title: Isolation and characterization of canine natural killer cells.
abstract: NK cells are non-T, non-B lymphocytes that kill target cells without previous activation. The immunophenotype and function of these cells in humans and mice are well defined, but canine NK cells remain incompletely characterized. Our objectives were to isolate and culture canine peripheral blood NK cells, and to define their immunophenotype and killing capability. PBMC were obtained from healthy dogs and T cells were depleted by immunomagnetic separation. The residual cells were cultured in media supplemented with IL-2, IL-15 or both, or with mouse embryonic liver (EL) feeder cells. Non-T, non-B lymphocytes survived and expanded in these cultures. IL-2 was necessary and sufficient for survival; the addition of IL-15 was necessary for expansion, but IL-15 alone did not support survival. Culture with EL cells and IL-2 also fostered survival and expansion. The non-T, non-B lymphocytes uniformly expressed CD45, MHC I, and showed significant cytotoxic activity against CTAC targets. Expression of MHC II, CD11/18 was restricted to subsets of these cells. The data show that cells meeting the criteria for NK cells in other species, i.e., non-T, non-B lymphocytes with cytotoxic activity, can be expanded from canine PBMC by T-cell depletion and culture with cytokines or feeder cells.
authors.affiliation: Department of Veterinary Clinical Sciences, College of Veterinary Medicine, University of Minnesota, 1352 Boyd Avenue, St. Paul, MN 55108, United States. helen.michael@nih.gov
authors.forename: Helen T
authors.initials: HT
authors.lastname: Michael
authors.forename: Daisuke
authors.initials: D
authors.lastname: Ito
authors.forename: Valarie
authors.initials: V
authors.lastname: McCullar
authors.forename: Bin
authors.initials: B
authors.lastname: Zhang
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Miller
authors.forename: Jaime F
authors.initials: JF
authors.lastname: Modiano
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002469:Cell Separation; D002478:Cells, Cultured; D004285:Dogs / Q000276:immunology*; D018189:Immunomagnetic Separation; D019409:Interleukin-15 / Q000494:pharmacology; D007376:Interleukin-2 / Q000494:pharmacology; D007694:Killer Cells, Natural / Q000276:immunology*
chemical_list: D019409:Interleukin-15; D007376:Interleukin-2
keywords: Canine; Interleukins; NK cells