EPSTEIN BARR VIRUS EXPRESSION IN NORMAL HUMAN EPITHELIUM

正常人上皮中的 Epstein Barr 病毒表达

• 批准号: 3446630
• 负责人: JOHN W SIXBEY
• 金额: $ 4.96万
• 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 1987-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3446630
• 关键词: Epstein Barr virus; Herpesviridae disease; cell differentiation; epithelium; genetic manipulation; genetic transcription; human subject; immunofluorescence technique; microorganism immunology; tissue /cell culture; viral carcinogenesis; virus antigen; virus genetics; virus infection mechanism

Identification of the primary cell type in which the Epstein-Barr virus (EBV) replicates is essential to understanding the biology of the virus, mode of transmission and pathogenesis. Virtually all we know about EBV comes froms studies of the virus in relation to lymphoid tissue. Yet, EBV is linked to three quite different diseases with involvement of not only lymphoid but also epithelial elements. The goal of this study is to define the effects of cell differentiation on EBV-epithelial cell interaction. Differences in membrane receptor expression, EBV-specific antigen expression, transcriptional patterns of the EBV genome, and virus productivity will be examined in both undifferentiated and differentiated epithelial cell subsets. These determinations will be made by indirect immunofluorescence techniques; electron microscopy; DNA-RNA cytohybridization with a new generation of cloned, biotinylated DNA probes; and by standard lymphocyte transformation assays. Studies of host cell control of infection are only now possible in this epithelial cell system with the establishment of techniques for long-term maintenance of primary explant cultures in an undifferentiated state and synchronized induction of cell differentiation. The problems imposed on EBV pathogenesis by the apparent lack of host cell receptors will also be addressed by infection and cocultivation of autologous lymphoid and epithelial cell populations. Evidence for transforming potential of EBV in epithelial cells will be sought in long-term cell cultures infected with wild-type EBV strains or transfected with either cloned EBV DNA fragments or cellular DNA from autologous lymphoblastoid cell cultures. Ease of cell passage, ability to grow in soft agar, and oncogenicity in nude mice will be assessed. Clinical correlates will be sought by cytohybridization analyses of benign lymphoepithelial lesions of the parotid and salivary glands as well as by delineation of selectively encoded regions of the EBV genome in oropharyngeal epithelial cells from various patient groups.

EB 病毒的原代细胞类型的鉴定 (EBV) 复制对于了解病毒的生物学至关重要， 传播方式和发病机制。 几乎我们所知道的关于 EBV 的一切 来自对病毒与淋巴组织相关的研究。 然而，EBV 与三种截然不同的疾病有关，不仅涉及 淋巴组织，也包括上皮细胞。 本研究的目的是确定细胞分化对 EBV-上皮细胞相互作用。 膜受体的差异 表达、EBV特异性抗原表达、转录模式 EBV 基因组和病毒生产力将在两个方面进行检查 未分化和分化的上皮细胞亚群。 这些 将通过间接免疫荧光技术进行测定； 电子显微镜；新一代 DNA-RNA 细胞杂交 克隆的生物素化 DNA 探针；并通过标准淋巴细胞转化 化验。 宿主细胞控制感染的研究现在才有可能 这种上皮细胞系统随着技术的建立 在未分化的环境中长期维持原代外植体培养物 细胞分化的状态和同步诱导。 明显缺乏宿主细胞给 EBV 发病机制带来的问题 受体也将通过感染和共培养来解决 自体淋巴和上皮细胞群。 证据 EBV 在上皮细胞中的转化潜力将在 感染野生型 EBV 株或转染的长期细胞培养物 使用克隆的 EBV DNA 片段或自体细胞 DNA 淋巴母细胞培养物。 细胞传代容易，生长能力强 将评估软琼脂和裸鼠的致癌性。 将通过良性肿瘤的细胞杂交分析来寻找临床相关性 腮腺和唾液腺的淋巴上皮病变以及 EBV基因组选择性编码区域的描绘 来自不同患者组的口咽上皮细胞。