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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 observe the effect of glycogen synthase kinase-3β inhibitor 4,6-disubstituted pyrrolopyrimidine (TWS119) on the proliferation and phenotype of γδT cells. Methods: Mononuclear cells were isolated from peripheral blood of healthy volunteers and cultured in complete medium of γδT cells. At day 1 and day 8, TWS119 (0 ~ 8.0μmol / L) was added to induce the culture. After 12 d of culture, the proliferation of γδT cells was measured by CCK-8 assay. The purity of γδT cells, the expression of CD45RA and CD62L were detected by flow cytometry. Results: TWS119 could activate Wnt / β-catenin signaling in γδT cells. With the increase of TWS119 concentration, the positive expression rates of CD45RA and CD62L on γδT cells gradually increased in TWS119 induction group on the first day, but inhibited the proliferation and purity of the cells. TWS119 could promote the expression of CD62L dose-dependently but had no effect on the expression of CD45RA in TWS119-induced culture group on the 8th day. At the same time, TWS119 could promote the proliferation and improve the purity of γδT cells in a certain concentration range. CONCLUSION: TWS119 can promote the proliferation of γδT cells and increase the purity of γδT cells in a certain concentration range, and activate the activation of γδT cells through Wnt / β-catenin signaling to obtain CD45RA + CD62L + γδT cells and CD62L + γδT cells.