IL-21对NK细胞表型和功能的影响  

The effect of IL-21 cytokine on NK cell makers and function

作　　者：李登瑞[1] 郭素敏[1] 李辉[1] 侯宏伟[1] 李晓霞[1] 刘欣燕[1] 杜杰杰 宋利超 高莉[2] 杨永辉[1] LI Deng - rui GUO Su - min LI Hui HOU Hong - wei LI Xiao -xia LIU Xin - yan DU Jie -jie SONG Li - chao GAO Li YANG Yong - hui(The Center of Lung Cancer Prevention, Hebei Province Chest Hospital ,Shijiazhuang 050041, China College of Life Science ,Hebei Normal University,Shijiazhuang,050024 ,China)

机构地区：[1]河北省胸科医院河北省肺癌防治中心,河北石家庄市050041 [2]河北师范大学生命科学学院,河北石家庄市050024

出　　处：《延安大学学报（医学科学版）》2017年第3期1-4,共4页Journal of Yan'an University:Medical Science Edition

基　　金：2012年国家自然科学基金(No:31101638);2013年河北省省级重大医学研究课题(No:ZD2013063);2013年河北省省级重大医学研究课题(No:ZD2013060);2014年省级科技计划项目(No:14277789D)

摘　　要：目的为了研究表达白细胞介素-21(IL-21)对自然杀伤细胞(NK)细胞的扩增的数量、表型和功能的影响。方法以人外周血单个核细胞(PBMC)为扩增培养对象,实验组加入IL-21,在体外培养条件下使NK细胞大量的扩增。采用流式细胞仪检测NK细胞表型。NK细胞作用到K562细胞,对NK细胞的杀伤性进行功能分析,^(51)Cr释放实验检测NK细胞对K562细胞杀伤水平的影响。结果 IL-21加入到PBMC后,培养体系经过23 d的刺激后,IL-21诱导PBMC扩增,CD56+CD16+CD3-细胞数量比对照组增加了2.6倍,对照组CD56+CD16+CD3-细胞的纯度为26.5%±1.7%,实验组扩增后是73.9%±2.1%。细胞毒实验表明,在NK效应细胞:K562靶细胞为10∶1和20∶1时,实验组明显高于对照组。结论本方法以单纯的添加细胞因子IL-21的方法,能够实现NK细胞体外扩增,建立了更优化的NK细胞体外扩增方法,且扩增的NK细胞对K562细胞的杀伤活性较好。本实验对于NK的大量扩增和临床应用具有一点的指导价值。Objective In order to study the influence of different culture conditions on activity of nature killer cells （NK）,whether joining IL-21 cytokine or not. Methods Peripheral blood mononuclear cells （PBMC） were extracted from human peripheral blood,and then were explanted by IL-21 cytokine stimulated. The number of NK cells increased by IL-21 cytokine. Immunophenotypic analysis of NK cell surface markers was performed by flow cytometry （FCM）.^（51）Cr release assay was employed to measure the specific lysis skilling of NK cell target K562 cells.Results We have used IL-21 added into the PBMC,an average of 2. 6 fold expansion of CD3-CD56 ＋ CD16 ＋cells was observed after 23 day of co-culture system. The NK cells population could proliferate more 26. 5% ± 1.7% after expansion in PBMC before expansion by FCM. The cytotoxical activity of NK cells which was induced by IL-21 cytokine was the strongest than induced by IL-6 cytokine alone. The expanded NK cells lysed 73. 9% ± 2. 1% of K562 targets in a 10： 1 and 20： 1 effectors to target ratio. In this case,the NK cell induced by IL-21 against tumor cells was more lethal. Conclusions PBMC based in vitro expansion of natural killer cells was set up by IL-21 cytokine. The cytotoxicity of NK cells was the strongest induced by IL-21 cytokine treated. These results had important guiding significance for the NK large number of amplification and used in clinical.

关 键 词：IL-21 自然杀伤细胞 扩增 51Cr释放实验 

分 类 号：R392.11[医药卫生—免疫学]