Molecules and Cells

, Volume 34, Issue 5, pp 481–493

Thymic epithelial requirement for γδ T cell development revealed in the cell ablation transgenic system with TSCOT promoter


  • Gwanghee Lee
    • Department of Cell Biology and PhysiologyWashington University School of Medicine
    • Department of Physiological ChemistryGenentech
  • Ki Yeon Kim
    • Department of Biological SciencesInha University
  • Cheong-Hee Chang
    • Department of Microbiology and ImmunologyThe University of Michigan Medical School
    • Department of Biological SciencesInha University

DOI: 10.1007/s10059-012-0246-4

Cite this article as:
Lee, G., Kim, K.Y., Chang, C. et al. Mol Cells (2012) 34: 481. doi:10.1007/s10059-012-0246-4


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 NTREGFP expressing TEC population. Therefore, these results support a division of labor within TEC subsets for the αβ and γδ lineage specification.


EGFPfetal thymic organ culturegamma delta T cellthymic epithelial celltransgenic mouse
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© The Korean Society for Molecular and Cellular Biology and Springer Netherlands 2012