本文選題：雷帕霉素 + 樹突狀細胞　； 參考：《昆明醫(yī)科大學》2015年碩士論文

【摘要】：研究目的：檢測雷帕霉素對樹突狀細胞的免疫表型和其細胞因子的分泌水平變化的影響,研究樹突狀細胞從未成熟到成熟的生理變化,為后期的樹突細胞在動物器官移植免疫耐受的作用研究提供實驗數(shù)據(jù)。研究方法：取6-8周大SD大鼠(雄性)脛骨和股骨,在無菌條件下沖洗出骨髓細胞,用ficoll法濾過去除部分紅細胞,所得的細胞在10%胎牛,GM-CSF(10ng/ml)和IL-4(5ng/ml) RPMI1640培養(yǎng)基內(nèi)培養(yǎng),隔天給予半換液,并且加濃度相同的G-CSF、IL-4,期間用光學顯微鏡觀察各組樹突狀細胞的外形變化。在培養(yǎng)到第4天時,A組加入雷帕霉素5ng/ml,B組加入雷帕霉素30ng/ml,C組加入雷帕霉素60ng/ml,D組不予處理。24小時后收集細胞培養(yǎng)上清液檢測四組之間IL-2、IL-6、IL-10和IL-12的分泌水平。離心收集細胞以流式細胞儀檢測不同組樹突狀細胞表面CD80、CD86、CD103和MH-Ⅱ的差異。研究結(jié)果：1.雷帕霉素對樹突細胞因子分泌水平的影響：收集培養(yǎng)7天的細胞上清液,以酶聯(lián)免疫吸附實驗的方法檢測4組的細胞因子分泌情況。實驗數(shù)據(jù)顯示雷帕霉素可以對樹突狀細胞的細胞因子的分泌量產(chǎn)生影響,而不同劑量的雷帕霉素對其影響也是不同的,在本實驗中5ng/ml和30ng/ml的劑量對其影響比較明顯。2.雷帕霉素對樹突狀細胞免疫表型的影響：在培養(yǎng)的第7天收集各組樹突狀細胞,以流式細胞儀檢測細胞表面分子的表達,實驗結(jié)果顯示雷帕霉素對處理后的樹突狀細胞表面分子的表達產(chǎn)生影響,其中CD80、CD86、MH-Ⅱ的表達明顯升高,而CD103的表達變化不是很明顯,其中30ng/ml組的表達所受影響最為明顯。結(jié)論：1.雷帕霉素對樹突狀細胞的細胞因子和表面分子免疫表型的分泌水平有影響。2.雷帕霉素的不同濃度對可影響樹突狀細胞的細胞因子和表面分子免疫表型產(chǎn)生的影響有所差異。3.雷帕霉素可能對樹突狀細胞的成熟過程有干預作用。
[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，

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