PLASMA MEMBRANE ALTERATIONS IN VIRAL HEPATITIS B

乙型病毒性肝炎中的质膜改变

基本信息

批准号：
3152147
负责人：
MICHAEL A GERBER
金额：
$ 10.94万
依托单位：
MOUNT SINAI SCHOOL OF MEDICINE OF CUNY
依托单位国家：
美国
项目类别：
财政年份：
1982
资助国家：
美国
起止时间：
1982-04-01 至 1988-06-30
项目状态：
已结题

项目摘要

Although significant advances in our knowledge of the moleular biology of hepatitis B virus (HBV) have been made, the mechanism of hepatocellular necrosis, the basic event in hepatitis B, remains unknown. The outer cell membrane probably represents the initial site of hepatocyte injury from immunologic reactions in hepatitis B. We postulate that the determinants, amount and structure of HBV antigens on the cell surface largely determine the interaction of infected hepatocytes with the host's immune response and that the expression of HBV antigens may be modulated by immune and non-immune factors. To understand the cellular and molecular basis of these interactions, we are studying 2 laboratory model systems of HBV infected cells: (1) human hepatocellular carcinoma cell lines (PLC/PRF/5 and Hep 3B) which resemble hepatocytes morphologically and functionally and are naturally infected with HBV; and (2) well defined subclones of NIH 3T3 mouse fibroblasts (4.10) which after transfection with cloned HBV DNA express all known HBV antigens (HBsAg, HBcAg, and HBeAg). Using poly- and monoclonal antibodies, fluorescence activated flow cytometry, quantitative radioimmunoassays and polypeptide analysis, we will define the HBV antigen(s) expressed, their amount, structure and relationship to host cell membrane proteins, particularly HLA class I and II antigens on the surface of cultured cells. Further, we will evaluate the effect of modulating agents, such as antibodies to HBV antigens and interferon-Gamma, on cell surface HBV antigens. Finally in a pilot study, we will explore the humoral and cellular immune response to HBV antigen positive subclones of 4.10 cells in NFS/N mice. These investigations may increase our understanding of the mechanism(s) whereby host immune factors lyse HBV-infected cells or modulate the expression of HBV antigens which may lead to virus persistence and chronic hepatitis B. This knowledge may result in a more rational approach to treatment of infected patients with interferon and prevention of hepatitis B by vaccination.

尽管我们对巨大生物学的了解有了重大进步肝炎病毒（HBV）已经制成了肝细胞的机制坏死是乙型肝炎的基本事件，仍然未知。外部细胞膜可能代表了肝细胞损伤的初始部位丙型肝炎中的免疫反应。我们假设决定因素，细胞表面上HBV抗原的数量和结构在很大程度上确定感染的肝细胞与宿主的免疫反应和 HBV抗原的表达可以通过免疫调节和非免疫因子。了解这些相互作用，我们正在研究HBV的2个实验室模型系统感染细胞：（1）人肝细胞癌细胞系（PLC/PRF/5 和Hep 3b），在形态和功能上类似于肝细胞天然感染了HBV；（2）NIH 3T3的明确定义的子克隆小鼠成纤维细胞（4.10）用克隆的HBV DNA转染后表达所有已知的HBV抗原（HBSAG，HBCAG和HBEAG）。使用poly-和单克隆抗体，荧光激活的流式细胞术，定量放射免疫测定和多肽分析，我们将定义HBV 抗原表达，其与宿主细胞的数量，结构和关系膜蛋白，尤其是表面的HLA I和II抗原培养的细胞。此外，我们将评估调制的效果诸如细胞上HBV抗原和干扰素伽马的抗体之类的药物表面HBV抗原。最后，在一项试点研究中，我们将探索对HBV抗原阳性亚克隆的体液和细胞免疫反应 NFS/N小鼠中的4.10个细胞。这些调查可能会增加我们对机制的理解宿主免疫因子裂解感染了HBV的细胞或调节 HBV抗原的表达可能导致病毒持久性和慢性丙型肝炎B。这种知识可能会导致更合理的方法治疗感染的干扰素患者和预防肝炎 b通过疫苗接种。