INITIATION OF CELL INFECTION WITH EPSTEIN-BARR VIRUS

EB病毒（EBV）是一种普遍存在的人类疱疹病毒， 传染性单核细胞增多症，与伯基特淋巴瘤有关， 与鼻咽癌的病因有关。 它 有助于淋巴增生综合征的发展， 被认为影响人类免疫缺陷的发病机制 病毒 通过内源性病毒募集正常细胞， 在免疫抑制发作期间增加的量， 与病毒诱导的病理学相关，可能有助于 恶化前状态的发展。 本报告的总体目标 一个建议是了解EB病毒如何进入它的两个目标，B 淋巴细胞和上皮细胞，并基于这样的前提， 阐明这一过程是如何发生的，对于合理设计至关重要 疾病的化学和生物抑制剂。 工作在以前的 奖励期间涉及病毒包膜糖蛋白之一，gp 85， EB病毒与淋巴细胞膜的融合，并提供了证据， 其他新的蛋白质，可能有助于早期事件， 感染 本提案的目标是继续研究 病毒进入和参与该过程的包膜蛋白， 描绘糖蛋白gp 85的功能结构域，并测试 推测gp 85是一种病毒融合蛋白。 有三个具体的 目标。 一是确定功能和结构/功能 GP 85的关系 将绘制一组单克隆抗体 蛋白质的不同区域以及它们的中和能力， 阻止病毒渗透将用于识别 特别重要的功能。 将表达gp 85的cDNA克隆 在牛痘病毒中产生足够的蛋白质用于抗体定位， 建立分子的取向和锚序列。 病毒体将用于制备gp 85的人工“缺失突变体”， 将努力获得足以分析的GP 85的量 其在插入脂质体后的功能。 第二个目标是 在生物化学和功能上表征三种新的EBV诱导的 膜蛋白 一种蛋白质将被测序，抗体将被 对衍生自候选开放阅读框的合成肽进行 把另外两个和病毒基因组对应起来 第三个目标是使用融合 测定EBV以检查影响内化的参数， 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.