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Construction of a Recombinant Human Adenovirus Expressing the ORF2 Antigen of HEV and Immunization of Mice by Mucosal System
Author(s): 
Pages: 324-328
Year: Issue:  3
Journal: ACTA ACADEMIAE MEDICINAE SINICAE

Keyword:  重组腺病毒戊型肝炎病毒疫苗;
Abstract: 目的构建可表达戊型肝炎病毒(hepatitis E virus,HEV)ORF2蛋白(112-660aa)的重组腺病毒Adasy-EORF2,探讨其经鼻腔和腹腔免疫小鼠后,诱导机体产生特异性体液免疫和粘膜免疫应答的能力,为研制戊型肝炎疫苗探索新的途径.方法经PCR扩增编码ORF2蛋白(112-660aa)的基因片段,将其连接于重组腺病毒Adasy系统的穿梭质粒pTrack-CMV上,与腺病毒基因组质粒pAdEasy-1共转化宿主菌BJ5183,经细菌内同源重组获得重组腺病毒基因组质粒,转染293细胞以产生、扩增重组腺病毒.将重组腺病毒以107pfu的剂量经鼻腔和腹腔免疫小鼠,通过ELISA检测血液IgG和粘膜IgA的应答情况.结果重组腺病毒经鼻腔免疫小鼠后可刺激机体产生特异性IgA、IgG免疫应答,IgG最高滴度达1:1000.经腹腔免疫可刺激小鼠产生IgG免疫应答,最高滴度可达1:10000,未检测到明显的IgA应答.结论表达HEV ORF2蛋白的重组腺病毒能刺激机体产生有效的免疫应答,具有成为新型戊型肝炎疫苗的潜力.
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