结核杆菌抗原诱导活化的人γδT细胞L-选择素的表达规律及调控机制

发布时间：2018-04-11 17:07

本文选题：L-选择素 + γδT细胞　；参考：《安徽医科大学》2005年硕士论文

【摘要】：目的 (1)探讨L-选择素(CD62L)在结核杆菌抗原(Mtb-Ag)诱导人γδT细胞表达的动力学特征；(2)观察不同细胞因子对活化人γδT细胞表达L-选择素的调节作用；(3)应用信号分子阻断剂初步探讨PKC，P13K，MAPK等信号途径在人γδT细胞活化过程中对L-选择素表达的调控作用。方法 (1)正常人外周血单个核细胞分别用Mtb-Ag和CD3单抗(CD3mAb)刺激并加IL-2培养扩增，分别获得Mtb-Ag激活的T细胞(MtbAT)(γδT细胞为主)和CD3mAb激活的T细胞(CD3AT)(αβT细胞为主)。(2)取新鲜PBMC和刺激后培养3～21天的γδT细胞和αβT细胞，用多色荧光抗体标记，在流式细胞仪上检测γδT细胞表面L-选择素的表达，并与αβT细胞作比较。(3)Mtb-Ag初次与再次刺激人γδT细胞分别培养7天，在培养同时分别按组加入IFN-γ、anti-IFN-γ、IL-12、anti-IL-12，并与培养的不同时间分别收集细胞，检测L-选择素表达情况。(4)培养6d～8d的MtbAT分别加入PMA、Ionomycin(IMN)、PMA+IMN培养3h、6h、12h、24h后，应用流式细胞术(FCM)分别检测L-选择素的表达率。(5)培养8天的γδT细胞分别加入Mtb-Ag、Rotterin+Mtb-Ag、PMA、Rotterin+PMA刺激4h，，收集细胞并检测L-选择素表达。(6)MtbAT或CD3AT预先用LY294002(P13K途径抑制剂)、SB203580(p38途径抑制剂)、
[Abstract]:Objective 1) to investigate the kinetic characteristics of L- selectin CD62L) on the expression of human 纬未 T cells induced by Mycobacterium tuberculosis antigen Mtb-Ag2) to observe the regulatory effect of different cytokines on the expression of L- selectin in activated human 纬未 T cells.To explore the regulation of L- selectin expression by PKC- P13K- MAPK signaling pathway in the activation of human 纬未 T cells.Methods 1) normal human peripheral blood mononuclear cells were stimulated with Mtb-Ag and CD3 monoclonal antibody CD3mAband cultured with IL-2.Mtb-Ag activated T cells (纬未 T cells) and CD3mAb activated T cells (CD3ATA) were harvested from fresh PBMC and cultured for 321 days after stimulation. The cells were labeled with polychromatic fluorescent antibody.The expression of L- selectin. 4) the MtbAT of 6d~8d was added to PMA Ionomycinia IMN for 3 h, 6 h, 12 h and 24 h, respectively.The expression rate of L- selectin was detected by flow cytometry (FCM). 纬未 T cells cultured for 8 days were added to Mtb-AgRotterin Mtb-AgPMA-Rotterin PMA for 4 h. The cells were collected and detected the expression of L- selectin. MtbAT or CD3AT were pretreated with LY294002(P13K pathway inhibitor SB203580p38.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392

【参考文献】