AR-A014418对γδT细胞增殖、凋亡及杀伤功能的影响  

Effect of AR-A014418 on human γδT cells growth,apoptosis and killing function in vitro

作　　者：郑璐[1] 孙蕾清[1] 陈复兴[1] 周遹 吕小婷[1] 陈玲[1] 李莹[1] 周忠海[1] 陈永强[1] ZHENG Lu;SUN Lei-Qing;CHEN Fu-Xing;ZHOU Yu;LV Xiao-Ting;CHEN Ling;LI Ying;ZHOU Zhong-Hai;CHEN Yong-Qiang(Department of Central Laboratory,97th Hospital of PLA,Xuzhou 221004,China)

机构地区：[1]解放军第97医院中心实验室,徐州221004

出　　处：《中国免疫学杂志》2019年第1期20-24,共5页Chinese Journal of Immunology

基　　金：南京军区医学科技创新研究基金(14MS032)资助

摘　　要：目的:观察糖原合酶激酶-3β(GSK-3β)选择性抑制剂AR-A014418对体外扩增的γδT细胞的细胞活性包括增殖、凋亡和杀伤功能的影响。方法:采用人淋巴细胞分离液分离健康人外周血中单个核细胞,在γδT细胞完全培养基中培养获得γδT细胞。用不同浓度的AR-A014418诱导培养γδT细胞72 h后,用流式细胞术检测各诱导处理组γδT细胞的增殖、凋亡和杀伤能力;用Western blot检测γδT细胞凋亡相关蛋白Bcl-2、Bax和Caspase-3表达情况。结果:AR-A014418浓度在0. 3～10μmol/L时能促进γδT细胞的增殖并抑制其凋亡。同时促凋亡蛋白Cleaved caspase-3和Bax表达逐渐减弱,而抗凋亡蛋白Bcl-2表达逐渐增强。尤其是在3μmol/L时,γδT细胞增殖活性最强,而且体外杀伤人肝癌HepG2细胞和人胃癌SGC-7901细胞活性也显著提高。结论:GSK-3β抑制剂AR-A014418在3μmol/L浓度时,可以抑制体外扩增γδT细胞的凋亡,并能增强其体外增殖和杀伤肿瘤活性,提示AR-A014418具有一定的免疫调节功能,为今后用于体外扩增γδT细胞提供实验依据。Objective:To investigate the effect of glycogen synthase kinase-3βselective inhibitor AR-A014418 on humanγδT cells growth,apoptosis and killing function in vitro.Methods:Peripheral blood mononuclear cell(PBMCs)was separated using Ficoll-Hypaque gradient centrifugation and cultivated inγδT complete medium in vitro.After co-cultured with various concentrations of AR-A014418 for 72 h,the proliferation,apoptosis and killing function in each group were determined by flow cytometric analysis.The protein expression was examined by Western blot analysis.Results:AR-A014418 at 0.3-10μmol/L could promote the growth ofγδT cells and enhance the cytotoxic activity ofγδT cells to HepG2 cells and SGC-7901,especially at 3μmol/L for 72h.Furthermore,AR-A014418 could upregulate the expression of the anti-apoptotic protein Bcl-2 and inhibition of pro-apoptotic protein cleaved caspase-3 and Bax expression.Conclusion:AR-A014418 at indicated concentrations can promote theγδT cells growth and enhance the cytotoxic activity ofγδT cells to HepG2 and SGC-7901 cells.These findings suggest that AR-A014418 can be a useful complementary agent for improvingγδT cell-based immunotherapy.

关 键 词：AR-A014418 ΓΔT细胞 增殖 凋亡 杀伤功能 

分 类 号：R392.12[医药卫生—免疫学] Q253[医药卫生—基础医学]