IMMUNOGENICITY OF DENGUE TYPE 4 VIRUS NONSTRUCTURAL PROTEIN NS1

登革热 4 型病毒非结构蛋白 NS1 的免疫原性

• 批准号: 3818274
• 负责人: B FALGOUT
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 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/3818274
• 关键词: antibody formation; dengue virus; enzyme linked immunosorbent assay; gene expression; genetic manipulation; glycoproteins; immunogenetics; laboratory mouse; molecular cloning; oligopeptides; protein engineering; protein sequence; vaccinia virus; virus antigen; virus envelope; virus genetics; virus protein

The nonstructural protein 1 (NS1) of dengue virus has been shown by others to induce protective immunity in mice. The use of NS1 for immunization against infection has one theoretical advantage over immunization with the other dengue protective antigen, the envelope glycoprotein (E). It is possible that antibodies against the envelope protein might cause immune enhancement of disease during subsequent infection by a heterotypic dengue virus. For this reason we initiated a study of the immunogenicity of dengue type 4 NS1. A series of overlapping oligopeptides that span the entire NS1 amino acid sequence were synthesized. In addition, we constructed recombinant vaccinia viruses for expression of: (i) the entire NS1, alone or in combination with other dengue products, or (ii) subfragments of NS1. Analysis of the binding of NS1 specific antibodies to the peptides by ELISA and immunoprecipitation of subfragments of NS1 indicated that the amino terminal third of the NS1 protein contains the major antigenic epitopes. Studies of the expression of NS1 by another series of vaccinia recombinants demonstrated that the hydrophobic sequence preceding NS1 is a functional signal whether it is located at the amino terminus or at internal position of an expressed polyprotein. In addition, the downstream NS2a sequence was found to be required in cis for the normal cleavage of the NS1/NS2a junction.

登革热病毒的非结构蛋白1(NS1)已被发现 由其他人诱导小鼠保护性免疫。NS1的用法 因为针对感染的免疫接种有一个理论上的优势 用另一种登革热保护性抗原过度免疫， 包膜糖蛋白(E)。有可能抗病毒的抗体 包膜蛋白可能导致疾病的免疫增强 在随后被异型登革热病毒感染期间。为 为此，我们发起了登革热免疫原性的研究。 类型4 ns1。一系列重叠的寡肽，跨越 合成了完整的NS1氨基酸序列。此外，我们 构建重组痘苗病毒用于表达：(I) 整个NS1单独或与其他登革热产品结合使用， 或(Ii)NS1亚片段。NS1的结合力分析 用酶联免疫吸附试验检测多肽的特异性抗体 NS1亚片段的免疫沉淀表明，该氨基 NS1蛋白末端的三分之一含有主要抗原性 表位。另一系列NS1基因表达的研究 痘苗病毒重组体证明了疏水序列 前面的ns1是一个功能信号，它是否位于 在表达的多蛋白的氨基端或内部位置。 此外，发现下游的NS2a序列是必需的 在顺式基因中，NS1/NS2a连接的正常切割。