Development of an inactivated rotavirus vaccine.

轮状病毒已被证明是发达国家和发展中国家婴幼儿严重腹泻的单一最重要的病因。我们计划(1)选择适合疫苗生产的人类轮状病毒菌株和细胞底物的最佳组合，(2)在大规模细胞培养系统中适应所选病毒，(3)建立纯化和灭活方法，以尝试开发灭活轮状病毒疫苗。我们选择了AU64n菌株，这是一种天然存在的单VP7基因替代重组物，因为该菌株先前被证明具有在儿童中流行的人类轮状病毒中普遍存在的VP7和VP4抗原。AU64n菌株从非洲绿猴肾细胞中分离出来，然后适应于Vero细胞克隆衍生物Vero CL-9中生长。感染滴度达107 pfu/ml，相当于病毒蛋白含量约为15 ug/ml。纯化后的病毒制剂回收率为20%，甲醛灭活后其免疫原性明显不降低。这项研究为研制灭活疫苗奠定了坚实的基础。

Rotavirus has been shown to be the single most important etiological agent of severe diarrhea of infants and young children in both developed and developing countries. We have planned (1) to select an optimal combination of a human rotaviurs strain and a cell substrate suitable for vaccine production, (2) to adapt the selected virus in a large scale cell culture system, and (3) to establish purification and inactivation methods in an attempt to develop an inactivated rotavirus vaccine. We have selected the AU64n strain, a naturally-occurring single VP7 gene substitution reassortant, because this strain was previously shown to possess VP7 and VP4 antigens prevalent in human rotaviruses circulating among children. The AU64n strain was isolated in African Green Monkey Kidney cells and then adapted to grow in Vero CL-9, a cloned-derivative of Vero cells. The infectious titer reached 107 pfu/ml which corresponds to ca 15 ug/ml of virus protein. Purified virus preparations were obtained at a recovery of 20% and inactivated with formaldehyde apparently without reducing its immunogenicity. This study founded a firm basis for the development of an inactivated vaccine.