ANTIGENIC AND MOLECULAR CHARACTERIZATION OF THE FUSION GLYCOPROTEIN OF RSV

RSV 融合糖蛋白的抗原和分子特征

• 批准号: 3792536
• 负责人: J A BEELER
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3792536
• 关键词: antigen antibody reaction; antiviral antibody; blocking antibody; enzyme linked immunosorbent assay; epitope mapping; genetic strain; glycoprotein structure; human tissue; immunity; laboratory rat; molecular biology; molecular site; monoclonal antibody; mutant; neutralizing antibody; protein structure function; respiratory syncytial virus; synthetic peptide; virus antigen; virus protein

Respiratory syncytial virus is the most common cause of lower respiratory tract disease in young children worldwide. The F glycoprotein of this virus is one of the important targets of a protective immune response; however, immunity is incomplete and repeated infections occur. Previously, a detailed map of the neutralization epitopes was developed based on the ability of a panel of murine anti-F monoclonal antibodies to neutralize 14 monoclonal antibody resistant mutant viruses of the A2 strain. The reactivity of the F monoclonal antibodies with the panel of F MARMs in ELISA was also assessed in order to corroborate the operational map previously developed and to demonstrate a direct correlation between resistance to neutralization and absence of antibody binding. In most cases MARMs did not bind the selecting antibody and verified the idea that escape from neutralization was due to specific mutations in the contact residues within the F epitope. In addition, 25 neutralizing and non-neutralizing F MAbs have been biotinylated and used in competitive binding studies to identify at least 6 antigenic sites on the F glycoprotein. These biotinylated MAbs were also used to develop an antigenic site blocking assay in order to analyze the immune response to specific regions on RSV-F following infection or immunization. Preliminary studies have shown that post- infection human and cotton rat sera that have similar neutralization titers, vary in the repetoire of their anti-F antibody response. During the coming year additional studies will be done to evaluate individual immune responses to the F glycoprotein following natural infection or immunization. As an extension of this work, the reactivity of F MAbs with synthetic peptides will be evaluated in order to develop an epitope specific site-directed ELISA assay.

呼吸道合胞病毒是最常见的原因， 呼吸道疾病的儿童全世界。的F 这种病毒的糖蛋白是一种重要的靶点之一， 保护性免疫反应;然而，免疫是不完全和重复的 发生感染。此前，一份详细的中和地图 表位是基于一组鼠抗F抗体的能力开发的。 单克隆抗体中和14种单克隆抗体耐药 A2株的突变病毒。F单克隆抗体的反应性 还评估了ELISA中F MARM组的抗体， 以证实以前制定的作战地图， 证明了中和抗性与 和不存在抗体结合。在大多数情况下， 筛选抗体，验证了逃避中和的想法 是由于特定的突变接触残基内的F 表位此外，25种中和和非中和F MAb具有 被生物素化，并用于竞争性结合研究，以确定 F糖蛋白上至少6个抗原位点。这些生物素化单克隆抗体 也用于开发抗原位点阻断试验， 分析对RSV-F上特定区域的免疫应答， 感染或免疫。初步研究表明， 感染人和棉鼠血清具有相似的中和作用 滴度在其抗F抗体应答的重复谱中变化。期间 明年将进行更多的研究， 自然感染后对F糖蛋白的免疫应答，或 次免疫 作为这项工作的延伸，F MAbs的反应性 为了开发表位， 特异性定点ELISA测定。