糖原合酶激酶-3β抑制剂TWS119对γδT细胞增殖及表型的影响  

Effect of glycogen synthase kinase-3β inhibitor TWS119 on proliferation and phenotypic characteristics of human γδT cells in vitro

作　　者：陈永强[1] 郑璐[1] 刘军权[1] 吕小婷[1] 周燏[1] 陈玲[1] 徐晶[1] 周忠海[1] 陈复兴[1] 

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》2015年第6期748-752,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抑制剂4,6-二取代吡咯并嘧啶（TWS119）对γδT细胞增殖和表型的影响。方法：从健康人外周血中分离单个核细胞,加入到γδT细胞完全培养基培养,分别于第1天和第8天时,加入TWS119（0~8.0μmol/L）诱导培养。培养到12 d后,用CCK-8法测定γδT细胞增殖能力;用流式细胞术检测γδT细胞纯度、CD45RA和CD62L的表达。结果：TWS119能够活化γδT细胞Wnt/β-catenin信号通路。在第1天加入TWS119诱导培养组中,随着TWS119浓度的增加,γδT细胞表面CD45RA和CD62L的阳性表达率逐渐升高,而对细胞的增殖及纯度呈抑制作用。在第8天加入TWS119诱导培养组中,TWS119能剂量依赖性促进CD62L的表达而对CD45RA表达无影响,同时在一定浓度范围能促进γδT细胞的增殖和提高纯度的作用。结论：TWS119在一定浓度范围内能促进γδT细胞增殖和提高γδT细胞纯度,并能活化γδT细胞Wnt/β-catenin信号通获得CD45RA＋CD62L＋γδT细胞和CD62L＋γδT细胞。Objective： To investigate the effect of glycogen synthase kinase-3β inhibitor TWS119 on hunman γδT cells growth and phenotypic characteristics. Methods： Using γδT medium to cultivate human peripheral blood γδT cells in vitro. After co-cultured with different concentrations of glycogen synthase kinase-3β（ GSK-3β） inhibitor 4,6-disubstituted pyrrolopyrimidine（ TWS119） for indicated time,growth curve and Wnt / β-catenin activation of in each group were determined by CCK-8 and Western blot assays. The CD62 L and CD45 RA expression the γδT cells were detected using flow cytometry. Results： Wnt / β-catenin pathway of γδT cells could activate by TWS119. In the first group,TWS119 could upregulate the expression of CD62 L and CD45 RA in dose dependent manner while inhibit the growth and ratio of γδT cells. In the second group,TWS119 could promote the growth and ratio of γδT cells with the increase in concentration from 0 μmol / L to 4. 0 μmol / L and decreased thereafter. Besides,TWS119 could promote the expression of CD62 L in a dose-dependent and had no effect on the CD45 RA. Conclusion： Human γδT cells isolated from peripheral blood treated with TWS119 gave rise to two subsets of CD45RA＋CD62L＋γδT cells and CD62L＋γδT cells.

关 键 词：TWS119 ΓΔT细胞 细胞增殖 表型 

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