INHIBITION OF EPSTEIN-BARR VIRUS RECEPTOR INTERACTION

人类感染爱泼斯坦-巴尔病毒导致显著的 发病和免疫抑制期间可导致致死性B细胞 淋巴组织增生性疾病 知识的快速积累 关于EBV/宿主细胞相互作用的最早期事件， 开发干扰EBV的抗病毒试剂的机会 与其细胞受体（CR2）结合。 该提案将审查 无论是重组CR2的可溶形式还是病毒附着蛋白， gp 350/220能够在体内阻断EBV的感染。 这些研究 将利用一种新开发的EBV诱导的 免疫缺陷小鼠淋巴组织增生性疾病（SCID）分析 可溶性CR2和gp 350/220的抗病毒特性以及合成的 肽GP 350/220类似物。 研究还计划检查EB病毒的分子基础 与CR2及其天然配体C3 dg的相互作用。 可溶性CR2 和gp 350/220进行系统的定点突变， 鉴定介导病毒-CR2相互作用的关键氨基酸残基。 用固定化蛋白质或在 溶液将用于评估 个体CR2和gp 350/220突变体。 CR2的二级结构和 将通过CD或NMR光谱分析gp 350/220突变体。 这些研究的长期目标是确定 CR2和CR2/gp 350/220和CR2/C3 dg的三维结构 使用X-射线晶体学或NMR光谱学对复合物进行分析。 溶液 配体/受体结构的合理设计应允许 阻断EBV结合而不干扰正常的 B细胞功能

Infection of humans by Epstein-Barr virus results in significant morbidity and during immunosuppression can result in fatal B cell lymphoproliferative disease. The rapid accumulation of knowledge concerning the earliest events in EBV/host cell interaction presents an opportunity to develop antiviral reagents which interfere with EBV attachment to its cellular receptor (CR2). This proposal will examine whether soluble forms of recombinant CR2 or the viral attachment protein, gp350/220, are capable of blocking infection of EBV in vivo. These studies will take advantage of a newly developed model of EBV-induced lymphoproliferative disease in the immunodeficient mouse (SCID) to analyze the antiviral properties of soluble CR2 and gp350/220 as well as synthetic peptide gp350/220 analogues. Studies are also planned to examine the molecular basis of EBV interaction with CR2 as well as with its natural ligand, C3dg. Soluble CR2 and gp350/220 will be subjected to systematic sitedirected mutagenesis to identify crucial amino acid residues which mediate virus-CR2 interaction. Quantitative binding assays carried out with immobilized proteins or in solution will be used to assess the relative functional activity of individual CR2 and gp350/220 mutants. The secondary structure of CR2 and gp350/220 mutants will be analyzed by CD or NMR spectroscopy. A long term goal of these studies is to determine the three-dimensional structure of CR2 and the CR2/gp350/220 and CR2/C3dg complexes using X-ray crystallography or NMR spectroscopy. The solution of the ligand/receptor structure should allow the rationale design of therapeutic agents which block EBV binding without interfering with normal B cell function.