【摘要】 目的 :观察神经酰胺对人γδT细胞和αβT细胞的诱导凋亡作用。 方法 :用结核杆菌低分子多肽抗原 (Mtb Ag)和CD3单抗 (CD3mAb)分别刺激正常人外周血单个核细胞 ,获得Mtb Ag激活的T细胞 (γδT细胞为主 )及CD3mAb激活的T细胞(αβT细胞为主 ) ;用不同浓度的神经酰胺 (C2 Cer)分别作用于γδT细胞和αβT细胞 ,培养 3h后收获细胞 ,应用Annexin V FITC/PI染色 ,流式细胞术检测不同亚群T细胞的凋亡 ,应用四甲基噻唑氮蓝 (MTT)比色法检测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αβT细胞比例 >90 % ;与对照组比较 ,低浓度的C2 Cer(≤ 1 0 μmol/L)对人γδT细胞和αβT细胞的诱导凋亡作用和细胞活性抑制作用均不明显 (P >0 .0 5 ) ;而高浓度的C2 Cer对γδT细胞和αβT细胞的诱导凋亡作用和细胞活性抑制作用存在显著差异 :1 0 0 μmol/L的C2 Cer诱导γδT细胞、αβT细胞凋亡比例分别增加 1 0 .4 % (P >0 .0 5 )及 73.5 % (P <0 .0 1 ) ,活细胞区 (R1区 )的细胞比例分别为对照组的 88.1 % (P >0 .0 5 )和7.8% (P <0 .0 1 ) ;而 5 0 0 μmol/L时诱导γδT细胞和αβT细胞凋亡比例分别增加 71 .9%和 73.8% (P <0 .0 1 ) ,R1区的细胞比例分别为对照组的更多还原

【Abstract】 Objective:To investigate the role of apoptosis of γδT cells and αβT cells induced by ceramide.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 to generate Mtb Ag activated T cells(γδT cells were predominent) and CD3mAb activated T cells(αβT cells were predominent).These two T cell subsets were treated with Ceramide(C2 Cer) at different concentration;after 3hours of cultivation,the apoptosis of different subsets of T cells was measured by flowcytometry(FCM),using Annexin V FITC/PI staining;and the availability of T cells was detected by methl thiazolyl tetrazolium(MTT) colorimetric assay.Results:The percentage of γδT cells in Mtb Ag activated T cells and αβT cells in CD3mAb activated T cells were 70% to 90% and >90%,respectively.The apoptosis and the inhibition of the availability of γδT cells and αβT cells induced by lower concentration of C2 Cer(≤10 μmol/L)appeared weak in contrast to their control.There were significant differences in the role of apoptosis of γδT cells and αβT cells induced by higher concentration of C2 Cer;the percentage of the apoptotic γδT cells and αβT cells induced by C2 Cer(100 μmol/L) increased by 10.4%( P > 0.05 ) and 73.5%( P <0.01) in contrast with their control,and the percentage of cells in the region live cells(R1) was 88.1%( P >0.05) and 7.8%( P <0.01) of the ones of their control;whereas the percentage of the apoptotic γδT cells and αβT cells induced by C2 Cer(500 μmol/L) increased by 71.9% and 73.8%( P <0.01) in contrast with their control,and the percentage of cells in R1 was 36.5% and 2.7%( P <0.01) of the ones of their control.Conclusions:The apoptosis of γδT cells and αβT cells can be induced by ceramide;it needs higher concentration of C2 Cer to induce the apoptosis of γδT cells than to induce the apoptosis of αβT cells.更多还原