The optimisation of Pseudotyped viruses for the characterisation of immune responses to equine influenza virus
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 in...
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MDPI
2016
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| Online Access: | https://eprints.nottingham.ac.uk/39687/ |
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| author | Scott, Simon D. Kinsley, Rebecca Temperton, Nigel Daly, Janet M. |
| author_facet | Scott, Simon D. Kinsley, Rebecca Temperton, Nigel Daly, Janet M. |
| author_sort | Scott, Simon D. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-14T19:39:18Z |
| format | Article |
| id | nottingham-39687 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:39:18Z |
| publishDate | 2016 |
| publisher | MDPI |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-396872020-05-04T18:25:51Z https://eprints.nottingham.ac.uk/39687/ The optimisation of Pseudotyped viruses for the characterisation of immune responses to equine influenza virus Scott, Simon D. Kinsley, Rebecca Temperton, Nigel Daly, Janet M. 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. MDPI 2016-12-15 Article PeerReviewed Scott, Simon D., Kinsley, Rebecca, Temperton, Nigel and Daly, Janet M. (2016) The optimisation of Pseudotyped viruses for the characterisation of immune responses to equine influenza virus. Pathogens, 5 (4). 68/1-68/8. ISSN 2076-0817 equine influenza; pseudotyped virus; neutralisation assay http://www.mdpi.com/2076-0817/5/4/68 doi:10.3390/pathogens5040068 doi:10.3390/pathogens5040068 |
| spellingShingle | equine influenza; pseudotyped virus; neutralisation assay Scott, Simon D. Kinsley, Rebecca Temperton, Nigel Daly, Janet M. The optimisation of Pseudotyped viruses for the characterisation of immune responses to equine influenza virus |
| title | The optimisation of Pseudotyped viruses for the characterisation of immune responses to equine influenza virus |
| title_full | The optimisation of Pseudotyped viruses for the characterisation of immune responses to equine influenza virus |
| title_fullStr | The optimisation of Pseudotyped viruses for the characterisation of immune responses to equine influenza virus |
| title_full_unstemmed | The optimisation of Pseudotyped viruses for the characterisation of immune responses to equine influenza virus |
| title_short | The optimisation of Pseudotyped viruses for the characterisation of immune responses to equine influenza virus |
| title_sort | optimisation of pseudotyped viruses for the characterisation of immune responses to equine influenza virus |
| topic | equine influenza; pseudotyped virus; neutralisation assay |
| url | https://eprints.nottingham.ac.uk/39687/ https://eprints.nottingham.ac.uk/39687/ https://eprints.nottingham.ac.uk/39687/ |