EPSTEIN-BARR VIRUS INFECTION OF MUCOSAL EPITHELIUM

粘膜上皮的 Epstein-Barr 病毒感染

• 批准号: 3197066
• 负责人: JOHN W SIXBEY
• 金额: $ 10.28万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-05-01 至 1995-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3197066
• 关键词: B lymphocyte; Epstein Barr virus; antiviral antibody; cell differentiation; epithelium; gene expression; genetic manipulation; genetic promoter element; genetically modified animals; laboratory mouse; latent virus infection; leukocyte activation /transformation; molecular cloning; nasopharyngeal neoplasms; oral leukoplakia; oral mucosa; polymerase chain reaction; secretory immune system; tissue /cell culture; viral carcinogenesis; virus DNA; virus genetics; virus infection mechanism; virus replication

The long term goal of the proposed research is to identify virologic, immunologic and cellular factors, unique to human mucosa, which contribute to EBV persistence and to mechanisms of pathogenesis. EBV's disease manifestations point to the importance of viral interactions in human mucosa: nasopharyngeal carcinoma and oral hairy leukoplakia (characterized by EBV latency and an aggressively replicative infection, respectively) are epithelial diseases. African Burkitt's lymphoma, with its unusual tissue distribution, is a malignancy of the mucosal-associated lymphoid tissue. Because EBV's two tissue tropisms suggest that the virus may take advantage of the close functional relations between epithelial and lymphoid elements of the common mucosal immune system, we propose the following aims to reach our long term goals: 1) Elucidation of the role of virus-specific IgA in EBV pathogenesis; 2) examination of the role of cellular factors in the regulation of the EBV life cycle in mucosal epithelium; 3) determination of molecular and biological characteristics of EBV variants in the epithelial tissue of oral hairy leukoplakia. Polyclonal human IgA to EBV as well as IgA monoclonal antibodies to the major glycoprotein of EBV (gp350) will be used to analyze "neutralization" of EBV infectivity for lymphocytes and concurrent IgA-enhanced viral entry into epithelial cells. The possibility of an immune-induced shift in EBV tissue tropisms has tremendous implications for immunization strategy as well as for general concepts of viral pathogenesis. The relation between state of cell differentiation and viral gene expression will be analyzed, first in epithelial tissues of transgenic mice by linking select EBV promoters to beta galactosidase coding sequences; second, in diverse populations of human B lymphocytes, by correlating markers of cell activation/differentiation with virus gene expression. These experiments will elucidate mechanisms of viral persistence and how virus may disseminate after primary infection. Finally, the ability of defective viral DNA to contribute to the high level of viral replication seen in hairy leukoplakia will be examined, first by determination of its structure with cloning and polymerase chain reaction analysis, then by use of novel culture techniques to test its in vitro biologic activity. The existence in nature of a defective virus particle with the ability to enhance, not inhibit, replication of the parent strain suggests mechanisms by which a persistent virus may move from latency to active replication.

拟议研究的长期目标是确定病毒学、 人类粘膜特有的免疫和细胞因素，有助于 EBV 持续性和发病机制。 EB病毒病 这些表现表明病毒相互作用在人类中的重要性 粘膜：鼻咽癌和口腔毛状白斑（特征为 分别由 EBV 潜伏期和积极复制感染引起） 上皮疾病。 非洲伯基特淋巴瘤及其不寻常的组织 分布，是粘膜相关淋巴组织的恶性肿瘤。 因为 EBV 的两种组织嗜性表明病毒可能会利用 上皮细胞和淋巴元件之间密切的功能关系 的共同粘膜免疫系统，我们提出以下目标以达到 我们的长期目标：1) 阐明病毒特异性 IgA 在 EB病毒发病机制； 2) 细胞因子在细胞因子中的作用的检查 粘膜上皮中 EBV 生命周期的调节； 3) 测定 上皮细胞中EBV变种的分子和生物学特征 口腔毛状白斑组织。 抗 EBV 的多克隆人 IgA 以及抗 EBV 的 IgA 单克隆抗体 EBV的主要糖蛋白（gp350）将用于分析“中和” EBV 对淋巴细胞的感染性和同时 IgA 增强的病毒进入的影响 进入上皮细胞。 EBV 免疫诱导变化的可能性 组织向性对免疫策略具有巨大影响，因为 以及病毒发病机制的一般概念。 之间的关系 将分析细胞分化和病毒基因表达的状态， 首先通过连接选择的 EBV 在转基因小鼠的上皮组织中 β半乳糖苷酶编码序列的启动子；其次，在多样化 通过关联细胞标记来分析人类 B 淋巴细胞群体 病毒基因表达的激活/分化。 这些实验 将阐明病毒持续存在的机制以及病毒如何 初次感染后传播。 最后，缺陷能力 病毒 DNA 有助于高水平的病毒复制 首先通过确定其结构来检查毛状白斑 通过克隆和聚合酶链反应分析，然后使用新型 培养技术测试其体外生物活性。 存在 本质上是有缺陷的病毒颗粒，具有增强能力，而不是 抑制亲本菌株的复制表明了一种机制 持久性病毒可能会从潜伏状态转移到主动复制状态。