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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 establish a allogeneic mixed lymphocyte reaction (MLR) system, the effects of low-dose Tk on cell proliferation were investigated by isotope 3 H-TdR incorporation, FACS, Real-time PCR, ELISA and Western blot. The results of FACS and isotope incorporation showed that low-dose Tk could significantly inhibit the proliferation of MLR by non-RIP. Real-time PCR and ELISA showed that the expression of IFN- 4, IL-9, IL-10, IL-24 expression increased; Western blot showed that Tk JNK phosphorylation increased. The above results indicate that Tk significantly inhibits the proliferation of human allogenic mixed lymphocytes, and its negative immune regulation may be mediated by a variety of cytokines, signaling pathways and T cell subsets.