Korean J Physiol Pharmacol.
2012 Jun;16(3):167-174. 10.4196/kjpp.2012.16.3.167.
Generation of CD2+CD8+ NK Cells from c-kit+ Bone Marrow Cells in Porcine
- Affiliations
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- 1Department of Physiology and Biosafety Research Institute, College of Veterinary Medicine, Chonbuk National University, Jeonju 561-756, Korea. jhkim1@chonbuk.ac.kr
- 2Department of Pathology, College of Veterinary Medicine, Chonbuk National University, Jeonju 561-756, Korea.
- 3Department of Laboratroy Animal Medicine, College of Veterinary Medicine, Chonbuk National University, Jeonju 561-756, Korea.
- 4Department of Parasitology, College of Veterinary Medicine, Chonbuk National University, Jeonju 561-756, Korea.
- 5Department of Veterinary Physiology, College of Veterinary Medicine, Seoul National University, Seoul 151-741, Korea.
- 6Department of Clinical Neurosciences, Neurology Unit, Addenbrooke's Hospital, University of Cambridge, Cambridge CB2 0QQ, UK.
- KMID: 1768008
- DOI: http://doi.org/10.4196/kjpp.2012.16.3.167
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.
MeSH Terms
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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
Cytokines
Granzymes
Hydrocortisone
Interleukin-15
Interleukin-7
Interleukins
Perforin
Stem Cell Factor
Tyrosine
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