中文摘要：

目的构建基于shRNA介导的中枢免疫耐受缺损模型。方法分离妊娠后13.5 d的胎鼠胸腺,除去淋巴细胞,获得原代胸腺基质细胞,利用慢病毒将小鼠TRAF6基因特异性shRNA慢病毒质粒（LV-T6-shRNA）导入原代胸腺基质细胞中,重新聚集这些胸腺基质细胞得到LV-T6-shRNA重塑胸腺,将此重塑胸腺移植入无胸腺的雌性小鼠肾脏荚膜下,饲养8周。结果慢病毒可有效转导LV-T6-shRNA慢病毒质粒入原代胸腺髓质上皮细胞中;移植LV-T6-shRNA重塑胸腺的小鼠饲养8周后,胸腺明显较小,成熟的胸腺髓质上皮细胞减少,脾脏肿大,脾脏中活化的T淋巴细胞增多,肺脏中出现淋巴细胞浸润等现象,这些表现型与TRAF6-/-小鼠的表现型相似。结论中枢免疫耐受缺损模型构建成功。

英文摘要：

The aim of this study is to establish a central tolerance-deficient model based on shRNA specific interference.Fetal thymic lobes were isolated from wild-type Balb/c-embryos 13.5 days after pregnancy,and then cultured for 4 days to eliminate lymphocytes.Thereafter,murine TRAF6-targeted lentiviral shRNA（LV-T6-shRNA） plasmid was introduced into the primary thymic stromal cells with lentivirus,which were then reassembled into reconstitute fetal thymus organ cultures（rTOC;LV-T6-shRNA rTOC） in vitro,and grafts of these rTOC under the renal capsule of athymic nude mice.These mice were bred for 8 weeks.After 8 weeks,LV-T6-shRNA rTOC grafted mice demonstrated smaller thymus and decreased mature medullary thymic epithelial cells（mTEC）,splenomegaly and increased percentage of activated CD4＋ T cells,as well as inflammatory infiltrates in lung,similar to those in TRAF6-/-mice and/or athymic nude mice grafted with TRAF6-/-thymus.These results suggest that central tolerance-deficient model is successfully established and will be helpful for further study.