MOLECULAR PATHOGENESIS 0F HAIRY LEUKOPLAKIA

本项目的重点是了解 通过表征爱泼斯坦巴尔病毒（EBV）的毛状白斑（HL） 基因表达在病变，检查个人的作用， 调节上皮分化的EBV基因产物，和 研究EB病毒基因在该病变中的表达调控， 与淋巴细胞相比。 我们取得了重大 为实现这些目标取得进展。 的标志 HL是EBV在分化上皮中的高水平复制。 因此，我们建议在未来五年， 包括上皮基因特征的研究 在HL中上调，并研究病毒和 细胞基因 迄今为止，对EBV的所有功能知之甚少， HL中的基因表达，几乎没有什么是已知的 上皮细胞基因的表达。 其目的是 建议描述HL中EBV基因表达的特征; 表征Hl中上调的细胞基因，以及 表征HL中表达的EBV基因产物的调节。 在接下来的五年里，我们利用我们构建的HlcDNA文库， 将继续扩大我们对EB病毒的了解， HL中表达的基因。 使用消减cDNA文库 从HL和临床正常的邻近组织中产生，我们将 还表征了HL中上调的细胞基因。 的 这些基因表达的拓扑学和特异性将被 在HL和对照组织中通过mRNA原位杂交研究。 研究细胞和病毒对EBV基因的调控 反式激活蛋白，我们将诱导蛋白质表达， 我们的HL cDNA文库 在文库中表达的蛋白质将被 用对应于启动子的放射性标记DNA进行探测 选择已知在HL中表达的EBV基因。 克隆编码 蛋白质将从文库中分离，并且特异性 候选的细胞和/或病毒DNA结合蛋白将 使用凝胶阻滞试验进行研究。 然后蛋白质会 研究它们对荧光素酶启动子的影响 报告人制度 HL的研究为表征EBV提供了独特的机会 上皮细胞感染。 上皮细胞的感染 在EBV的生命周期中的重要作用，并了解 EB病毒与细胞基因在发病机制中的相互作用 对人类语言的理解具有更广泛的意义， EBV相关病变，包括鼻咽癌 和非霍奇金淋巴瘤

The focus of this project is to understand the pathogenesis of hairy leukoplakia (HL) by characterizing Epstein Barr virus (EBV) gene expression in the lesion, examining the role of individual EBV gene products in modulating epithelial differentiation, and studying regulation of EBV gene expression in this lesion for comparison with that of lymphocytes. We have made significant progress toward achieving each of these aims. The hallmark of HL is high level EBV replication in differentiated epithelium. In the next fiev years,we therefore propose to extend these studies to include characterization of epithelial genes upregulated in HL, and to study the interaction between viral and cellular genes. To date, relatively little is known about the repertoire of EBV gene expression within HL, and almost nothing is known of epithelial gene expression in the lesion. The aims of this proposal are to characterize EBV gene expression in HL; to characterize cellular genes upregulated in Hl, and to characterize the regulation of EBV gene products expressed in HL. In the next five years, using an HlcDNA library we generated, we will continue to expand our knowledge of the repertoire of EBV genes expressed in HL. Using a subtraction cDNA library generated from HL and clinically normal adjacent tissues, we will also characterize cellular genes upregulated in HL. The topography and specificity of expression of these genes will be studied in HL and control tissues by mRNA in situ hybridization. To study the regulation of EBV genes by cellular and viral transactivating proteins, we will induce protein expression by our HL cDNA library. Proteins expressed in the library will be probed with radiolabeled DNA corresponding to promoters of selected EBV genes known to be expressed in HL. Clones encoding the proteins will be isolated from the library, and specificity of the candidate cellular and/or viral DNA binding proteins will be studied using gel retardation assays. The proteins will then be expressed to study their effect on the promoter in luciferase reporter system. Studies of HL offer a unique opportunity to characterize EBV infection of epithelium. Infection of epithelium plays an important role in the EBV live cycle, and understanding the interaction between EBV and cellular genes in the pathogenesis of HL has broader implications for understanding of other EBV-associated lesions as well, including nsaopharyngeal cancer and non-Hodgkin's lymphoma.