Schim van der Loeff Ina, Hsu Lih-Yun, Saini Manoj, Weiss Art, Seddon Benedict
Division of Immune Cell Biology, Medical Research Council National Institute for Medical Research, London NW7 1AA, United Kingdom; and.
Rosalind Russell and Ephraim P. Engleman Rheumatology Research Center, Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, CA 94143.
J Immunol. 2014 Sep 15;193(6):2873-80. doi: 10.4049/jimmunol.1400858. Epub 2014 Aug 4.
Survival of naive T cells requires engagement of TCR with self-peptide major histocompatibility Ags. The signaling pathways required to transmit this survival signal are poorly understood. In this study, we asked whether the tyrosine kinase Zap70 is required to transmit survival signals in naive CD8 T cells. In the absence of Zap70 expression, thymic development is completely blocked. Using a tetracycline-inducible Zap70 transgene (TetZap70), thymic development of Zap70-deficient TCR transgenic F5 mice was restored. Feeding mice doxycycline to induce Zap70 expression resulted in repopulation of the peripheral naive compartment. Zap70 transgene expression was then ablated by withdrawal of doxycycline. Survival of Zap70-deficient naive CD8 T cells depended on host environment. In hosts with a replete T cell compartment, naive T cells died rapidly in the absence of Zap70 expression. In lymphopenic hosts, Zap70-deficient T cells survived far longer, in an IL-7-dependent manner, but failed to undergo lymphopenia-induced proliferation. Analyzing mixed bone marrow chimeras revealed that intact Zap70-dependent signaling was important for integration of recent thymic emigrants into the mature naive compartment. Finally, we asked whether adaptor function conferred by Zap70 tyrosines 315 and 319 was necessary for transmission of homeostatic TCR signals. This was done by analyzing F5 mice expressing mutant Zap70 in which these residues had been mutated to alanines (Zap70(YYAA)). Inducible Zap70 expression rescued thymic development in F5 TetZap70 Zap70(YYAA) mice. However, in the absence of wild-type Zap70 expression, the Zap70(YYAA) mutant failed to transmit either survival or proliferative homeostatic signals.
初始T细胞的存活需要TCR与自身肽主要组织相容性抗原结合。传递这种存活信号所需的信号通路尚不清楚。在本研究中,我们探究了酪氨酸激酶Zap70是否是在初始CD8 T细胞中传递存活信号所必需的。在缺乏Zap70表达的情况下,胸腺发育完全受阻。利用四环素诱导型Zap70转基因(TetZap70),恢复了Zap70缺陷型TCR转基因F5小鼠的胸腺发育。给小鼠喂食强力霉素以诱导Zap70表达,导致外周初始细胞库重新填充。然后通过停用强力霉素消除Zap70转基因表达。Zap70缺陷型初始CD8 T细胞的存活取决于宿主环境。在T细胞库充足的宿主中,初始T细胞在缺乏Zap70表达时迅速死亡。在淋巴细胞减少的宿主中,Zap70缺陷型T细胞存活时间长得多,以IL-7依赖的方式,但未能经历淋巴细胞减少诱导的增殖。分析混合骨髓嵌合体表明,完整的Zap70依赖信号对于将近期胸腺迁出细胞整合到成熟初始细胞库中很重要。最后,我们探究了Zap70酪氨酸315和319赋予的衔接蛋白功能对于稳态TCR信号传递是否必要。这是通过分析表达突变型Zap70的F5小鼠来完成的,其中这些残基已突变为丙氨酸(Zap70(YYAA))。可诱导的Zap70表达挽救了F5 TetZap70 Zap70(YYAA)小鼠的胸腺发育。然而,在缺乏野生型Zap70表达时,Zap70(YYAA)突变体未能传递存活或增殖的稳态信号。