title: Alcelaphine herpesvirus 1 glycoprotein B: recombinant expression and antibody recognition.
abstract: The gammaherpesvirus alcelaphine herpesvirus 1 (AlHV-1) causes fatal malignant catarrhal fever (MCF) in susceptible species including cattle, but infects its reservoir host, wildebeest, without causing disease. Pathology in cattle may be influenced by virus-host cell interactions mediated by the virus glycoproteins. Cloning and expression of a haemagglutinin-tagged version of the AlHV-1 glycoprotein B (gB) was used to demonstrate that the AlHV-1-specific monoclonal antibody 12B5 recognised gB and that gB was the main component of the gp115 complex of AlHV-1, a glycoprotein complex of five components identified on the surface of AlHV-1 by immunoprecipitation and radiolabelling. Analysis of AlHV-1 virus particles showed that the native form of gB was detected by mAb 12B5 as a band of about 70 kDa, whilst recombinant gB expressed by transfected HEK293T cells appeared to be subject to additional cleavage and incomplete post-translational processing. Antibody 12B5 recognised an epitope on the N-terminal furin-cleaved fragment of gB on AlHV-1 virus particles. It could be used to detect recombinant and virus-expressed gB on western blots and on the surface of infected cells by flow cytometry, whilst recombinant gB was detected on the surface of transfected cells by immunofluorescence. Recombinant gB has potential as an antigen for ELISA detection of MCF virus infection and as a candidate vaccine antigen.
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Midlothian, EH26 0PZ, UK.|The Roslin Institute, University of Edinburgh, Midlothian, EH25 9RG, UK.
authors.forename: Inga
authors.initials: I
authors.lastname: Dry
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Midlothian, EH26 0PZ, UK.
authors.forename: Helen
authors.initials: H
authors.lastname: Todd
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Midlothian, EH26 0PZ, UK.
authors.forename: David
authors.initials: D
authors.lastname: Deane
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Midlothian, EH26 0PZ, UK.
authors.forename: Ann
authors.initials: A
authors.lastname: Percival
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Midlothian, EH26 0PZ, UK.
authors.forename: Kevin
authors.initials: K
authors.lastname: Mclean
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Midlothian, EH26 0PZ, UK.
authors.forename: Neil F
authors.initials: NF
authors.lastname: Inglis
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Midlothian, EH26 0PZ, UK.
authors.forename: Erin D T
authors.initials: ED
authors.lastname: Manson
authors.affiliation: School of Veterinary Medicine and Science, Nottingham University, Sutton Bonnington, Leicestershire, LE12 5RD, UK.
authors.forename: David M
authors.initials: DM
authors.lastname: Haig
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Midlothian, EH26 0PZ, UK.|Division of Medicine, Royal Free Campus, University College London, Rowland Hill Street, London, NW3 2PF, UK.
authors.forename: Shilpa
authors.initials: S
authors.lastname: Nayuni
authors.affiliation: Microbiology and Immunology, LSU Health Sciences Center, Shreveport, LA, USA.
authors.forename: Lindsey M
authors.initials: LM
authors.lastname: Hutt-Fletcher
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Midlothian, EH26 0PZ, UK.
authors.forename: Dawn M
authors.initials: DM
authors.lastname: Grant
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Midlothian, EH26 0PZ, UK.
authors.forename: Kathryn
authors.initials: K
authors.lastname: Bartley
authors.affiliation: Department of Infection Biology, University of Liverpool, Liverpool, L3 5RF, UK.
authors.forename: James P
authors.initials: JP
authors.lastname: Stewart
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Midlothian, EH26 0PZ, UK. george.russell@moredun.ac.uk.
authors.forename: George C
authors.identifier: http://orcid.org/0000-0002-1951-3215
authors.initials: GC
authors.lastname: Russell
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000276:immunology* / Q000302:isolation & purification; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis*; D018142:Gammaherpesvirinae / Q000737:chemistry / Q000276:immunology*; D006023:Glycoproteins / Q000032:analysis / Q000276:immunology*; D047468:Immunoprecipitation; D008304:Malignant Catarrh / Q000175:diagnosis*; D008565:Membrane Proteins / Q000032:analysis / Q000276:immunology; D011863:Radioimmunoassay; D015678:Viral Structural Proteins / Q000032:analysis / Q000276:immunology*; D014771:Virion / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D006023:Glycoproteins; D008565:Membrane Proteins; D015678:Viral Structural Proteins