可溶性Tim-3体外增强免疫应答的作用及机制研究

发布时间：2018-01-13 10:21

本文关键词：可溶性Tim-3体外增强免疫应答的作用及机制研究　出处：《军事医学》2017年01期 　论文类型：期刊论文

【摘要】：目的评价重组人(hTim-3)-Fc融合蛋白在调节免疫应答中的作用及其机制研究。方法分别将重组hTim-3-Fc融合蛋白与人巨噬细胞系U937细胞、T淋巴细胞系Jurkat细胞及正常人外周血细胞共培养,采用ELISA、实时荧光定量PCR(RT-PCR)和蛋白质免疫印迹(Western印迹)方法检测免疫相关细胞因子水平的变化,探讨重组hTim-3-Fc融合蛋白对免疫细胞活性的影响并阐明机制。结果重组hTim-3-Fc融合蛋白可显著提高免疫细胞活性,表现为U937中IL-8分泌水平,Jurkat细胞IL-2分泌水平,以及人外周血中IL-2和IFN-γ分泌水平显著升高;在mRNA水平,发现重组hTim-3-Fc融合蛋白能显著提高上述细胞中Ⅰ型干扰素(包括IFN-α2、IFN-β1)的表达。在分子机制方面,重组hTim-3-Fc融合蛋白可增强T细胞(Jurkat)及巨噬细胞(U937)中stat-1的磷酸化水平。结论制备的重组人Tim-3-Fc融合蛋白在体外能显著增强免疫细胞活性,有望成为免疫抑制性疾病新的免疫干预药物。
[Abstract]:Objective to evaluate the recombinant human hTim-3). The role and mechanism of -FC fusion protein in regulating immune response. Methods Recombinant hTim-3-Fc fusion protein was transfected into human macrophage cell line U937. T-lymphocytes were co-cultured with Jurkat cells and normal peripheral blood cells, and ELISA was used. The changes of immune-associated cytokines were detected by real-time fluorescence quantitative PCR- RT-PCR and Western blot. To explore the effect of recombinant hTim-3-Fc fusion protein on immune cell activity and elucidate the mechanism. Results the recombinant hTim-3-Fc fusion protein could significantly improve the immune cell activity. The level of IL-8 secretion in U937 and the levels of IL-2 and IFN- 纬 in human peripheral blood were significantly increased. At the mRNA level, it was found that the recombinant hTim-3-Fc fusion protein could significantly increase the expression of interferon type I (including IFN- 伪 2, IFN- 尾 1) in these cells. Recombinant hTim-3-Fc fusion protein enhanced T cell Jurkat and macrophage U937). Conclusion the recombinant human Tim-3-Fc fusion protein can significantly enhance the activity of immune cells in vitro. It is expected to be a new immune intervention drug for immunosuppressive diseases.
【作者单位】：温州医科大学基础医学院;军事医学科学院基础医学研究所免疫学研究室;安徽医科大学;
【基金】：国家自然科学基金面上资助项目(81471540) 北京市自然科学基金重点资助项目(7141007)
【分类号】：R392
【正文快照】： T细胞免疫球蛋白及黏蛋白3(T cell immuno-globulin and mucin-domain containing-3,Tim-3)是表达于活化的Th1、Th17、巨噬细胞、树突状细胞等表面的免疫调控分子,隶属于Tim家族成员,通过与其配体半乳凝素9(galectin-9,Gal-9)结合诱导细胞凋亡,或通过其他机制抑制T效应细胞的

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