MOLECULAR PATHOGENESIS 0F HAIRY LEUKOPLAKIA

本项目的重点是了解糖尿病的发病机制。 EB病毒特征的毛状白斑(HL) 基因在病变中的表达，考察个体的作用 EBV基因产物在调节上皮分化中的作用 研究EBV基因在本病皮损中的表达规律 与淋巴细胞比较。我们已经取得了重大进展 在实现这些目标方面取得了进展。的标志 HL是分化上皮中高水平的EBV复制。 因此，在未来的FIEV年，我们建议延长这些 包括上皮基因特征的研究 在HL中表达上调，并研究病毒与HL之间的相互作用 细胞基因。 到目前为止，人们对EB病毒的谱系知之甚少。 HL中的基因表达，几乎什么都不知道 皮损中上皮基因的表达。这样做的目的是 建议对HL中EBV基因的表达进行表征； 鉴定在h1中上调的细胞基因，并 描述在HL中表达的EBV基因产物的调控。 在接下来的五年里，使用我们创建的HlcDNA文库，我们 将继续扩大我们对EBV曲目的了解 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 也描述了HL中上调的细胞基因。这个 这些基因表达的地形和特异性将是 用原位杂交技术对HL组织和对照组织进行了研究。 细胞和病毒对EBV基因调控的研究 反式激活蛋白质，我们将通过以下方式诱导蛋白质表达 我们的HL基因文库。在文库中表达的蛋白质将是 用与启动子相对应的放射性标记DNA进行探针 选定的已知在HL中表达的EBV基因。克隆编码 这些蛋白质将从文库中分离出来，并具有特异性 候选细胞和/或病毒DNA结合蛋白的 用凝胶滞留试验进行研究。然后蛋白质就会 表达以研究其对荧光素酶启动子的影响 记者制度。 对HL的研究为确定EBV的特征提供了一个独特的机会 上皮性感染。上皮细胞感染起着重要作用 在EBV生命周期中的重要作用，并理解 EB病毒与细胞基因在发病机制中的相互作用 对理解其他疾病有更广泛的影响 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.