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 利用蛋白质研究人类VP4抗原多态性 轮状病毒株。 VP4 的三种不同血清型和一种亚型 在 17 种人类轮状病毒株中鉴定出含有 五种不同 VP7 血清型的代表。 交叉结果 VP8 抗血清的免疫沉淀和中和试验表明 VP4 的 VP8 裂解亚基包含主要抗原位点 负责通过 VP4 血清型特异性中和轮状病毒。 用人VP4的VP8裂解亚基免疫小鼠 轮状病毒 KU、DS-1、1076 或 K8 株诱导的多克隆抗体 中和了 VP4 亚基来源的轮状病毒。 然而， 迄今为止获得的大多数VP4单克隆抗体都已识别 异源菌株。 我们将尝试生产血清型特异性的 使用“马赛克病毒”产生的针对 VP4 的单克隆抗体 (MAb) 昆虫细胞与表达VP6的重组杆状病毒的共感染， 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.