高表达IDO的树突细胞及色氨酸代谢产物3-HAA抑制小鼠小肠移植急性排斥反应的研究

发布时间：2018-06-27 02:30

本文选题：吲哚胺2 + 3-双加氧酶　；参考：《天津医科大学》2015年硕士论文

【摘要】：本文旨在研究吲哚胺2,3-双加氧酶(IDO)抑制小鼠小肠移植排斥反应的作用机制。通过合成携带IDO序列的载体转染获得高表达IDO的小鼠树突状细胞(IDO+DC),结合色氨酸下游产物3-羟基邻氨基苯甲酸(3-HAA),作用于体外受体来源的T细胞及小鼠小肠移植模型,探索IDO抑制小鼠小肠移植免疫反应的具体原理和机制。目的：研究IDO+DC和3-HAA的抑制小鼠小肠移植排斥作用的机制。方法：DC用含IDO序列的腺病毒载体转染,获得高表达IDO的树突状细胞。获取受体脾CD4+T淋巴细胞,分别与供体DC、DC+3-HAA、IDO+DC和IDO+DC+3-HAA进行混合培养,使用MTS、FCM以及ELISA等方法,全面检测T淋巴细胞的增殖、凋亡、向Treg转化及细胞因子环境变化情况。将IDO+DC及3-HAA输注小肠移植受体小鼠(Balb\c),比较各组小鼠生存时间、抑制小肠组织情况、脾脏Treg亚群、血液细胞因子变化。结果：细胞实验中IDO+DC口3-HAA使T淋巴细胞的增殖受明显抑制而凋亡增加,Treg亚群上调,混合培养体系内IFN-γ, TGF-β,IL-2口IL-10较空白对照组增加。动物实验中,IDO+DC及3-HAA均能增加移植小鼠生存时间、减轻移植小肠组织损伤、上调Treg亚群,且二者效应有叠加作用。结论：联合应用IDO+DC及3-HAA合用和单独给予其中一种干预因素相比,对T细胞功能拥有更强的抑制效应。IDO通过色氨酸代谢与色氨酸产物积累,直接途径抑制T淋巴细胞增殖、诱导T淋巴细胞凋亡,间接途径诱导Treg产生,共同抑制小鼠小肠抑制排斥反应。
[Abstract]:The aim of this study was to investigate the mechanism of indoleamine 2o 3-dioxygenase (IDO) inhibiting rejection of small bowel transplantation in mice. Mouse dendritic cells (IDO DC) with high expression of IDO were obtained by synthesizing the vector carrying IDO sequence and binding 3-hydroxyo-aminobenzoic acid (3-HAA), a downstream product of tryptophan, to act on receptor derived T cells in vitro and small intestine transplantation model in mice. Objective: to explore the mechanism and mechanism of IDO inhibiting immune response of small bowel transplantation in mice. Aim: to study the mechanism of IDO DC and 3-HAA inhibiting the rejection of small bowel transplantation in mice. Methods dendritic cells with high expression of IDO were obtained by transfection with adenovirus vector containing IDO sequence. The recipient spleen CD4 T lymphocytes were obtained and cultured with donor DC 3-HAAZIDO DC and IDO DC 3-HAA, respectively. The proliferation, apoptosis, Treg transformation and cytokine environmental changes of T lymphocytes were detected by MTS- FCM and Elisa. The survival time, inhibition of intestinal tissue, spleen Treg subsets and changes of blood cytokines were compared in IDO-DC and 3-HAA recipient mice (Balb\ c),). Results: in the cell experiment, the proliferation of T lymphocytes was significantly inhibited and apoptosis was increased by 3-HAA in IDO DC, and IFN- 纬 and TGF- 尾 IL-2 were increased in mixed culture system as compared with those in control group. In animal experiments, both IDO DC and 3-HAA could increase the survival time of transplanted mice, alleviate the injury of small intestine and up-regulate the Treg subsets, and the two effects were superimposed. Conclusion: the combination of IDO DC and 3-HAA has a stronger inhibitory effect on T cell function. IDO directly inhibits T lymphocyte proliferation through tryptophan metabolism and tryptophan product accumulation. T-lymphocyte apoptosis was induced, Treg production was induced by indirect pathway, and small intestinal rejection was inhibited in mice.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R656.7

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