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Generation of Vα14 NKT cells in vitro from hematopoietic precursors residing in bone marrow and peripheral blood
Author(s) -
Shimamura Michio,
Kobayashi Kumi,
Watanabe Hiroko,
Huang YiYing,
Okamoto Naoki,
Kanie Osamu,
Goji Hiroshi,
Kobayashi Masumi
Publication year - 2004
Publication title -
european journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.272
H-Index - 201
eISSN - 1521-4141
pISSN - 0014-2980
DOI - 10.1002/eji.200324579
Subject(s) - natural killer t cell , bone marrow , haematopoiesis , biology , immunology , stromal cell , spleen , t cell , microbiology and biotechnology , stem cell , cancer research , immune system
Abstract We previously reported the generation of Vα14 invariant TCR + (Vα14i) 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 Vα14i NKT cell differentiation in the culture of hematopoietic precursors residing in bone marrow or peripheralblood. 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–α‐galactosylceramide tetramer, indicating that Vα14i 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.