Generation of functional NKT cells in vitro from embryonic stem cells bearing rearranged invariant Valpha14-Jalpha18 TCRalpha gene

Blood. 2010 Jan 14;115(2):230-7. doi: 10.1182/blood-2009-04-217729. Epub 2009 Nov 6.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CD24 Antigen / immunology
  • CD24 Antigen / metabolism
  • Calcium-Binding Proteins
  • Cell Differentiation / immunology*
  • Cell Nucleus / immunology*
  • Cell Nucleus / metabolism
  • Cells, Cultured
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / immunology*
  • Embryonic Stem Cells / metabolism
  • Hyaluronan Receptors / immunology
  • Hyaluronan Receptors / metabolism
  • Intercellular Signaling Peptides and Proteins / immunology
  • Intercellular Signaling Peptides and Proteins / metabolism
  • Interferon-gamma / immunology
  • Interferon-gamma / metabolism
  • Interleukin-4 / immunology
  • Interleukin-4 / metabolism
  • Mice
  • Mice, Inbred ICR
  • Mice, Knockout
  • Natural Killer T-Cells / cytology
  • Natural Killer T-Cells / immunology*
  • Natural Killer T-Cells / metabolism
  • Nuclear Transfer Techniques
  • Receptors, Antigen, T-Cell, alpha-beta / immunology*
  • Receptors, Antigen, T-Cell, alpha-beta / metabolism
  • Receptors, Notch / immunology
  • Receptors, Notch / metabolism
  • Time Factors

Substances

  • CD24 Antigen
  • Calcium-Binding Proteins
  • Dlk1 protein, mouse
  • Hyaluronan Receptors
  • Intercellular Signaling Peptides and Proteins
  • Receptors, Antigen, T-Cell, alpha-beta
  • Receptors, Notch
  • Interleukin-4
  • Interferon-gamma