Generation of Valpha14 NKT cells in vitro from hematopoietic precursors residing in bone marrow and peripheral blood.
We previously reported the generation of Valpha14 invariant TCR+ (Valpha14i) NK1.1+ natural killer T (NKT) cells in the cytokine-activated suspension culture of murine fetal liver cells. In this study, we attempted to apply this finding to the induction of Valpha14i NKT cell differentiation in the culture of hematopoietic precursors residing in bone marrow or peripheral blood. Preferential generation of NKT cells was found in the culture of Thy-1(+)-depleted bone marrow cells in the presence of culture supernatant from Con A-stimulated spleen T cells and a combination of recombinant IL-3, IL-4, IL-7 and GM-CSF. NKT cell development from peripheral blood hematopoietic precursors was induced when they were cultured on stromal cell monolayers prepared from Thy-1(+)-depleted bone marrow or fetal liver cells, suggesting that certain environments derived from hematopoietic organs are required for the induction of NKT cells from precursors in vitro. A significant fraction of NKT cells generated in the culture were positive for staining with CD1-alpha-galactosylceramide tetramer, indicating that Valpha14i NKT cells were the major subset among the NKT cells. The present methods for obtaining NKT cells in the culture of bone marrow or peripheral blood cells are applicable to the treatment of patients suffering from diseases with numerical and functional disorders of NKT cells.
Shimamura
Michio
M
Developmental Immunology Unit, Mitsubishi Kagaku Institute of Life Sciences, Tokyo, Japan.
Kobayashi
Kumi
K
Developmental Immunology Unit, Mitsubishi Kagaku Institute of Life Sciences, Tokyo, Japan.
Watanabe
Hiroko
H
Developmental Immunology Unit, Mitsubishi Kagaku Institute of Life Sciences, Tokyo, Japan.
Huang
Yi-Ying
YY
Developmental Immunology Unit, Mitsubishi Kagaku Institute of Life Sciences, Tokyo, Japan.
Okamoto
Naoki
N
Developmental Immunology Unit, Mitsubishi Kagaku Institute of Life Sciences, Tokyo, Japan.
Kanie
Osamu
O
Glycotechnology Unit, Mitsubishi Kagaku Institute of Life Sciences, Tokyo, Japan.
Goji
Hiroshi
H
Developmental Immunology Unit, Mitsubishi Kagaku Institute of Life Sciences, Tokyo, Japan.
Kobayashi
Masumi
M
Developmental Immunology Unit, Mitsubishi Kagaku Institute of Life Sciences, Tokyo, Japan.
D000818:Animals; D018949:Antigens, CD1 / Q000235:genetics / Q000502:physiology; D001773:Blood Cells / Q000166:cytology*; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D007135:Immunoglobulin Variable Region / Q000032:analysis*; D007694:Killer Cells, Natural / Q000166:cytology* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009369:Neoplasms / Q000276:immunology; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000032:analysis*; D017154:Stromal Cells / Q000502:physiology; D016176:T-Lymphocyte Subsets / Q000166:cytology* / Q000276:immunology
D018949:Antigens, CD1; D004268:DNA-Binding Proteins; D007135:Immunoglobulin Variable Region; C499308:Rag2 protein, mouse; D016693:Receptors, Antigen, T-Cell, alpha-beta; C064655:V(D)J recombination activating protein 2