Oxidative Stress Induces Mitochondrial Compromise in CD4 T Cells From Chronically HCV-Infected Individuals.
Oxidative Stress Induces Mitochondrial Compromise in CD4 T Cells From Chronically HCV-Infected Individuals.
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DOI：
10.3389/fimmu.2021.760707
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发表时间：
2021
影响因子：
7.3
通讯作者：
Yao ZQ
中科院分区：
文献类型：
--
作者：
Schank M;Zhao J;Wang L;Nguyen LNT;Cao D;Dang X;Khanal S;Zhang J;Zhang Y;Wu XY;Ning S;Gazzar ME;Moorman JP;Yao ZQ
我们之前已经证明慢性丙型肝炎病毒（HCV）感染可诱导DNA损伤和T细胞过度氧化应激的免疫功能障碍。此外，有证据表明，丙型肝炎病毒有助于增加对代谢紊乱的易感性。然而，HCV感染损害CD4 T细胞代谢的潜在机制尚不清楚。在这项研究中，我们用流式细胞术评估了慢性hcv感染者和健康受试者的CD4 T细胞的线粒体质量和细胞内和线粒体活性氧（ROS）的产生，用实时qPCR评估了线粒体DNA （mtDNA）的含量，用海马分析仪评估了细胞呼吸，用液相色谱-质谱法（LC-MS）评估了线粒体定位蛋白失调。线粒体质量减少，细胞内和线粒体ROS增加，线粒体主要调节因子过氧化物酶体增殖物激活受体1α (PGC-1α)和线粒体转录因子A （mtTFA）表达下调，氧化应激增加，线粒体DNA拷贝数减少。重要的是，CRISPR/ cas9介导的mtTFA敲低会损害细胞呼吸并减少mtDNA拷贝数。此外，在HCV-CD4 T细胞中，负责介导氧化应激、细胞凋亡和mtDNA维持的蛋白质显著改变。这些结果表明，线粒体功能在HCV-CD4 T细胞中受到损害，可能是通过一些线粒体调节蛋白的失调。
We have previously shown that chronic Hepatitis C virus (HCV) infection can induce DNA damage and immune dysfunctions with excessive oxidative stress in T cells. Furthermore, evidence suggests that HCV contributes to increased susceptibility to metabolic disorders. However, the underlying mechanisms by which HCV infection impairs cellular metabolism in CD4 T cells remain unclear. In this study, we evaluated mitochondrial mass and intracellular and mitochondrial reactive oxygen species (ROS) production by flow cytometry, mitochondrial DNA (mtDNA) content by real-time qPCR, cellular respiration by seahorse analyzer, and dysregulated mitochondrial-localized proteins by Liquid Chromatography-Mass Spectrometry (LC-MS) in CD4 T cells from chronic HCV-infected individuals and health subjects. Mitochondrial mass was decreased while intracellular and mitochondrial ROS were increased, expressions of master mitochondrial regulators peroxisome proliferator-activated receptor 1 alpha (PGC-1α) and mitochondrial transcription factor A (mtTFA) were down-regulated, and oxidative stress was increased while mitochondrial DNA copy numbers were reduced. Importantly, CRISPR/Cas9-mediated knockdown of mtTFA impaired cellular respiration and reduced mtDNA copy number. Furthermore, proteins responsible for mediating oxidative stress, apoptosis, and mtDNA maintenance were significantly altered in HCV-CD4 T cells. These results indicate that mitochondrial functions are compromised in HCV-CD4 T cells, likely via the deregulation of several mitochondrial regulatory proteins.