EXPRESSION OF ROTAVIRUS OUTER CAPSID PROTEIN VP4 BY A BACULOVIRUS RECOMBINANT

轮状病毒外壳蛋白VP4是一种主要的独立保护性蛋白 抗原。一个代表症状性人VP4基因的cDNA克隆 轮状病毒KU株(VP7血清1型)或DS-1株(VP7血清2型)或 构建了无症状人轮状病毒1076株(VP7血清2型) 并将其插入杆状病毒表达载体中 多面体启动子。由此产生的重组体表达了适当的 真正的VP4蛋白。用这些VP4蛋白免疫豚鼠 开发了特异性抗体，中和了病毒的传染性 产生免疫VP4的轮状病毒。这些抗血清是 然后在中和试验中使用，以确定 人轮状病毒VP4抗原性多态。三个截然不同的 确定了VP4外衣壳蛋白的血清型和一个亚型 在此前被归类为5株的17株人轮状病毒中 不同的VP7血清型。在大多数情况下，VP4血清型是分离的 与VP7血清型无关。 在此前已测出VP4基因序列的10株轮状病毒株中， VP4血清型的归属完全一致。 根据VP4的氨基酸同源性进行中和和分类。 因此，表现出VP4氨基酸同源性的轮状病毒为或 经中和鉴定，大毒株属于相同的VP4血清型。 最后，获得了血清型特异性结构域被定位的证据 在VP4的VP8裂解亚基上。

Rotavirus outer capsid protein VP4 is a major independent protective antigen. A cDNA clone representing the VP4 gene of symptomatic human rotavirus strain KU (VP7 serotype 1) or DS-1 (VP7 serotype 2) or asymptomatic human rotavirus strain 1076 (VP7 serotype 2) was constructed and inserted into a baculovirus expression vector under the control of the polyhedrin promoter. The resulting recombinants expressed the appropriate authentic VP4 protein. Guinea pigs immunized with these VP4 proteins developed specific antibodies which neutralized infectivity of the rotavirus from which the immunizing VP4 was derived. These antisera were then used in neutralization tests to define the extent and distribution of VP4 antigenic polymorphism among human rotaviruses. Three distinct serotypes and one subtype of the VP4 outer capsid protein were identified among 17 human rotavirus strains that had previously been assigned to 5 distinct VP7 serotypes. For the most part, VP4 serotype segregated independently of VP7 serotype. Among the 10 rotavirus strains whose VP4 gene was previously sequenced, there was complete concordance between assignment of VP4 serotype by neutralization and classification according to VP4 amino acid homology. Thus, rotaviruses that exhibited a VP4 amino acid homology of 89% or greater belonged to the same VP4 serotype as determined by neutralization. Finally, evidence was obtained that the serotype-specific domain is located on the VP8 cleavage subunit of VP4.