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Expansion of Vα24+Vβ11+ NKT cells from cord blood mononuclear cells using IL‐15, IL‐7 and Flt3‐L depends on monocytes

Human Vα24+Vβ11+ NKT cells are a unique T cell population specifically and potently activated by α‐galactosylceramide (αGalCer; KRN7000) presented by CD1d. Here, we present a simple and efficient method for expanding Vα24+Vβ11+ NKT cells from human cord blood mononuclear cells (CBMNC) using αGalCer in the presence of interleukin (IL)‐15, IL‐7 and Flt3‐L. The addition of αGalCer from day 0, compared to its addition from day 8 or day 15, induced a greater expansion of NKT cells. The maximal expansion of NKT cells was observed after 15 days (2300‐fold). Thereafter, the number of NKT cells decreased slowly, a decrease that was correlated with the diminution of CD1d‐positive cells. NKT cell proliferation induced by αGalCer was not observed when CD1d‐expressing monocytes were depleted from CBMNC, whereas B cell and dendritic cell depletions had no effect. Expanded NKT cells were CD4+CD8– and secreted both IL‐4 and IFN‐γ. In this system, CD3+ T cells and CD3–CD56+ NK cells were also expanded. However, the expansion of NKT cells had no significant functional effect on T and NK cells. This expansion method of CBMNC‐derived NKT cells is simple and may be helpful for clinical use.