Expansion of human V alpha 24 super(+) NKT cells by repeated stimulation with KRN7000

• Published in: Journal of immunological methods 2004-02, Vol.285 (2), p.197-214
• Main Authors: Rogers, PR, Matsumoto, A, Naidenko, O, Kronenberg, M, Mikayama, T, Kato, S
• Format: Article
• Language: English
• ISSN: 0022-1759
• DOI: 10.1016/j.jim.2003.12.003

Changes in V alpha 24 super(+)V beta 11 super(+) NKT cell number and function are associated with human autoimmune diseases and cancer. Restoration of this corresponding NKT cell population in mice or in vivo activation with alpha -galactosylceramide (KRN7000) can prevent or reduce tumor growth and autoimmunity. Although the therapeutic value of these natural killer T (NKT) cells in man remains to be determined, large numbers of functional antigen-specific NKT cells can be expanded in vitro. We show that V alpha 24 super(+)V beta 11 super(+) human NKT cells are expanded by repeated stimulation with KRN7000, unfractionated donor peripheral blood mononuclear cells (PBMC), and recombinant human interleukin-2 (rhIL-2). NKT cells were expanded continuously for more than 2 months with a potential yield of >10 super(12) cells. The expanded NKT cells retained their CD4 super(+) or CD4 super(-) phenotype after restimulation and were functional as shown by cytokine secretion, killing of antigen-pulsed target cells, and activation of NK cell cytotoxicity. This expansion method may be useful for proof-of-concept studies involving adoptive transfer of ex vivo-expanded NKT cells as a new therapeutic option for cancer and autoimmune diseases.