The College of Life Sciences, Division of Cell Signalling and Immunology, The University of Dundee, Scotland, UK.
Biochem J. 2012 Mar 15;442(3):649-59. doi: 10.1042/BJ20111700.
PKD (protein kinase D) 2 is a serine/threonine kinase activated by diacylglycerol in response to engagement of antigen receptors in lymphocytes. To explore PKD2 regulation and function in TCR (T-cell antigen receptor) signal transduction we expressed TCR complexes with fixed affinity for self antigens in the T-cells of PKD2-null mice or mice deficient in PKD2 catalytic activity. We also developed a single cell assay to quantify PKD2 activation as T-cells respond to developmental stimuli or engagement of α/β TCR complexes in vivo. Strikingly, PKD2 loss caused increases in thymic output, lymphadenopathy and splenomegaly in TCR transgenic mice. The precise magnitude and timing of PKD2 activation during T-cell development is thus critical to regulate thymic homoeostasis. PKD2-null T-cells that exit the thymus have a normal transcriptome, but show a limited and abnormal transcriptional response to antigen. Transcriptional profiling reveals the full consequences of PKD2 loss and maps in detail the selective, but critical, function for PKD2 in signalling by α/β mature TCR complexes in peripheral T-cells.
蛋白激酶 D2(PKD2)是一种丝氨酸/苏氨酸激酶,可被二酰基甘油激活,以响应淋巴细胞抗原受体的结合。为了探索 PKD2 在 T 细胞抗原受体(TCR)信号转导中的调节和功能,我们在 PKD2 缺失小鼠或缺乏 PKD2 催化活性的小鼠的 T 细胞中表达了具有固定亲和力的 TCR 复合物。我们还开发了一种单细胞测定法来定量 PKD2 的激活,以检测 T 细胞对发育刺激或体内 α/β TCR 复合物的结合的反应。令人惊讶的是,PKD2 的缺失导致 TCR 转基因小鼠的胸腺输出增加、淋巴结病和脾肿大。因此,PKD2 在 T 细胞发育过程中的激活的精确幅度和时间对于调节胸腺稳态至关重要。离开胸腺的 PKD2 缺失 T 细胞具有正常的转录组,但对抗原的反应有限且异常。转录谱分析揭示了 PKD2 缺失的全部后果,并详细绘制了 PKD2 在周围 T 细胞中成熟的 α/β TCR 复合物信号转导中的选择性但关键功能。