Thymic epithelial requirement for γδ T cell development revealed in the cell ablation transgenic system with TSCOT promoter.
Thymic epithelial requirement for γδ T cell development revealed in the cell ablation transgenic system with TSCOT promoter.
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DOI：
10.1007/s10059-012-0246-4
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发表时间：
2012-11
影响因子：
3.8
通讯作者：
Kim, Moon Gyo
中科院分区：
文献类型：
--
作者：
Lee, Gwanghee;Kim, Ki Yeon;Chang, Cheong-Hee;Kim, Moon Gyo
为了研究胸腺上皮细胞(TEC)亚群在T细胞发育中的作用，我们建立了一个新的转基因系统，能够诱导细胞特异性的消融，并使用双顺反子细菌硝基还原酶和EGFP基因标记TEC亚群。转基因小鼠的两个不同长度的TSCOT启动子，分别命名为3.1T-NE和9.1T-NE，驱动EGFP在TEC中的表达。在成体中，EGFP的表达定位于髓质，TSCOT启动子较小，启动子大小为3.1kb，而皮质中的EGFP表达维持在9.1kb的启动子，提示在两个启动子中可能存在TEC特异性的顺式元件和隔室特异性的顺式元件。在呋喃妥因、甲硝唑等前体药物作用下，硝基还原酶诱导的细胞死亡具有特异性，无旁观者死亡。细胞死亡的程度取决于前药物在细胞和胎儿胸腺器官培养(FTOCs)中的剂量。根据EPCAM、MHCII、CDR1和/或UEA-1的表达水平分析胎儿胸腺基质细胞群。不同亚群的EGFP表达模式不同，表明每个TEC亚群的TSCOT启动子活性不同。FTOCs的前药治疗减少了胸腺细胞的总数和亚群。在两个转基因品系中，都有一个CD4+CD8+双阳性细胞群高度易感。令人惊讶的是，仅在9.1T-NE胸腺中γδT细胞数量明显减少，这表明它们需要表达TEC群的NTREGFP。因此，这些结果支持αβ和γδ谱系规范在TEC子集内的分工。
In order to investigate the role of thymic epithelial cell (TEC) subsets during T-cell development, we established a new transgenic system, enabling inducible cell-specific ablation as well as marking the TEC subsets using bicistronic bacterial nitroreductase and EGFP genes. Two different lengths of the TSCOT promoter in transgenic mice, named 3.1T-NE and 9.1T-NE, drive EGFP expression into TECs. In adult life, EGFP expression was located in the medulla with a smaller 3.1 kb TSCOT promoter, while it was maintained in the cortex with a 9.1 kb promoter, suggesting putative TEC specific as well as compartment specific cis elements within two promoters. Nitroreductase induced cell death was specific without bystander killing upon the treatment of prodrugs such as nitrofurantoin and metronidazol. The degree of cell death was dependent on the dose of the prodrug in the cell and the fetal thymic organ cultures (FTOCs). Fetal thymic stromal populations were analyzed based on the expression levels of EpCAM, MHCII, CDR1 and/or UEA-1. EGFP expression patterns varied among subsets indicating the differential TSCOT promoter activity in each TEC subset. Prodrug treatment in FTOCs reduced the numbers of total and subsets of thymocytes. A CD4+CD8+ double positive cell population was highly susceptible in both transgenic lines. Surprisingly, there was a distinct reduction in γδ T cell population only in the 9.1T-NE thymus, indicating that they require a NTR-EGFP expressing TEC population. Therefore, these results support a division of labor within TEC subsets for the αβ and γδ lineage specification.
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DOI：
10.1073/pnas.1010890108
发表时间：
2011-02-22
影响因子：
11.1
作者：
Barbee, Susannah D.;Woodward, Martin J.;Hayday, Adrian C.
通讯作者：
Hayday, Adrian C.
影响因子：
4.4
作者：
Alves, Nuno L.;Huntington, Nicholas D.;Di Santo, James P.
通讯作者：
Di Santo, James P.
影响因子：
9.8
作者：
Ahn, Sejin;Lee, Gwanghee;Kim, Moon Gyo
通讯作者：
Kim, Moon Gyo
影响因子：
4.3
作者：
Bowlus, CL;Ahn, J;Gruen, JR
通讯作者：
Gruen, JR
影响因子：
2.2
作者：
Gray, DHD;Chidgey, AP;Boyd, RL
通讯作者：
Boyd, RL