HCV in Alcoholics

酗酒者中的丙型肝炎病毒

• 批准号: 7911255
• 负责人: Jack R Wands
• 金额: $ 4.63万
• 依托单位: RHODE ISLAND HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7911255
• 关键词: Adoptive Transfer; Alcohol abuse; Animals; Antibodies; Antigen Presentation; Antigen Targeting; Antigens; Antiviral Agents; Antiviral Response; Biological Models; CD8-Positive T-Lymphocytes; CD8B1 gene; Cell physiology; Cell surface; Cells; Cholesterol; Chronic; Chronic Hepatitis C; Cirrhosis; Cytotoxic T-Lymphocytes; DNA delivery; Dendritic Cells; Development; Endocytosis; Ethanol; Experimental Animal Model; Experimental Models; Functional disorder; Gene Expression; Generations; Genetic; Grant; Granulocyte-Macrophage Colony-Stimulating Factor; Hepatitis C; Hepatitis C virus; IL2 gene; Immune response; Immune system; Immunization; Immunologics; In Vitro; Incidence; Infection; Investigation; Liver; Liver diseases; Mediating; Medical; Modeling; Molecular Profiling; Mus; Peptides; Plasmids; Play; Primary carcinoma of the liver cells; Proteins; RNA; Research Personnel; Risk; Role; Spermine; Spleen; Stimulus; Structural Protein; T-Lymphocyte; Testing; Transfection; Viral; Viral Antigens; Viral Proteins; Virus Diseases; alcohol effect; alcohol exposure; base; cell type; chronic alcohol ingestion; cytokine; cytotoxic; feeding; genetic immunization strategies; improved; in vivo; interest; mannose receptor; novel strategies; problem drinker; programs; receptor mediated endocytosis; uptake; virus core; virus infection mechanism

DESCRIPTION (provided by applicant): Chronic hepatitis C (HCV) infection is a major cause of liver disease, cirrhosis, and hepatocellular carcinoma (HCC) in alcoholics. Although it has been established that chronic alcoholics have a high incidence of CV infection, the reasons for these high rates are unknown but may partially relate to the effects of ethanol on the humoral and cellular immune responses to viral structural and non-structural proteins. Past studies revealed that chronic ethanol feeding in a murine model has a profound inhibitory effect on the generation of viral-specific CD4+ and CD8+ T-cell activity to HCV core and NS5 proteins used as the immunogens. More important, this inhibitory effect of ethanol was completely reversed by co-immunization with an IL2 or GM- CSF expression plasmids following genetic immunization suggesting that dendritic cells (DCs) could be involved in these abnormal immune responses and thus, there may not be intrinsic abnormalities in CD4+ and CD8+ cells per se. We have established an experimental model system to study the effects of chronic ethanol consumption on DC function generated in vivo under controlled conditions. This approach allowed us to characterize ethanol's effect on DCs with respect to cell surface markers, cytokine profiles, antigen presentation ability, and endocytosis capacity. Furthermore, we evaluated the subsequent role of DC dysfunction in the generation of cytotoxic T-lymphocytes (CTL) immune responses following genetic immunization with a plasmid expressing NS5 protein by using adoptive transfer of syngeneic DCs. In vivo generation of DCs combined with syngeneic DC transfer and DMA-based immunization revealed that ethanol induced dysfunction of DCs is the major factor responsible for the reduce cellular immune response to HCV. This finding may have relevance to persistent HCV infection in alcoholics and identifies a critical cell type that is at risk for ethanol effects. The central hypothesis in this proposal is that altered DC function is one of the major immunologic changes produced by ethanol and there may be differential effects of chronic ethanol on DC subpopulations which subsequently impairs the cellular immune response necessary for viral clearance. Therefore, we would like to develop and implement strategies that increase the ability of the ethanol consuming host to generate better anti-viral responses. We plan to do the following: Specific Aim 1: Evaluate the role of DCs on immune responses to HCV in chronic ethanol-fed mice. Specific Aim 2: Explore two new approaches of specific viral antigen targeting to activate DCs in vivo during chronic ethanol exposure. Specific Aim 3. Characterize the possible genetic mechanisms responsible for DC dysfunction produced by ethanol. These investigations open new avenues to enhance antiviral immune response in the setting of chronic ethanol consumption by improving DC function.

描述（由申请人提供）：慢性丙型肝炎（HCV）感染是酒精中毒中肝病，肝硬化和肝细胞癌（HCC）的主要原因。尽管已经确定慢性酗酒者的CV感染发生率很高，但这些高率的原因尚不清楚，但可能部分与乙醇对病毒结构和非结构性蛋白质的体液和细胞免疫反应有关。过去的研究表明，鼠模型中的慢性乙醇喂养对病毒特异性CD4+和CD8+ T细胞活性的产生具有深远的抑制作用，并将其用于HCV核心和用作免疫原子的NS5蛋白。更重要的是，通过与遗传免疫后的IL2或GM-CSF表达质粒共免疫完全逆转了乙醇的这种抑制作用，这表明树突状细胞（DCS）可能与这些异常免疫反应有关，因此可能与CD4+和CD8+细胞中的固有异常相关。我们已经建立了一个实验模型系统，以研究慢性乙醇消耗对受控条件下体内生成的直流功能的影响。这种方法使我们能够表征乙醇对DC在细胞表面标记，细胞因子谱，抗原表现能力和内吞作用能力方面的影响。此外，我们评估了直流功能障碍在遗传免疫后用质粒表达NS5蛋白的遗传免疫后，DC功能障碍在生成细胞毒性T-淋巴细胞（CTL）免疫反应中的作用。在体内生成DC结合了同步DC转移和基于DMA的免疫，表明乙醇诱导的DC功能障碍是负责减少细胞免疫反应HCV的主要因素。这一发现可能与酗酒者中持续的HCV感染有关，并确定有乙醇作用风险的关键细胞类型。该提议中的中心假设是，DC功能的改变是乙醇产生的主要免疫学变化之一，并且慢性乙醇对DC亚群的影响可能会差异，随后会损害病毒清除所需的细胞免疫反应。因此，我们想制定和实施策略，以提高消费乙醇宿主产生更好的抗病毒反应的能力。我们计划执行以下操作：具体目标1：评估DC在慢性乙醇喂养小鼠中对HCV免疫反应的作用。特定目标2：探索两种特定病毒抗原靶向的新方法，以在慢性乙醇暴露期间激活体内DC。具体目的3。表征导致乙醇产生的直流功能障碍的可能的遗传机制。这些研究开放了新的途径，以通过改善直流功能来增强慢性乙醇消耗的抗病毒免疫反应。