两种工艺制备双价肾综合征出血热灭活疫苗的安全性及免疫原性评价  

Safety and immunogenicity of bivalent inactivated hemorrhagic fever with renal syndrome vaccine prepared by two processes

作　　者：张峰典 张颖 李文娟 梅可佳 张朔 孙超 孙宏亮 ZHANG Feng-dian;ZHANG Ying;LI Wen-juan;MEI Ke-jia;ZHANG Shuo;SUN Chao;SUN Hong-liang(Jilin Provincial Center for Drug Inspection,Changchun 130062,Jilin Province,China)

机构地区：[1]吉林省药品审核查验中心,吉林长春130062 [2]长春生物制品研究所有限责任公司,吉林长春130012

出　　处：《中国生物制品学杂志》2022年第2期129-132,137,共5页Chinese Journal of Biologicals

基　　金：吉林省省级医药健康产业发展专项基金项目(20160203008YY).

摘　　要：目的对两种不同工艺制备的双价肾综合征出血热灭活疫苗的安全性及免疫原性进行评价。方法分别采用细胞工厂、原10 L转瓶工艺培养肾综合征病毒(hemorrhagic fever with renal syndrome virus,HFRSV)Ⅰ型和Ⅱ型,经病毒收获、灭活、超滤浓缩、纯化及除菌过滤后制备单价疫苗原液,将Ⅰ型、Ⅱ型单价疫苗原液按1∶1等体积混合,采用透射电子显微镜观察病毒形态,12%SDS-PAGE分析蛋白相对分子质量。采用家兔刺激试验和豚鼠主动过敏性试验对两种工艺制备的各3批疫苗进行安全性评价,并对疫苗进行小鼠免疫保护性试验。结果两种工艺分别制备了3批疫苗原液,经透射电镜观察可见100 nm左右的粗糙圆形病毒颗粒,12%SDS-PAGE分析可见相对分子质量分别为70000~73000和约50000的蛋白条带,病毒形态及蛋白条带大小均与HFRSV一致。两种工艺制备的疫苗家兔注射部位经肉眼观察未见刺激性,豚鼠主动过敏试验中也未出现过敏性反应;疫苗免疫保护指数均大于1∶10。结论细胞工厂工艺和原有10 L转瓶工艺制备的双价肾综合征出血热灭活疫苗均具有良好的安全性及免疫原性。Objective To evaluate the safety and immunogenicity of bivalent inactivated hemorrhagic fever with renal syndrome vaccine prepared by two processes.Methods Hemorrhagic fever with renal syndrome virus(HFRSV)typesⅠandⅡwere cultured by cell factory and original 10 L rotating bottle process separately.The monovalent bulk of vaccine was prepared by virus harvest,inactivation,concentration by ultrafiltration,purification,and sterilization by filtration.The monovalent bulks of typesⅠandⅡat an equal volume were mixed,observed for morphology of virus by transmission electron microscopy,and determined for relative molecular mass by 12%SDS-PAGE.The vaccine prepared by two processes,three batches for each,were evaluated for safety by stimulation test in rabbits and active allergic test in guinea pigs,and for immune protection in mice.Results Rough round virus particles a size of about 100 nm were observed by transmission electron microscopy in the prepared bu lks by two processes.12%SDS-PAGE showed protein bands with relative molecular masses of 70000~73000 and 50000 respectively.Both the morphology and protein band size were consistent with those of HFRSV.No stimulation was observed visually in injection site of vaccine prepared by two processes in rabbits,and no allergic reactions in guinea pigs.The immune protection indexes of vaccines were more than 1∶10.Conclusion Both the bivalent inactivated renal syndrome haemorrhagic fever vaccines prepared by cell factory and original 10 L rotating bottle process showed good safety and immunogenicity.

关 键 词：双价肾综合征出血热灭活疫苗 细胞工厂 转瓶 安全性 免疫原性 

分 类 号：R392[医药卫生—免疫学]