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.

journal_name

Arch Virol

journal_title

Archives of virology

authors

Dry I,Todd H,Deane D,Percival A,Mclean K,Inglis NF,Manson ED,Haig DM,Nayuni S,Hutt-Fletcher LM,Grant DM,Bartley K,Stewart JP,Russell GC

doi

10.1007/s00705-015-2701-y

subject

Has Abstract

pub_date

2016-03-01 00:00:00

pages

613-9

issue

3

eissn

0304-8608

issn

1432-8798

pii

10.1007/s00705-015-2701-y

journal_volume

161

pub_type

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