title: Efficient generation of vesicular stomatitis virus (VSV)-pseudotypes bearing morbilliviral glycoproteins and their use in quantifying virus neutralising antibodies.
abstract: Morbillivirus neutralising antibodies are traditionally measured using either plaque reduction neutralisation tests (PRNTs) or live virus microneutralisation tests (micro-NTs). While both test formats provide a reliable assessment of the strength and specificity of the humoral response, they are restricted by the limited number of viral strains that can be studied and often present significant biological safety concerns to the operator. In this study, we describe the adaptation of a replication-defective vesicular stomatitis virus (VSVΔG) based pseudotyping system for the measurement of morbillivirus neutralising antibodies. By expressing the haemagglutinin (H) and fusion (F) proteins of canine distemper virus (CDV) on VSVΔG pseudotypes bearing a luciferase marker gene, neutralising antibody titres could be measured rapidly and with high sensitivity. Further, by exchanging the glycoprotein expression construct, responses against distinct viral strains or species may be measured. Using this technique, we demonstrate cross neutralisation between CDV and peste des petits ruminants virus (PPRV). As an example of the value of the technique, we demonstrate that UK dogs vary in the breadth of immunity induced by CDV vaccination; in some dogs the neutralising response is CDV-specific while, in others, the neutralising response extends to the ruminant morbillivirus PPRV. This technique will facilitate a comprehensive comparison of cross-neutralisation to be conducted across the morbilliviruses.
authors.affiliation: MRC-University of Glasgow Centre for Virus Research, Garscube Estate, Glasgow G61 1QH, United Kingdom. Electronic address: nicola.logan@glasgow.ac.uk.
authors.forename: Nicola
authors.initials: N
authors.lastname: Logan
authors.affiliation: MRC-University of Glasgow Centre for Virus Research, Garscube Estate, Glasgow G61 1QH, United Kingdom. Electronic address: elizabeth.mcmonagle@glasgow.ac.uk.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: McMonagle
authors.affiliation: MRC-University of Glasgow Centre for Virus Research, Garscube Estate, Glasgow G61 1QH, United Kingdom. Electronic address: angiedrew@hotmail.co.uk.
authors.forename: Angharad A
authors.initials: AA
authors.lastname: Drew
authors.affiliation: Royal Veterinary College, University of London, London NW1 0TU, United Kingdom. Electronic address: emi.a.takahashi@gmail.com.
authors.forename: Emi
authors.initials: E
authors.lastname: Takahashi
authors.affiliation: Veterinary Diagnostic Services, University of Glasgow, Garscube Estate, Glasgow G61 1QH, United Kingdom. Electronic address: mike.mcdonald@glasgow.ac.uk.
authors.forename: Michael
authors.initials: M
authors.lastname: McDonald
authors.affiliation: The Pirbright Institute, Pirbright, Surrey GU24 0NF, United Kingdom. Electronic address: michael.baron@pirbright.ac.uk.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Baron
authors.affiliation: Wildlife Conservation Society, Bronx, NY, USA; Boyd Orr Centre for Population and Ecosystem Health, Institute of Biodiversity Animal Health and Comparative Medicine, University of Glasgow, Glasgow G12 8QQ, United Kingdom. Electronic address: mgilbert@wcs.org.
authors.forename: Martin
authors.initials: M
authors.lastname: Gilbert
authors.affiliation: Boyd Orr Centre for Population and Ecosystem Health, Institute of Biodiversity Animal Health and Comparative Medicine, University of Glasgow, Glasgow G12 8QQ, United Kingdom. Electronic address: sarah.cleaveland@glasgow.ac.uk.
authors.forename: Sarah
authors.initials: S
authors.lastname: Cleaveland
authors.affiliation: Boyd Orr Centre for Population and Ecosystem Health, Institute of Biodiversity Animal Health and Comparative Medicine, University of Glasgow, Glasgow G12 8QQ, United Kingdom. Electronic address: daniel.haydon@glasgow.ac.uk.
authors.forename: Daniel T
authors.initials: DT
authors.lastname: Haydon
authors.affiliation: MRC-University of Glasgow Centre for Virus Research, Garscube Estate, Glasgow G61 1QH, United Kingdom. Electronic address: margaret.hosie@glasgow.ac.uk.
authors.forename: Margaret J
authors.initials: MJ
authors.lastname: Hosie
authors.affiliation: MRC-University of Glasgow Centre for Virus Research, Garscube Estate, Glasgow G61 1QH, United Kingdom. Electronic address: brian.willett@glasgow.ac.uk.
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Willett
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood; D000914:Antibodies, Viral / Q000097:blood*; D002522:Chlorocebus aethiops; D003429:Cross Reactions; D004217:Distemper Virus, Canine; D004285:Dogs; D057809:HEK293 Cells; D006801:Humans; D018103:Morbillivirus / Q000276:immunology*; D009500:Neutralization Tests; D018107:Peste-des-petits-ruminants virus; D014709:Vero Cells; D014721:Vesicular stomatitis Indiana virus; D014760:Viral Fusion Proteins / Q000276:immunology*
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D014760:Viral Fusion Proteins
keywords: CDV; Morbillivirus; Neutralisation; PPRV; Pseudotype