Generation of CD2+CD8+ NK Cells from c-kit+ Bone Marrow Cells in Porcine

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 exclusi...

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Main Authors:	Lim, Kyu Hee, Han, Ji-Hui, Roh, Yoon Seok, Kim, Bumseok, Kwon, Jung-Kee, You, Myoung Jo, Han, Ho Jae, Ejaz, Sohail, Kang, Chang-Won, Kim, Jong-Hoon
Format:	Online
Language:	English
Published:	The Korean Physiological Society and The Korean Society of Pharmacology 2012
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3394918/

id	pubmed-3394918
recordtype	oai_dc
spelling	pubmed-33949182012-07-16 Generation of CD2+CD8+ NK Cells from c-kit+ Bone Marrow Cells in Porcine Lim, Kyu Hee Han, Ji-Hui Roh, Yoon Seok Kim, Bumseok Kwon, Jung-Kee You, Myoung Jo Han, Ho Jae Ejaz, Sohail Kang, Chang-Won Kim, Jong-Hoon Original Article 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. The Korean Physiological Society and The Korean Society of Pharmacology 2012-06 2012-06-26 /pmc/articles/PMC3394918/ /pubmed/22802697 http://dx.doi.org/10.4196/kjpp.2012.16.3.167 Text en Copyright © 2012 The Korean Physiological Society and The Korean Society of Pharmacology http://creativecommons.org/licenses/by-nc/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Lim, Kyu Hee Han, Ji-Hui Roh, Yoon Seok Kim, Bumseok Kwon, Jung-Kee You, Myoung Jo Han, Ho Jae Ejaz, Sohail Kang, Chang-Won Kim, Jong-Hoon
spellingShingle	Lim, Kyu Hee Han, Ji-Hui Roh, Yoon Seok Kim, Bumseok Kwon, Jung-Kee You, Myoung Jo Han, Ho Jae Ejaz, Sohail Kang, Chang-Won Kim, Jong-Hoon Generation of CD2+CD8+ NK Cells from c-kit+ Bone Marrow Cells in Porcine
author_facet	Lim, Kyu Hee Han, Ji-Hui Roh, Yoon Seok Kim, Bumseok Kwon, Jung-Kee You, Myoung Jo Han, Ho Jae Ejaz, Sohail Kang, Chang-Won Kim, Jong-Hoon
author_sort	Lim, Kyu Hee
title	Generation of CD2+CD8+ NK Cells from c-kit+ Bone Marrow Cells in Porcine
title_short	Generation of CD2+CD8+ NK Cells from c-kit+ Bone Marrow Cells in Porcine
title_full	Generation of CD2+CD8+ NK Cells from c-kit+ Bone Marrow Cells in Porcine
title_fullStr	Generation of CD2+CD8+ NK Cells from c-kit+ Bone Marrow Cells in Porcine
title_full_unstemmed	Generation of CD2+CD8+ NK Cells from c-kit+ Bone Marrow Cells in Porcine
title_sort	generation of cd2+cd8+ nk cells from c-kit+ bone marrow cells in porcine
description	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.
publisher	The Korean Physiological Society and The Korean Society of Pharmacology
publishDate	2012
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3394918/
_version_	1611542519727783936

Generation of CD2+CD8+ NK Cells from c-kit+ Bone Marrow Cells in Porcine

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