呼吸道合胞病毒样颗粒疫苗制备及免疫效果研究  

Preparation and protective efficacy of respiratory syncytial virus-like particles vaccine in Balb/c mice

作　　者：孙苏静 谷宏婧 陈锐[1] 段跃强[1] 王希良[1] 杨鹏辉[1] 

机构地区：[1]军事医学科学院微生物流行病研究所,北京100071

出　　处：《免疫学杂志》2014年第5期387-392,共6页Immunological Journal

基　　金：国家十二五"863"重大课题(SS2012AA020905)

摘　　要：目的利用昆虫表达系统表达以流感病毒基质蛋白M1为骨架制备的嵌合呼吸道合胞病毒(RSV)融合蛋白F和表面糖蛋白G蛋白的呼吸道合胞病毒样颗粒(RSV-F/G VLPs)疫苗候选株,并滴鼻免疫Balb/c小鼠,评价其免疫原性与免疫保护性。方法通过基因重组技术制备以流感基质蛋白M1为骨架,并嵌合RSV F/G蛋白的RSV-F/G VLPs,经超滤浓缩、蔗糖密度梯度离心纯化VLP,滴鼻免疫Balb/c小鼠,检测血清抗体IgG、IgG1、IgG2a,黏膜sIgA效价以及细胞因子IL-4、INF-γ等,评价候选疫苗的免疫原性;二免后2周RSV A2株攻毒,通过检测小鼠体质量变化,肺病毒载量及肺病理切片来评价候选疫苗的免疫保护性。结果成功制备了RSV-F VLPs、RSV-G VLPs疫苗抗原,动物实验结果证实,候选疫苗株RSV-F/G VLPs可刺激机体产生特异性Th1型细胞免疫及黏膜免疫;攻毒实验结果表明候选疫苗株RSV-F/G VLPs针对RSV A2株的攻击可以产生一定的保护效果。结论成功制备了RSV-F VLPs、RSV-G VLPs候选疫苗抗原,并通过动物实验证明RSV-F/G VLPs具有良好的免疫原性和免疫保护性,为发展新型RSV疫苗提供实验依据。Respiratory syncytial virus （RSV） is the most major viral pathogens causing of respiratory infection in infants and elderly, and there is no licensed vaccine. This study was designed to prepare virus-like particles （VLPs） consisting of RSV fusing protein （17） or RSV attachment protein （G） and influenza virus matrix protein（M1） by Bac-to-Bac baculovirus expression system, and characterize the immunogenicity and vaccine efficacy of the RSV-F/G VLPs in a mouse model. After purification and confirmed by Western blot and electron microscopy, the RSV-F/G VLPs was used to immunize 4-6 week female Balb/c mice intranasally. Specific viral antibody responses against RSV were observed in the mice. We further studied active immunity, antibodies （IgG, IgG1, IgG2a and sIgA） and cytokines （IL -4 and IFN -γ） response against RSV-A2 viruses in Balb/c mice, and found mice immunized with RSV-F/G VLPs showed robust viral neutralizing antibodies in vitro and significantly decreased lung virus loads in vivo after live RSV-A2 challenge. This study demonstrates that VLP vaccination provides effective orotection a^ainst RSV infection and should be further confirmed in cotton mouse and monkey model.

关 键 词：呼吸道合胞病毒 病毒样颗粒 疫苗 

分 类 号：R392.11[医药卫生—免疫学]