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本研究探讨细胞因子诱导的杀伤性细胞(CIK)通过NKG2D受体和配体相互作用杀伤血液肿瘤细胞的机制。用含有rhIL-2,抗CD3抗体,IFN-γ的培养液培养健康人的外周血单个核细胞(PBMNC)2周后,用流式细胞仪分析CIK细胞的细胞亚群以及细胞表面NK细胞受体,同时检测血液肿瘤细胞株表面NKG2D配体的表达水平。CAM标记靶细胞,用流式细胞仪检测CIK细胞对血液肿瘤细胞的杀伤作用。结果表明,大部分CIK细胞为CD3+细胞(97.85±1.95%),CD3+CD8+细胞和CD3+CD56+细胞的比率较培养前明显升高(P<0.001;P=0.033);约86%的CIK细胞表达NKG2D受体,几乎不表达CD158a,CD158b和NCR受体;血液肿瘤细胞株U266、K562和Daudi均表达一定水平的NKG2D配体,CIK细胞对这3种血液肿瘤细胞株均具有较高的杀伤作用,这种杀伤作用可以被抗NKG2D抗体部分抑制(U266 52.67±4.63%vs 32.67±4.81%,P=0.008;K562 71.67±4.91%vs 50.33±4.91%,P=0.007;Daudi 68.67±5.04 vs 52.67±2.60%,P=0.024)。结论:大多数的CIK细胞表达NKG2D受体,NKG2D受体和配体的相互作用可能是CIK细胞杀伤血液肿瘤细胞的作用机制之一。
This study explored the mechanism by which cytokine-induced killer cells (CIKs) kill hematological tumor cells through the interaction of NKG2D receptors with ligands. Peripheral blood mononuclear cells (PBMNCs) of healthy individuals were cultured for 2 weeks in a culture medium containing rhIL-2, anti-CD3 antibody, and IFN-γ, and the cell subpopulation of CIK cells and NK cell surface were analyzed by flow cytometry At the same time, the expression of NKG2D ligand on the surface of hematological tumor cell lines was detected. CAM labeled target cells, cytotoxicity of CIK cells on hematological tumor cells was detected by flow cytometry. The results showed that the majority of CIK cells were CD3 + cells (97.85 ± 1.95%), the ratio of CD3 + CD8 + cells and CD3 + CD56 + cells was significantly higher than that before culture (P <0.001; P = 0.033) NKG2D receptor was expressed with little expression of CD158a, CD158b and NCR receptors. The hematological tumor cell lines U266, K562 and Daudi all expressed certain level of NKG2D ligand, and CIK cells had higher killing effect on all three hematological tumor cell lines (U266 52.67 ± 4.63% vs 32.67 ± 4.81%, P = 0.008; K562 71.67 ± 4.91% vs 50.33 ± 4.91%, P = 0.007; Daudi 68.67 ± 5.04 vs 52.67 ± 2.60%, P = 0.024). CONCLUSION: Most CIK cells express NKG2D receptor. The interaction between NKG2D receptor and ligand may be one of the mechanisms of CIK cell killing of hematological tumor cells.