IFN-β1a对多发性硬化树突状细胞共刺激分子表达和成熟状态影响的研究

发布时间：2018-05-13 23:23

本文选题：树突状细胞 + 多发性硬化　；参考：《广西医科大学》2009年硕士论文

【摘要】： 目的研究多发性硬化(MS)患者外周血单核细胞诱导树突状细胞(DC)的体外培养扩增和鉴定;从分子水平分析DC在MS病理免疫中的作用;探讨IFN-β1a对患者DC共刺激分子、成熟状态的影响。方法设立MS组(10例)和健康对照组(8人),抽取外周血20ml,采用梯度密度离心法提取单核细胞。在体外经rhGM-CSF 100ng/mL和rhlL-4 50ng/mL联合培养诱导成为DC,用LPS刺激成熟,经IFN-β1a (rebif)处理,于培养的第九天用倒置相差显微镜和电镜进行形态学鉴定;流式细胞仪检测HLA-DR、CD80、CD86、CD83表达水平。结果第5天体外培养的DC由贴壁状态变为悬浮毛刺状细胞,第9天为形态不规则的毛刺状,呈现典型DC形态;流式细胞仪检测健康人成熟DC的各项表型CD80、CD86、CD83、HLA-DR表达百分率分别为(73.2±11)%、(90.4±5.2) %、(81.4±10.3) %和(91.3±5.9) %。MS组DC表达CD83、CD86明显低于健康对照组(p0.05)。两组间CD80、HLA-DR的表达无显著差异性(p0.05)。经IFN-β1a处理后MS组DC表达CD83、CD86均显著上调(p 0.05),而DC表型HLA-DR和CD80表达显著下调(p 0.05)。结论1、利用rhGM-CSF联合rhIL-4诱导人外周血细胞单核细胞,能得到大量纯度较高的DC,是一种比较简便经济的DC培养方法,有一定的临床应用价值;2、MS患者单核细胞来源的DC表面CD86表达降低,成熟度受损,在MS的发病中具有重要作用;3、IFN-β1a通过下调DC的共刺激分子CD80表达,上调CD86表达,调整细胞处理和提呈抗原能力等方面发挥免疫调节作用,是其治疗MS的重要机制之一。
[Abstract]:Objective to study the proliferation and identification of dendritic cells (DC) induced by peripheral blood monocytes in patients with multiple sclerosis (MS) in vitro, to analyze the role of DC in MS pathological immunity at the molecular level, and to explore the costimulatory molecules of DC induced by IFN- 尾 1a. The effect of maturity. Methods MS group (n = 10) and healthy control group (n = 8) were used to extract 20 ml peripheral blood and to extract monocytes by gradient density centrifugation. After co-cultured with rhGM-CSF 100ng/mL and rhlL-4 50ng/mL, DCS was induced to mature with LPS, then treated with IFN- 尾 1a rebif, morphological identification was performed by inverted phase contrast microscope and electron microscope on the 9th day of culture, and CD86 CD83 expression level of HLA-DRN CD80 was detected by flow cytometry. Results DC cultured in vitro on the 5th day changed from adherent state to suspending hairy cells, and on the 9th day it was irregular burr shape, which showed typical DC morphology. The percentage of HLA-DR expression in all phenotypes of mature DC was 73.2 卤110.4 卤5.2) and 91.4 卤10.3% and 91.3 卤5.9%, respectively. The expression of CD83p86 in DC in MS group was significantly lower than that in healthy control group (p0.05). There was no significant difference in the expression of HLA-DR between the two groups (p 0.05). After treatment with IFN- 尾 _ 1a, the expression of CD83-, CD86 in DC of MS group was significantly up-regulated, while the expression of HLA-DR and CD80 in DC was significantly down-regulated. Conclusion 1. RhGM-CSF combined with rhIL-4 can induce monocytes of human peripheral blood cells to obtain a large number of DCs with high purity, which is a simple and economical method for DC culture. The expression of CD86 on DC surface is decreased and the maturity is impaired. It plays an important role in the pathogenesis of MS. IFN- 尾 1a up-regulates the expression of CD86 by down-regulating the expression of CD80, a costimulatory molecule of DC. Regulating the ability of cell processing and presenting antigen plays an important role in the treatment of MS.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392

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