title: Influence of bovine respiratory syncytial virus F glycoprotein N-linked glycans on in vitro expression and on antibody responses in BALB/c mice.
abstract: Bovine respiratory syncytial virus (BRSV) is an etiological component of the bovine respiratory tract disease complex. Infection with BRSV following vaccination, or re-infection following natural infection is common since protection is incomplete. The objectives of this study were to create plasmid DNA constructs encoding single or multiple N-glycosylation-site deletion BRSV fusion (F) proteins, and evaluate their expression in cell culture, and potential to induce anti-BRSV F antibody responses in BALB/c mice. Four plasmid DNAs were constructed, each encoding 1-4 N-glycosylation-site deletions: Gly4, Gly2/4, Gly1/2/4 and Gly1/2/3/4. Each of the N-glycosylation-site deletion BRSV F proteins were expressed in COS-7 cells following transfection with plasmid DNA. Inoculation of BALB/c mice with plasmid DNA, resulted in a significant anti-BRSV F IgG response to the wild-type (WT) F and glycosylation-site deletion protein Gly2/4. Gly2/4 elicited a higher antibody titer than the fully glycosylated WT F protein. Significant neutralizing antibody titers were detected following immunization with the Gly2/4 plasmid DNA. These glycosylation-site deletion BRSV F proteins will be useful to characterize the effects of glycosylation on immunogenicity in the natural host, and may lead to a new approach for the generation of BRSV vaccines.
authors.affiliation: Department of Veterinary and Biomedical Sciences, University of Nebraska, East Campus Loop and Fair Street, Lincoln, NE 68583-0905, USA.
authors.forename: Holly A
authors.initials: HA
authors.lastname: Klink
authors.forename: Ryan P
authors.initials: RP
authors.lastname: Brady
authors.forename: Christina L
authors.initials: CL
authors.lastname: Topliff
authors.forename: Kent M
authors.initials: KM
authors.lastname: Eskridge
authors.forename: Subramaniam
authors.initials: S
authors.lastname: Srikumaran
authors.forename: Clayton L
authors.initials: CL
authors.lastname: Kelling
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D002417:Cattle; D002460:Cell Line; D002522:Chlorocebus aethiops; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006031:Glycosylation; D007074:Immunoglobulin G / Q000097:blood; D051379:Mice; D008807:Mice, Inbred BALB C; D023421:Models, Animal; D009500:Neutralization Tests; D011134:Polysaccharides / Q000737:chemistry / Q000276:immunology*; D011499:Protein Processing, Post-Translational; D018112:Respiratory Syncytial Virus, Bovine / Q000276:immunology*; D019444:Vaccines, DNA / Q000276:immunology; D014760:Viral Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D007074:Immunoglobulin G; D011134:Polysaccharides; D019444:Vaccines, DNA; D014760:Viral Fusion Proteins; D014765:Viral Vaccines