| 人CD83基因转染细胞株的构建及其功能初步研究 |
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| 引用本文： | 陈礼文,高超,沈双,仲维学,朱一蓓,张学光. 人CD83基因转染细胞株的构建及其功能初步研究[J]. 免疫学杂志, 2011, 0(6): 482-485 |
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| 作者姓名： | 陈礼文 高超 沈双 仲维学 朱一蓓 张学光 |
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| 作者单位： | 苏州大学医学生物技术研究所;安徽医学高等专科学校;江苏省盐城市中心血站;苏州大学附属第一医院临床免疫室; |
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| 基金项目： | 国家自然科学基金项目(30700728); 安徽高校省级自然科学研究项目(KJ2011B072) |
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| 摘 要： | 目的构建人CD83全长编码基因转染293细胞,并探讨其对单核细胞的作用。方法 RT-PCR方法从成熟人树突状细胞(DC)扩增出CD83 cDNA,插入pIRES2-EGFP载体,脂质体转染293细胞,筛选出稳定表达目的基因细胞株;从人外周血单个核细胞(PBMC)中磁珠分选单核细胞,LPS刺激的同时加入293/CD83细胞或293/mock,24 h后检测细胞活化状态以及培养上清中TNF-α水平。结果经流式细胞术反复检测绿色荧光蛋白(GFP)报告基因及CD83表达水平,筛选获得获得一株稳定表达膜型CD83分子的基因转染细胞株293/CD83;体外实验显示293/CD83可促进LPS刺激的单核细胞活化水平和TNF-α分泌量。结论成功构建表达人CD83分子的293细胞株,293/CD83对LPS刺激的单核细胞有协同刺激作用。
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| 关 键 词： | 树突状细胞 转染 免疫调控 细胞活化 TNF-α |
Construction of the transfected cell line expressing human CD83 and its function study |
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| CHEN Liwen,GAO Chao,SHEN Shuang,ZHONG Weixue,ZHU Yibei,ZHANG Xueguang Institute of Medical Biotechnology,Soochow University,Suzhou ,China. Construction of the transfected cell line expressing human CD83 and its function study[J]. Immunological Journal, 2011, 0(6): 482-485 |
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| Authors: | CHEN Liwen GAO Chao SHEN Shuang ZHONG Weixue ZHU Yibei ZHANG Xueguang Institute of Medical Biotechnology Soochow University Suzhou China |
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| Affiliation: | CHEN Liwen,GAO Chao,SHEN Shuang,ZHONG Weixue,ZHU Yibei,ZHANG Xueguang Institute of Medical Biotechnology,Soochow University,Suzhou 215007,China |
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| Abstract: | To construct the 293 transfectant expressing human CD83 molecule and explore its effects on monocytes,the gene encoding human CD83 was amplified by RT-PCR from mature dendritic cells,and then the target gene fragment was inserted into pIRES2-EGFP vector.The recombinant vector was transfected into 293 cells with Lipofect 2000 and the stable cell line expressing human CD83 was established by GFP reporter assay and CD83 expression.After purified from PBMC by MACS sorting,monocytes were stimulated with LPS in t... |
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