title: Replication kinetics of neurovirulent versus non-neurovirulent equine herpesvirus type 1 strains in equine nasal mucosal explants.
abstract: Equine herpesvirus type 1 (EHV-1) is the causative agent of equine herpes myeloencephalopathy, of which outbreaks are reported with increasing frequency throughout North America and Europe. This has resulted in its classification as a potentially emerging disease by the US Department of Agriculture. Recently, it was found that a single nucleotide polymorphism (SNP) in the viral DNA polymerase gene (ORF30) at aa 752 (N-->D) is associated with the neurovirulent potential of EHV-1. In the present study, equine respiratory mucosal explants were inoculated with several Belgian isolates typed in their ORF30 as D(752) or N(752), to evaluate a possible difference in replication in the upper respiratory tract. In addition, to evaluate whether any observed differences could be attributed to the SNP associated with neurovirulence, the experiments were repeated with parental Ab4 (reference neurovirulent strain), parental NY03 (reference non-neurovirulent strain) and their N/D revertant recombinant viruses. The salient findings were that EHV-1 spreads plaquewise in the epithelium, but plaques never cross the basement membrane (BM). However, single EHV-1-infected cells could be observed below the BM at 36 h post-inoculation (p.i.) for all N(752) isolates and at 24 h p.i. for all D(752) isolates, and were identified as monocytic cells and T lymphocytes. Interestingly, the number of infected cells was two to five times higher for D(752) isolates compared with N(752) isolates at every time point analysed. Finally, this study showed that equine respiratory explants are a valuable and reproducible model to study EHV-1 neurovirulence in vitro, thereby reducing the need for horses as experimental animals.
authors.affiliation: Department of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium.
authors.forename: Annelies P
authors.initials: AP
authors.lastname: Vandekerckhove
authors.affiliation: Department of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium.
authors.forename: S
authors.initials: S
authors.lastname: Glorieux
authors.affiliation: Department of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium.
authors.forename: A C
authors.initials: AC
authors.lastname: Gryspeerdt
authors.affiliation: Department of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium.
authors.forename: L
authors.initials: L
authors.lastname: Steukers
authors.affiliation: Department of Physiology and Biometrics, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium.
authors.forename: L
authors.initials: L
authors.lastname: Duchateau
authors.affiliation: Institut für Virologie, Freie Universitaet Berlin, Philippstrasse 13, 10115 Berlin, Germany.
authors.forename: N
authors.initials: N
authors.lastname: Osterrieder
authors.affiliation: Department of Physiology and Biometrics, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium.|Department of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium.
authors.forename: G R
authors.initials: GR
authors.lastname: Van de Walle
authors.affiliation: Department of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium.
authors.forename: H J
authors.initials: HJ
authors.lastname: Nauwynck
mesh_terms: D000818:Animals; D001530:Belgium; D004259:DNA-Directed DNA Polymerase / Q000235:genetics* / Q000502:physiology; D004861:Herpesvirus 1, Equid / Q000254:growth & development* / Q000302:isolation & purification / Q000472:pathogenicity*; D006736:Horses / Q000821:virology*; D009000:Monocytes / Q000821:virology; D020125:Mutation, Missense; D009297:Nasal Mucosa / Q000821:virology*; D009924:Organ Culture Techniques; D013601:T-Lymphocytes / Q000821:virology; D013997:Time Factors; D014764:Viral Proteins / Q000235:genetics / Q000502:physiology; D014774:Virulence; D037521:Virulence Factors / Q000235:genetics* / Q000502:physiology; D014779:Virus Replication
chemical_list: D014764:Viral Proteins; D037521:Virulence Factors; D004259:DNA-Directed DNA Polymerase