人CD200基因转染未成熟树突状细胞体外诱导调节性T细胞的实验研究
发布时间：2018-11-27 21:24
【摘要】： CD200是近年来新发现的免疫球蛋白超家族成员,研究证明CD200与其受体相互作用,可以下调髓样细胞的活性、减轻自身免疫性疾病的症状、诱导免疫耐受等作用。1995年,Sakaguchi等发现了CD4~+CD25~+调节性T细胞(Tr),调节性T细胞可能是机体以“主动”方式来维持自身稳定的主要途径。Tr对免疫反应具有抑制效应,在体外增殖能力低,在免疫病理、移植物耐受、阻止自身免疫反应和维持机体免疫平衡方面都有一定作用。 本研究用脂质体转染的方法,将pcDNA3-CD200重组质粒转染到外周血分离的未成熟树突状细胞(imDC)内,用荧光标记的抗人CD200抗体作用后,经流式细胞仪监测CD200的表达情况。再用经转染CD200基因的未成熟DC诱导脐血淋巴细胞产生CD4~+CD25~+调节性T细胞(Tr),经流式细胞仪检测调节性T细胞的表型,为进一步探讨CD200基因诱导机体免疫耐受机制的研究奠定基础。 结果表明:pcDNA3-CD200重组质粒转染到未成熟DC后,CD200分子在DC表面大量表达,转染CD200基因的未成熟DC诱导的产生CD4~+CD25~+调节性T细胞(Tr)明显高于对照组;同时证明调节性T细胞(Tr)表面CD4~+CD25~+分子的表达与其表面FOXP3分子的表达在统计学上没有显著性差异。 本研究的意义在于:建立稳定表达CD200分子的未成熟DC体系,转染CD200基因的未成熟DC可以诱导产生更多的Tr,提示诱生Tr是CD200诱导免疫耐受的途径之一;同时,在调节性T细胞中FOXP3的表达与CD4~+CD25~+分子的表达在统计学上差异没有显著性,且CD200分子可以诱生更多的Tr,从而推断CD200基因具有诱导FOXP3分子表达的作用,为CD200诱导免疫耐受机制的进一步研究奠定了基础。
[Abstract]:CD200 is a newly discovered member of immunoglobulin superfamily in recent years. It has been proved that the interaction between CD200 and its receptors can down-regulate the activity of myeloid cells, relieve the symptoms of autoimmune diseases, induce immune tolerance and so on. Sakaguchi et al found that CD4~ CD25~ regulatory T cell (Tr), regulatory T cell may be the main way for the body to maintain its own stability in an "active" manner. Tr has inhibitory effect on immune response, low proliferative ability in vitro and immunopathology. Graft tolerance, blocking autoimmune responses and maintaining immune balance play a role. In this study, pcDNA3-CD200 recombinant plasmid was transfected into (imDC) of immature dendritic cells isolated from peripheral blood by liposome transfection, and the expression of CD200 was monitored by flow cytometry after using fluorescent labeled anti-human CD200 antibody. Immature DC transfected with CD200 gene was used to induce CD4~ CD25~ regulatory T cell (Tr), in cord blood lymphocytes. Flow cytometry was used to detect the phenotype of regulatory T cells. It lays a foundation for further study on the mechanism of immune tolerance induced by CD200 gene. The results showed that after transfection of pcDNA3-CD200 recombinant plasmid into immature DC, CD200 molecules were expressed on the surface of DC. (Tr) of CD4~ CD25~ regulatory T cells induced by immature DC transfected with CD200 gene was significantly higher than that of control group. At the same time, there was no statistical difference between the expression of CD4~ CD25~ molecule on regulatory T cell (Tr) surface and that of FOXP3 molecule on regulatory T cell surface. The significance of this study lies in the establishment of immature DC system expressing CD200 molecules stably. Immature DC transfected with CD200 gene can induce more Tr, to induce Tr as one of the pathways of CD200 induced immune tolerance. At the same time, there was no statistically significant difference between the expression of FOXP3 and CD4~ CD25~ in regulatory T cells, and CD200 could induce more Tr, thus inferred that CD200 gene could induce the expression of FOXP3. It lays a foundation for the further study of the mechanism of immune tolerance induced by CD200.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R392
本文编号：2362074
[Abstract]:CD200 is a newly discovered member of immunoglobulin superfamily in recent years. It has been proved that the interaction between CD200 and its receptors can down-regulate the activity of myeloid cells, relieve the symptoms of autoimmune diseases, induce immune tolerance and so on. Sakaguchi et al found that CD4~ CD25~ regulatory T cell (Tr), regulatory T cell may be the main way for the body to maintain its own stability in an "active" manner. Tr has inhibitory effect on immune response, low proliferative ability in vitro and immunopathology. Graft tolerance, blocking autoimmune responses and maintaining immune balance play a role. In this study, pcDNA3-CD200 recombinant plasmid was transfected into (imDC) of immature dendritic cells isolated from peripheral blood by liposome transfection, and the expression of CD200 was monitored by flow cytometry after using fluorescent labeled anti-human CD200 antibody. Immature DC transfected with CD200 gene was used to induce CD4~ CD25~ regulatory T cell (Tr), in cord blood lymphocytes. Flow cytometry was used to detect the phenotype of regulatory T cells. It lays a foundation for further study on the mechanism of immune tolerance induced by CD200 gene. The results showed that after transfection of pcDNA3-CD200 recombinant plasmid into immature DC, CD200 molecules were expressed on the surface of DC. (Tr) of CD4~ CD25~ regulatory T cells induced by immature DC transfected with CD200 gene was significantly higher than that of control group. At the same time, there was no statistical difference between the expression of CD4~ CD25~ molecule on regulatory T cell (Tr) surface and that of FOXP3 molecule on regulatory T cell surface. The significance of this study lies in the establishment of immature DC system expressing CD200 molecules stably. Immature DC transfected with CD200 gene can induce more Tr, to induce Tr as one of the pathways of CD200 induced immune tolerance. At the same time, there was no statistically significant difference between the expression of FOXP3 and CD4~ CD25~ in regulatory T cells, and CD200 could induce more Tr, thus inferred that CD200 gene could induce the expression of FOXP3. It lays a foundation for the further study of the mechanism of immune tolerance induced by CD200.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R392
【参考文献】
相关期刊论文 前2条
1 侯治富,郭楠,高申,郑德明,卜丽莎,高嵩,才华;人外周血树突状细胞的诱导与鉴定[J];吉林大学学报(医学版);2005年05期
2 李浩威,段连宁;树突状细胞诱导T细胞免疫耐受的机理[J];国外医学(免疫学分册);2003年05期
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