Please use this identifier to cite or link to this item: http://hdl.handle.net/1893/22759
Appears in Collections:Aquaculture Journal Articles
Peer Review Status: Refereed
Title: Expression of immunogenic structural proteins of cyprinid herpesvirus 3 in vitro assessed using immunofluorescence
Authors: Monaghan, Sean
Thompson, Kimberly
Bron, James
Bergmann, Sven
Jung, Tae
Aoki, Takashi
Muir, Katherine
Dauber, Malte
Reiche, Sven
Chee, Diana
Chong, Shin M
Chen, Jing
Adams, Alexandra
Contact Email: s.j.monaghan@stir.ac.uk
Issue Date: 8-Jan-2016
Publisher: BioMed Central
Citation: Monaghan S, Thompson K, Bron J, Bergmann S, Jung T, Aoki T, Muir K, Dauber M, Reiche S, Chee D, Chong SM, Chen J & Adams A (2016) Expression of immunogenic structural proteins of cyprinid herpesvirus 3 in vitro assessed using immunofluorescence, Veterinary Research, 47, Art. No.: 8.
Abstract: Cyprinid herpesvirus 3 (CyHV-3), also called koi herpesvirus (KHV), is the aetiological agent of a fatal disease in carp and koi (Cyprinus carpioL.), referred to as koi herpesvirus disease. The virus contains at least 40 structural proteins, of which few have been characterised with respect to their immunogenicity. Indirect immunofluorescence assays (IFAs) using two epitope-specific monoclonal antibodies (MAbs) were used to examine the expression kinetics of two potentially immunogenic and diagnostically relevant viral antigens, an envelope glycoprotein and a capsid-associated protein. The rate of expression of these antigens was determined following a time-course of infection in two CyHV-3 susceptible cell lines. The results were quantified using an IFA, performed in microtitre plates, and image analysis was used to analyse confocal micrographs, enabling measurement of differential virus-associated fluorescence and nucleus-associated fluorescence from stacks of captured scans. An 8-tenfold increase in capsid-associated protein expression was observed during the first 5days post-infection compared to a ≤2-fold increase in glycoprotein expression. A dominant protein of ~100kDa reacted with the capsid-associated MAb (20F10) in western blot analysis. This band was also recognised by sera obtained from carp infected with CyHV-3, indicating that this capsid-associated protein is produced in abundance during infection in vitro and is immunogenic to carp. Mass spectrometry carried out on this protein identified it as a previously uncharacterised product of open reading frame 84. This abundantly expressed and immunogenic capsid-associated antigen may be a useful candidate for KHV serological diagnostics.
Type: Journal Article
URI: http://hdl.handle.net/1893/22759
DOI Link: http://dx.doi.org/10.1186/s13567-015-0297-6
Rights: © 2016 Monaghan et al. Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Affiliation: Aquaculture
Aquaculture
Aquaculture
Friedrich-Loeffler Institut
Gyeongsang National University
Tokyo University of Fisheries
Aquaculture
Friedrich-Loeffler Institut
Friedrich-Loeffler Institut
University of Stirling
Agri-Food and Veterinary Authority of Singapore
Agri-Food and Veterinary Authority of Singapore
Aquaculture

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