Alcelaphine herpesvirus 1 glycoprotein B: recombinant expression and antibody recognition

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 vir...

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Bibliographic Details
Published in:Archives of virology 2016-03, Vol.161 (3), p.613-619
Main Authors: Dry, Inga, Todd, Helen, Deane, David, Percival, Ann, Mclean, Kevin, Inglis, Neil F, Manson, Erin D. T, Haig, David M, Nayuni, Shilpa, Hutt-Fletcher, Lindsey M, Grant, Dawn M, Bartley, Kathryn, Stewart, James P, Russell, George C
Format: Article
Language:English
Subjects:
Alcelaphine herpesvirus 1
Animals
Antibodies, Monoclonal - immunology
Antibodies, Monoclonal - isolation & purification
Antibodies, Viral - immunology
Antibodies, Viral - isolation & purification
Antigens
Biomedical and Life Sciences
Biomedicine
Cattle
Cattle Diseases - diagnosis
enzyme-linked immunosorbent assay
epitopes
Fever
flow cytometry
fluorescent antibody technique
Gammaherpesvirinae - chemistry
Gammaherpesvirinae - immunology
Gammaherpesvirus
Genomes
Glycoproteins
Glycoproteins - analysis
Glycoproteins - immunology
Herpes viruses
Immunoprecipitation
Infections
Infectious Diseases
Malignant Catarrh - diagnosis
malignant catarrhal fever
Medical Microbiology
Membrane Proteins - analysis
Membrane Proteins - immunology
Monoclonal antibodies
Original Article
Polypeptides
precipitin tests
Radioimmunoassay
Vaccines
Viral Structural Proteins - analysis
Viral Structural Proteins - immunology
virion
Virion - chemistry
Virology
viruses
Western blotting
wildebeest
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Summary: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.
ISSN:0304-8608
1432-8798
DOI:10.1007/s00705-015-2701-y