Studying longitudinal neutralising antibody levels against Equid herpesvirus 1 in experimentally infected horses using a novel pseudotype based assay
Infection with equid herpesvirus 1 (EHV-1), a DNA virus of the Herpesviridae family represents a significant welfare issue in horses and a great impact on the equine industry. During EHV-1 infection, entry of the virus into different cell types is complex due to the presence of twelve glycoproteins...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2024-01-01
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| Series: | Virus Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0168170223002241 |
| _version_ | 1827739891790774272 |
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| author | Cecilia Di Genova Gabrielle Sutton Romain Paillot Nigel Temperton Stéphane Pronost Simon D. Scott |
| author_facet | Cecilia Di Genova Gabrielle Sutton Romain Paillot Nigel Temperton Stéphane Pronost Simon D. Scott |
| author_sort | Cecilia Di Genova |
| collection | DOAJ |
| description | Infection with equid herpesvirus 1 (EHV-1), a DNA virus of the Herpesviridae family represents a significant welfare issue in horses and a great impact on the equine industry. During EHV-1 infection, entry of the virus into different cell types is complex due to the presence of twelve glycoproteins (GPs) on the viral envelope. To investigate virus entry mechanisms, specific combinations of GPs were pseudotyped onto lentiviral vectors. Pseudotyped virus (PV) particles bearing gB, gD, gH and gL were able to transduce several target cell lines (HEK293T/17, RK13, CHO-K1, FHK-Tcl3, MDCK I & II), demonstrating that these four EHV-1 glycoproteins are both essential and sufficient for cell entry. The successful generation of an EHV-1 PV permitted development of a PV neutralisation assay (PVNA). The efficacy of the PVNA was tested by measuring the level of neutralising serum antibodies from EHV-1 experimentally infected horses (n = 52) sampled in a longitudinal manner. The same sera were assessed using a conventional EHV-1 virus neutralisation (VN) assay, exhibiting a strong correlation (r = 0.82) between the two assays. Furthermore, PVs routinely require -80 °C for long term storage and a dry ice cold-chain during transport, which can impede dissemination and utilisation in other stakeholder laboratories. Consequently, lyophilisation of EHV-1 PVs was conducted to address this issue. PVs were lyophilised and pellets either reconstituted immediately or stored under various temperature conditions for different time periods. The recovery and functionality of these lyophilised PVs was compared with standard frozen aliquots in titration and neutralisation tests. Results indicated that lyophilisation could be used to stably preserve such complex herpesvirus pseudotypes, even after weeks of storage at room temperature, and that reconstituted EHV-1 PVs could be successfully employed in antibody neutralisation tests. |
| first_indexed | 2024-03-11T03:13:32Z |
| format | Article |
| id | doaj.art-7f303ac92f9146d3a5c4e5c216d23865 |
| institution | Directory Open Access Journal |
| issn | 1872-7492 |
| language | English |
| last_indexed | 2024-03-11T03:13:32Z |
| publishDate | 2024-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Virus Research |
| spelling | doaj.art-7f303ac92f9146d3a5c4e5c216d238652023-11-18T04:28:07ZengElsevierVirus Research1872-74922024-01-01339199262Studying longitudinal neutralising antibody levels against Equid herpesvirus 1 in experimentally infected horses using a novel pseudotype based assayCecilia Di Genova0Gabrielle Sutton1Romain Paillot2Nigel Temperton3Stéphane Pronost4Simon D. Scott5Viral Pseudotype Unit, Medway School of Pharmacy, Universities of Kent and Greenwich, Chatham Maritime, Kent ME4 4 TB, United Kingdom; Animal and Plant Health Agency (APHA), Weybridge, Surrey KT15 3NB, United KingdomLABÉO Frank Duncombe, 14280 Saint-Contest, France; BIOTARGEN, Normandie Univ, UNICAEN, 14000 Caen, France; Université de Montréal, H3C 3J7 Montreal, Quebec, CanadaLABÉO Frank Duncombe, 14280 Saint-Contest, France; BIOTARGEN, Normandie Univ, UNICAEN, 14000 Caen, France; School