Fernandez David R, Telarico Tiffany, Bonilla Eduardo, Li Qing, Banerjee Sanjay, Middleton Frank A, Phillips Paul E, Crow Mary K, Oess Stefanie, Muller-Esterl Werner, Perl Andras
Division of Rheumatology, Department of Medicine, State University of New York, Syracuse, NY 13210, USA.
J Immunol. 2009 Feb 15;182(4):2063-73. doi: 10.4049/jimmunol.0803600.
Persistent mitochondrial hyperpolarization (MHP) and enhanced calcium fluxing underlie aberrant T cell activation and death pathway selection in systemic lupus erythematosus. Treatment with rapamycin, which effectively controls disease activity, normalizes CD3/CD28-induced calcium fluxing but fails to influence MHP, suggesting that altered calcium fluxing is downstream or independent of mitochondrial dysfunction. In this article, we show that activity of the mammalian target of rapamycin (mTOR), which is a sensor of the mitochondrial transmembrane potential, is increased in lupus T cells. Activation of mTOR was inducible by NO, a key trigger of MHP, which in turn enhanced the expression of HRES-1/Rab4, a small GTPase that regulates recycling of surface receptors through early endosomes. Expression of HRES-1/Rab4 was increased in CD4(+) lupus T cells, and in accordance with its dominant impact on the endocytic recycling of CD4, it was inversely correlated with diminished CD4 expression. HRES-1/Rab4 overexpression was also inversely correlated with diminished TCRzeta protein levels. Pull-down studies revealed a direct interaction of HRES-1/Rab4 with CD4 and TCRzeta. Importantly, the deficiency of the TCRzeta chain and of Lck and the compensatory up-regulation of FcepsilonRIgamma and Syk, which mediate enhanced calcium fluxing in lupus T cells, were reversed in patients treated with rapamcyin in vivo. Knockdown of HRES-1/Rab4 by small interfering RNA and inhibitors of lysosomal function augmented TCRzeta protein levels in vitro. The results suggest that activation of mTOR causes the loss of TCRzeta in lupus T cells through HRES-1/Rab4-dependent lysosomal degradation.
持续性线粒体超极化(MHP)和增强的钙通量是系统性红斑狼疮中异常T细胞活化和死亡途径选择的基础。雷帕霉素治疗可有效控制疾病活动,使CD3/CD28诱导的钙通量正常化,但对MHP无影响,这表明钙通量改变是线粒体功能障碍的下游事件或与之无关。在本文中,我们表明,作为线粒体跨膜电位传感器的哺乳动物雷帕霉素靶蛋白(mTOR)在狼疮T细胞中的活性增加。mTOR的激活可被NO诱导,NO是MHP的关键触发因素,进而增强了HRES-1/Rab4的表达,HRES-1/Rab4是一种小GTP酶,通过早期内体调节表面受体的再循环。HRES-1/Rab4在CD4(+)狼疮T细胞中的表达增加,并且由于其对CD4内吞再循环的主要影响,它与CD4表达减少呈负相关。HRES-1/Rab4的过表达也与TCRzeta蛋白水平降低呈负相关。下拉实验揭示了HRES-1/Rab4与CD4和TCRzeta之间的直接相互作用。重要的是,在用雷帕霉素进行体内治疗的患者中,狼疮T细胞中介导增强钙通量的TCRzeta链、Lck的缺陷以及FcepsilonRIgamma和Syk的代偿性上调均得到逆转。在体外,通过小干扰RNA敲低HRES-1/Rab4和抑制溶酶体功能可增加TCRzeta蛋白水平。结果表明,mTOR的激活通过HRES-1/Rab4依赖的溶酶体降解导致狼疮T细胞中TCRzeta的丢失。