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.

项目概要 严重酒精性肝炎（SAH）是酒精性肝病（ALD）中最严重的一种。长期酗酒 滥用与肺炎、感染（例如艾滋病毒、丙型肝炎病毒和结核病）的易感性增加有关， 并伴有慢性炎症。过去三十年对人类和动物模型的研究表明 ALD 可能是由酗酒对适应性免疫系统的影响引起的。发现长期酗酒 减少 B 细胞数量，减少抗原特异性抗体反应，并增加 B 细胞的产生 针对肝脏自身抗原和氧化损伤副产物的自身抗体。此外，增加的水平 IgG、IgA 和 IgE 同种型免疫球蛋白以及针对肝抗原的自身抗体的产生 在 SAH 患者中观察到。因此，我们假设患者体内产生自身抗体 SAH 会导致肝损伤。为了充分检验这一假设，需要鉴定这些抗原 被 SAH 肝脏中沉积的抗体识别是了解 SAH 的重要基石 疾病的病因学。此外，其中一些抗体可能是通过交叉反应产生的 针对肠道细菌的抗体。该提案的目标是确定这些的起源 SAH 患者体内的 SAH 特异性抗体，并表征其功能和对肝损伤的影响。 我们将充分利用从 SAH 患者的移植肝脏中采集的生物样本， 以及供体肝脏和诊断为其他肝脏疾病的肝脏，并采用尖端蛋白质 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 从 SAH 患者的移植肝脏或血液中提取抗体； 2) 分析抗体特征 SAH 患者的细菌抗原； 3) 表征从 SAH 肝脏中提取的抗体的功能 使用基于细胞的模型； 4) 检查酒精滥用引起的自身抗体是否导致同种异体肝移植 肝移植后酒精复发患者的损伤。我们相信这个项目将使我们能够 确定新的治疗靶点并开发新的治疗方法来干预和预防 SAH。