title: Equus caballus major histocompatibility complex class I is an entry receptor for equine herpesvirus type 1.
abstract: In this study, Equus caballus major histocompatibility complex class I (MHC-I) was identified as a cellular entry receptor for the alphaherpesvirus equine herpesvirus type 1 (EHV-1). This novel EHV-1 receptor was discovered using a cDNA library from equine macrophages. cDNAs from this EHV-1-susceptible cell type were inserted into EHV-1-resistant B78H1 murine melanoma cells, these cells were infected with an EHV-1 lacZ reporter virus, and cells that supported virus infection were identified by X-Gal (5-bromo-4-chloro-3-indolyl-beta-d-galactopyranoside) staining. Positive cells were subjected to several rounds of purification to obtain homogeneous cell populations that were shown to be uniformly infected with EHV-1. cDNAs from these cell populations were amplified by PCR and then sequenced. The sequence data revealed that the EHV-1-susceptible cells had acquired an E. caballus MHC-I cDNA. Cell surface expression of this receptor was verified by confocal immunofluorescence microscopy. The MHC-I cDNA was cloned into a mammalian expression vector, and stable B78H1 cell lines were generated that express this receptor. These cell lines were susceptible to EHV-1 infection while the parental B78H1 cells remained resistant to infection. In addition, EHV-1 infection of the B78H1 MHC-I-expressing cell lines was inhibited in a dose-dependent manner by an anti-MHC-I antibody.
authors.affiliation: Department of Biology and Marine Biology, University of North Carolina Wilmington, 601 S. College Rd., Wilmington, NC 28403, USA.
authors.forename: Brian M
authors.initials: BM
authors.lastname: Kurtz
authors.forename: Lauren B
authors.initials: LB
authors.lastname: Singletary
authors.forename: Sean D
authors.initials: SD
authors.lastname: Kelly
authors.forename: Arthur R
authors.initials: AR
authors.lastname: Frampton
mesh_terms: D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry; D015870:Gene Expression; D015723:Gene Library; D017930:Genes, Reporter; D004861:Herpesvirus 1, Equid / Q000502:physiology*; D015395:Histocompatibility Antigens Class I / Q000502:physiology*; D006736:Horses / Q000821:virology*; D008264:Macrophages; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D011991:Receptors, Virus / Q000502:physiology*; D017422:Sequence Analysis, DNA; D013194:Staining and Labeling / Q000379:methods; D014162:Transfection; D053586:Virus Internalization; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D015395:Histocompatibility Antigens Class I; D011991:Receptors, Virus; D001616:beta-Galactosidase