人外周血树突状细胞的诱导及鉴定
发布时间：2018-10-08 12:13
【摘要】：树突状细胞(dendritic cell, DC)是已知体内功能最强,唯一能活化初始型T 细胞的专职抗原递呈细胞(antigen presenting cell, APC),其数量、成熟状态可决定机体免疫状态,故对其深入的研究具有重要理论和实际意义。DC 具有体外可操作性,因此获取一定量有功能的DC 是深入研究其生物特性的关键。 本研究目的为在体外建立一个稳定的从人外周血单核细胞诱导未成熟及成熟DC 的培养方法,经流式细胞术检测细胞表面分子的表达情况并应用MTT 法对培养细胞的生物学功能进行测定,从而确保获得具有一定形态和功能特征的DC,为进一步探讨DC 在免疫耐受及抗肿瘤方面作用奠定基础。 研究结果:本研究建立了诱导外周血单核细胞获得不同发育阶段DC的方法,获得较大量,纯度较高的CD83+DC。与以往报道有所不同,经流式细胞术检测高表达CD83分子的DC仍有CD14分子的表达。经过再次混合淋巴细胞反应发现具有中度表达共刺激分子的未成熟DC在体外可以诱导同一来源的T细胞免疫低应答。 通过本研究我们建立了稳定的体外DC 培养体系,为移植免疫耐受研究和抗肿瘤研究深入发展提供了理想的基因转移靶细胞。
[Abstract]:Dendritic cell (dendritic cell, DC) is known to have the strongest function in vivo. The only specialized antigen-presenting cell (antigen presenting cell, APC),) that activates the initial T cell is the number of (antigen presenting cell, APC), and the mature state determines the immune state of the body. Therefore, the in-depth study of DC has important theoretical and practical significance. DC can be operated in vitro, so obtaining a certain amount of functional DC is the key to further study its biological characteristics. The aim of this study was to establish a stable culture method for inducing immature and mature DC from human peripheral blood monocytes in vitro. The expression of cell surface molecules was detected by flow cytometry and the biological function of cultured cells was determined by MTT assay. So as to ensure that DC, with certain morphological and functional characteristics lay a foundation for further study of the role of DC in immune tolerance and antitumor. Results: in this study, we established a method to induce peripheral blood monocytes to obtain DC at different developmental stages. A large number of CD83 DC. with high purity were obtained. Different from previous reports, DC with high expression of CD83 still expressed CD14 by flow cytometry. After remixed lymphocyte reaction it was found that immature DC with moderate expression of costimulatory molecules could induce low immune response of T cells from the same source in vitro. Through this study, we established a stable culture system of DC in vitro, which provides an ideal target cell for gene transfer for the further development of transplantation immune tolerance and antitumor research.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392
本文编号：2256706
[Abstract]:Dendritic cell (dendritic cell, DC) is known to have the strongest function in vivo. The only specialized antigen-presenting cell (antigen presenting cell, APC),) that activates the initial T cell is the number of (antigen presenting cell, APC), and the mature state determines the immune state of the body. Therefore, the in-depth study of DC has important theoretical and practical significance. DC can be operated in vitro, so obtaining a certain amount of functional DC is the key to further study its biological characteristics. The aim of this study was to establish a stable culture method for inducing immature and mature DC from human peripheral blood monocytes in vitro. The expression of cell surface molecules was detected by flow cytometry and the biological function of cultured cells was determined by MTT assay. So as to ensure that DC, with certain morphological and functional characteristics lay a foundation for further study of the role of DC in immune tolerance and antitumor. Results: in this study, we established a method to induce peripheral blood monocytes to obtain DC at different developmental stages. A large number of CD83 DC. with high purity were obtained. Different from previous reports, DC with high expression of CD83 still expressed CD14 by flow cytometry. After remixed lymphocyte reaction it was found that immature DC with moderate expression of costimulatory molecules could induce low immune response of T cells from the same source in vitro. Through this study, we established a stable culture system of DC in vitro, which provides an ideal target cell for gene transfer for the further development of transplantation immune tolerance and antitumor research.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392
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