IL-10基因修饰的大鼠树突状细胞表型分析及生物学特性的研究

The phenotypes and biological characteristics of interleukin-10 gene modified dendritic cell

目的研究IL-10基因修饰后的大鼠树突状细胞(DC)的表型及其生物学特性。方法以含IL-10基因的重组腺病毒载体体外转染大鼠骨髓来源的DC,Western blot测定转染后各组DC中IL-10蛋白的表达,流式细胞仪检测各组DC表面抗原CD83、CD86分子的表达情况,混合淋巴细胞反应法测定各组DC刺激同种异体T细胞增殖的能力。结果 IL-10基因修饰组DC可检测到IL-10高表达,表面抗原CD83、CD86低表达,其刺激T淋巴细胞增殖水平较其他各组低。结论 IL-10基因修饰的DC可有效的表达有功能的IL-10,为研究IL-10修饰的DC诱导同种异体移植免疫耐受奠定了基础。

In this research,we aim to investigate the phenotypes and biological characteristics of interleukin-10 gene modified dendritic cell（DC）.The DCs were trained six days,then randomly divided into three groups： the first group was trained with IL-10-GFP-Adenovirus as IL-10-DC,the second group was trained with GFP-Adenovirus as GFP-DC,and the third group was trained without adenovirus as 12-DC.The three groups were continued to train until the twelfth day.Then the protein of IL-10 was detected by Western blotting,the phenotypes of CD83 and CD86 were detected by flow cytometry,and the proliferation activity was assayed with proliferation experiment.The result of Western blotting showed that IL-10 was more effective expressed in IL-10-DC than that in the other two groups,but the phenotypes of CD83 and CD86 demonstrated the lowest expression in IL-10-DC group in three groups.Flow cytometry indicated that the proliferation activity of allogeneic T cell in IL-10-DC group was the lowest in all three groups.Above results imply that IL-10 could be effective expressed in IL-10 gene modified DC,and the IL-10 gene modified DC was in favor of the allotransplantation.