title: Vaccinia virus-expressed bovine ephemeral fever virus G but not G(NS) glycoprotein induces neutralizing antibodies and protects against experimental infection.
abstract: Two related glycoproteins (G and G(NS)) encoded in the bovine ephemeral fever virus (BEFV) genome were expressed from recombinant vaccinia viruses (rVV). Both proteins were detected in lysates of rVV-infected cells by labelling with D-[6-3H]glucosamine or by immuno-blotting. The recombinant G protein (mol. mass 79 kDa) appeared slightly smaller than the native G protein but reacted with monoclonal antibodies directed against all defined neutralizing antigenic sites (G1, G2, G3a, G3b and G4). The recombinant G(NS) protein (mol. mass 90kDa) was identical in size to the native G(NS) protein and failed to react by immuno-fluorescence with anti-G protein monoclonal or poly-clonal antibodies. Antisera raised in rabbits against rVV-G or rVV-G(NS) both reacted strongly by immuno-fluorescence and immuno-electron microscopy with BEFV-infected cells. The G protein was localized intracellularly in the endoplasmic reticulum/Golgi complex and at the cell surface associated with budding and mature virus particles. The G(NS) protein also localized intracellularly in the endoplasmic reticulum/Golgi complex; however, at the cell surface it was associated with amorphous structures and not with budding or mature virions. Rabbits vaccinated with rVV-G developed high levels of antibodies which neutralized BEFV grown in either mammalian or insect cells. Cattle vaccinated with rVV-G also produced neutralizing antibodies and were protected against experimental BEFV infection. In contrast, rVV-G(NS) vaccinated rabbits and cattle failed to produce neutralizing antibodies and, after challenge, BEFV was isolated from two-thirds of the vaccinated cattle.
authors.affiliation: CSIRO Australian Animal Health Laboratory, Geelong, Victoria, Australia.
authors.forename: C
authors.initials: C
authors.lastname: Hertig
authors.forename: A D
authors.initials: AD
authors.lastname: Pye
authors.forename: A D
authors.initials: AD
authors.lastname: Hyatt
authors.forename: S S
authors.initials: SS
authors.lastname: Davis
authors.forename: S M
authors.initials: SM
authors.lastname: McWilliam
authors.forename: H G
authors.initials: HG
authors.lastname: Heine
authors.forename: P J
authors.initials: PJ
authors.lastname: Walker
authors.forename: D B
authors.initials: DB
authors.lastname: Boyle
mesh_terms: D000330:Aedes / Q000166:cytology; D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis* / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D006224:Cricetinae; D004279:DNA, Viral; D004810:Ephemeral Fever / Q000276:immunology / Q000517:prevention & control*; D018115:Ephemeral Fever Virus, Bovine / Q000235:genetics / Q000276:immunology* / Q000648:ultrastructure; D005822:Genetic Vectors / Q000235:genetics / Q000276:immunology; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D011817:Rabbits; D014614:Vaccines, Synthetic / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014616:Vaccinia virus / Q000235:genetics / Q000276:immunology; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014765:Viral Vaccines / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D004279:DNA, Viral; D006023:Glycoproteins; D014614:Vaccines, Synthetic; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins; D014765:Viral Vaccines; C077307:BEFV G protein, Bovine ephemeral fever virus; C077308:GNS protein, Bovine ephemeral fever virus