【摘要】 目的　观察不同浓度的乙醇及二甲基亚砜 (DMSO)对人γδT细胞和αβT细胞凋亡与细胞活性的影响。方法　分别用结核杆菌低分子多肽抗原 (Mtb Ag)、CD3单抗(CD3mAb)刺激正常人外周血单个核细胞 ,分别获得Mtb Ag激活的T细胞 (αβT细胞为主 )及CD3mAb激活的T细胞(γδT细胞为主 ) ;用不同浓度的乙醇及二甲基亚砜分别作用于γδT细胞和γδT细胞 4、2 4h后 ,应用Annexin V FITC/PI染色 ,流式细胞术检测不同T细胞亚群的凋亡 ,应用四唑盐(MTT)法检测T细胞的活性。结果　Mtb Ag激活的T细胞中γδT细胞比例可达 70 %～ 90 % ;CD3mAb激活的T细胞中γδT细胞比例 >90 % ;与对照组相比 ,除浓度为 1 0 %及 2 0 %的DMSO作用于γδT细胞 2 4h的诱导凋亡作用明显 (P <0 0 5 )及浓度为 2 0 %的DMSO作用于γδT细胞 4h对细胞活性抑制明显 (P <0 0 5 )外 ,低浓度乙醇及DMSO(≤ 2 0 % )对人γδT细胞和γδT细胞的诱导凋亡作用及对细胞活性的抑制作用均无显著性 (P >0 0 5 ) ;与对照组相比 ,除 5 0 %乙醇作用于γδT细胞 4h及 5 0 %DMSO作用于γδT细胞 4h诱导细胞凋亡作用不明显 (P >0 0 5 )外 ,高浓度乙醇和DMSO(5 0 % )对γδT细胞和γδT细胞的诱导凋亡作用及对细胞活性的抑制作用均增加 (P <0 0 5 )。结论?更多还原

【Abstract】 Objective To investigate effects of ethanol and dimethyl sulfoxide (DMSO) on the apoptosis and the availability of γδT cells and αβT cells. Methods The peripheral blood mononuclear lymphocytes of healthy donors were stimulated respectively with the low molecular peptide antigen derived from Mycobacterium tuberculosis antigen (Mtb-Ag) and CD3mAb, cultured in IL-2 containing media, to generate Mtb-Ag activated T cells (γδT cells were predominant ) and CD3mAb activated T cells (γδT cells were predominant ). These 2 T cell subsets were treated with ethanol and DMSO at different concentration, after 4 h and 24 h of cultivation. The apoptosis of different subsets of T cells were measured by flowcytometry (FCM) using Annexin-V-FITC/PI staining, and the availability of T cells were detected by MTT assay. Results The percentage of γδT cells in Mtb-Ag activated T cells and γδT cells in CD3mAb activated T cells were 70%～90% and >90%, respectively. Besides 1.0% and 2.0% DMSO in culture for 24 h induced profoundl apoptosis (P<0.05) on γδT cells, and 2.0% DMSO in culture for 4 h inhibited significantly the availability of γδT cells (P<0.05), the apoptosis and the inhibition of the availability of γδT cells and γδT cells induced by lower concentration of ethanol and DMSO(≤2.0%)appeared to be weak (P>0.05) in contrast to its control. With the exception of inapparentl apoptosis (P>0.05) of γδT cells induced by 5.0% ethanol in culture for 4 h and γδT cells induced by 5.0% DMSO in culture for 4 h, the apoptosis and the inhibition of the availability of γδT cells and γδT cells induced by higher concentration of ethanol and DMSO(5.0%)appeared to be significantly increased (P<0.05) as compared with its control. Conclusions Ethanol and DMSO can induce the apoptosis of γδT cells and γδT cells, and inhibit the availability of T cells.更多还原