TGFβ1基因转染对大鼠树突状细胞分子表型和细胞因子分泌的影响

The effects of TGFβ1 gene transfection on the molecular phenotype and cytokine secretion of rat bone marrow-derived dendritic cells

目的:研究转化生长因子β1(TGFβ1)基因转染大鼠树突状细胞(DC)表面分子表达以及细胞因子分泌的影响。方法:构建TGFβ1重组真核表达质粒TGFβ1-pcDNA3,以脂质体Am ine介导转染大鼠骨髓DC,经过W estern b lot、RT-PCR、免疫细胞化学以及免疫荧光染色检测转染效率,应用FACS和免疫细胞化学等方法检测TGFβ1-DC分子表型以及细胞因子分泌的变化。结果:结果显示TGFβ1基因转染后可得到一定数量稳定的TGFβ1-DC,并获得有效TGFβ1的mRNA和蛋白表达。并发现TGFβ1-DC表面RT1B、B7-2、ICAM-1和OX62等分子表达均明显低于对照组,TGFβ1-DC分泌的Th2型细胞因子TGFβ1和IL-10的表达明显高于反义TGFβ1转染对照组,而Th1型细胞因子IL-12的合成则受到一定程度的下调。结论:TGFβ1基因转染可调节大鼠骨髓DC分子表型和细胞因子的分泌,增强DC的免疫耐受性,在器官移植治疗中有应用前景。

Objective：To implore the effects of TGFβ1 gene transfecion on the molecular phenotypes expression and the cytokine secretion of rat bone marrow-derived dendritic cells（BM-DC）. Methods ：The recombinant mammalian expressive plasmid TGFβ1- pcDNA3 was constructed and transduced into the rat bone marrow-derived DCs miedated by Lipofectine Amine. The expression of TGFβ1 was detected by Western blot, RT-PCR, Northern blot and fluorescence immunecytochemistry. The expressions of surface molecules and the secretion of cytokines were assayed by FACS and Immunecytechemistry. Results：TGFβ1 gene could be transduced into BM-DC successfully and obtained substantial protein production. The expression levels of the cell surface molecules, such as RT-1B, B7-2, ICAM-1 and OX62 in the TGFβ1-DC group were significantly lower than those in the antisense TGFβ1 group and the pcDNA3 control group. The production of Th2 cytokines, such as TGFβ1 and IL-10 was significantly higher than those in control groups, and Thl cytokine IL-12 was decreased to a certain extent. Conclusion：The results suggested that TGFβ1 gene transfection could modulate the molecule phenotype and cytokine secretion of bone marrow-derived DC and enhance the tolerogenecity of DCs. The experiments convinced the prospect of therapeutic application of DCs in organ transplantation therapeutics.