引用本文:何云锋,吴小候,李家兵,陈 刚,张 尧,唐亚雄.ROR-γt干扰腺病毒载体抑制Th17细胞功能[J].重庆医科大学学报,2014,38(6):790~792
ROR-γt干扰腺病毒载体抑制Th17细胞功能
Construction of ROR-γt interference adenovirus and its influence on Th17 cell function
DOI:
中文关键词:  腺病毒  ROR-γt  质粒  辅助性T 细胞17
英文关键词:adenovirus  ROR-γt  plasmid  T helper cell 17
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作者单位
何云锋,吴小候,李家兵,陈 刚,张 尧,唐亚雄 重庆医科大学附属第一医院泌尿外科重庆 400016 
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中文摘要:
      目的:设计和构建ROR-γt特异性干扰腺病毒载体,观察其对大鼠脾脏来源的辅助性T 细胞17(T helper cell 17,Th17)功能的影响。方法:设计ROR-γt编码区干扰序列,DNA聚合酶作用下形成双链序列,与线性化pSES-HUS质粒连接。转入BJ5183,转染病毒包装体系293细胞系,收集细胞,得到病毒液。反复感染4次得到高滴度pSES-HUS-ROR-γt病毒液。将其感染大鼠脾脏来源的Th17细胞,检测其对Th17细胞功能的影响。结果:成功构建和包装pSES-HUS-ROR-γt干扰腺病毒液,Th17细胞感染pSES-HUS-ROR-γt干扰病毒后,其分泌细胞因子白介素(interleukin,IL)-17、IL-6和TNF-γ水平下降。结论:pSES-HUS-ROR-γt干扰腺病毒能够有效抑制Th17细胞功能,可能对肾移植慢性排斥反应具有抑制或者延缓作用。
英文摘要:
      Objective:To construct the ROR-γt interference adenovirus,to transfect the ROR-γt interference adenovirus into T helper cell 17(Th17 cell) and to detect the change of Th17 cell was after being affected by ROR-γt interference adenovirus. Methods: ROR-γt interference sequences were designed to form a double chain sequence by DNA polymerase and to connect with the linearized pSES-HUS plasmid. pSES-HUS-ROR-γt plasmid was transfected into BJ5183 by electroporation,and BJ5183 was transfected into the 293 virus package system. The ROR-γt interference adenovirus vector was achieved and named as pSES-HUS-ROR-γt. pSES-HUS-ROR-γt was infected the Th17 cell which was isolated from the spleen of rat. The changes of IL-17,IL-6 and TNF-γ were detected by ELISA. Results:pSES-HUS-ROR-γt interference adenovirus vector was successfully constructed. Th17 cytokines(IL-17,IL-6 and TNF-γ) were decreased after Th17 being infected with pSES-HUS-ROR-γt interference adenovirus. Conclusion: Th17 cell function can be inhibited by pSES-HUS-ROR-γt interference adenovirus vector,and the adenovirus vector has the potential treatment function in the chronic rejection of renal transplantion.
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