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Alcelaphine herpesvirus 1 glycoprotein B: recombinant expression and antibody recognition

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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.

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Acknowledgments

This work was supported by the Scottish Government (MRI/09504); BBSRC (BBS/B/00468; BB/H008950) and the UK Department for International Development. We are grateful to the Moredun Research Institute Bioservices Division for their valuable contribution to these studies.

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Author notes

  1. Inga Dry

    Present address: The Roslin Institute, University of Edinburgh, Midlothian, EH25 9RG, UK

  2. Shilpa Nayuni

    Present address: Division of Medicine, Royal Free Campus, University College London, Rowland Hill Street, London, NW3 2PF, UK

Authors and Affiliations

  1. Moredun Research Institute, Pentlands Science Park, Midlothian, EH26 0PZ, UK

    Inga Dry, Helen Todd, David Deane, Ann Percival, Kevin Mclean, Neil F. Inglis, Erin D. T. Manson, Shilpa Nayuni, Dawn M. Grant, Kathryn Bartley & George C. Russell

  2. School of Veterinary Medicine and Science, Nottingham University, Sutton Bonnington, Leicestershire, LE12 5RD, UK

    David M. Haig

  3. Microbiology and Immunology, LSU Health Sciences Center, Shreveport, LA, USA

    Lindsey M. Hutt-Fletcher

  4. Department of Infection Biology, University of Liverpool, Liverpool, L3 5RF, UK

    James P. Stewart

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  1. Inga Dry
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  2. Helen Todd
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Correspondence to George C. Russell.

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Dry, I., Todd, H., Deane, D. et al. Alcelaphine herpesvirus 1 glycoprotein B: recombinant expression and antibody recognition. Arch Virol 161, 613–619 (2016). https://doi.org/10.1007/s00705-015-2701-y

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  • DOI: https://doi.org/10.1007/s00705-015-2701-y

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