EPSTEIN-BARR VIRUS EXPRESSION IN NORMAL HUMAN EPITHELIUM

上皮细胞为EBV允许细胞的鉴定 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诱导的现象 细菌的生长转化、持久性和复活 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 EB病毒在上皮细胞中的激活及细胞表达 分化抗原；(2)检测潜伏EBV的作用 感染在诱导上皮细胞过度增殖中的作用 分析表皮中淋巴细胞与上皮细胞的相互作用 关于EB病毒的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 具有体外分化能力的原代外植体培养。至 细胞成熟阶段与EBV抗原表达相关， 我们将使用单抗来对抗细胞分化抗原。 (角蛋白、总蛋白)。病毒之间的功能相互关系 一过性上皮细胞骨架的蛋白质和元素 在细胞分化过程中表达的蛋白将通过 免疫电子显微镜和显微注射。的表达 EB病毒潜伏膜蛋白在细胞中的重要作用 转化，将在上皮细胞中进行检查。在目标2下， EB病毒诱导小鼠皮肤基底上皮细胞增殖的能力 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 在子宫颈上。EB病毒在疾病病因学中作用的证据 通过杂交分析获得上皮细胞的增殖 已知可诱导细胞转化的EBV基因产物； 利用EBV末端结构分析细胞克隆性； 增殖细胞中EB病毒DNA组织的检测 (整合、上体、线形)。在目标3中，EBV的能力 潜伏感染的表皮中的B细胞将重新激活 通过两种细胞类型的共培养和通过 宫颈炎性浸润物的细胞杂交。这个 B细胞内源性传播EBV的问题将进一步 通过分析EBV分离株，从分离的 用于限制性片段长度多态的解剖位置。 与淋巴细胞不同，整个病毒周期-从潜伏期到 复制--在上皮细胞中明显存在。这些研究将 提供有关EBV病理生物学和细胞调控的新信息 病毒基因的表达。

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.