Department of Microbiology, Chung-Ang University College of Medicine, Seoul 156-756.
Biomol Ther (Seoul). 2012 May;20(3):320-5. doi: 10.4062/biomolther.2012.20.3.320.
Rotavirus and hepatitis A virus (HAV) spread by the fecal-oral route and infections are important in public health, especially in developing countries. Here, two antigenic epitopes of the HAV polyprotein, domain 2 (D2) and domain 3 (D3), were recombined with rotavirus VP7, generating D2/VP7 and D3/VP7, cloned in a baculovirus expression system, and expressed in Spodoptera frugiperda 9 (Sf9) insect cells. All were highly expressed, with peak expression 2 days post-infection. Western blotting and ELISA revealed that two chimeric proteins were antigenic, but only D2/VP7 was immunogenic and elicited neutralizing antibody responses against rotavirus and HAV by neutralization assay, implicating D2/VP7 as a multivalent subunit-vaccine Candidate for preventing both rotavirus and HAV infections.
轮状病毒和甲型肝炎病毒(HAV)通过粪-口途径传播,感染在公共卫生中非常重要,尤其是在发展中国家。在这里,HAV 多蛋白的两个抗原表位,结构域 2(D2)和结构域 3(D3)与轮状病毒 VP7 重组,生成 D2/VP7 和 D3/VP7,克隆在杆状病毒表达系统中,并在 S 型粉纹夜蛾 9( Sf9)昆虫细胞中表达。所有这些都高度表达,感染后 2 天达到峰值表达。Western blotting 和 ELISA 显示两种嵌合蛋白具有抗原性,但只有 D2/VP7 具有免疫原性,并通过中和试验诱导针对轮状病毒和 HAV 的中和抗体反应,表明 D2/VP7 是一种多价亚单位疫苗候选物,可预防轮状病毒和 HAV 感染。