权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

胸腺内T細胞分化に関与する上皮細胞表面分子の解析

胸腺内 T 细胞分化中涉及的上皮细胞表面分子分析

基本信息

批准号：
07807039
负责人：
TAKEUCHI Tamotsu
金额：
$ 1.34万
依托单位：
National Institute of Neuroscience, National Center of Neurology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07807039/
关键词：
Thymic epithelial cells HS9 N14

项目摘要

Previously we reported a novel thymic stromal cell antigen, HS9, as a potent molecule participating in intrathymic T cell development. HS9 antigen is expressed on thymic stromal cells especially in the cortex but out on thymocytes. In the present study, we isolated and characterized a novel cDNA,N14, encoding HS9 antigen. Sequencing analysis of N14cDNA has revealed it to be a nocel one without any significant homology to previously reported functional molecules. COS7 cells transfected with expression vectors harboring N14 cDNA became reactive with HS9-specific mAb. Northern blot analysis and in situ hybridization revealed that several tissues, which are positive for HS9 mAb, expressed N14 mRNA.In order to examine the role of this molecule in T cell development, transgenic mice were generated. In situ hybridization study showed that the transgene was significantly over-expressed on both cortical and medullar thymic stromal cells but not on thymocytes. Flowcytometric analysis showed that the percentage of mature CD4^-CD8^+ or CD4^+CD8^- thymocytes of transgenic mice were approximately twice and triple, respectively, than control littermates. Moreover, substantial CD4^+CD8^+ thymocytes appeared to be TCRhigh as comparable to peripheral T cells. Histological examination revealed that trangenic mice had thin cortex and relatively developed medulla. These data indicate the critical role of the N14 gene in T cell development.

此前，我们报道了一种新的胸腺基质细胞抗原HS9，它是参与胸腺内T细胞发育的有效分子。HS9抗原表达于胸腺基质细胞，尤其是皮质，但不表达于胸腺细胞。在本研究中，我们分离并鉴定了编码HS9抗原的新基因N14。序列分析表明，它是一个新的基因，与以前报道的功能分子没有任何显著的同源性。转染N14基因表达载体的COS7细胞可与HS9特异性单抗发生反应。Northern印迹分析和原位杂交显示，HS9 mAb阳性的几个组织表达N14mRNA.为了研究该分子在T细胞发育中的作用，建立了转基因小鼠.原位杂交研究表明，该基因在皮质和髓质胸腺基质细胞上均有显着高表达，但在胸腺细胞上无明显表达。流式细胞仪分析表明，转基因小鼠成熟的CD4^-CD8^+或CD4^+CD8^-胸腺细胞百分比分别约为对照小鼠的两倍和三倍。此外，大量的CD4+CD8+胸腺细胞似乎具有与外周T细胞相当的高TCR。组织学检查显示转基因小鼠皮质较薄，髓质相对发达。这些数据表明了N14基因在T细胞发育中的关键作用。