Alcelaphine herpesvirus 1 glycoprotein B: recombinant expression and antibody recognition
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.
Neil F. Inglis
Erin D. T. Manson
David M. Haig
Lindsey M. Hutt-Fletcher
Dawn M. Grant
James P. Stewart
George C. Russell
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.
|Journal Article Type||Article|
|Journal||Archives of Virology|
|Peer Reviewed||Peer Reviewed|
|Institution Citation||Dry, I., Todd, H., Deane, D., Percival, A., Mclean, K., Inglis, N. F., …Russell, G. C. (2016). Alcelaphine herpesvirus 1 glycoprotein B: recombinant expression and antibody recognition. Archives of Virology, 161(3), 613-619. doi:10.1007/s00705-015-2701-y|
|Keywords||HEK293T Cell Malignant Catarrhal Fever Transfected HEK293T Cell Furin Cleavage Monoclonal Antibody 12B5|