CYTOKINES AND HUMAN AUTOIMMUNE LIVER DISEASE

细胞因子和人类自身免疫性肝病

• 批准号: 3456598
• 负责人: Sheri M. Krams
• 金额: $ 9.35万
• 依托单位: CALIFORNIA PACIFIC MED CTR RES INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-09-01 至 1998-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3456598
• 关键词: autoantibody; autoimmune disorder; cytokine; cytotoxic T lymphocyte; eosinophil; esterase; hepatitis; histocompatibility antigens; human subject; immunocytochemistry; in situ hybridization; interferon gamma; interleukin 1; interleukin 4; interleukin 5; interleukin 6; liver transplantation; natural killer cells; polymerase chain reaction; pore forming protein; primary biliary cirrhosis; relapse /recurrence; transplant rejection; tumor necrosis factor alpha; western blottings

Primary biliary cirrhosis (PBC) and autoimmune hepatitis (AI-CAH) are two chronic diseases of the liver, believed to be autoimmune in nature, with etiologies and pathogenesis that remain unknown. These disorders affect predominantly women, mostly in the prime of their life. For example, over 90% of patients with PBC are women between the ages of 20-70. Liver transplantation has proven to be an effective treatment for both AI-CAH and PBC. However, disease recurrence remains an important concern. The goal of this project is to elucidate the role of cytokines and cytotoxic mediators in autoimmune liver disease. Specifically, we propose to: 1) characterize the cytokine profiles within the liver of patients with PBC and AI-CAH 2) identify the cytotoxic mediators 3) characterize the functional properties of lymphoid cells that have infiltrated the liver and 4) apply this information to establish immunological parameters that distinguish disease recurrence from allograft rejection. Molecular methodologies will be utilized to determine and compare the cytokine arrays in normal liver and liver explant tissue from patients with PBC and AI-CAH. RNA obtained from liver tissue will be reverse transcribed, and analyzed for cytokine gene expression using semi- quantitative PCR. In situ hybridization and immunohistochemical techniques will identify the cellular source of cytokine message within the liver. evidence for specific cellular pathways of liver tissue injury will be obtained by immunohistochemical techniques and Western blot hybridization analysis for serine esterases, perforin, eosinophil cytotoxic proteins and autoantibodies. Direct in vitro examination of infiltrating cells recovered from the PBC and AI-CAH liver will establish the diversity of T cells within the infiltrate, the cytotoxic activity and specificity of T lymphocytes, the frequency of specific cytokine producing cells, the arrays of intrahepatic autoantibodies, and the presence of non-specific effector cells including NK cells and eosinophils. The effect of recombinant cytokines on MHC antigen expression by target tissue will be tested on epithelial and hepatocyte-derived cell lines. Finally, patterns of cytokine production in PBC and AI-CAH will be compared with these of- patients who have received liver transplants because of autoimmune liver disease in an effort to distinguish disease recurrence from allograft rejection. Taken together, these studies will provide important mechanistic insights into these two enigmatic autoimmune diseases as well as clarify the issue of disease recurrence post-orthotopic transplantation.

原发性胆道肝硬化（PBC）和自身免疫性肝炎（AI-CAH）为两个 肝脏的慢性疾病，被认为是自身免疫的， 病因和发病机理仍然未知。 这些疾病会影响 主要是妇女，主要是一生的巅峰时期。例如，结束 90％的PBC患者是20-70岁的女性。 肝 事实证明，移植是AI-CAH的有效治疗方法 和PBC。但是，疾病复发仍然是一个重要的问题。 该项目的目的是阐明细胞因子和 自身免疫性肝病中的细胞毒性介质。 具体来说，我们建议 TO：1）表征患者肝脏内的细胞因子谱 使用PBC和AI-CAH 2）识别细胞毒性介质3）表征 渗入的淋巴样细胞的功能特性 肝脏和4）应用此信息以建立免疫学参数 将疾病复发与同种异体移植排斥区分开。 分子方法将用于确定和比较 正常肝脏和肝外植体组织中的细胞因子阵列 与PBC和AI-CAH。 从肝组织获得的RNA将反向 转录并使用半因子基因表达进行分析 定量PCR。原位杂交和免疫组织化学技术 将识别肝脏内细胞因子消息的细胞来源。 肝组织损伤的特定细胞途径的证据将是 通过免疫组织化学技术和Western印迹杂交获得 分析丝氨酸酯酶，穿孔蛋白，嗜酸性粒细胞细胞毒性蛋白和 自动抗体。直接在体外检查浸润细胞 从PBC和AI-CAH肝脏中回收的将确定T的多样性 浸润中的细胞，t的细胞毒性活性和特异性 淋巴细胞，特异性细胞因子的频率， 肝内自身抗体的阵列和非特异性的存在 效应细胞在内，包括NK细胞和嗜酸性粒细胞。效果 重组细胞因子对目标组织的MHC抗原表达的重组细胞因子将是 在上皮和肝细胞衍生的细胞系上进行了测试。最后，模式 将PBC和AI-CAH中的细胞因子产生与这些相比 因自身免疫性肝而接受肝移植的患者 疾病以区分疾病复发与同种异体移植 拒绝。 综上所述，这些研究将提供重要的机械见解 进入这两种神秘的自身免疫性疾病，并澄清该问题 疾病复发后正常移植。