lck-Driven Cre Expression Alters T Cell Development in the Thymus and the Frequencies and Functions of Peripheral T Cell Subsets
lck-Driven Cre Expression Alters T Cell Development in the Thymus and the Frequencies and Functions of Peripheral T Cell Subsets
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DOI：
10.4049/jimmunol.1600827
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发表时间：
2016-09-15
影响因子：
4.4
通讯作者：
Rottenberg, Martin E.
中科院分区：
文献类型：
--
作者：
Carow, Berit;Gao, Yu;Rottenberg, Martin E.
使用噬菌体衍生的 Cre 重组酶的条件基因打靶广泛应用于小鼠功能基因研究。在 lck 基因启动子控制下的 Cre 转基因小鼠用于研究 loxP 靶向基因在 T 细胞发育和功能中的作用。在本文中，我们展示了在近端 lck 启动子下表达 Cre 的小鼠（lck-cre(+) 小鼠）的细胞结构显着减少了 65%，gamma delta 相对于 alpha beta T 细胞优先发育，以及胸腺中 IL-7R 的表达增加。 CD4/CD8双阴性细胞向双阳性细胞的转变被阻断，lck-cre(+)双阳性细胞更容易发生凋亡，并表现出更高水平的Cre表达。重要的是，lck-cre(+) 小鼠的脾脏和淋巴结中初始 T 细胞的数量减少。相比之下，gγδT细胞、CD44(+)CD62L(-)效应T细胞和Foxp3(+)调节性T细胞的频率升高，分泌IFN-γ的CD4(+)和CD8(+)T细胞的频率也升高。对 332 篇使用 lck-cre(+) 小鼠删除 floxed 基因的文章进行的文献调查表明，结果在统计上受到所用对照（lck-cre(+) 或 lck-cre(-)）的影响，如果使用 lck-cre(-) 对照，则更常见地类似于本文中描述的 lck-cre(+) 表型。总而言之，在解释已发表的结果并使用 lck-cre(+) 菌株正确控制目标基因删除时应小心。
Conditional gene targeting using the bacteriophage-derived Cre recombinase is widely applied for functional gene studies in mice. Mice transgenic for Cre under the control of the lck gene promoter are used to study the role of loxP-targeted genes in T cell development and function. In this article, we show a striking 65% reduction in cellularity, preferential development of gamma delta versus alpha beta T cells, and increased expression of IL-7R in the thymus of mice expressing Cre under the proximal lck promoter (lck-cre(+) mice). The transition from CD4/CD8 double-negative to double-positive cells was blocked, and lck-cre(+) double-positive cells were more prone to apoptosis and showed higher levels of Cre expression. Importantly, numbers of naive T cells were reduced in spleens and lymph nodes of lck-cre(+) mice. In contrast, frequencies of g gamma delta T cells, CD44(+)CD62L(-) effector T cells, and Foxp3(+) regulatory T cells were elevated, as was the frequency of IFN-gamma-secreting CD4(+) and CD8(+) T cells. A literature survey of 332 articles that used lck-cre(+) mice for deletion of floxed genes indicated that results are statistically influenced by the control used (lck-cre(+) or lck-cre(-)), more frequently resembling the lck-cre(+) phenotype described in this article if lck-cre(-) controls were used. Altogether, care should be taken when interpreting published results and to properly control targeted gene deletions using the lck-cre(+) strain.