title: Equine ELISPOT assay to study secretion of IFNγ and IL-4 from peripheral blood mononuclear cells.
abstract: Human and mouse immune system cells are the most frequently used specimens in ELISPOT assays. In an effect to expand the application of ELISPOT assay to other species, we developed matched antibody pairs for ready-to-use kits designed for studying the frequency of equine IFNγ- and IL-4-secreting peripheral blood mononuclear cells (PBMCs). Equine PBMCs were stimulated with either concanavalin A (Con A) or calcium ionomycin mixed with phorbol 12-myristate 13-acetate (CaI + PMA). We found that Con A, in general, had a more profound stimulating effect than CaI + PMA on IL-4 secretion, whereas both stimulatory and inhibitory effects were observed on IFNγ secretion. Our data demonstrate a large dynamic range in IFNγ and IL-4 secretion among different donors, which may reflect animal health and serve as a valuable diagnostic marker.
authors.affiliation: R&D Systems, Inc., Minneapolis, MN, USA.
authors.forename: Jodi
authors.initials: J
authors.lastname: Hagen
authors.forename: Chris
authors.initials: C
authors.lastname: Hartnett
authors.forename: Jeffrey P
authors.initials: JP
authors.lastname: Houchins
authors.forename: Steeve
authors.initials: S
authors.lastname: Giguère
authors.forename: Alexander E
authors.initials: AE
authors.lastname: Kalyuzhny
mesh_terms: D000818:Animals; D002118:Calcium / Q000494:pharmacology; D003208:Concanavalin A / Q000494:pharmacology; D058501:Enzyme-Linked Immunospot Assay / Q000662:veterinary*; D006736:Horses / Q000097:blood* / Q000276:immunology*; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism*; D015847:Interleukin-4 / Q000276:immunology / Q000378:metabolism*; D015759:Ionomycin / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D003208:Concanavalin A; D015847:Interleukin-4; D015759:Ionomycin; C016658:phorbolol myristate acetate; D007371:Interferon-gamma; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium