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

项目 3 - 外植肝脏的蛋白质组学分析及自身抗原的表征

• 批准号: 10560561
• 负责人: Heng Zhu
• 金额: $ 32.55万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10560561
• 关键词: ADCC Assay; Acceleration; Acute; Adaptive Immune System; Adrenal Cortex Hormones; Affect; Alcohol abuse; Alcohol consumption; Alcoholic Hepatitis; Alcoholic Liver Diseases; Animal Model; Antibodies; Antibody Response; Antigens; Autoantibodies; Autoantigens; B-Lymphocytes; Bacteria; Bacterial Antigens; Bioinformatics; Biological Assay; Biopsy Specimen; Blood; Cell Separation; Cells; Chronic; Circulation; Cross Reactions; Deposition; Development; Diagnosis; Disease; Escherichia coli; Etiology; Generations; Goals; HIV; HIV/HCV; Hepatitis C virus; Hepatocyte; Human; IgE; Immunofluorescence Immunologic; Immunoglobulin A; Immunoglobulin G; Immunoglobulins; Immunohistochemistry; Individual; Infection; Inflammation; Inflammatory; Injury; Liver; Liver diseases; Microarray Analysis; Modeling; Molecular; Mycobacterium tuberculosis; Patients; Pneumonia; Predisposition; Prevention; Production; Protein Microchips; Proteins; Proteome; Proteomics; Relapse; Research Personnel; Sampling; Serum; Stains; T-Lymphocyte; Testing; Therapeutic Intervention; Tissues; Transplant Recipients; Transplantation; Tuberculosis; Virus Diseases; alcohol relapse; chronic alcohol ingestion; cohort; cytokine; expectation; gut bacteria; human model; interdisciplinary approach; liver allograft; liver biopsy; liver injury; liver transplantation; mortality; new therapeutic target; novel therapeutics; oxidative damage; relapse patients

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.

项目摘要 重度酒精性肝炎（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（例如，HIV、HCV和结核病）， 和慢性炎症。过去30年的人类和动物模型研究表明， 酒精性肝病可能是由于酒精滥用对适应性免疫系统的影响而引起的。慢性酒精滥用被发现 减少B细胞的数量，降低抗原特异性抗体应答，并增加 抗肝脏自身抗原的自身抗体和氧化损伤的副产品。此外， IgG、伊加和IgE同种型的免疫球蛋白和针对肝抗原的自身抗体的产生， 在SAH患者中观察到。因此，我们假设患者体内自身抗体的产生 会导致肝损伤为了充分验证这一假设，这些抗原的鉴定 由SAH肝脏中沉积的抗体识别是理解SAH的关键基石。 疾病的病因。此外，这些抗体中的一些很可能是由交叉反应产生的。 针对肠道细菌的抗体。这项提案的目的是确定这些问题的起源， SAH特异性抗体在SAH患者中的作用及其对肝损伤的影响。 我们将充分利用从SAH患者的肝脏中收集的生物标本， 以及捐赠的肝脏和诊断为其他肝脏疾病的肝脏， 微阵列技术实现了四个特定目的：1）识别特异性识别的自身抗原 通过从SAH患者的肝脏或血液中提取的抗体; 2）描绘抗体特征 SAH患者血清中抗细菌抗原的抗体; 3）SAH肝组织中抗体的功能鉴定 使用基于细胞的模型;和4）检查酒精滥用诱导的自身抗体是否导致肝移植 肝移植术后酒精复发患者的损伤。我们相信，这个项目将使我们能够 确定新治疗靶点并开发用于干预和预防SAH的新疗法。