Aghajani Katayoun, Keerthivasan Shilpa, Yu Yu, Gounari Fotini
Division of Rheumatology and Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, Illinois, USA.
Genesis. 2012 Dec;50(12):908-13. doi: 10.1002/dvg.22052. Epub 2012 Sep 12.
After thymic emigration CD4-T-cells continue to differentiate into multiple effector and suppressor sublineages in peripheral lymphoid organs. In vivo analysis of peripheral CD4-T-cell differentiation has relied on animal models with targeted gene mutations. These are expressed either constitutively or conditionally after Cre mediated recombination. Available Cre transgenic strains to specifically target T-cells act at stages of thymocyte development that precede thymic selection. Tracing gene functions in CD4-T-cell development after thymic exit becomes complicated when the targeted gene is essential during thymic development. Other approaches to conditionally modify gene functions in peripheral T-cells involve infection of in vitro activated cells with Cre expressing lenti-, retro-, or adenoviruses, which precludes in vivo analyses. To study molecular mechanisms of peripheral CD4-T-cell differentiation in vivo and in vitro we generated transgenic mice expressing a tamoxifen inducible Cre recombinase (CreER(T2) ) under the control of the CD4 gene promoter. We show here that in CD4CreER(T2) mice Cre is inducibly and selectively activated in CD4-T-cells. Tamoxifen treatment both in vivo and in vitro results in efficient recombination of loci marked by LoxP sites. Moreover, this strain shows no abnormalities related to transgene insertion. Therefore it provides a valuable tool for studying gene function during differentiation of naïve peripheral CD4-T-cells into effector or suppressor sub-lineages.
在胸腺迁出后,CD4 + T细胞在外周淋巴器官中继续分化为多种效应和抑制亚群。对外周CD4 + T细胞分化的体内分析依赖于具有靶向基因突变的动物模型。这些突变在Cre介导的重组后组成性或条件性表达。可用于特异性靶向T细胞的Cre转基因品系作用于胸腺选择之前的胸腺细胞发育阶段。当靶向基因在胸腺发育过程中至关重要时,追踪胸腺退出后CD4 + T细胞发育中的基因功能就变得复杂。其他在外周T细胞中条件性修饰基因功能的方法涉及用表达Cre的慢病毒、逆转录病毒或腺病毒感染体外激活的细胞,这排除了体内分析。为了研究体内和体外外周CD4 + T细胞分化的分子机制,我们构建了在CD4基因启动子控制下表达他莫昔芬诱导型Cre重组酶(CreER(T2))的转基因小鼠。我们在此表明,在CD4CreER(T2)小鼠中,Cre在CD4 + T细胞中被诱导性和选择性地激活。体内和体外的他莫昔芬处理导致由LoxP位点标记的基因座的有效重组。此外,该品系未显示与转基因插入相关的异常。因此,它为研究幼稚外周CD4 + T细胞分化为效应或抑制亚群过程中的基因功能提供了一个有价值的工具。