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目的研究趋化因子受体-7(chemokine receptor-7,CCR7)重组腺病毒体外转染对小鼠未成熟树突状细胞(immature dendritic cells,imDCs)成熟和迁移功能的影响。方法小鼠骨髓经梯度离心及rmIL-4、rmGM-CSF诱导培养未成熟树突状细胞;CCR7重组腺病毒经增强离心法转染未成熟树突状细胞,观察CCR7重组腺病毒对未成熟树突状细胞的形态、细胞表型及趋化功能的影响。结果扫描电镜显示CCR7腺病毒转染后细胞树枝状突起较未成熟DC增多,即轻微地诱使imDCs向成熟转化,但IL-10可抑制成熟;流式结果表明转染后细胞表型符合典型imDCs的特征;Western blot证实CCR7重组腺病毒转染imDCs后可增强CCR7的表达;体外趋化实验显示转染后imDCs体外迁移百分比由4.2%增加到36.1%。结论CCR7重组腺病毒转染imDCs后,其仍保留未成熟特性,CCR7蛋白表达增强,体外迁移能力明显增强。
Objective To investigate the effects of in vitro transfection of recombinant adenovirus carrying chemokine receptor-7 (CCR7) on the maturation and migration of mouse immature dendritic cells (imDCs). Methods Immature dendritic cells were induced by gradient centrifugation and rmIL-4, rmGM-CSF in bone marrow of mice. Immature dendritic cells were transfected with CCR7 recombinant adenovirus by enhanced centrifugation. The effects of CCR7 recombinant adenovirus on immature tree Effect of dendritic cell morphology, cell phenotype and chemotactic function. Results The results of scanning electron microscopy showed that the dendrites of CCR7 cells transfected with CCR7 had more dendrites than those of immature DCs, that is, imDCs were slightly induced to mature, but IL-10 inhibited the maturation. The flow cytometry showed that the phenotype of cells transfected with CCR7 was typical imDCs. The expression of CCR7 was enhanced by transfection of imDCs with CCR7 recombinant adenovirus by Western blot. In vitro chemotaxis assay showed that the percentage of in vitro migration of imDCs increased from 4.2% to 36.1% after transfection. CONCLUSION: After transfection with imDCs, the CCR7 recombinant adenovirus still retains its immature characteristics and enhances the expression of CCR7 protein in vitro.