NKG2D在细胞因子诱导的杀伤性细胞(CIK)抗血液肿瘤细胞的作用  

Roles of NKG2D in Cytokine-induced Killer(CIK) against Hematological Malignant Cells Lines

作　　者：何金媛[1] 贾祝霞[1] 蔡晓辉[2] 韩文敏[1] 肖溶[1] 马玲娣[3] 卢绪章[2] 周民[1] 陈宝安[2] 

机构地区：[1]常州市第二人民医院血液科,江苏常州213003 [2]东南大学医学院附属中大医院血液科,江苏南京210009 [3]常州市第二人民医院中心实验室,江苏常州213003

出　　处：《中国实验血液学杂志》2013年第6期1380-1384,共5页Journal of Experimental Hematology

基　　金：中国博士后科学基金(编号:20110491337);江苏省博士后基金(编号:11010112C);东南大学博士后启动基金(编号:1124000246);国家自然科学基金(编号:81101647);常州市卫生局重大科技项目(编号:ZD201010)

摘　　要：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 was purposed to investigate the CIK cell cytotoxicity to hematological malignant cell lines by interaction NKG2D receptors and corresponding ligands. The CIK cells was expanded from healthy individual with interferon （IFN） γ, CD3 monoclonal antibodies （mAb） and interleukin-2 （IL-2 ）. The subset of lymphocyte and the expression of NK cell receptors on CIK cells was detected by flow cytometry; NKG2D ligand expression on hematological malignant cell lines was also analyzed by flow cytometry, the calcein acetoxymethyl ester （CAM） was used for labeling target ceils, then the cytotoxicity of CIK cells to hematological malignant cell lines was detected by flow cytometry. The results showed that most of CIK cells expressed CD3 （97.85 ± 1.95% ）, CD3 ＋ CD8 ＋ ceils and CD3 ＋ CD56 ＋ cells increased significantly as compared with un-cultured cells （ P 〈 0. 001 ; P = 0. 033 ）. About 86% CIK cells expressed NKG2D receptor but no other NK receptors such as CD158a, CD158b and NCR. Different levels of NKG2D ligands were detected in hematological maligant cell lines U266, K562 and Daudi. CIK cells showed high cytotoxicity to these three different cell lines, and this cytotoxicity was partially blocked by treating CIK cells with anti-NKG2D antibody （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）. It is concluded that most of CIK cells express NKG2D receptor, interaction of NKG2D-NKG2D ligands may be one of the mechanisms, by which CIK cells kill hematological malignant cells.

关 键 词：NKG2D受体 NKG2D配体 CIK细胞 血液肿瘤细胞 

分 类 号：R730.51[医药卫生—肿瘤] R392.12[医药卫生—临床医学]