Ethanol modulate central and peripheral immune responses via HMGB1, IL-1Beta, and other Immune Signaling Molecules

项目总结/摘要 酒精使用障碍是可预防死亡的第三大原因，占 每年约有90，000例可预防的死亡（Danaei et al 2009）。近50% 死亡是继发于与炎症和异常 免疫激活，如败血症和酒精性肝病（Mokdad et al 2000）。 最近的研究发现，酒精激活了先天免疫系统， CNS和外周。因此，了解酒精对免疫系统的影响 是至关重要的，以形成一个基础的发现，导致理解这两个 酒精中毒和许多酒精相关疾病的病理学。我们发现 最近，酒精通过“主调节器”激活先天免疫系统， 先天免疫HMGB 1在这项提案中，我们的目的是调查酒精的影响， 通过HMGB 1及其伴随物对中枢和外周免疫激活的影响 细胞因子IL-10。使用体外细胞培养系统（目标1）、体内实验（目标2-3） 在人类酗酒者、烧伤患者和酒精性肝炎患者中的评估 (Aim 4），我们希望获得对酒精免疫影响的了解贯穿始终 身体我将获得新的实验室经验与真实的时间PCR，酶联 免疫吸附测定，免疫共沉淀，蛋白质印迹，脑内AAV 5 病毒注射、质谱、免疫细胞化学和活体荧光成像。 我们还研究了这种病理学中细胞间通讯的新机制， 酒精诱导的促炎微粒释放。此外，我们调查了 PTEN/PI 3 K/Akt/mTOR通路在酒精免疫效应中的作用及研究 这一途径的抑制剂，可能是新的治疗选择。我们的初步数据 强烈表明酒精引起中枢和外周HMGB 1和IL-1 β释放 通过PTEN/PI 3 K/Akt/mTOR连接机制在微粒中表达。该项目还 将大量的职业发展与职业和研究指导相结合， 顶级研究人员和临床医生，学习新的实验技术， 课程工作，以提高临床/翻译专业知识。共同导师富尔顿T.船员们， 博士和布鲁斯凯恩斯医学博士提供了基础科学，临床研究， 职业发展辅导，这将有助于确保 候选人是一个顶级的独立翻译研究员。

Project Summary/Abstract Alcohol use disorders are third leading cause of preventable death, accounting for approximately 90,000 preventable deaths each year (Danaei et al 2009). Nearly 50% deaths are secondary to diseases that are associated with inflammation and aberrant immune activation such as sepsis and alcoholic liver disease (Mokdad et al 2000). Recent discoveries find that alcohol activates the innate immune system in both the CNS and periphery. Thus, understanding the effects of alcohol on the immune system is critical to form a foundation of discovery leading to the understanding of both the pathology of alcoholism and numerous alcohol-associated diseases. We have found recently that alcohol activates the innate immune system through the ‘master regulator’ of innate immunity, HMGB1. In this proposal we aim to investigate the effects of alcohol on central and peripheral immune activation through HMGB1 and its companion cytokine IL-. Using in vitro cell culture systems (Aim 1), in vivo experiments (Aim 2-3) and assessments in human alcoholics, burn patients and alcoholic hepatitis patients (Aim 4), we hope to gain an understanding of the immune effects of alcohol throughout the body. I will gain new laboratory experience with real time-PCR, enzyme-linked immunosorbant assays, co-immunoprecipitation, western blotting, intracerebral AAV5 viral injections, mass spectrometry, immunocytochemistry, and live fluorescent imaging. We also investigate a novel mechanism of cell-cell communication in this pathology, alcohol-induced release of pro-inflammatory microparticles. Further, we investigate the role of the PTEN/PI3K/Akt/mTOR pathway in the immune effects of alcohol, and study inhibitors of this pathway which may be novel therapeutic options. Our preliminary data strongly suggest that alcohol causes central and peripheral HMGB1 and IL-1 release in microparticles through a PTEN/PI3K/Akt/mTOR linked mechanism. This project also incorporates substantial career development with career and research mentoring from top tier researchers and clinicians, the learning of new experimental techniques, and course work to enhance clinical/translational expertise. Co-mentors Fulton T. Crews, PhD and Bruce Cairns MD provide a depth of basic science, clinical research, and career development mentorship that will help ensure the successful transition of the candidate to being a top independent translational researcher.