Project 3-Proteomic Analysis of Explanted Livers with characterization of Autoantigens

项目总结 重型酒精性肝炎(SAH)是酒精性肝病(ALD)中最严重的一种。慢性酒精 滥用与肺炎、感染(如艾滋病毒、丙型肝炎和结核病)的易感性增加有关， 并伴有慢性炎症。过去30年在人类和动物模型中的研究表明， ALD可能是酒精滥用对适应性免疫系统的影响所致。长期酗酒被发现 减少B细胞数量，降低抗原特异性抗体反应，增加 抗肝脏自身抗原和氧化损伤副产物的自身抗体。此外，更高水平的 Ig G、Ig A和Ig E亚型免疫球蛋白和抗肝抗原自身抗体的产生 已在SAH患者中观察到。因此，我们假设患者体内产生的自身抗体 SAH会对肝脏造成损害。为了充分检验这一假设，识别这些抗原 被沉积在SAH肝脏中的抗体识别是了解SAH的关键踏脚石 疾病的病因学。此外，其中一些抗体很可能是通过交叉反应而产生的。 针对肠道细菌的抗体。这项提议的目标是确定这些 SAH患者中的SAH特异性抗体，并表征其功能和对肝脏损害的贡献。 我们将充分利用从SAH患者的移植肝脏中收集的生物检疫试剂，如 以及供体肝脏和被诊断为其他肝病的肝脏，并使用尖端蛋白质 微阵列技术实现四个具体目标：1)识别特定识别的自身抗原 通过从SAH患者的移植肝脏或血液中提取抗体；2)轮廓抗体特征 SAH患者对细菌抗原的作用；3)从SAH肝脏提取的抗体的功能 使用基于细胞的模型；以及4)检查酒精滥用诱导的自身抗体是否导致同种异体肝移植 肝移植术后酒精复吸患者的损伤。我们相信，这个项目的成功将使我们能够 确定新的治疗靶点，开发干预和预防SAH的新疗法。

PROJECT SUMMARY Severe alcoholic hepatitis (SAH) is the most severe form of the alcoholic liver diseases (ALD). Chronic alcohol abuse is associated with increased susceptibility to pneumonia, infections (e.g., HIV, HCV, and Tuberculosis), and with chronic inflammation. Studies in the past thirty years in both humans and animal models revealed that ALD is likely caused by impact of alcohol abuse on adaptive immune system. Chronic alcohol abuse was found to reduce the number of B cells, decrease antigen-specific antibody responses, and increase the production of autoantibodies against liver autoantigens and byproducts of oxidative damage. Moreover, increased levels of immunoglobulin of IgG, IgA, and IgE isotypes and production of autoantibodies against liver antigens have been observed in patients with SAH. Therefore, we hypothesize that generation of autoantibodies in patients with SAH contributes to liver damage. To fully test this hypothesis, the identification of those antigens recognized by antibodies deposited in SAH livers is a crucial stepping-stone towards understanding the etiology of the diseases. In addition, it is likely that some of these antibodies are created from cross-reacting antibodies directed against bacteria in the gut. The goal of this proposal is to determine the origins of these SAH-specific antibodies in patients with SAH and characterize their function and contribution to liver damage. We will take full advantage of the biospecimens collected from the explanted livers of patients with SAH, as well as donor livers and livers diagnosed for other liver diseases, and employ the cutting-edge protein microarray technology to achieve four Specific Aims: 1) Identify autoantigens that are specifically recognized by antibodies extracted from explanted livers or blood of the patients with SAH; 2) Profile antibody signatures to bacterial antigens in patients with SAH; 3) Characterize functions of the antibodies extracted from SAH livers using a cell-based model; and 4) Examine whether alcohol abuse-induced autoantibodies cause liver allograft injury in alcohol relapse patients after liver transplantation. We believe that of this project will allow us to identify novel therapeutic targets and develop novel therapy for intervention and prevention of SAH.