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机构地区:[1]上海交通大学医学院,上海市免疫学研究所,上海200025
出 处:《现代免疫学》2012年第2期94-98,共5页Current Immunology
基 金:国家自然基金项目(30972691,30700766);上海市青年科技启明星人才计划A类(10QA1405900);上海市卫生系统优秀青年人才培养计划(XYQ2011015);上海市科委基础重点资助项目(11JC1410802)
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。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 3 H-TdR incorporation. Moreover, flow cytometry assayed Tk-induced immunosuppression is not caused by ribosome-inactivating protein. By Real-time quantitative PCR and ELISA as say, 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.
关 键 词:天花粉蛋白 混合淋巴细胞增殖反应 细胞因子 负向调节
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