EPSTEIN-BARR VIRUS EXPRESSION IN NORMAL HUMAN EPITHELIUM

正常人上皮中的 Epstein-Barr 病毒表达

• 批准号: 3177274
• 负责人: JOHN W SIXBEY
• 金额: $ 19.26万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-12-01 至 1996-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3177274
• 关键词: B lymphocyte; Burkitt's lymphoma; Epstein Barr virus; Herpesviridae disease; athymic mouse; cell differentiation; epithelium; gene deletion mutation; gene expression; genetic manipulation; genetic markers; genetic transcription; host organism interaction; human subject; immunoelectron microscopy; immunofluorescence technique; latent virus infection; microorganism immunology; molecular cloning; monoclonal antibody; nasopharyngeal neoplasms; nucleic acid hybridization; nucleic acid probes; nucleic acid sequence; oncogenic virus; oral mucosa; polymerase chain reaction; tissue /cell culture; viral carcinogenesis; virus DNA; virus antigen; virus cytopathogenic effect; virus genetics; virus infection mechanism; virus replication

The long term goal of the proposed research is to identify viral and cellular factors, unique to human mucosa, which contribute to Epstein-Barr virus (EBV) pathogenesis and to mechanisms of persistence. EBV-associated diseases 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 cell diseases. African Burkitt's lymphoma, with its unusual tissue distribution, is a malignancy of the mucosal-associated lymphoid tissue. Utilizing information gained in the current grant period which suggests for the first time the existence of multiple EBV variants with distinct biologic differences and tissue tropisms, we propose the following aims to reach our long-term goals: 1) Determination of the molecular structure of lytic EBV; 2) Evaluation of the distribution of transforming and lytic variants within epithelial and lymphoid components of the mucosal system; 3) Elucidation of mechanisms for EBV persistence in the epithelial components of the mucosa. Based on the known biologic activity of a spontaneous laboratory mutant deleted for the EBV nuclear antigen 2 (EBNA2) encoding gene, we postulate that similar wild type variants represent the "missing link' in our understanding of EBV pathogenic mechanisms. By molecular cloning and polymerase chain reaction analysis, we will delineate (under aim 1) the genetic structure of EBNA2 deleted virus taken directly from throat washings, thus avoiding the selection bias inherent with passage in lymphocyte culture. Demonstration of deletions and potential gene rearrangements will provide an indication of altered biologic activity. In aim #2, use of select genotypic markers amenable to PCR analysis will allow us to track EBV variants through tissue compartments of the mucosal system, identify tissue tropisms, and locate sites for potential interaction between strains. Finally, we will use EBV strains, isogenic except for their EBNA2 gene, to analyze in experimentally infected epithelium transcription from EBV's putative transforming genes. In infected human epithelial cell implants maintained in nude mice, we will relate patterns of gene expression to the EBNA2 deletion mutant's ability to sustain long-term infection in the absence of a viable B cell component.

长期 拟议研究的目标是识别病毒， 细胞因子，人类粘膜特有的，有助于 EB病毒（EBV）的发病机制和持久性。 EB病毒相关疾病指出了病毒相互作用在 人粘膜：鼻咽癌和口腔毛状白斑。 （特征为EBV潜伏期和侵袭性复制感染， 分别）是上皮细胞疾病。 非洲伯基特淋巴瘤 其不寻常的组织分布，是一种恶性肿瘤， 粘膜相关淋巴组织。 利用在《公约》中获得的信息， 目前的赠款期间，这表明第一次存在的 具有明显生物学差异和组织学特征的多种EBV变异体 根据我们的取向，我们提出以下目标来实现我们的长期目标：1） 裂解性EBV的分子结构的测定; 2）裂解性EBV的分子结构的评价; 上皮内转化和裂解变体的分布， 粘膜系统的淋巴成分; 3）阐明机制 EB病毒在粘膜上皮成分中的持久性。 基于自发实验室的已知生物活性 缺失EBV核抗原2（EBNA 2）编码基因的突变体，我们 假设类似的野生型变异体代表了 我们对EB病毒致病机制的了解。 通过分子克隆， 聚合酶链反应分析，我们将描绘（根据目标1） 直接从咽喉中提取的EBNA 2缺失病毒的遗传结构 洗涤，从而避免选择偏见固有的通道， 淋巴细胞培养 缺失和潜在基因的证明 重排将提供改变的生物活性的指示。 在目标#2中，使用适合于PCR分析的选择基因型标记将 使我们能够通过粘膜的组织隔室追踪EBV变异体， 系统，识别组织向性，并定位潜在的 菌株间的相互作用 最后，我们将使用EBV毒株， 除了他们的EBNA 2基因， 上皮细胞从EBV的假定转化基因转录。 在 感染的人上皮细胞移植物在裸鼠中维持，我们将 将基因表达模式与EBNA 2缺失突变体的能力相关联 在缺乏存活B细胞的情况下维持长期感染 成分