雷帕霉素对树突细胞免疫表型影响的初步研究

发布时间：2018-04-29 02:22

  本文选题：雷帕霉素 + 树突状细胞　； 参考：《昆明医科大学》2015年硕士论文

【摘要】：研究目的：检测雷帕霉素对树突状细胞的免疫表型和其细胞因子的分泌水平变化的影响,研究树突状细胞从未成熟到成熟的生理变化,为后期的树突细胞在动物器官移植免疫耐受的作用研究提供实验数据。研究方法：取6-8周大SD大鼠(雄性)胫骨和股骨,在无菌条件下冲洗出骨髓细胞,用ficoll法滤过去除部分红细胞,所得的细胞在10%胎牛,GM-CSF(10ng/ml)和IL-4(5ng/ml) RPMI1640培养基内培养,隔天给予半换液,并且加浓度相同的G-CSF、IL-4,期间用光学显微镜观察各组树突状细胞的外形变化。在培养到第4天时,A组加入雷帕霉素5ng/ml,B组加入雷帕霉素30ng/ml,C组加入雷帕霉素60ng/ml,D组不予处理。24小时后收集细胞培养上清液检测四组之间IL-2、IL-6、IL-10和IL-12的分泌水平。离心收集细胞以流式细胞仪检测不同组树突状细胞表面CD80、CD86、CD103和MH-Ⅱ的差异。研究结果：1.雷帕霉素对树突细胞因子分泌水平的影响：收集培养7天的细胞上清液,以酶联免疫吸附实验的方法检测4组的细胞因子分泌情况。实验数据显示雷帕霉素可以对树突状细胞的细胞因子的分泌量产生影响,而不同剂量的雷帕霉素对其影响也是不同的,在本实验中5ng/ml和30ng/ml的剂量对其影响比较明显。2.雷帕霉素对树突状细胞免疫表型的影响：在培养的第7天收集各组树突状细胞,以流式细胞仪检测细胞表面分子的表达,实验结果显示雷帕霉素对处理后的树突状细胞表面分子的表达产生影响,其中CD80、CD86、MH-Ⅱ的表达明显升高,而CD103的表达变化不是很明显,其中30ng/ml组的表达所受影响最为明显。结论：1.雷帕霉素对树突状细胞的细胞因子和表面分子免疫表型的分泌水平有影响。2.雷帕霉素的不同浓度对可影响树突状细胞的细胞因子和表面分子免疫表型产生的影响有所差异。3.雷帕霉素可能对树突状细胞的成熟过程有干预作用。
[Abstract]:Objective: to investigate the effects of rapamycin on the immunophenotype and cytokine secretion of dendritic cells, and to study the physiological changes of dendritic cells from mature to mature. To provide experimental data for the study of the role of dendritic cells in animal organ transplantation immune tolerance. Methods: the tibia and femur of SD rats (male) aged 6-8 weeks were harvested, bone marrow cells were washed out under aseptic conditions, and some red blood cells were removed by ficoll method. The cells were cultured in 10% fetal bovine GM-CSF 10 ng / ml and IL-4 5 ng / ml RPMI1640 medium. At the same concentration of G-CSF IL-4, the morphological changes of dendritic cells in each group were observed by optical microscope. At the 4th day after culture, the supernatant of cell culture was collected to detect the levels of IL-2, IL-6, IL-6, IL-10 and IL-12 in group A and group B with rapamycin 30 ng / ml, group C and group C added rapamycin 60 ng / ml ~ (-1), respectively, after 24 hours of treatment, the supernatants of cell culture were collected to detect the secretion of IL-10 and IL-12 between the four groups. The difference of CD80 CD86, CD103 and MH- 鈪，

本文编号：1817961