Expression and immunogenic characterization of recombinant gp350 for developing a subunit vaccine against Epstein-Barr virus

Abstract

Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus that is linked to the development of various malignancies. There is an urgent need for effective vaccines against EBV. EBV envelope glycoprotein gp350 is an attractive candidate for a prophylactic vaccine. This study was undertaken to produce the truncated (codons 1-443) gp350 protein (gp350(1-443)) in Pichia pastoris and evaluate its immunogenicity. The gp350(1-443) protein was expressed as a secretory protein with an N-terminal His-tag in P. pastoris and purified through Ni-NTA chromatography. Immunization with the recombinant gp350(1-443) could elicit high levels of gp350(1-443)-specific antibodies in mice. Moreover, gp350(1-443)-immunized mice developed strong lymphoproliferative and Th1/Th2 cytokine responses. Furthermore, the recombinant gp350(1-443) could stimulate CD4(+) and CD8(+) T cell responses in vaccinated mice. Collectively, these findings demonstrated that the yeast-expressed gp350(1-443) retained strong immunogenicity. This study will provide a useful source for developing EBV subunit vaccine candidates.

Recent advances in the production of recombinant subunit vaccines inPichia pastoris

“…The secretory form of truncated gp350 (gp350 1-443 , codons 1-443) has the potential to be developed into EBV subunit vaccine. 63 Moreover, the recombinant protein was able to induce higher levels of specific antibodies than synthetic gp350 peptides. 64 EBV nuclear antigen 1 (EBNA1) is thought to be a promising antigen for EBV therapeutic vaccines.…”

Sneaking Out for Happy Hour: Yeast-Based Approaches to Explore and Modulate Immune Response and Immune Evasion

“…Indeed, EBV is recognized as an important vaccine target, in particular for cancer prevention and several efforts has been undertaken, most of them being focused on the EBV glycoprotein gp350 which is the most abundant glycoprotein on the virus and on virus-infected cells and the main target of naturally occurring neutralizing antibodies [23,160]. This way the yeast Pichia pastoris , which is a popular workhorse for production of recombinant proteins, has been exploited to express truncated forms of EBNA1 or gp350, both as secretory proteins with an N-terminal histidine tag that allow their purification [161,162]. In both cases, the truncated EBV proteins retained good immunogenicity and could therefore represent a useful source for developing EBV vaccine candidates.…”

Evaluation of glycoprotein Ov8 as a potential antigen for an OvHV-2-specific diagnostic assay

“…ORF Ov8 is localized in the genome between ORFs 50 and 52 [ 12 ]. In some gammaherpesviruses, genes localized between ORFs 50 and 52, such as Epstein-Barr virus gp350/220, Kaposi’s sarcoma-associated herpes virus K8.1, and Murid herpesvirus-4 gp150, encode immunogenic glycoproteins that have been used as target antigens for serological assays [ 14 – 17 ]. In this report, we tested the hypothesis that Ov8 is antigenic and induces OvHV-2-specific antibodies.…”