【摘要】 建立同种异体混合淋巴细胞反应(MLR)体系,采用同位素3 H-TdR掺入、FACS、Real-time PCR、ELISA、Westernblot等方法研究低剂量Tk抑制细胞增殖反应的效果及作用机制。FACS及同位素掺入检测结果显示,低剂量Tk通过非RIP的方式显著抑制MLR的增殖;Real-time PCR及ELISA结果表明,天花粉抑制的增殖体系中较对照组IFN-γ表达水平降低,IL-4、IL-9、IL-10、IL-24表达水平增强;Western blot显示,Tk使得JNK磷酸化增强。以上结果说明Tk显著抑制人同种异体混合淋巴细胞增殖反应,其负向免疫调节作用可能由多种细胞因子、信号通路及T细胞亚群的介导参与。更多还原

【Abstract】 To elucidate the mechanisms underlying the Trichosanthin-induced immunosuppression and its potential clinical application,human allogeneic mixed lymphocyte reaction(MLR) was established,we found that MLR was significantly inhibited with a suppression rate of 66.8% at a Tk concentration 1 μg/ml by 3H-TdR incorporation.Moreover,flow cytometry assayed Tk-induced immunosuppression is not caused by ribosome-inactivating protein.By Real-time quantitative PCR and ELISA assay,our data showed that there are differential patterns of the cytokines secretion in the presence of 1 μg/ml Tk or not,the expression of cytokines IL-4,IL-9,IL-10,IL-24 were increased in the presence of 1 μg/ml Tk,while production of IFN-γ decreased.Tk can also significantly up-regulate and activated c-Jun N-terminal kinase(JNK),Phospho-JNK of allogeneic MLR by Western blotting.更多还原