IMMUNOGENICITY OF DENGUE TYPE 4 VIRUS NONSTRUCTURAL PROTEIN NS1

登革病毒的非结构蛋白1（NS 1）已被证实 通过其他方法诱导小鼠的保护性免疫。 使用NS 1 对感染进行免疫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）。 有可能是针对 包膜蛋白可能引起疾病的免疫增强 在随后的异型登革热病毒感染期间。 为 因此，我们开始研究登革热的免疫原性， 4型NS 1。 一系列重叠的寡肽， 合成了完整的NS 1氨基酸序列。 另外我们 构建的重组牛痘病毒用于表达：（i） 整个NS 1，单独或与其它登革热产品组合， 或（ii）NS 1的亚片段。 NS 1结合分析 - 通过ELISA检测针对肽的特异性抗体， NS 1亚片段的免疫沉淀表明， NS 1蛋白的末端三分之一含有主要抗原 表位 通过另一系列的细胞因子研究NS 1的表达， 牛痘重组体证明， 前面的NS 1是一个功能信号，无论它是否位于 氨基末端或在表达的多聚蛋白的内部位置。 此外，发现下游NS 2a序列是必需的。 对于NS 1/NS 2a连接的正常切割顺式。

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.