MONOCLONAL ANTIBODIES DIRECTED TO VP8 SUBUNIT OF VP4

此前，针对重组外衣壳轮状病毒VP4的抗血清 用蛋白质研究人类VP4抗原性多态性 轮状病毒株。VP4的三个不同血清型和一个亚型 从17株人轮状病毒中鉴定出含有 五种不同VP7血清型的代表。交叉试验的结果- VP8抗血清的免疫沉淀和中和分析表明 VP4的VP8裂解亚基含有主要的抗原位点 通过VP4负责轮状病毒的血清型特异性中和。 人VP4 VP8裂解亚单位免疫小鼠的研究 轮状病毒KU、DS-1、1076或K8株诱导的多克隆抗体 中和产生VP4亚单位的轮状病毒。然而， 到目前为止获得的大多数VP4单抗都已识别 异源菌株。我们将尝试产生特定于血清类型的 用产生的花叶病毒产生的抗VP4的单抗 表达VP6的重组杆状病毒与昆虫细胞共感染 VP7和VP4基因。

Previously, antisera against recombinant outer capsid rotavirus VP4 proteins were used to study the VP4 antigenic polymorphism among human rotavirus strains. Three distinct serotypes and one subtype of the VP4 protein were identified among 17 human rotavirus strains that contained representatives of five different VP7 serotypes. Results from cross- immunoprecipitation and neutralization assays with VP8 antisera suggest that the VP8 cleavage subunit of VP4 contains the major antigenic sites responsible for the serotype-specific neutralization of rotavirus via VP4. Immunization of mice with the VP8 cleavage subunit of VP4 of the human rotavirus KU, DS-1, 1076, or K8 strain induced polyclonal antibodies that neutralized the rotavirus from which the VP4 subunit was derived. However, most of the VP4-monoclonal antibodies obtained so far have recognized heterologous strains. We will attempt to produce serotype-specific monoclonal antibodies (MAbs) against VP4 using "mosaic virus" produced by coinfection of insect cells with baculovirus recombinant expressing VP6, VP7, and VP4 genes.