title: A novel tetrameric gp350 1-470 as a potential Epstein-Barr virus vaccine.
abstract: Infectious mononucleosis and B-cell transformation in response to infection with Epstein-Barr virus (EBV) is dependent upon binding of the EBV envelope glycoprotein gp350 to CD21 on B-cells. Gp350-specific antibody comprises most of the EBV neutralizing activity in the serum of infected patients, making this protein a promising target antigen for a prophylactic EBV vaccine. We describe a novel, tetrameric gp350-based vaccine that exhibits markedly enhanced immunogenicity relative to its monomeric counterpart. Plasmid DNA was constructed for synthesis, within transfected CHO cells, of a tetrameric, truncated (a.a. 1-470) gp350 protein (gp350(1-470)). Tetrameric gp350(1-470) induced ≈ 20-fold higher serum titers of gp350(1-470)-specific IgG and >19-fold enhancements in neutralizing titers at the highest dose, and was >25-fold more immunogenic on a per-weight basis than monomeric gp350(1-470). Further, epidermal immunization with plasmid DNA encoding gp350(1-470) tetramer induced 8-fold higher serum titers of gp350(1-470)-specific IgG relative to monomer. Tetrameric gp350(1-470) binding to human CD21 was >24-fold more efficient on a per-weight basis than monomer, but neither tetramer nor monomer mediated polyclonal human B-cell activation. Finally, the introduction of strong, universal tetanus toxoid (TT)-specific CD4+ T-cell epitopes into the tetrameric gp350(1-470) had no effect on the gp350(1-470)-specific IgG response in naïve mice, and resulted in suppressed gp350(1-470)-specific IgG responses in TT-primed mice. Collectively, these data suggest that tetrameric gp350(1-470) is a potentially promising candidate for testing as a prophylactic EBV vaccine, and that protein multimerization, using the approach described herein, is likely to be clinically relevant for enhancing the immunogenicity of other proteins of vaccine interest.
authors.affiliation: Department of Pathology, Uniformed Services University of Health Sciences, Bethesda, MD 20814, USA.
authors.forename: Xinle
authors.initials: X
authors.lastname: Cui
authors.forename: Zhouhong
authors.initials: Z
authors.lastname: Cao
authors.forename: Goutam
authors.initials: G
authors.lastname: Sen
authors.forename: Gouri
authors.initials: G
authors.lastname: Chattopadhyay
authors.forename: Deborah H
authors.initials: DH
authors.lastname: Fuller
authors.forename: James T
authors.initials: JT
authors.lastname: Fuller
authors.forename: Dustin M
authors.initials: DM
authors.lastname: Snapper
authors.forename: Andrew L
authors.initials: AL
authors.lastname: Snow
authors.forename: James J
authors.initials: JJ
authors.lastname: Mond
authors.forename: Clifford M
authors.initials: CM
authors.lastname: Snapper
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology; D020031:Epstein-Barr Virus Infections / Q000517:prevention & control*; D053061:Herpes Zoster Vaccine / Q000276:immunology*; D004854:Herpesvirus 4, Human / Q000276:immunology*; D006801:Humans; D007114:Immunization; D051379:Mice; D010957:Plasmids; D014162:Transfection; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D000914:Antibodies, Viral; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D018984:Epitopes, T-Lymphocyte; D053061:Herpes Zoster Vaccine; D014763:Viral Matrix Proteins