Saxena Mansi, Busca Aurelia, Holcik Martin, Kumar Ashok
Department of Biochemistry, Microbiology and Immunology, Children's Hospital of Eastern Ontario, University of Ottawa, Ottawa, Ontario K1H 8L1, Canada;
Research Institute, Children's Hospital of Eastern Ontario, University of Ottawa, Ottawa, Ontario K1H 8L1, Canada; and.
J Immunol. 2016 May 1;196(9):3754-67. doi: 10.4049/jimmunol.1402379. Epub 2016 Mar 11.
Monocytes and macrophages are important HIV reservoirs, as they exhibit marked resistance to apoptosis upon infection. However, the mechanism underlying resistance to apoptosis in these cells is poorly understood. Using HIV-viral protein R-52-96 aa peptide (Vpr), we show that primary monocytes and THP-1 cells treated with Vpr are highly susceptible to mitochondrial depolarization, but develop resistance following stimulation with bacterial DNA or CpG oligodeoxynucleotide. We have shown that Vpr-induced mitochondrial depolarization is mediated by TNFR-associated factor-1 (TRAF-1) and TRAF-2 degradation and subsequent activation of caspase-8, Bid, and Bax. To provide the mechanism governing such resistance to mitochondrial depolarization, our results show that prior stimulation with CpG oligodeoxynucleotide or Escherichia coli DNA prevented: 1) TRAF-1/2 downregulation; 2) activation of caspase-8, Bid, and Bax; and 3) subsequent mitochondrial depolarization and release of apoptosis-inducing factor and cytochrome c Furthermore, this protection was mediated by upregulation of antiapoptotic protein (c-IAP-2) through calmodulin-dependent kinase-II activation. Thus, c-IAP-2 may prevent Vpr-mediated mitochondrial depolarization through stabilizing TRAF-1/2 expression and sequential inhibition of caspase-8, Bid, and Bax.
单核细胞和巨噬细胞是重要的HIV储存库,因为它们在感染后对细胞凋亡表现出显著抗性。然而,这些细胞中抗细胞凋亡的潜在机制仍知之甚少。使用HIV病毒蛋白R-52-96氨基酸肽(Vpr),我们发现用Vpr处理的原代单核细胞和THP-1细胞极易发生线粒体去极化,但在用细菌DNA或CpG寡脱氧核苷酸刺激后会产生抗性。我们已经表明,Vpr诱导的线粒体去极化是由肿瘤坏死因子受体相关因子-1(TRAF-1)和TRAF-2降解以及随后的半胱天冬酶-8、Bid和Bax激活介导的。为了揭示这种对线粒体去极化抗性的机制,我们的结果表明,事先用CpG寡脱氧核苷酸或大肠杆菌DNA刺激可防止:1)TRAF-1/2下调;2)半胱天冬酶-8、Bid和Bax激活;3)随后的线粒体去极化以及凋亡诱导因子和细胞色素c的释放。此外,这种保护作用是通过钙调蛋白依赖性激酶-II激活上调抗凋亡蛋白(c-IAP-2)来介导的。因此,c-IAP-2可能通过稳定TRAF-1/2表达并依次抑制半胱天冬酶-8、Bid和Bax来防止Vpr介导的线粒体去极化。