摘要
目的检测人白细胞介素10(hIL-10)基因在大鼠骨髓来源的未成熟树突状细胞(immature dendritic cell,imDC)中的表达。方法实验分4组:mDC组、imDC组、空载体组及hIL-10基因转染组。用hIL-10真核表达载体(pIRES2-EGFP-hIL-10)转染大鼠骨髓来源的imDC,应用ELISA、Western blot、流式细胞术及混合淋巴细胞反应(MLR)检测hIL-10在各组中的表达。结果荧光显微镜观察显示pIRES2-EGFP-hIL-10质粒转染imDC后可见绿色荧光;ELISA检测hIL-10基因转染组的hIL-10表达明显高于mDC组、imDC组及空载体组;经Westen blot检测hIL-10基因转染组有hIL-10蛋白表达;流式细胞术显示hIL-10基因转染组hIL-10基因转染后imDC表面分子CD86、CD80与其他三组相比略呈低表达;转染后混合淋巴细胞反应(MLR)显示,hIL-10转染组的imDC对T细胞的增殖反应均弱于imDC组和空载体组(P=0.024和P=0.033)。结论外源性hIL-10基因成功导入大鼠imDC并表达。
Objective To investigate the expression of hIL-10 gene in dendritic cells from bone marrow stem cells of rats. Methods The experiment was divided into four groups:mDC group, imDC group, empty vector group and IL-10 gene transfection group. The plasmids of pIRES2-EGFP-hIL-10 were transfected into imDC to detect expression of hIL-10 gene by ELISA, Western blot, flow cytometry and MLR. Results After transfection,green fluorescence was observed in imDC under fluorescence microscope. The result of ELISA showed that the expression of hIL-10 in hIL-10 group was significantly higher than that in the other groups(P = 0. 004,0. 004 and 0.003). Western blot result showed the positive expression of IL-10 protein in IL-10 gene transfection group. CD86 and CD80 expression on the imDC surface in IL-10 gene transfection group was lower than in the other groups by flow cytometry. The result of MLR demonstrated that the imDC induced lower T-cell in hIL-10 group than in imDC group and empty vector group( P = 0.024,P = 0.033 ). Conclusion The extrinsic source gene is successfully transfected into imDC and could be expressed.
出处
《山西医科大学学报》
CAS
2009年第10期888-892,共5页
Journal of Shanxi Medical University
基金
福建省自然科学基金资助项目(C0710023)
福建医科大学第一临床学院学科带头人培养对象及青年骨干教师培养专项基金资助项目(JXK200709)