Aire对胸腺髓质上皮细胞异位基因表达影响的研究

Study of promiscuous genes expression in mTECs influenced by Aire

目的探讨自身免疫调节因子(Autoimmune regulator,Aire)对胸腺髓质上皮细胞(Medullary thymic epithelial cells,mTECs)异位基因表达的影响。方法原代培养高纯度C57BL/6小鼠胸腺上皮细胞,脂质体法转染pEGFPC3/Aire质粒,RT-PCR法检测mTECs表达Aire及3种常见异位基因mRNA的水平。结果(1)体外培养9天后胸腺上皮细胞占85%左右,转染pEGFPC3/Aire质粒可获得较高转染效率。(2)mTECs中Aire和异位基因表达随培养时间逐渐减弱,至第11天都完全消失。(3)Aire能在mTECs培养早期促进异位基因短期表达。(4)胸腺细胞可延长Aire-mTECs上异位基因表达时间。结论(1)Aire对mTECs异位基因表达有明显的促进作用;(2)胸腺细胞是Aire和异位基因上游激活信号的重要来源。

Objective：To investigate the effects of autoimmune regulator（Aire） to promiscuous genes expression in medullary thymic epithelial cells（mTECs）. Methods：Highly purified mTECs of C57BL/6 mice were primarily cultured, pEGFPC3/Aire plasmid was transfectcd with liposome. The RT-PCR assay was used to investigate the mRNA expression level of Aire and three common promiscuous genes in mTECs. Results： （1）After nine days of culture, the purity of thymic epithelial cells could be about 85% ,when higher efficiency of transfection with pEGFPC3/Aire into mTECs was achieved. （2） The expression levels of Aire and promiscuous genes in mTECs descended gradually and disappeared at the 11th day of culture. （3）Only at the earlier period of culture could Aire promote promiscuous genes expression to a short term. （4）Co-culture with thymocytes could prolong the promiscuous genes expression time in Aire-mTECs. Conclusion： （ 1 ） There is obvious promotion of Aire to promiscuous genes expression in mTECs. （2） Thymocytes are important for upstream activating signals to Aire and promiscuous genes.