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浙江大学学报(医学版)  2011, Vol. 40 Issue (5): 475-481    DOI: 10.3785/j.issn.1008-9292.2011.05.003
专题报道     
体外抗体功能修饰抑制性KIR对人NK细胞的影响
吴功强1,2 ,赵妍敏1 ,黄河1 ,来晓瑜1
1.浙江大学医学院附属第一医院骨髓移植中心、血液研究所,浙江 杭州 310003; 2.义乌市中心医院血液科,浙江 义乌 322000
Effects of blocking inhibitory KIR receptors on cytotoxic activity of human NK cells in vitro
WU Gong-qiang 1,2,ZHAO Yan-min1,HUANG He1,LAI Xiao-yu1
1.Bone Marrow Transplantation Center,Institute of Hematology,The First Affiliated Hospital,Zhejiang University School of Medicine,Hangzhou 310003,China; 2.The Central Hospital of Yiwu City,Yiwu 322000,China
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摘要: 目的:检测抗体封闭抑制性受体KIR2DL1和KIR2DL2/2DL3对人NK细胞功能的影响。
方法:应用Rosettesep人NK分选试剂盒分选外周血NK细胞,观察抗体封闭KIR2DL1和KIR2DL2/2DL3受体后,NK细胞对不同白血病细胞(人肿瘤细胞株NB4、Raji和K-562)及同种异体的成熟树突状细胞(DC)的杀伤活性;检测混合淋巴细胞反应中NK细胞对T淋巴细胞增殖的影响;检测细胞因子TGF-β1的表达水平。
结果:任何一个抗体封闭KIR后,NK细胞杀伤NB4细胞活性均增强,并与抗体浓度呈现量效关系(P<0.05),2个抗体联用能增强NK细胞的杀伤活性;对Raji细胞的杀伤活性则改变较小;NK细胞对K-562细胞具有很强的杀伤活性,但经抗体封闭后,这种活性并没有提高。经抗体封闭KIR后,NK细胞对成熟DC的杀伤活性明显增强,并与抗体浓度呈现量效关系(2.20%±1.10% vs 37.59%±5.06%),各组间比较差异有统计学意义(P<0.05)。经抗体封闭后的NK细胞在混合淋巴细胞反应中能明显提高对T细胞的增殖抑制(77.85%±8.31% vs 43.05%±5.95%,P<0.05),上清液中细胞因子TGF-β1的含量增加。
结论:以抗体修饰抑制性KIR受体有助于提高NK细胞杀伤肿瘤及同种异体成熟DC的能力,活化的NK细胞通过分泌TGF-β1,抑制T细胞的增殖、活化。
关键词: 杀伤细胞天然受体KIR2DL1 受体KIR2DL2 受体KIR2DL3 抗体单克隆 淋巴细胞培养试验混合 树突细胞 T淋巴细胞    
Abstract: Objective: To investigate the effect of blocking the inhibitory receptors KIR2DL1 and KIR2DL2/2DL3 with monoclonal antibody on cytotoxic activity of human NK cells.
Methods: Human peripheral blood NK cells were isolated by Rosettesep NK sorting kit.The cytotoxic activity of NK cells against human leukemia NB4,K-562,Raji cells and allogeneic mature or dendritic cells (DCs) was detected before or after KIR2DL1 and KIR2DL2/2DL3 were blocked.The effect of NK cells on T lymphocyte proliferation was detected by mixed lymphocyte reaction and TGF-β1 concentration in culture supernatant was measured.
Results: The cytotoxicity of NK cells to NB4 cells was augmented with increasing concentration of the antibody.Combination of both antibodies enhanced killing activity of NK cells.NK cells had strong cytotoxicity to K-562 cells,but were not enhanced by the blockade of inhibitory receptors.The cytotoxicity to Raji cells was not evidently augmented.The cytotoxicity of NK cells to mature DC was enhanced remarkably with the increase of concentration of the antibodies(2.20%±1.10% vs 37.59%±5.06%,P<0.05).In mixed lymphocyte reaction,the blockade of two antibodies enhanced the inhibition effect of NK cells on T cell proliferation(77.85%±8.31% vs 43.05%±5.95%,P<0.05)and the content of TGF-β1 in the supernatant was increased.
Conclusion: The cytotoxic effects of human NK cells against target cells were significantly enhanced with the blockade of inhibitory KIR receptor; and the cytokine TGF-β1 secreted by NK cells further inhibits T cells proliferation.
Key words: Killer cell,natural    Receptors,KIR2DL1; Receptors,KIR2DL2; Receptors,KIR2DL3; Antibodies,monoclonal; Lymphocyte culture test,mixed; Dendritic cells; T-lymphocytes
出版日期: 2011-09-25
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引用本文:

吴功强;赵妍敏;黄河;来晓瑜. 体外抗体功能修饰抑制性KIR对人NK细胞的影响[J]. 浙江大学学报(医学版), 2011, 40(5): 475-481.

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https://www.zjujournals.com/med/CN/Y2011/V40/I5/475

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