肿瘤坏死因子-α诱导蛋白-8样分子2在调节性T细胞中的表达  

The expression of tumor necrosis factor-α induced protein 8 like-2 in CD4^+CD25^+ regulatory T cells

作　　者：栾樱译[1] 姚咏明[2] 

机构地区：[1]南方医科大学基础医学院,广东广州510515 [2]解放军总医院第一附属医院全军烧伤研究所,北京100048

出　　处：《感染．炎症．修复》2011年第2期69-72,I0002,共5页Infection Inflammation Repair

基　　金：国家自然科学基金资助(30901561);国家重点基础研究发展计划项目(2005CB522602)

摘　　要：目的:观察肿瘤坏死因子-α诱导蛋白-8样分子2(TIPE2)在CD4^+CD25^+调节性T细胞(CD4^+CD25^+Treg)中的表达。方法:免疫磁珠法分离正常BALB/C小鼠脾脏CD4^+CD25^+Tregs,流式细胞术鉴定CD4^+CD25^+Treg的纯度。激光共聚焦荧光法检测Treg细胞中TIPE2的分布,并进行初步定位;进一步采用逆转录-聚合酶链反应(RT-PCR)和Western blot技术分别从基因和蛋白水平检测Treg细胞中TIPE2表达。结果:免疫磁珠分选法得到的CD4^+CD25^+Tregs纯度在92%以上,台盼蓝染色显示细胞活性大于97%。Western blot证实Treg细胞中存在清晰TIPE2条带,分子质量为21kD;采用RT-PCR技术在Treg细胞中检测到147 bp大小的特异性TIPE2目的基因条带。结论:TIPE2可表达于小鼠CD4^+CD25^+Treg细胞。Objective：To investigate the expression of tumor necrosis factor-α induced protein 8 like-2 （TIPE2） in CD4^＋CD25^＋ regulatory T cells （CD4^＋CD25^＋ Tregs）. Methods： CD4^＋CD25^＋ Tregs were isolated from the spleens of male BALB/C mice by magnetic beads, and the purity of these cells was determined by flow cytometry. The present study was designed to determine TIPE2 expression in Tregs by confocal microscopy analysis, Western blot and reverse transcription-polymerase chain reaction （RT-PCR） analysis, respectively. Results： Purity of CD4^＋CD25^＋ Tregs was greater than 92%. The expression of TIPE2 was detected by confocal microscopy, and it was a cytoplasmic protein expressed in CD4^＋CD25^＋ Tregs. To confirm the expression of TIPE2, it was detected by Western blot analysis using specific TIPE2 antibody, and a clear band with a molecular mass of approximately 21 kD from CD4^＋CD25^＋ Tregs was found. Moreover, to determine the gene expression of TIPE2, total RNA was extracted from CD4^＋CD25^＋ Tregs and RT-PCR was performed, a band of the size of 147 bp was noted as expected. Conclution： T1PE2 appears to be a cytoplasmic protein expressed in CD4^＋ CD25^＋ Tregs.

关 键 词：肿瘤坏死因子-α诱导蛋白-8样分子2 CD4^+CD25^+调节性T细胞免疫 

分 类 号：R392.11[医药卫生—免疫学]