叉头状转录因子Foxp3重组腺病毒的构建及鉴定
发布时间：2018-11-15 09:38
【摘要】：Foxp3是叉头样转录因子(Forkhead transcription factor)P亚家族的新成员。目前研究证实该基因特异性表达于固有CD4~+CD25~+调节性T细胞,并在该细胞的发育和功能维持中起重要作用,是CD4~+CD25~+调节性T细胞特异性标志物之一,并作为转录抑制因子参与免疫应答的调控。Foxp3包含叉头状结构域、C_2H_2锌指结构及亮氨酸拉链等保守序列,在与靶基因启动子和某些蛋白质结合上起着重要作用。 近年来,人们通过构建Foxp3重组逆转录病毒、Foxp3转基因模型等方法对该基因的作用机制及临床应用展开了广泛的研究。实验发现Foxp3的异位表达在体外可诱导与固有CD4~+CD25~+调节性T细胞表型及作用相似的调节性T细胞的产生;诱导产生的调节性T细胞在体内可直接或间接抑制效应性T淋巴细胞、B淋巴细胞及某些抗原提呈细胞(如树突状细胞)等的增殖、分化和免疫应答,诱导体内免疫耐受的产生,对治疗某些自身免疫病及移植排斥反应有一定效果;随着研究的深入,该基因在肿瘤免疫的作用也越来越受到关注,有文献报道在某些肿瘤组织及周围淋巴结等处发现Foxp3的表达,为肿瘤逃逸机制的研究以及新型靶向制剂的开发提供了新思路。 为更深入研究Foxp3的作用机制及诱导免疫耐受方面的应用,本实验利用AdEasy~(TM)XL系统构建携带Foxp3的重组腺病毒,并行初步鉴定。方法:从小鼠胸腺组织获取约1.3kb大小的目的基因,克隆至穿梭载体pShuttle-IRES-hrGFP-1
[Abstract]:Foxp3 is a new member of (Forkhead transcription factor) P subfamily. At present, it has been confirmed that this gene is specifically expressed in inherent CD4~ CD25~ regulatory T cells and plays an important role in the development and functional maintenance of the cells. It is one of the specific markers of CD4~ CD25~ regulatory T cells. As a transcription suppressor, Foxp3 contains some conserved sequences, such as fork head domain, zinc finger structure of C_2H_2 and leucine zipper, which play an important role in binding to target gene promoter and some proteins. In recent years, the mechanism and clinical application of Foxp3 recombinant retrovirus and Foxp3 transgenic model have been extensively studied. It was found that the heterotopic expression of Foxp3 could induce the production of regulatory T cells similar to that of CD4~ CD25~ regulatory T cells in vitro. In vivo, regulatory T cells can directly or indirectly inhibit the proliferation, differentiation and immune response of effector T lymphocytes, B lymphocytes and some antigen-presenting cells (such as dendritic cells). The induction of immune tolerance in vivo is effective in the treatment of some autoimmune diseases and transplant rejection. With the development of the research, the role of the gene in tumor immunity has been paid more and more attention. It has been reported that the expression of Foxp3 is found in some tumor tissues and surrounding lymph nodes. It provides a new idea for the study of the mechanism of tumor escape and the development of new targeted preparations. In order to further study the mechanism of Foxp3 and its application in inducing immune tolerance, the recombinant adenovirus carrying Foxp3 was constructed by AdEasy~ (TM) XL system and identified. Methods: the target gene of about 1.3kb size was obtained from mouse thymus tissue and cloned into shuttle vector pShuttle-IRES-hrGFP-1.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R346
本文编号：2332931
[Abstract]:Foxp3 is a new member of (Forkhead transcription factor) P subfamily. At present, it has been confirmed that this gene is specifically expressed in inherent CD4~ CD25~ regulatory T cells and plays an important role in the development and functional maintenance of the cells. It is one of the specific markers of CD4~ CD25~ regulatory T cells. As a transcription suppressor, Foxp3 contains some conserved sequences, such as fork head domain, zinc finger structure of C_2H_2 and leucine zipper, which play an important role in binding to target gene promoter and some proteins. In recent years, the mechanism and clinical application of Foxp3 recombinant retrovirus and Foxp3 transgenic model have been extensively studied. It was found that the heterotopic expression of Foxp3 could induce the production of regulatory T cells similar to that of CD4~ CD25~ regulatory T cells in vitro. In vivo, regulatory T cells can directly or indirectly inhibit the proliferation, differentiation and immune response of effector T lymphocytes, B lymphocytes and some antigen-presenting cells (such as dendritic cells). The induction of immune tolerance in vivo is effective in the treatment of some autoimmune diseases and transplant rejection. With the development of the research, the role of the gene in tumor immunity has been paid more and more attention. It has been reported that the expression of Foxp3 is found in some tumor tissues and surrounding lymph nodes. It provides a new idea for the study of the mechanism of tumor escape and the development of new targeted preparations. In order to further study the mechanism of Foxp3 and its application in inducing immune tolerance, the recombinant adenovirus carrying Foxp3 was constructed by AdEasy~ (TM) XL system and identified. Methods: the target gene of about 1.3kb size was obtained from mouse thymus tissue and cloned into shuttle vector pShuttle-IRES-hrGFP-1.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R346
【参考文献】
相关期刊论文 前2条
1 丁涵露,吴雄飞,高闻达,贺伟峰,张晓容,吴军;重组AdPD-L1腺病毒的构建表达及鉴定[J];第三军医大学学报;2005年12期
2 陈祖兵,陶剑平,梁力建,刘晓平,胡文杰,李绍强;腺病毒介导的T-bet基因转染诱导Th1型淋巴细胞分化[J];中国病理生理杂志;2005年06期
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