Generation of CD2+CD8+ NK Cells from c-kit+ Bone Marrow Cells in Porcine.
10.4196/kjpp.2012.16.3.167
- Author:
Kyu Hee LIM
1
;
Ji Hui HAN
;
Yoon Seok ROH
;
Bumseok KIM
;
Jung Kee KWON
;
Myoung Jo YOU
;
Ho Jae HAN
;
Sohail EJAZ
;
Chang Won KANG
;
Jong Hoon KIM
Author Information
1. Department of Physiology and Biosafety Research Institute, College of Veterinary Medicine, Chonbuk National University, Jeonju 561-756, Korea. jhkim1@chonbuk.ac.kr
- Publication Type:In Vitro ; Original Article
- Keywords:
Hematopoietic stem cells;
Natural killer cells;
Differentiation;
Cytokines;
Cytotoxicity
- MeSH:
Adoptive Transfer;
Blotting, Western;
Bone Marrow;
Bone Marrow Cells;
Cytokines;
Granzymes;
Hematopoietic Stem Cells;
Hydrocortisone;
Interleukin-15;
Interleukin-7;
Interleukins;
K562 Cells;
Killer Cells, Natural;
Perforin;
Stem Cell Factor;
Stromal Cells;
Transplantation, Heterologous;
Tyrosine
- From:The Korean Journal of Physiology and Pharmacology
2012;16(3):167-174
- CountryRepublic of Korea
- Language:English
-
Abstract:
Natural killer (NK) cells provide one of the initial barriers of cellular host defense against pathogens, in particular intracellular pathogens. Because bone marrow-derived hematopoietic stem cells (HSCs), lymphoid protenitors, can give rise to NK cells, NK ontogeny has been considered to be exclusively lymphoid. Here, we show that porcine c-kit+ bone marrow cells (c-kit+ BM cells) develop into NK cells in vitro in the presence of various cytokines [interleukin (IL)-2, IL-7, IL-15, IL-21, stem cell factor (SCF), and fms-like tyrosine kinase-3 ligand (FLT3L)]. Adding hydrocortisone (HDC) and stromal cells greatly increases the frequency of c-kit+ BM cells that give rise to CD2+CD8+ NK cells. Also, intracellular levels of perforin, granzyme B, and NKG2D were determined by RT-PCR and western blotting analysis. It was found that of perforin, granzyme B, and NKG2D levels significantly were increased in cytokine-stimulated c-kit+ BM cells than those of controls. And, we compared the ability of the cytotoxicity of CD2+CD8+ NK cells differentiated by cytokines from c-kit+ BM cells against K562 target cells for 28 days. Cytokines-induced NK cells as effector cells were incubated with K562 cells as target in a ratio of 100:1 for 4 h once a week. In results, CD2+CD8+ NK cells induced by cytokines and stromal cells showed a significantly increased cytotoxicity 21 days later. Whereas, our results indicated that c-kit+ BM cells not pretreated with cytokines have lower levels of cytotoxicity. Taken together, this study suggests that cytokines-induced NK cells from porcine c-kit+ BM cells may be used as adoptive transfer therapy if the known obstacles to xenografting (e.g. immune and non-immune problems) were overcome in the future.
