中文摘要：

目的观察应用西罗莫司联合可溶性肿瘤坏死因子受体Ⅰ（sTNFRI）-IgGFc基因转染对大鼠骨髓源性树突状细胞（DC）形态、表型、功能和分化的影响及抑制DC成熟的作用。方法获取和培养Wistar大鼠骨髓源性DC。实验分5组，未成熟DC组（imDC组），成熟DC组（miDC组），西罗莫司组（经西罗莫司处理的DC），sTNFRI基因转染组（转染sTNFRI-IgGFc基因的DC）与联合处理组（经西罗莫司处理和sTNFRI基因转染的DC）。采用流式细胞技术检测各组IX；表面分子MHCⅡ、CD80、CD86的表达情况，进行单向式混合淋巴细胞反应后进行MTT实验检测各组IX；活化T淋巴细胞能力，采用酶联免疫吸附试验试剂盒检测各组IX；培养上清液中白细胞介素12（IL-12）、γ干扰素（IFN-7）及sTNFRI-IgGFc蛋白的表达状况。结果培养第10天，mDC组IX；表面MHCⅡ、CDS0、CD86的表达量显著高于其他4组（P〈0．05）；联合处理组较imDC组CD86的表达显著降低（P〈0．05）。mDC组DC刺激T淋巴细胞增殖能力显著高于其他4组（P〈0．05），联合处理组显著低于imDC组、mDC组和西罗莫司组（P〈0．05）。mDC组培养液中IL-12和INF-γ的含量显著高于其他4组（P〈0．05），而联合处理组较其他4组培养液中IL-12和INF-y的水平显著降低（P〈0．05）。结论西罗莫司和sTNFRI-IgGFc基因修饰均能抑制DC的成熟，二者联合应用具有协同效应，较单独使用具有更强地抑制DC成熟的作用。

英文摘要：

Objective To explore the morphology, cell phenotype and cell function in dendritic cells （DCs） derived from bone marrow after treatment with Sirolimus or Sirolimus combined with sTNFRI-IgGFc gene segment transfection. Method DCs were divided into 5 groups （imDCs, mDCs, Rapa-DCs, sTNFRI-DCs and Rapa-sTNFRI-DCs） according to different interventions. The expresson of MHC-Ⅱ, CD80 and CD86 was detected by flow eytometry. T cell proliferation of the mixed lymphocyte reaction was evaluated by MTT method. The levels of IL-12, IFN-γ and sTNFRI-IgGFc were determined by ELISA. Result On the day 10, the flow cytometry showed that the expression levels of MHC-II, CD80 and CD86 on the cell surface in Sirolimus group were significantly higher than the other groups （P〈0. 05）. The expression level of CD86 in Rapa-TNFRI group was significantly lower than in imDC group （P〈0. 05）. MTT results demonstrated that T ceil proliferation ability in Sirolimus group, sTNFRI group and Rapa-sTNFRI group were reduced as compared with mDC group （P〈0. 05）. The ELISA results revealed that the levels of IL-12 and INF-γ in mDC group were significantly higher than other groups （P〈0. 05）. The levels of IL-12 and INF-γ in Rapa-sTNFRI group were significantly lower than other groups （P〈0. 05）. Conclusion Sirolimus combined with modified sTNFRHgGFc gene could synergistically inhibit maturation of DCs more effectively than Sirolimus or modified sTNFRI-IgGFc gene used alone.