【摘要】 目的 :研究神经酰胺对结核杆菌低分子多肽抗原 (Mtb)诱导的γδT细胞活化及凋亡作用。方法 :用结核杆菌低分子多肽抗原刺激正常人PBMC ,并用磁珠阳性分选法获得高纯度的γδT细胞。通过MTT试验观察神经酰胺、鞘磷脂酶以及神经酰胺合成酶抑制剂FumonisinB1对γδT细胞的活化作用 ;FACS检测其对γδT细胞的凋亡作用。结果 :Mtb刺激获得的γδT细胞可达 73 % ,磁珠分选后可高达 98% ;高浓度神经酰胺及鞘磷脂酶能抑制γδT细胞的增殖 ,而出现凋亡 ,FumonisinB1则对γδT细胞的增殖及凋亡无明显影响。结论 :鞘磷脂水解产生的神经酰胺对γδT细胞有致凋亡作用更多还原

【Abstract】 Objective:To study the role of ceramide in the activation and apoptosis of γδT cells induced by low molecular peptide antigen of Mycobacterium tuberculosis.Methods:The low molecular peptide antigen dervied from Mycobacterium tuberculosis(Mtb) was applied to stimulate the peripheral blood mononuclear lymphoctyes of healthy donors.γδT lymphocytes coated by TCR γ/δ Hapten antibody in combination with MACS anti Hapten microbeads FITC were separated with a LS + positive selection MACS separator.γδT cells induced by ceramide,Sphingomyelinase and Fumonisin B1 were detected for their activation by MTT assays and for their apoptosis by flow cytometry.Results:The purity of the γδT lymphocytes stimulated by Mtb and sorted by MACS were up to 73% and 98% respectively. The high concentration of C2 Ceramide and sphingomyelinase were able to inhibit proliferation and to enhance apoptosis of γδT lymphocytes.Conclusion:Ceramide hydrolyzed from sphingomyeline can induced apoptosis of γδT cells.更多还原