title: Yeast-secreted bovine herpesvirus type 1 glycoprotein D has authentic conformational structure and immunogenicity.
abstract: Bovine herpesvirus-1 (BHV-1) glycoprotein D (gD), an envelope glycoprotein, engenders mucosal and systemic immunity protecting cattle from viral infection. Production of gD with authentic immunogenicity is required for a subunit vaccine. We placed the truncated BHV-1 gD gene, lacking its putative transmembrane and cytoplasmic domains, under the control of the methanol-inducible AOX1 promoter in the yeast Pichia pastoris. Truncated BHV-1 gD (tgD) was efficiently secreted into the culture medium as a 68 kDa protein using either the yeast alpha prepro or native BHV-1 gD signal sequences. The yeast-secreted tgD had N-linked glycosylation and appears to have authentic conformational structure and immunogenicity based on the following observations A panel of monoclonal antibodies recognizing five neutralizing epitopes reacted with yeast tgD. Sera from yeast tgD-immunized mice immunoprecipitated native BHV-1 gD and neutralized BHV-1 infection in vitro. Yeast tgD competitively blocked all reaction between native gD and monospecific gD polyclonal sera from cattle. Based on these data, yeast-derived BHV-1 tgD is an excellent candidate for a subunit vaccine.
authors.affiliation: Department of Animal Health and Biomedical Sciences, University of Wisconsin-Madison 53706, USA.
authors.forename: X
authors.initials: X
authors.lastname: Zhu
authors.forename: S
authors.initials: S
authors.lastname: Wu
authors.forename: G J
authors.initials: GJ
authors.lastname: Letchworth
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D001667:Binding, Competitive; D002417:Cattle; D002460:Cell Line; D005822:Genetic Vectors; D006031:Glycosylation; D007242:Herpesvirus 1, Bovine / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D010843:Pichia / Q000235:genetics*; D011487:Protein Conformation; D021382:Protein Sorting Signals / Q000737:chemistry; D014614:Vaccines, Synthetic / Q000235:genetics / Q000276:immunology; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014765:Viral Vaccines / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D021382:Protein Sorting Signals; D014614:Vaccines, Synthetic; D014764:Viral Proteins; D014765:Viral Vaccines; C046998:bovine herpesvirus type-1 glycoproteins