title: Ex vivo expansion of canine cytotoxic large granular lymphocytes exhibiting characteristics of natural killer cells.
abstract: Canine NK cells still are not well-characterized due to the lack of information concerning specific NK cell markers and the fact that NK cells are not an abundant cell population. In this study, we selectively expanded the canine cytotoxic large granular lymphocytes (CLGLs) that exhibit morphologic, genetic, and functional characteristics of NK cells from normal donor PBMCs. The cultured CLGLs were characterized by a high proportion of CD5(dim) expressing cells, of which the majority of cells co-expressed CD3 and CD8, but did not express TCRαβ and TCRγδ. The phenotype of the majority of the CLGLs was CD5(dim)CD3(+)CD8(+) TCRαβ(-)TCRγδ(-)CD4(-)CD21(-)CD11c(+/-)CD11d(+/-)CD44(+). The expression of mRNAs for NK cell-associated receptors (NKG2D, NKp30, NKp44, Ly49, perforin, and granzyme B) were highly upregulated in cultured CLGLs. Specifically, NKp46 was remarkably upregulated in the cultured CLGLs compared to PBMCs. The mRNAs for the NKT-associated iTCRα gene in CLGLs was present at a basal level. The cytotoxic activity of the CLGLs against canine NK cell-sensitive CTAC cells was remarkably elevated in a dose-dependent manner, and the CLGLs produced large amounts of IFN-γ. The antitumor activity of CLGLs extended to different types of canine tumor cells (CF41.Mg and K9TCC-pu-AXC) without specific antigen recognition. These results are consistent with prior reports, and strongly suggest that the selectively expanded CLGLs represent a population of canine NK cells. The results of this study will contribute to future research on canine NK cells as well as NK cell-based immunotherapy.
authors.affiliation: Department of Companion & Laboratory Animal Science, Kongju National University, Yesan, Chungnam, 340-702, Republic of Korea.
authors.forename: Dong-Jun
authors.initials: DJ
authors.lastname: Shin
authors.forename: Ji-Yun
authors.initials: JY
authors.lastname: Park
authors.forename: Youn-Young
authors.initials: YY
authors.lastname: Jang
authors.forename: Je-Jung
authors.initials: JJ
authors.lastname: Lee
authors.forename: Youn-Kyung
authors.initials: YK
authors.lastname: Lee
authors.forename: Myung-Geun
authors.initials: MG
authors.lastname: Shin
authors.forename: Ji-Youn
authors.initials: JY
authors.lastname: Jung
authors.forename: William E
authors.initials: WE
authors.lastname: Carson
authors.forename: Duck
authors.initials: D
authors.lastname: Cho
authors.forename: Sang-Ki
authors.initials: SK
authors.lastname: Kim
mesh_terms: D000818:Animals; D019002:CD56 Antigen / Q000235:genetics; D045744:Cell Line, Tumor; D003602:Cytotoxicity, Immunologic; D004285:Dogs / Q000276:immunology*; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000096:biosynthesis; D007694:Killer Cells, Natural / Q000276:immunology*; D055650:NK Cell Lectin-Like Receptor Subfamily A / Q000235:genetics; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics
chemical_list: D019002:CD56 Antigen; D055650:NK Cell Lectin-Like Receptor Subfamily A; D016693:Receptors, Antigen, T-Cell, alpha-beta; D007371:Interferon-gamma
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