Abstract
Objective To analyse the effects of IL-15 on the expression of NKG2D and the cytotoxicity of cytokine-induced killer(CIK) cells against human esophagus carcinoma cell EC9706 in vitro.Methods Peripheral blood mononuclear cells were isolated from healthy donors then divided into two groups: the control group(cells were cultured in the presence of IFN-γ,anti-CD3 antibody and IL-2) and IL-15 group(cells were cultured in the presence of IFN-γ,anti-CD3 antibody,IL-2 and 10 ng/ml IL-15).Phenotypic characteristics of CIK cells and the expression of NKG2D were analyzed by flow cytometry.After 14 days culture,cytotoxicity of CIK cells against EC9706 cells was measured by using a standard LDH releasing assay.Effector cells were added to target cells at E∶T radios of 20∶1,30∶1.In blocking experiments,NKG2D monoclonal antibody was added to CIK cells for 15 min before plating at 30∶1 E∶T ratio.Results NKG2D molecules were significantly up-regulated during the culture period on CD3+ cells(the CIK cells population) and the CD56+cells.The differences of NKG2D expressions were significant between IL-15 group and the control group(P0.05).Cytotoxicity of CIK cells treated by the IL-15 group was higher than that of the control group,both at 20∶1 E∶T ratios and at 30∶1 E∶T ratios(P0.05).When the NKG2D molecules on CIK cell ular membrance were blocked by the NKG2D monoclonal antibody,the cytolytic activity in the control group cells and IL-15 group cells was significantly inhibited.Conclusion IL-15 up-regulated the expression of NKG2D on CIK cells,which enhanced the NKG2D mediated cytotoxicity against EC9706 cells.CIK cells played a role through the NKG2D molecules.
Published Version
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