EPSTEIN-BARR VIRUS EXPRESSION IN NORMAL HUMAN EPITHELIUM

正常人上皮中的 Epstein-Barr 病毒表达

• 批准号: 3177272
• 负责人: JOHN W SIXBEY
• 金额: $ 15.04万
• 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 1991-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3177272
• 关键词: B lymphocyte; Epstein Barr virus; Herpesviridae disease; athymic mouse; cell differentiation; colchicine; epithelium; genetic manipulation; genetic transcription; human subject; immunoelectron microscopy; immunofluorescence technique; keratin; latent virus infection; membrane proteins; microorganism immunology; molecular cloning; monoclonal antibody; nucleic acid hybridization; nucleic acid probes; tissue /cell culture; viral carcinogenesis; virus DNA; virus antigen; virus genetics; virus infection mechanism; virus replication

Identification of the epithelial cell as the cell permissive for EBV replication in the oropharynx has reordered our perception of EBV biology. We will reexamine the phenomena of EBV-induced growth transformation, persistance and reactivation in the context of epithelial cell-lymphocyte interactions. To this end, our specific aims are: (1) to determine the relationship between EBV activation in epithelium and cellular expression of differentiation antigens; (2) to examine the role of latent EBV infection in the induction of epithelial hyperproliferation; (3) to analyses lymphocyte-epithelial cell interactions in the epidermis with regard to EBV reactivation and intercellular exchange of virus. Laboratory and wild-type EBV will be used to infect epithelial primary explant cultures capable of in vitro differentiation. To correlate stage of cell maturation with EBV antigen expression, we will use monoclonal antibodies to cell differentiation antigens (keratin, involucrin). Functional interrelationships between viral proteins and elements of epithelial cytoskeleton transiently expressed during cell differentiation will be examined by immunoelectron microscopy and microinjection. Expression of the EBV latent membrane protein, important in cell transformation, will be examined in epithelium. Under aim #2, EBV's ability to induce proliferation of basal epithelial cells in a manner analogous to B cells immortalization will be examined in epithelial grafts in nude mice and in hyperplastic epithelial lesions of the uterine cervix. Evidence for an etiologic role of EBV in epithelial proliferation will be obtained by hybridization analysis for EBV gene products known to induce cell transformation; analysis of cell clonality using the structure of EBV termini; and determination of EBV DNA organization in proliferating cells (integrated, episomal, linear). In aim #3, the ability of EBV to reactivate in latently infected B cells in the epidermis will be analysed by cocultivation of the two cell types and by cytohybridization to inflammatory infiltrates of the cervix. The issue of enogenous spread of EBV by B cells will be further addressed by analysis of EBV isolates, shed from separate anatomical sites, for restriction fragment length polymorphisms. Unlike in lymphocytes, the entire viral cycle - from latency to replication - is manifest in epithelial cells. These studies will provide new information on EBV pathobiology and cell regulation of the viral gene expression.

鉴定上皮细胞为 EBV 允许细胞 口咽部的复制改变了我们对 EBV 的认知 生物学。 我们将重新审视EBV引起的现象 生长转化、持久性和重新激活 上皮细胞-淋巴细胞相互作用的背景。 为此， 我们的具体目标是：（1）确定之间的关系 上皮中的 EBV 激活和细胞表达 分化抗原； (2)考察潜伏性EBV的作用 感染诱导上皮过度增殖； (3) 至 分析表皮中淋巴细胞-上皮细胞的相互作用 关于 EBV 重新激活和细胞间交换 病毒。 实验室和野生型 EBV 将用于感染上皮细胞 能够体外分化的原代外植体培养物。 到 将细胞成熟阶段与 EBV 抗原表达相关联， 我们将使用细胞分化抗原的单克隆抗体 （角蛋白、外皮）。 病毒之间的功能相互关系 瞬时上皮细胞骨架的蛋白质和元件 细胞分化期间表达的检测将通过 免疫电子显微镜和显微注射。 表达 EBV 潜伏膜蛋白，在细胞中很重要 转化，将在上皮中进行检查。 根据目标#2， EBV 能够诱导基底上皮细胞增殖 类似于 B 细胞永生化的方式将在 裸鼠和增生性上皮病变中的上皮移植物 子宫颈。 EBV 病因学作用的证据 通过杂交分析获得上皮增殖情况 已知可诱导细胞转化的 EBV 基因产物； 使用 EBV 末端结构分析细胞克隆性；和 增殖细胞中 EBV DNA 组织的测定 （积分、游离、线性）。 在目标 3 中，EBV 能够 表皮中潜伏感染的 B 细胞会重新激活 通过两种细胞类型的共培养进行分析 子宫颈炎症浸润的细胞杂交。 这 B细胞对EBV的内源性传播问题将进一步得到解决 通过分析 EBV 分离株来解决这个问题， 解剖位点，用于限制性片段长度多态性。 与淋巴细胞不同，整个病毒周期 - 从潜伏期到 复制 - 在上皮细胞中表现出来。 这些研究将 提供有关 EBV 病理学和细胞调控的新信息 病毒基因表达。