■慢性肝感染对循环中抗原非特异性免疫细胞的影响仍然知之甚少。我们报道了HCV感染的肝硬化患者外周CD8T细胞的持续总体功能亢进。大量CD8T细胞中的基因表达模式是否与HCV感染中肝纤维化的严重程度相关尚不清楚。
■来自初始治疗的血液CD8T细胞的RNA测序,HCV感染的个体具有轻微(MetavirF0-1≤7.0kPa)或晚期纤维化或肝硬化(F4≥12.5kPa),在直接抗病毒治疗之前和之后,已执行。通过流式细胞术评估CD8T细胞功能。
■在DAA前患者的CD8T细胞中,与轻度纤维化相比,基因本体分析和基因集富集分析确定了与细胞功能和代谢相关的差异基因表达,包括上调的Hedgehog(Hh)信号,IFN-α,-γ,TGF-β反应基因,凋亡,顶端表面途径,磷脂酶信号,磷脂酰胆碱/肌醇活性,和第二信使介导的信号。相比之下,与核过程相关的通路中的基因,RNA转运,细胞骨架动力学,cMyc/E2F法规,氧化磷酸化,和mTOR信号,减少了。Hh信号通路是肝硬化中最高的特征基因集,其中标志基因GLI1和PTCH1排名很高。与最小纤维化相比,Smo依赖性Hh信号传导的抑制消除了来自晚期慢性HCV感染患者的刺激CD8T细胞中IFN-γ和穿孔素的表达。DAA后的CD8T细胞基因表达谱与DAA前的表达谱仍然聚集在一起,并且在晚期和最小纤维化之间存在差异。表明病毒清除后很长时间基因表达的持续扰动。
对慢性HCV感染中的大量CD8T细胞基因表达的这种分析提示在肝硬化状态下CD8T细胞池的大量重编程。肝硬化中Hh信号的增加可能导致慢性HCV感染中观察到的全身性CD8T细胞功能亢进。了解免疫细胞功能障碍的持久性可能有助于减轻HCV清除后仍然存在的临床挑战,更广泛地说,改善严重肝病患者的长期预后.
UNASSIGNED: The impact of chronic hepatic infection on antigen non-specific immune cells in circulation remains poorly understood. We reported lasting global hyperfunction of peripheral CD8 T cells in HCV-infected individuals with cirrhosis. Whether gene expression patterns in bulk CD8 T cells are associated with the severity of liver fibrosis in HCV infection is not known.
UNASSIGNED: RNA sequencing of blood CD8 T cells from treatment naïve, HCV-infected individuals with minimal (Metavir F0-1 ≤ 7.0 kPa) or advanced fibrosis or cirrhosis (F4 ≥ 12.5 kPa), before and after direct-acting antiviral therapy, was performed. CD8 T cell function was assessed by flow cytometry.
UNASSIGNED: In CD8 T cells from pre-DAA patients with advanced compared to minimal fibrosis, Gene Ontology analysis and Gene Set Enrichment Analysis identified differential gene expression related to cellular function and metabolism, including upregulated Hedgehog (Hh) signaling, IFN-α, -γ, TGF-β response genes, apoptosis, apical surface pathways, phospholipase signaling, phosphatidyl-choline/inositol activity, and second-messenger-mediated signaling. In contrast, genes in pathways associated with nuclear processes, RNA transport, cytoskeletal dynamics, cMyc/E2F regulation, oxidative phosphorylation, and mTOR signaling, were reduced. Hh signaling pathway was the top featured gene set upregulated in cirrhotics, wherein hallmark genes GLI1 and PTCH1 ranked highly. Inhibition of Smo-dependent Hh signaling ablated the expression of IFN-γ and perforin in stimulated CD8 T cells from chronic HCV-infected patients with advanced compared to minimal fibrosis. CD8 T cell gene expression profiles post-DAA remained clustered with pre-DAA profiles and disparately between advanced and minimal fibrosis, suggesting a persistent perturbation of gene expression long after viral clearance.
UNASSIGNED: This analysis of bulk CD8 T cell gene expression in chronic HCV infection suggests considerable reprogramming of the CD8 T cell pool in the cirrhotic state. Increased Hh signaling in cirrhosis may contribute to generalized CD8 T cell hyperfunction observed in chronic HCV infection. Understanding the lasting nature of immune cell dysfunction may help mitigate remaining clinical challenges after HCV clearance and more generally, improve long term outcomes for individuals with severe liver disease.