He Jin-Yuan, Jia Zhu-Xia, Cai Xiao-Hui, Han Wen-Min, Xiao Rong, Ma Ling-Di, Lu Xu-Zhang, Zhou Min, Chen Bao-An
Department of Hematology, Changzhou Mumicipal People's Hospital, Changzhou 213003,Jiangsu Province, China.
Central Laboratory, Changzhou Mumicipal People's Hospital, Changzhou 213003,Jiangsu Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2013 Dec;21(6):1380-4. doi: 10.7534/j.issn.1009-2137.2013.06.003.
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 cells, 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(+) cells 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 malignant 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.
本研究旨在通过NKG2D受体及其相应配体的相互作用，探讨CIK细胞对血液系统恶性肿瘤细胞系的细胞毒性作用。CIK细胞由健康个体外周血单个核细胞，经γ干扰素（IFN）γ、CD3单克隆抗体（mAb）及白细胞介素-2（IL-2）体外诱导培养扩增获得。采用流式细胞术检测CIK细胞淋巴细胞亚群及NK细胞受体的表达；采用流式细胞术分析血液系统恶性肿瘤细胞系NKG2D配体的表达，用羧基荧光素二醋酸盐琥珀酰亚胺酯（calcein acetoxymethyl ester，CAM）标记靶细胞，通过流式细胞术检测CIK细胞对血液系统恶性肿瘤细胞系的细胞毒性作用。结果显示，多数CIK细胞表达CD3（97.85±1.95%），与未培养细胞相比，CD3(+)CD8(+)细胞及CD3(+)CD56(+)细胞显著增加（P<0.001；P=0.033）。约86%的CIK细胞表达NKG2D受体，而不表达其他NK细胞受体如CD158a、CD158b及自然细胞毒性受体（NCR）。在血液系统恶性肿瘤细胞系U266、K562及Daudi中检测到不同水平的NKG2D配体表达。CIK细胞对这三种不同细胞系均显示出高细胞毒性，且这种细胞毒性作用可被抗NKG2D抗体部分阻断（U266：52.67±4.63%比32.67±4.81%，P=0.008；K562：71.67±4.91%比50.33±4.91%，P=0.007；Daudi：68.67±5.04比52.67±2.60%，P=0.024）。结论：多数CIK细胞表达NKG2D受体，NKG2D-NKG2D配体相互作用可能是CIK细胞杀伤血液系统恶性肿瘤细胞的机制之一。
