CCR7基因转染对小鼠未成熟树突状细胞成熟及迁移功能的影响.docVIP

CCR7基因转染对小鼠未成熟树突状细胞成熟及迁移功能的影响 辛海明，彭毅志，郭 浩，袁志强 （第三军医大学西南医院全军烧伤研究所，创伤、烧伤与复合伤国家重点实验室，重庆400038） 摘要：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 cellsimDCs）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腺病毒转染树枝状突起较未成熟DC增多，即轻微地诱使imDC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imDC的特征；Western lot证实重组腺病毒转染imDC后可增强CCR7的表达；体外趋化实验显示转染后imDC体外迁移由4.2%增加到36.1%。结论 CCR7重组腺病毒转染imDC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。 关键词：CCR7；未成熟树突状细胞；重组腺病毒；免疫耐受ransfection of ccr7 recombinant adenovirus enhances maturization and migration function of murine immature dendritic cells XIN Hai-ming, PENG Yi-zhi, GUO Hao, YUAN Zhi-qiang (State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Burns, Southwest Hospital, Third Military Medical University, Chongqing 400038, China) Abstract： Objective To investigate the matur and migration function of murine immature dendritic cells (DCs) after transfection with chemokine receptor-7 (ccr7) recombinant adenovirus in vitro. Methods Bone marrow cells were isolated from health Balb/c mice and then the mononuclear cells were screened with gradient centrifugation. Immature dendritic cells were induced in the presence of recombinant murine interleukin-4 and granulocyte macrophage colony-stimulating factor. Recombinant adenovirus vector encoding full-length murine ccr7 cDNA was transfected into the immature dendritic cells by centrifugal force method. Then the morphology, cell phenotype, CCR7 expression and migration in the immature dendritic cells were observed after the transfection with ccr7 recombinant adenovirus by scanning electron microscopy, flow cytometry, Western blot analysis and Transwell cell migration assay . Results The recombinant adenovirus slightly induced imDCs to transform into mature DCs which was observed by scanning electron microscopy, but IL-10 inhibited the process; Flow cytometry indicated the cell phenotype of the