PMID- 37931881
OWN - NLM
STAT- MEDLINE
DCOM- 20231216
LR  - 20240520
IS  - 1872-7492 (Electronic)
IS  - 0168-1702 (Print)
IS  - 0168-1702 (Linking)
VI  - 339
DP  - 2024 Jan 2
TI  - Studying longitudinal neutralising antibody levels against Equid herpesvirus 1 in 
      experimentally infected horses using a novel pseudotype based assay.
PG  - 199262
LID - S0168-1702(23)00224-1 [pii]
LID - 10.1016/j.virusres.2023.199262 [doi]
LID - 199262
AB  - 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  degrees 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.
CI  - Copyright (c) 2023. Published by Elsevier B.V.
FAU - Di Genova, Cecilia
AU  - Di Genova C
AD  - Viral 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 Kingdom.
FAU - Sutton, Gabrielle
AU  - Sutton G
AD  - LABEO Frank Duncombe, 14280 Saint-Contest, France; BIOTARGEN, Normandie Univ, 
      UNICAEN, 14000 Caen, France; Universite de Montreal, H3C 3J7 Montreal, Quebec, 
      Canada.
FAU - Paillot, Romain
AU  - Paillot R
AD  - LABEO 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 Kingdom.
FAU - Temperton, Nigel
AU  - Temperton N
AD  - Viral Pseudotype Unit, Medway School of Pharmacy, Universities of Kent and 
      Greenwich, Chatham Maritime, Kent ME4 4 TB, United Kingdom.
FAU - Pronost, Stephane
AU  - Pronost S
AD  - LABEO Frank Duncombe, 14280 Saint-Contest, France; BIOTARGEN, Normandie Univ, 
      UNICAEN, 14000 Caen, France.
FAU - Scott, Simon D
AU  - Scott SD
AD  - Viral Pseudotype Unit, Medway School of Pharmacy, Universities of Kent and 
      Greenwich, Chatham Maritime, Kent ME4 4 TB, United Kingdom. Electronic address: 
      s.d.scott@kent.ac.uk.
LA  - eng
PT  - Journal Article
DEP - 20231117
PL  - Netherlands
TA  - Virus Res
JT  - Virus research
JID - 8410979
RN  - 0 (Antibodies, Viral)
RN  - 0 (Antibodies, Neutralizing)
RN  - 0 (Glycoproteins)
SB  - IM
MH  - Humans
MH  - Animals
MH  - Horses
MH  - *Herpesvirus 1, Equid/genetics
MH  - HEK293 Cells
MH  - Antibodies, Viral
MH  - Antibodies, Neutralizing
MH  - *Herpesviridae Infections/veterinary
MH  - Glycoproteins
MH  - *Horse Diseases
MH  - *Herpesvirus 4, Equid/genetics
PMC - PMC10694342
OTO - NOTNLM
OT  - Equid herpesvirus 1
OT  - Lentiviral pseudotype virus
OT  - Neutralisation assay
OT  - Serology
COIS- Declaration of Competing Interest The authors declare that they have no known 
      competing financial interests or personal relationships that could have appeared 
      to influence the work reported in this paper.
EDAT- 2023/11/07 00:42
MHDA- 2023/12/17 13:19
PMCR- 2023/11/17
CRDT- 2023/11/06 19:17
PHST- 2023/08/24 00:00 [received]
PHST- 2023/11/02 00:00 [revised]
PHST- 2023/11/03 00:00 [accepted]
PHST- 2023/12/17 13:19 [medline]
PHST- 2023/11/07 00:42 [pubmed]
PHST- 2023/11/06 19:17 [entrez]
PHST- 2023/11/17 00:00 [pmc-release]
AID - S0168-1702(23)00224-1 [pii]
AID - 199262 [pii]
AID - 10.1016/j.virusres.2023.199262 [doi]
PST - ppublish
SO  - Virus Res. 2024 Jan 2;339:199262. doi: 10.1016/j.virusres.2023.199262. Epub 2023 
      Nov 17.