Isolation and characterization of canine natural killer cells

Helen T. Michael, Daisuke Ito, Valarie McCullar, Bin Zhang, Jeffrey S. Miller, Jaime F. Modiano

Research output: Contribution to journalArticlepeer-review

36 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)211-217
Number of pages7
JournalVeterinary immunology and immunopathology
Volume155
Issue number3
DOIs
StatePublished - Sep 15 2013

Bibliographical note

Funding Information:
We thank Megan Duckett for expert technical help. We would like to acknowledge the assistance of the Flow Cytometry Core Facility of the Masonic Cancer Center, a comprehensive cancer center designated by the National Cancer Institute, supported in part by grant P30 CA77598 from the National Institutes of Health of the United States Public Health Service . This project was supported in part by a University of Minnesota Companion Animal Grant and by the University of Minnesota Animal Cancer Care and Research Program/Comparative Oncology Research Fund .

Keywords

  • Canine
  • Interleukins
  • NK cells

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