Zhang Peirui, Gu Hongjing, Bian Chengrong, Liu Na, Li Zhiwei, Duan Yueqiang, Zhang Shaogeng, Wang Xiliang, Yang Penghui
302 Military Hospital, Beijing 100039, PR China.
Beijing Institute of Microbiology and Epidemiology, State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, PR China.
J Gen Virol. 2014 Sep;95(Pt 9):1886-1891. doi: 10.1099/vir.0.064105-0. Epub 2014 Jun 9.
Respiratory syncytial virus (RSV) is the most common cause of respiratory infection in infants and the elderly, and no vaccine against this virus has yet been licensed. Here, we report a recombinant PR8 influenza virus with the RSV fusion (F) protein epitopes of the subgroup A gene inserted into the influenza virus non-structural (NS) gene (rFlu/RSV/F) that was generated as an RSV vaccine candidate. The rescued viruses were assessed by microscopy and Western blotting. The proper expression of NS1, the NS gene product, and the nuclear export protein (NEP) of rFlu/RSV/F was also investigated using an immunofluorescent assay. The rescued virus replicated well in the MDCK kidney cell line, A549 lung adenocarcinoma cell line and CNE-2Z nasopharyngeal carcinoma cell line. BALB/c mice immunized intranasally with rFlu/RSV/F had specific haemagglutination inhibition antibody responses against the PR8 influenza virus and RSV neutralization test proteins. Furthermore, intranasal immunization with rFlu/RSV/F elicited T helper type 1-dominant cytokine profiles against the RSV strain A2 virus. Taken together, our findings suggested that rFlu/RSV/F was immunogenic in vivo and warrants further development as a promising candidate vaccine.
呼吸道合胞病毒(RSV)是婴儿和老年人呼吸道感染最常见的病因,目前尚无针对该病毒的疫苗获得许可。在此,我们报告了一种重组PR8流感病毒,其将A亚组基因的RSV融合(F)蛋白表位插入流感病毒非结构(NS)基因(rFlu/RSV/F),该病毒作为RSV疫苗候选株构建而成。通过显微镜检查和蛋白质印迹法对拯救出的病毒进行评估。还使用免疫荧光测定法研究了rFlu/RSV/F的NS基因产物NS1和核输出蛋白(NEP)的正确表达情况。拯救出的病毒在MDCK肾细胞系、A549肺腺癌细胞系和CNE-2Z鼻咽癌细胞系中复制良好。用rFlu/RSV/F经鼻免疫BALB/c小鼠,可产生针对PR8流感病毒的特异性血凝抑制抗体反应以及针对RSV中和试验蛋白的反应。此外,用rFlu/RSV/F经鼻免疫可引发针对RSV A2株病毒的1型辅助性T细胞主导的细胞因子谱。综上所述,我们的研究结果表明,rFlu/RSV/F在体内具有免疫原性,作为一种有前景的候选疫苗值得进一步研发。