Expansion of human Valpha24+ NKT cells by repeated stimulation with KRN7000.
Changes in Valpha24+Vbeta11+ 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 Valpha24+Vbeta11+ 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(12) cells. The expanded NKT cells retained their CD4+ or CD4- 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.
Research Laboratory, Gemini Science, Inc., 10355 Science Center Drive, San Diego, CA, 92121, USA. progers@liai.org
Paul R
PR
Rogers
Atsushi
A
Matsumoto
Olga
O
Naidenko
Mitchell
M
Kronenberg
Toshifumi
T
Mikayama
Shinichiro
S
Kato
D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D015704:CD4 Antigens / Q000378:metabolism; D016827:CD8 Antigens / Q000378:metabolism; D002478:Cells, Cultured; D003602:Cytotoxicity, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005699:Galactosylceramides / Q000494:pharmacology*; D006133:Growth Substances / Q000032:analysis / Q000494:pharmacology; D006801:Humans; D007376:Interleukin-2 / Q000032:analysis / Q000494:pharmacology; D019169:Jurkat Cells; D007694:Killer Cells, Natural / Q000187:drug effects* / Q000276:immunology*; D008213:Lymphocyte Activation / Q000276:immunology*; D011948:Receptors, Antigen, T-Cell / Q000276:immunology; D011994:Recombinant Proteins / Q000494:pharmacology
D000276:Adjuvants, Immunologic; D015704:CD4 Antigens; D016827:CD8 Antigens; D005699:Galactosylceramides; D006133:Growth Substances; D007376:Interleukin-2; D011948:Receptors, Antigen, T-Cell; D011994:Recombinant Proteins; C103873:KRN 7000