Skip to main content

Research Repository

See what's under the surface

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.

Authors

Inga Dry

Helen Todd

David Deane

Ann Percival

Kevin Mclean

Neil F. Inglis

Erin D. T. Manson

David M. Haig

Shilpa Nayuni

Lindsey M. Hutt-Fletcher

Dawn M. Grant

Kathryn Bartley

James P. Stewart

George C. Russell



Contributors

I Dry
Researcher

H Todd
Researcher

D Deane
Researcher

A Percival
Researcher

K Mclean
Researcher

NF Inglis
Researcher

ED Manson
Researcher

DM Haig
Researcher

S Nayuni
Researcher

LM Hutt-Fletcher
Researcher

DM Grant
Researcher

K Bartley
Researcher

JP Stewart
Researcher

GC. Russell
Researcher

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.

Journal Article Type Article
Publication Date 2016-03
Journal Archives of Virology
Print ISSN 0304-8608
Electronic ISSN 1432-8798
Publisher BMC
Peer Reviewed Peer Reviewed
Volume 161
Issue 3
Pages 613-619
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
DOI https://doi.org/10.1007/s00705-015-2701-y
Keywords HEK293T Cell Malignant Catarrhal Fever Transfected HEK293T Cell Furin Cleavage Monoclonal Antibody 12B5
Publisher URL https://link.springer.com/article/10.1007%2Fs00705-015-2701-y




Downloadable Citations