Next Article in Journal
Detection of Campylobacter jejuni in Lizard Faeces from Central Australia Using Quantitative PCR
Next Article in Special Issue
Evolution and Divergence of H3N8 Equine Influenza Viruses Circulating in the United Kingdom from 2013 to 2015
Previous Article in Journal
Varicella-Zoster Virus Infectious Cycle: ER Stress, Autophagic Flux, and Amphisome-Mediated Trafficking
Previous Article in Special Issue
How to Meet the Last OIE Expert Surveillance Panel Recommendations on Equine Influenza (EI) Vaccine Composition: A Review of the Process Required for the Recombinant Canarypox-Based EI Vaccine
Article Menu

Export Article

Open AccessArticle
Pathogens 2016, 5(4), 68; doi:10.3390/pathogens5040068

The Optimisation of Pseudotyped Viruses for the Characterisation of Immune Responses to Equine Influenza Virus

1
Viral Pseudotype Unit, School of Pharmacy, University of Kent, Central Avenue, Chatham Maritime ME4 4TB, UK
2
School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington LE12 5RD, UK
*
Author to whom correspondence should be addressed.
Academic Editor: Lawrence S. Young
Received: 1 September 2016 / Revised: 20 November 2016 / Accepted: 4 December 2016 / Published: 15 December 2016
(This article belongs to the Special Issue Equine Influenza)
View Full-Text   |   Download PDF [1186 KB, uploaded 15 December 2016]   |  

Abstract

Pseudotyped viruses (PVs) produced by co-transfecting cells with plasmids expressing lentiviral core proteins and viral envelope proteins are potentially powerful tools for studying various aspects of equine influenza virus (EIV) biology. The aim of this study was to optimise production of equine influenza PVs. Co-transfection of the HAT protease to activate the haemagglutinin (HA) yielded a higher titre PV than TMPRSS2 with the HA from A/equine/Richmond/1/2007 (H3N8), whereas for A/equine/Newmarket/79 (H3N8), both proteases resulted in equivalent titres. TMPRSS4 was ineffective with the HA of either strain. There was also an inverse relationship between the amount of protease-expression plasmids and the PV titre obtained. Interestingly, the PV titre obtained by co-transfection of a plasmid encoding the cognate N8 NA was not as high as that generated by the addition of exogenous neuraminidase (NA) from Clostridium perfringens to allow the release of nascent PV particles. Finally, initial characterisation of the reliability of PV neutralisation tests (PVNTs) demonstrated good intra-laboratory repeatability. In conclusion, we have demonstrated that equine influenza PV production can be readily optimised to provide a flexible tool for studying EIV. View Full-Text
Keywords: equine influenza; pseudotyped virus; neutralisation assay equine influenza; pseudotyped virus; neutralisation assay
Figures

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

Scifeed alert for new publications

Never miss any articles matching your research from any publisher
  • Get alerts for new papers matching your research
  • Find out the new papers from selected authors
  • Updated daily for 49'000+ journals and 6000+ publishers
  • Define your Scifeed now

SciFeed Share & Cite This Article

MDPI and ACS Style

Scott, S.D.; Kinsley, R.; Temperton, N.; Daly, J.M. The Optimisation of Pseudotyped Viruses for the Characterisation of Immune Responses to Equine Influenza Virus. Pathogens 2016, 5, 68.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Pathogens EISSN 2076-0817 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top