was added at the C-terminus (Figure 1A)
We showed that the Fc portion led to the dimerization of the normally monomeric gp[350]

Summary

Introduction

Epstein–Barr virus (EBV), known as human herpesvirus 4, is a highly prevalent virus from the Herpesviridae γ subfamily (1). EBV is the first virus reported to cause tumorigenesis in infected humans (1–3). It is widely spread among human populations, and over 95% of adults carry latent EBV worldwide (4, 5). EBV establishes lifelong latency in infected individuals. The latent status of EBV is closely linked to the development and progression of many diseases, such as infectious mononucleosis (IM), nasopharyngeal carcinoma (NPC), posttransplant lymphoproliferative disease, EBV-related gastric carcinoma, and Hodgkin lymphoma (7–14). Latent EBV infection was recently shown to accelerate the decay of childhood measles and rubella vaccine responses (15). Development of a successful EBV vaccine is difficult due to (i) the lack of appropriate animal models for evaluation of infection, (ii) the poor protection of previous vaccine candidates, and (iii) inadequate investment in vaccine development (16–18)

Methods

Results

Conclusion