INITIATION OF CELL INFECTION WITH EPSTEIN-BARR VIRUS

爱泼斯坦-巴尔病毒(EBV)是一种普遍存在的人类疱疹病毒，引起 传染性单核细胞增多症，与Burkitt淋巴瘤有关， 与鼻咽癌的病因学有关。它 有助于淋巴增生性综合征和IS的发展 影响人类免疫缺陷发病机制的思考 病毒。内源性病毒对正常细胞的招募，产生于 在免疫抑制期间增加剂量，是非常重要的 与病毒诱导的病理相关，并可能有助于 癌前状态的发展。这样做的总体目标是 建议是了解EBV是如何进入其两个目标的，B 淋巴细胞和上皮细胞，并基于这样的前提 澄清这一过程是如何发生的对于合理的设计是至关重要的 化学和生物的疾病抑制剂。在上一版本中工作 获奖期涉及病毒包膜糖蛋白之一gp85，在 EB病毒与淋巴细胞膜的融合为临床诊断提供依据 其他可能对早期事件起作用的新蛋白质 感染。当前提案的目标是继续研究 病毒进入和参与这一过程的包膜蛋白，以 勾画糖蛋白gp85的功能结构域，并检测 假设gp85是一种病毒融合蛋白。有三个具体的 目标。第一是确定功能和结构/功能 Gp85的相互关系。一组单抗将被绘制成 与蛋白质的不同区域以及它们中和和 阻止病毒渗透将用于识别以下区域 具有特殊的功能重要性。将表达gp85的cdna克隆。 在痘苗病毒中制造足够的蛋白质用于抗体图谱和 建立分子的取向和锚定序列(S)。 病毒小体将被用来制造gp85和gp85的人工“缺失突变体” 将努力获得足够数量的gp85进行分析。 将其插入脂质体后即可发挥作用。第二个目标是 EB病毒诱导的三种新型病毒的生化和功能表征 膜蛋白。一种蛋白质将被测序，抗体将被测序 由候选开放阅读框衍生的合成肽制成 将另外两个基因映射到病毒基因组。第三个目标是使用核聚变 EBV检测，以检查影响内化的参数和 确定不同包膜蛋白对进入的相对重要性 转化为B细胞和上皮细胞。

Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus that causes infectious mononucleosis, is associated with Burkitt's lymphoma and has been implicated in the etiology of nasopharyngeal carcinoma. It contributes to the development of lymphoproliferative syndromes and is thought to influence the pathogenesis of the human immunodeficiency virus. Recruitment of normal cells by endogenous virus, produced in increased amounts during episodes of immunosuppression, is of great relevance to virus induced pathology and probably contributes to development of the premalignant state. The overall objective of this proposal is to understand how EBV enters its two targets, the B lymphocyte and the epithelial cell and is based on the premise that clarification of how this process occurs is critical to rational design of chemical and biologic inhibitors of disease. Work in the previous award period implicated one of the virus envelope glycoproteins, gp85, in fusion of EBV with the lymphocyte membrane and provided evidence for additional novel proteins that may contribute to early events in infection. The goals of the current proposal are to continue study of virus entry and the envelope proteins involved in the process, to delineate functional domains of glycoprotein gp85, and to test the hypothesis that gp85 is a virus fusion protein. There are three specific aims. The first is to determine the function and structure/function relationships of gp85. A panel of monoclonal antibodies will be mapped to different regions of the protein and their ability to neutralize and block virus penetration will be used to identify regions that are of particular functional importance. A cDNA clone of gp85 will be expressed in vaccinia virus to make enough protein for the antibody mapping and to establish the orientation and anchor sequence(s) of the molecule. Virosomes will be used to make artificial "deletion mutants" of gp85 and an effort will be made to obtain quantities of gp85 sufficient to analyze its functions after insertion into liposomes. The second aim is to characterize biochemically and functionally the three novel EBV-induced membrane proteins. One protein will be sequenced and antibodies will be made to synthetic peptides derived from candidate open reading frames to map the other two to the viral genome. the third aim is to use a fusion assay for EBV to examine parameters that influence internalization and to determine the relative importance of different envelope proteins to entry into B cells and epithelial cells.