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研究小鼠重组腺病毒载体Ad-mIL-10及其转染的树突状细胞(dendritic cell,DC)对小鼠哮喘模型气道炎症的影响,探讨其相关的发病机制。用基因工程的方法构建小鼠IL-10腺病毒重组体Ad-mIL-10,并转染小鼠骨髓来源的DC。以卵白蛋白(OVA)腹腔注射致敏小鼠并雾化吸入激发的方法制作小鼠哮喘模型。第一次腹腔OVA致敏后静脉给予Ad-mIL-10、Ad-EGFP、Ad-EGFP转染的DC或Ad-mIL-10转染的DC,并设立哮喘模型制作对照组,观察各组小鼠血液、肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中的CD3+T细胞中的IL-10+IL-4-的Tr细胞及IL-10+IL-4+的Th2细胞比例变化,ELISA方法测定外周血及肺泡灌洗液中的IFN-γ、IL-4、IL-10水平,以观察Th1、Th2细胞的数量、功能变化以及IL-10的变化。结果显示Ad-mIL-10、Ad-EGFP、Ad-EGFP-DC对哮喘模型小鼠体内Tr细胞和Th2细胞的数量及肺部炎症局部的Th1及Th2细胞功能无明显影响;Ad-mIL-10转染的DC可以诱导哮喘模型小鼠体内IL-4-IL-10+的Tr细胞的增加及IL-4+IL-10+的Th2细胞的减少,并抑制哮喘模型小鼠肺部Th2细胞的功能,但对Th1细胞的功能无明显影响。提示单独应用Ad-mIL-10静脉注射产生高浓度的IL-10并不能诱导Tr细胞分化和抑制Th2细胞的激活。Ad-mIL-10转染的DC可以诱导原始T细胞向Tr细胞方向分化,并抑制Th2细胞的数量和功能。
To investigate the effect of mouse recombinant adenovirus vector Ad-mIL-10 and its transfected dendritic cell (DC) on airway inflammation in mouse asthma model and to explore its related pathogenesis. Recombinant mouse IL-10 adenovirus Ad-mIL-10 was constructed by gene engineering and transfected into mouse bone marrow-derived DCs. A mouse asthma model was established by intraperitoneal injection of ovalbumin (OVA) into sensitized mice and inhalation challenge. After the first intraperitoneal OVA sensitization, Ad-mIL-10, Ad-EGFP, Ad-EGFP-transfected DCs or Ad-mIL-10 transfected DCs were intravenously administrated and asthma models were established to make a control group. Tr-IL-10 + IL-4-dependent Tr cells and Th2 cell ratio of IL-10 + IL-4 + in CD3 + T cells in bronchoalveolar lavage fluid (BALF) The levels of IFN-γ, IL-4 and IL-10 in peripheral blood and bronchoalveolar lavage fluid were measured to observe the changes of Th1 and Th2 cells, their function and IL-10. The results showed that there was no significant effect of Ad-mIL-10, Ad-EGFP and Ad-EGFP-DC on the number of Tr cells and Th2 cells and the function of Th1 and Th2 cells in lung inflammation in asthmatic mice. The transfected DCs can induce the increase of IL-4-IL-10 + Tr cells and the decrease of Th2 cells of IL-4 + IL-10 + in asthmatic mice and inhibit the expression of Th2 cells Function, but no significant effect on the function of Th1 cells. Tip Ad-mIL-10 alone intravenously to produce high concentrations of IL-10 can not induce Tr cell differentiation and inhibit Th2 cell activation. Ad-mIL-10-transfected DCs induced the differentiation of naive T cells toward Tr cells and inhibited the number and function of Th2 cells.