从健康人外周血单个核细胞中分离出高纯度的CD3^-CD56^+CD16^+NK细胞对肝癌细胞的杀伤作用分析  

Analysis of killing effect of high purity CD3^-CD56^+CD16^+ NK cells isolated from peripheral blood mononuclear cells of healthy people against human hepatocellular carcinoma cell lines

作　　者：卢铨 陈浩 沙卫红 王启仪 LU Quan;CHEN Hao;SHA Weihong;WANG Qiyi(Shantou University Medical College, Shantou 515041;Department of Gastroenterology, Guangdong General Hospital and Guangdong Academy of Medical Sciences, China)

机构地区：[1]汕头大学医学院,广东汕头515041 [2]广东省人民医院广东省医学科学院消化内科

出　　处：《胃肠病学和肝病学杂志》2019年第9期978-982,共5页Chinese Journal of Gastroenterology and Hepatology

基　　金：国家自然科学基金(81300279,81741067);广东省自然科学基金(2016A030313815,2017A030313464);广东省科技计划项目(2015A020212029);广州市科技计划项目(201707010419,201804010050)

摘　　要：目的观察经IL-2/IL-15/SCF诱导的CD3^-CD56^+CD16^+ NK细胞体外扩增能力、NKG2D表达变化及其对肝癌细胞的杀伤作用。方法应用免疫磁珠分选法(MACS)从健康人外周血单个核细胞中分离出高纯度的CD3^-CD56^+CD16^+ NK细胞。采用细胞计数法、流式细胞术免疫表型分析和CCK-8等方法比较IL-2/IL-15/SCF对NK细胞扩增倍数、NKG2D表达及对HepG2、SMMC-7721肝癌细胞株杀伤活性的影响;同时观察封闭NKG2D受体对CD3^-CD56^+CD16^+ NK细胞杀伤活性的影响。结果 IL-2/IL-15/SCF培养14 d后,CD3^-CD56^+CD16^+ NK细胞扩增倍数、NKG2D表达以及对HepG2和SMMC-7721的杀伤活性显著增强,与对照组、IL-2组和IL-15组相比,差异均有统计学意义(P<0.05)。NKG2D单抗可以下调其对HepG2细胞的杀伤效应,但对SMMC-7721的杀伤无明显影响。结论 IL-2/IL-15/SCF诱导后CD3^-CD56^+CD16^+ NK细胞对肝癌细胞的杀伤活性增强,NKG2D途径是NK细胞杀伤肝癌细胞的重要机制之一。Objective To investigate the effects of IL-2, IL-15 and stem cell factor(SCF) on the efficiency of expansion, expression of NKG2D and the cytotoxcity of CD3^-CD56^+CD16^+ NK cells against human hepatocellular carcinoma cell lines. Methods The CD3^-CD56^+CD16^+ NK cells purified by the sorting method of magnetic microbeads activated cells sorting(MACS) were expanded in the presence of IL-2 or IL-15 or SCF with or without anti-NKG2D monoclonal antibody. The efficiency of expansion, expression of NKG2D and cytotoxcity of CD3^-CD56^+CD16^+ NK cells against HepG2, SMMC-7721 were determined by trypan blue staining, flow cytometry and CCK-8 assay, respectively. Results The efficiency of expansion, expression of NKG2D and cytotoxcity of CD3^-CD56^+CD16^+ NK cells treated with IL-2 or IL-15 or SCF were significantly increased than those in other groups. Most interestingly, the cytotoxic effects of CD3^-CD56^+CD16^+ NK cells on HepG2 cells were significantly reduced by the pretreatment with anti-NKG2D monoclonal antibody(P<0.05). However, the treatment with the same antibody had little influence on the cytotoxicity of CD3^-CD56^+CD16^+ NK cells against SMMC-7721 cells. Conclusion IL-2 or IL-15 or SCF enhances the efficiency of expansion, expression of NKG2D and cytotoxicity of CD3^-CD56^+CD16^+ NK cells against human hepatocellular carcinoma cell lines. NKG2D plays an important role in mediating the cytotoxicity of NK cells on specific types of tumor cells.

关 键 词：自然杀伤细胞 肝癌 NKG2D 白细胞介素2 白细胞介素15 细胞治疗 

分 类 号：R735.7[医药卫生—肿瘤]