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

【摘要】：研究目的：檢測(cè)雷帕霉素對(duì)樹突狀細(xì)胞的免疫表型和其細(xì)胞因子的分泌水平變化的影響,研究樹突狀細(xì)胞從未成熟到成熟的生理變化,為后期的樹突細(xì)胞在動(dòng)物器官移植免疫耐受的作用研究提供實(shí)驗(yàn)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(xì)胞,用ficoll法濾過去除部分紅細(xì)胞,所得的細(xì)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(xué)顯微鏡觀察各組樹突狀細(xì)胞的外形變化。在培養(yǎng)到第4天時(shí),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(shí)后收集細(xì)胞培養(yǎng)上清液檢測(cè)四組之間IL-2、IL-6、IL-10和IL-12的分泌水平。離心收集細(xì)胞以流式細(xì)胞儀檢測(cè)不同組樹突狀細(xì)胞表面CD80、CD86、CD103和MH-Ⅱ的差異。研究結(jié)果：1.雷帕霉素對(duì)樹突細(xì)胞因子分泌水平的影響：收集培養(yǎng)7天的細(xì)胞上清液,以酶聯(lián)免疫吸附實(shí)驗(yàn)的方法檢測(cè)4組的細(xì)胞因子分泌情況。實(shí)驗(yàn)數(shù)據(jù)顯示雷帕霉素可以對(duì)樹突狀細(xì)胞的細(xì)胞因子的分泌量產(chǎn)生影響,而不同劑量的雷帕霉素對(duì)其影響也是不同的,在本實(shí)驗(yàn)中5ng/ml和30ng/ml的劑量對(duì)其影響比較明顯。2.雷帕霉素對(duì)樹突狀細(xì)胞免疫表型的影響：在培養(yǎng)的第7天收集各組樹突狀細(xì)胞,以流式細(xì)胞儀檢測(cè)細(xì)胞表面分子的表達(dá),實(shí)驗(yàn)結(jié)果顯示雷帕霉素對(duì)處理后的樹突狀細(xì)胞表面分子的表達(dá)產(chǎn)生影響,其中CD80、CD86、MH-Ⅱ的表達(dá)明顯升高,而CD103的表達(dá)變化不是很明顯,其中30ng/ml組的表達(dá)所受影響最為明顯。結(jié)論：1.雷帕霉素對(duì)樹突狀細(xì)胞的細(xì)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(duì)可影響樹突狀細(xì)胞的細(xì)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(duì)樹突狀細(xì)胞的成熟過程有干預(yù)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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