Generation of functional NKT cells in vitro from embryonic stem cells bearing rearranged invariant Valpha14-Jalpha18 TCRalpha gene.
Establishment of a system with efficient generation of natural killer T (NKT) cells from embryonic stem (ES) cells would enable us to identify the cells with NKT-cell potential and obtain NKT cells with desired function. Here, using cloned ES (NKT-ES) cells generated by the transfer of nuclei from mature NKT cells, we have established a culture system that preferentially developed functional NKT cells and also identified early NKT progenitors, which first appeared on day 11 as a c-kit(+) population in the cocultures on OP9 cells with expression of Notch ligand, delta-like1 (OP9/Dll-1) and became c-kit(lo/-) on day 14. Interestingly, in the presence of Notch signals, NKT-ES cells differentiated only to thymic CD44(lo) CD24(hi) NKT cells producing mainly interleukin-4 (IL-4), whereas NKT cells resembling CD44(hi) CD24(lo) liver NKT cells producing mainly interferon gamma (IFN-gamma) and exhibiting strong adjuvant activity in vivo were developed in the switch culture starting at day 14 in the absence of Notch. The cloned ES culture system offers a new opportunity for the elucidation of the molecular events on NKT-cell development and for the establishment of NKT-cell therapy.
Watarai
Hiroshi
H
Laboratory for Immune Regulation, RIKEN Research Center for Allergy and Immunology, Kanagawa, Japan.
Rybouchkin
Andrei
A
Hongo
Naomi
N
Nagata
Yuko
Y
Sakata
Sakura
S
Sekine
Etsuko
E
Dashtsoodol
Nyambayar
N
Tashiro
Takuya
T
Fujii
Shin-ichiro
S
Shimizu
Kanako
K
Mori
Kenji
K
Masuda
Kyoko
K
Kawamoto
Hiroshi
H
Koseki
Haruhiko
H
Taniguchi
Masaru
M
D000818:Animals; D051927:CD24 Antigen / Q000276:immunology / Q000378:metabolism; D002135:Calcium-Binding Proteins; D002454:Cell Differentiation / Q000276:immunology*; D002467:Cell Nucleus / Q000276:immunology* / Q000378:metabolism; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D018960:Hyaluronan Receptors / Q000276:immunology / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000276:immunology / Q000378:metabolism; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D015847:Interleukin-4 / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008813:Mice, Inbred ICR; D018345:Mice, Knockout; D055611:Natural Killer T-Cells / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D053652:Nuclear Transfer Techniques; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000276:immunology* / Q000378:metabolism; D051880:Receptors, Notch / Q000276:immunology / Q000378:metabolism; D013997:Time Factors
D051927:CD24 Antigen; D002135:Calcium-Binding Proteins; C512925:Dlk1 protein, mouse; D018960:Hyaluronan Receptors; D036341:Intercellular Signaling Peptides and Proteins; D016693:Receptors, Antigen, T-Cell, alpha-beta; D051880:Receptors, Notch; D015847:Interleukin-4; D007371:Interferon-gamma