of Equine and Veterinary Physiotherapy, Writtle University College, Writtle, Chelmsford, Essex CM1 3RR, United KingdomViral Pseudotype Unit, Medway School of Pharmacy, Universities of Kent and Greenwich, Chatham Maritime, Kent ME4 4 TB, United KingdomLABÉO Frank Duncombe, 14280 Saint-Contest, France; BIOTARGEN, Normandie Univ, UNICAEN, 14000 Caen, FranceViral Pseudotype Unit, Medway School of Pharmacy, Universities of Kent and Greenwich, Chatham Maritime, Kent ME4 4 TB, United Kingdom; Corresponding author.Infection with equid herpesvirus 1 (EHV-1), a DNA virus of the Herpesviridae family represents a significant welfare issue in horses and a great impact on the equine industry. During EHV-1 infection, entry of the virus into different cell types is complex due to the presence of twelve glycoproteins (GPs) on the viral envelope. To investigate virus entry mechanisms, specific combinations of GPs were pseudotyped onto lentiviral vectors. Pseudotyped virus (PV) particles bearing gB, gD, gH and gL were able to transduce several target cell lines (HEK293T/17, RK13, CHO-K1, FHK-Tcl3, MDCK I & II), demonstrating that these four EHV-1 glycoproteins are both essential and sufficient for cell entry. The successful generation of an EHV-1 PV permitted development of a PV neutralisation assay (PVNA). The efficacy of the PVNA was tested by measuring the level of neutralising serum antibodies from EHV-1 experimentally infected horses (n = 52) sampled in a longitudinal manner. The same sera were assessed using a conventional EHV-1 virus neutralisation (VN) assay, exhibiting a strong correlation (r = 0.82) between the two assays. Furthermore, PVs routinely require -80 °C for long term storage and a dry ice cold-chain during transport, which can impede dissemination and utilisation in other stakeholder laboratories. Consequently, lyophilisation of EHV-1 PVs was conducted to address this issue. PVs were lyophilised and pellets either reconstituted immediately or stored under various temperature conditions for different time periods. The recovery and functionality of these lyophilised PVs was compared with standard frozen aliquots in titration and neutralisation tests. Results indicated that lyophilisation could be used to stably preserve such complex herpesvirus pseudotypes, even after weeks of storage at room temperature, and that reconstituted EHV-1 PVs could be successfully employed in antibody neutralisation tests.http://www.sciencedirect.com/science/article/pii/S0168170223002241Equid herpesvirus 1Lentiviral pseudotype virusSerologyNeutralisation assay |
| spellingShingle | Cecilia Di Genova Gabrielle Sutton Romain Paillot Nigel Temperton Stéphane Pronost Simon D. Scott Studying longitudinal neutralising antibody levels against Equid herpesvirus 1 in experimentally infected horses using a novel pseudotype based assay Virus Research Equid herpesvirus 1 Lentiviral pseudotype virus Serology Neutralisation assay |
| title | Studying longitudinal neutralising antibody levels against Equid herpesvirus 1 in experimentally infected horses using a novel pseudotype based assay |
| title_full | Studying longitudinal neutralising antibody levels against Equid herpesvirus 1 in experimentally infected horses using a novel pseudotype based assay |
| title_fullStr | Studying longitudinal neutralising antibody levels against Equid herpesvirus 1 in experimentally infected horses using a novel pseudotype based assay |
| title_full_unstemmed | Studying longitudinal neutralising antibody levels against Equid herpesvirus 1 in experimentally infected horses using a novel pseudotype based assay |
| title_short | Studying longitudinal neutralising antibody levels against Equid herpesvirus 1 in experimentally infected horses using a novel pseudotype based assay |
| title_sort | studying longitudinal neutralising antibody levels against equid herpesvirus 1 in experimentally infected horses using a novel pseudotype based assay |
| topic | Equid herpesvirus 1 Lentiviral pseudotype virus Serology Neutralisation assay |
| url | http://www.sciencedirect.com/science/article/pii/S0168170223002241 |
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