title: Equine monocyte-derived macrophage cultures and their applications for infectivity and neutralization studies of equine infectious anemia virus.
abstract: Equine infectious anemia virus (EIAV) has been shown to infect cells of monocyte/macrophage lineage. These primary cells are intrinsically difficult to obtain, to purify and to culture in vitro for extended periods of time. As a result, most in vitro studies concerning this lentivirus make use of primary equine fibroblasts or transformed canine or feline cell lines. We describe methods that yield reproducibly pure cultures of equine blood monocytes from peripheral blood mononuclear cells. The in vitro differentiation of these cells into mature equine macrophage was verified using various cytochemical staining methods. The equine monocyte-derived macrophage (MDM) cultures were found to replicate cell-adapted and field strains of EIAV more efficiently than cultures of fully differentiated equine splenic macrophage. Having established reproducible and fully differentiated cultures of equine macrophage, in vitro assays of virus infectivity and serum neutralization were developed using the in vivo target cell of EIAV. These procedures, while developed for the EIAV system, should be equally useful for in vitro cultures of other macrophage-tropic pathogens of horses.
authors.affiliation: Department of Molecular Genetics and Biochemistry, School of Medicine, University of Pittsburgh, Pennsylvania 15261, USA.
authors.forename: M R
authors.initials: MR
authors.lastname: Raabe
authors.forename: C J
authors.initials: CJ
authors.lastname: Issel
authors.forename: R C
authors.initials: RC
authors.lastname: Montelaro
mesh_terms: D000367:Age Factors; D000818:Animals; D002460:Cell Line; D005260:Female; D006736:Horses; D004860:Infectious Anemia Virus, Equine / Q000472:pathogenicity / Q000502:physiology*; D008264:Macrophages / Q000166:cytology* / Q000502:physiology / Q000821:virology; D009000:Monocytes / Q000166:cytology; D009500:Neutralization Tests / Q000662:veterinary*; D015203:Reproducibility of Results; D013154:Spleen / Q000166:cytology; D013997:Time Factors; D014779:Virus Replication