DEVELOPMENT OF RECOMBINANT BCG-VP8 VACCINES

轮状病毒是婴幼儿严重腹泻的主要原因。 发达国家和发展中国家的儿童。两个外壳 轮状病毒蛋白，VP7和VP4，与诱导 已在实验动物中显示的中和抗体 与抵抗疾病联系在一起。血清型特异性是 主要与VP7有关。然而，最近的研究阐明了 人轮状病毒VP4的抗原性关系 比VP7基因更少的多态。人类轮状病毒株是 与症状性感染相关，并表现出VP7特异性 血清1、2、3、4或9的每一个都具有相似的VP4，由 中和试验。本研究的目的是：(I)使用VP8 VP4亚基构建卡介苗-VP8重组体及其活性评价 诱导特异性轮状病毒中和抗体的实验研究 动物，以及(Ii)利用这样获得的信息 口服人轮状病毒疫苗的研制。 代表轮状病毒外衣壳蛋白VP8亚基的cDNA 人轮状病毒KU株VP4基因在三种不同卡介苗中的克隆 向量。在这些系统中表达的VP8亚单位在 用高免抗血清中存在的抗体进行免疫印迹分析： (I)人轮状病毒Wa株，(Ii)表达KU-VP4，和(Iii) 表达了VP4裂解亚基KU-VP8。这些观察结果表明 VP8亚单位以抗原性正确的形式通过 重组体为正品VP8。

Rotavizuses are the major cause of severe diarrhea in infants and young children in both developed and developing countries. Two outer capsid rotavirus proteins, VP7 and VP4, are associated with the induction of neutralizing antibodies which have been shown in experimental animals to be associated with resistance to illness. Serotype specificity is associated primarily with VP7. However, recent studies have elucidated the antigenic relationships of the VP4 of human rotaviruses which appear to be less polymorphic than VP7. Human rotavirus strains that are associated with symptomatic infection and that exhibit VP7 specificity of serotype 1, 2, 3, 4 or 9 each possess a similar VP4 as determined by neutralization assay. Objectives of this study were: (i) to use the VP8 subunit of VP4 to develop a BCG-VP8 recombinant and evaluate its ability to induce specific rotavirus neutralizing antibodies in experimental animals, and (ii) to utilize the information thus obtained for the development of a human rotavirus vaccine for oral administration. The cDNA representing the VP8 subunit of outer capsid rotavirus protein VP4 from the human rotavirus KU strain was cloned in three different BCG vectors. The VP8 subunit expressed in these systems was recognized in an immunoblot assay by antibodies present in hyperimmune antiserum to: (i) human rotavirus strain Wa, (ii) expressed KU-VP4, and (iii) expressed KU-VP8, a cleavage subunit of VP4. These observations suggest that the VP8 subunit was expressed in an antigenically correct form by the recombinant as authentic VP8.