NK细胞与CIK细胞体外诱导扩增及其活性比较  

Induction and expansion of NK and CIK cells in vitro and comparison of their activities

作　　者：赵杨祉[1] 崔久嵬[1] 徐东升[1] 牛超[1] 金浩范[1] 梁婷婷[1] 李红[1] 王冠军[1] 李薇[1] 

机构地区：[1]吉林大学第一医院肿瘤中心,吉林长春130021

出　　处：《吉林大学学报（医学版）》2013年第3期477-481,共5页Journal of Jilin University：Medicine Edition

基　　金：教育部科研基金资助课题(311015);吉林省财政厅科研基金资助课题(3D5116523428);吉林省科技厅科研基金资助课题(20100749)

摘　　要：目的:探讨体外诱导和扩增NK细胞和CIK细胞的方法,比较二者的增殖能力及其对肿瘤细胞的杀伤活性。方法:提取肿瘤患者外周血单个核细胞(PBMCs),在细胞因子诱导下培养NK细胞和CIK细胞。观察2种细胞的形态学变化,采用流式细胞术检测CD3-CD56+NK细胞和CD3+CD56+CIK细胞比例,计算细胞扩增倍数;采用ELISA法检测NK细胞和CIK细胞分泌干扰素-γ(IFN-γ)的水平;采用CCK-8法检测NK和CIK细胞对K562细胞株的杀伤活性。结果:经过14d体外诱导培养后,NK细胞扩增(160.00±12.15)倍,CIK细胞扩增(110.00±15.48)倍,NK细胞的扩增能力强于CIK细胞(P<0.05);NK和CIK细胞分泌IFN-γ量分别为(4 227.75±731.94)和(525.96±304.84)μg.L-1,NK细胞分泌IFN-γ的能力强于CIK细胞(P<0.01);诱导扩增后NK和CIK细胞对K562细胞株具有较强的杀伤活性,随着效靶比的升高杀伤活性增强。在效靶比为25∶1时,NK细胞和CIK细胞对K562细胞的杀伤率分别为65.35%和57.68%,NK细胞的杀伤活性强于CIK细胞(P<0.01)。结论:在体外成功建立了NK细胞和CIK细胞的诱导扩增体系,NK细胞的扩增能力、IFN-γ分泌水平以及其对K562细胞株的杀伤作用均强于CIK细胞。Objective To explore the methods to induce and expand the NK and CIK cells in vitro and to compare the proliferation abilities and anti-tumor activities of NK and CIK cells.Methods Peripheral blood mononuclear cells（PBMCs） were separated from the peripheral blood of cancer patients.The PBMCs were induced by cytokines to culture NK and CIK cells.The morphological changes of NK and CIK cells were observed.The percentages of CD3-CD56＋NK and CD3＋CD56＋ CIK cells were detected by flow cytometry.And the expansion folds of NK and CIK cells were calculated.The levels of interferon-γ（IFN-γ） secreted by NK and CIK cells were detected by ELISA method.And the killing activities of NK and CIK cells against K562 cell strain were detected by CCK-8 method.Results After induction and expansion in vitro for 14 d,the NK cells expanded（160.00±12.15） folds,and the CIK cells expanded（110.00±15.48） folds.The expansion ability of the NK cells was higher than that of the CIK cells（P0.05）.The secretion levels of IFN-γ in the supernatant of NK and CIK culture system raised to（4 227.75±731.94） and（525.96±304.84） μg·L-1,respectively.The secretion level of IFN-γ of the NK cells was higher than that of the CIK cells（P0.01）.After induction and expansion,the cytotoxicities of NK and CIK cells against K562 cells were enhanced with the increase of effector-target ratio.The killing rates of NK and CIK cells against K562 cells were 65.35% and 57.68% at 25∶1 effector-target ratio,respectively.The killing activity of NK cells was higher than that of CIK cells at the same effector-target ratio（P0.01）.Conclusion The induction and expansion system of NK and CIK cells in vitro is established successfully,and the expansion capability,the secretion level of IFN-γ,and the cytotoxicity of the NK cells are higher than those of the CIK cells.

关 键 词：自然杀伤细胞 细胞因子诱导的杀伤细胞 杀伤活性 

分 类 号：R733[医药卫生—肿瘤]