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 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 maturization 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 fulllength murine CCR7 eDNA was transfected into the immature dendritic ceils 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 adenovirns 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 cells after transfection were corresponded to the typical characteristics of imDCs; Western blotting confirmed the CCR7 expression was enhanced in the transfected imDCs; The migration percentage of transfected imDCs was improved from 4.2% to 36.1%. Conclusion The imDCs transfected with CCR7 recombinant adenovirus show the morphology and cell phenotype are consistent with the typical characteristics of imDCs ; but presenting enhanced CCR7 expression and obviously improved ability of migration.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2009年第17期1629-1632,共4页
Journal of Third Military Medical University
基金
国家自然科学基金(30672173)
全军医药卫生科研基金专项课题(08Z026)~~
关键词
CCR7
未成熟树突状细胞
重组腺病毒
免疫耐受
chemokine receptor 7
immature dendritic cell
recombinant adenovirns
immune tolerance
