HCV, Alcohol and Host Defense

HCV、酒精和宿主防御

• 批准号: 7114949
• 负责人: Gyongyi Szabo
• 金额: $ 38.82万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-15 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7114949
• 关键词: CD14 molecule; CHO cells; RNase protection assay; alcoholism /alcohol abuse; chemokine; clinical research; dendritic cells; flow cytometry; gel mobility shift assay; genetically modified animals; hepatitis C virus; host organism interaction; human subject; immunoprecipitation; inflammation; laboratory mouse; leukocyte activation /transformation; macrophage; monocyte; polymerase chain reaction; protein structure function; toll like receptor; transfection; virus protein; western blottings; CD14 molecule; CHO cells; RNase protection assay; alcoholism /alcohol abuse; chemokine; clinical research; dendritic cells; flow cytometry; gel mobility shift assay; genetically modified animals; hepatitis C virus; host organism interaction; human subject; immunoprecipitation; inflammation; laboratory mouse; leukocyte activation /transformation; macrophage; monocyte; polymerase chain reaction; protein structure function; toll like receptor; transfection; virus protein; western blottings

DESCRIPTION (provided by applicant): Alcohol use accelerates the progression of liver disease in chronic hepatitis C (HCV) infection; however, the interactions between alcohol and HCV have not yet been explored. Chronic HCV infection is associated with reduced HCV-specific immune responses, and alcohol use suppresses antigen-specific T cell activation via inhibition of monocyte and dendritic cell (DC) antigen presentation. Both chronic alcohol consumption and chronic HCV infection are independently associated with activation of inflammatory pathways that mediate hepatocellular injury. Thus, we hypothesize that alcohol, HCV interacts to activate non-specific pro-inflammatory cascades in chronic HCV infection, and this process may contribute to progression of liver damage. Based on our preliminary results, we propose that the pattern recognition receptor, toll-like receptor 2 (TLR2), mediates cellular activation by HCV core and NS3 proteins. We further postulate that alcohol may increase inflammatory cell activation by interacting with TLR2-mediated pathways. Our preliminary results show that myeloid DC (dendritic cells), the most potent antigen presenting cell type, have reduced T-cell activation capacity in patients with chronic HCV infection and this can be further decreased by alcohol treatment of DCs. Thus, we propose that alcohol contributes to the persistence of chronic infection in HCV by inhibiting functional maturation of dendritic cells, thereby decreasing antigen-specific T cell activation and viral recognition. The Specific Aims of this proposal are: 1. To investigate interactions between HCV- and alcohol-induced inflammatory pathways at the protein and mRNA levels and on activation of the nuclear regulatory kB/Rel pathway in normal monocytes and in patients with chronic HCV infection with and without alcohol use or chronic in vitro alcohol treatment. 2. To investigate the pattern recognition receptor, TLR2, as a putative receptor in cell activation by HCV proteins. This will be accomplished by testing cellular activation by HCV proteins in TLR2-transfected CHO and HEK cells and in macrophages of TLR2-deficient mice. The cooperation of TLR2 with other components of the TLR2 receptor complex required for successful transmission of HCV core and NS3 mediated cell activation will be identified by selectively investigating the roles of CD14, TLR 1, and TLR6. 3. To evaluate the interactions between alcohol and HCV proteins on TLR2-mediated cell activation pathways by determining the effects of acute and chronic alcohol treatment on TLR2 expression, gene activation, and TLR2-mediated monocyte activation from normal controls and chronic HCV infected patients. Investigation of downstream elements involved in TLR2-mediated intracellular signaling after stimulation with core and NS3 proteins and their modulation by alcohol will be tested. 4. To delineate the role of TLR2-mediated signaling by HCV core and NS3 proteins in the reduced dendritic cell differentiation in HCV infected patients and to evaluate the effect of alcohol on TLR2-mediated signals in DC development. Results from these studies should reveal molecular and cellular mechanisms by which alcohol interacts with HCV infection to accelerate disease persistence and progression.

描述（由申请人提供）：饮酒加速了慢性丙型肝炎（HCV）感染中肝病的进展；但是，尚未探索酒精与HCV之间的相互作用。 慢性HCV感染与HCV特异性免疫反应降低有关，酒精使用通过抑制单核细胞和树突状细胞（DC）抗原表现来抑制抗原特异性T细胞活化。 慢性酒精消耗和慢性HCV感染都与介导肝细胞损伤的炎症途径的激活有关。 因此，我们假设酒精，HCV相互作用以激活慢性HCV感染中的非特异性促炎级联反应，并且此过程可能有助于肝脏损伤的进展。 基于我们的初步结果，我们建议模式识别受体Toll样受体2（TLR2）介导HCV核心和NS3蛋白的细胞激活。 我们进一步假设酒精可以通过与TLR2介导的途径相互作用来增加炎症细胞的激活。 我们的初步结果表明，髓样DC（树突状细胞）是最有效的抗原细胞类型，在慢性HCV感染患者的T细胞激活能力下降低了，并且通过DC的酒精治疗可以进一步降低这一点。 因此，我们建议酒精通过抑制树突状细胞的功能成熟，从而降低抗原特异性T细胞活化和病毒识别，从而有助于HCV中慢性感染的持续性。 该提案的具体目的是：1。研究蛋白质和mRNA水平上HCV和酒精诱导的炎症途径之间的相互作用，以及正常单核细胞中核调节性KB/RER途径的激活以及在慢性HCV感染的患者中，无酒精使用或慢性酒精使用或慢性酒精治疗。 2。研究模式识别受体TLR2，作为HCV蛋白细胞激活中的假定受体。这将通过在TLR2转染的CHO和HEK细胞以及TLR2缺乏小鼠的巨噬细胞中测试HCV蛋白的细胞激活来实现。 通过选择性研究CD14，TLR 1和TLR6的作用，可以鉴定出成功传播HCV核心和NS3介导的细胞活化所需的TLR2受体复合物的其他成分与TLR2受体复合物的合作。 3。通过确定急性和慢性酒精治疗对TLR2表达，基因激活和TLR2介导的单核细胞激活的影响，从正常对照和慢性HCV感染患者中确定急性和慢性酒精治疗对TLR2表达，基因激活以及TLR2介导的单核细胞激活TLR2介导的细胞激活途径的相互作用。用核心和NS3蛋白刺激后，研究参与TLR2介导的细胞内信号传导及其通过酒精调节后的细胞内信号传导的研究。 4。为了描述HCV核心和NS3蛋白在HCV感染患者中降低的树突状细胞分化中TLR2介导的信号传导的作用，并评估酒精对DC发育中TLR2介导的信号的影响。 这些研究的结果应揭示酒精与HCV感染相互作用以加速疾病持久性和进展的分子和细胞机制。