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Background and Objective: Cytokine-induced killer (CIK) cells are usually generated from peripheral blood mononuclear cells (PBMCs) with the stimulation of IL-2 in vitro.However, the concomitant expanded Regulatory T (Treg) cells and Treg cells related cytokines in CIK cells significantly decreased the cytotoxicity of CIK cells.Materials and methods: In the present study, we respectively generated IL-2-CIK cells or IL-15-CIK cells using the same PBMCs and monitored the phenotype during the process of culture.Then, we measured the IL-2-CIK cells and IL-15-CIK cells-mediated cytotoxicity against human leukemia cells.Finally, the investigation of cytokines expression was performed to determine the potential mechanism of IL-2-and IL-15-CIK cells.Results: Compared with IL-2-CIK cells, the percentage of CD3+CD56+ cells was significantly increased in IL-15-CIK cells, but the expression of Treg cells and IL-35 was significantly decreased in IL-15-CIK cells.Meanwhile, the in vitro cytotoxicity against human myeloid leukemia cells of IL-15-CIK cells was significantly augmented compared with IL-2-CIK cells.Conclusions: These data suggest that IL-15 improves the cytotoxicity of CIK cells against leukemia cells by up-regulating CD3+CD56+ cells and down-regulating Treg cells as well as IL-35.