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：2018-06-10 07:11

本文选题：树突状细胞 + 调节性T细胞　；参考：《吉林大学》2006年硕士论文

【摘要】：90年代中期以来,大量研究发现一类具有免疫调节功能的细胞亚群,如CD4+CD25+T细胞、Tr1细胞和Th3细胞等[1-3],尽管这些细胞的来源和分化目前还不是很明确,但是根据其功能特点可将其统称为调节T细胞(regulatory T cell,Tr)。人和啮齿类动物体内已经成功分离纯化出Tr细胞,其抑制或调节功能在体内外实验中得到证实。因Tr细胞在感染免疫、肿瘤免疫及器官移植耐受的诱导中具有广泛的应用前景,目前已成为免疫耐受研究中的一个新热点。本研究利用体外培养人外周血单核细胞来源的未成熟树突状细胞(immature dendritic cell, iDC)诱导脐血淋巴细胞产生Tr细胞,经流式细胞仪测定其细胞表型及细胞内细胞因子,应用MTT法测定诱导细胞的生物学功能,为进一步探讨未成熟DC诱导免疫耐受的机制奠定基础。结果表明:细胞因子联合诱导人外周血单核细胞可获得较大量、纯度较高的不同发育阶段的DC。未成熟DC与脐血淋巴细胞混合培养可诱导Tr细胞的生成,其细胞内IL-10的分泌量明显增加,CD4/CD25分子的表达与阴性对照组相比无统计学差异。经混合淋巴细胞反应发现Tr能抑制同种异体淋巴细胞的增殖以及降低细胞毒性T细胞(CTL)的抗肿瘤效应。本研究意义在于:我们通过建立的DC体外培养体系,诱导Tr细胞,为Tr细胞在感染免疫、肿瘤免疫和移植耐受的深入研究和临床应用提供了理论依据。
[Abstract]:Since the mid-1990s, a large number of studies have found a class of immunomodulatory cell subsets, such as CD4 CD25 T cells, Tr1 cells and Th3 cells [1-3], although the origin and differentiation of these cells are not yet clear. However, according to its functional characteristics, it can be called regulatory T cell Trus. Tr cells have been successfully isolated and purified from human and rodent, and their inhibitory or regulatory functions have been confirmed in vitro and in vivo. Tr cells may be widely used in the induction of infection immunity, tumor immunity and organ transplantation tolerance. In this study, immature dendritic cells (DCs) derived from human peripheral blood monocytes (PBMC) were used to induce Tr cells from cord blood lymphocytes. The cell phenotypes and intracellular cytokines were measured by flow cytometry, and the biological functions of induced cells were determined by MTT assay. The results showed that the combination of cytokines induced human peripheral blood mononuclear cells (PBMC) to obtain a large number of DCs with high purity at different developmental stages. Immature DC and umbilical cord blood lymphocytes co-cultured could induce Tr cells to produce Tr cells. The secretion of IL-10 increased significantly the expression of CD4 / CD25 molecules in immature DCs and umbilical cord blood lymphocytes was not significantly different from that in negative control group. Tr can inhibit the proliferation of allogeneic lymphocytes and reduce the antitumor effect of cytotoxic T cell line CTL. The significance of this study is that we can induce Tr cells through the established DC culture system in vitro. It provides a theoretical basis for the further study and clinical application of Tr cells in infection immunity, tumor immunity and transplantation tolerance.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392

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