两种不同培养工艺制备森林脑炎灭活疫苗的免疫原性及安全性评价

目的评价细胞工厂工艺制备森林脑炎灭活疫苗的免疫原性及安全性。方法利用10 L转瓶和细胞工厂两种工艺在地鼠肾细胞上培养森林脑炎病毒(tick-borne encephalitis virus,TBEV),收获病毒液,经病毒灭活、超滤浓缩、柱层析纯化、除菌过滤后,制备森林脑炎灭活疫苗。采用透射电子显微镜及SDS-PAGE对病毒进行形态观察及鉴定。将两种工艺共制备的6批疫苗免疫小白鼠,评价疫苗的免疫原性;通过家兔刺激试验及豚鼠主动过敏性试验评价疫苗的安全性。结果两种工艺共制备的6批疫苗原液,透射电镜下观察均可见约为50 nm的圆形病毒颗粒,SDS-PAGE分析均可见相对分子质量分别为50 000和40 000~45 000的蛋白条带,病毒形态及蛋白条带大小均与TBEV相符;免疫保护指数均大于1. 0×10~5,符合《中国药典》三部(2015版)标准;家兔刺激试验对注射部位无刺激性,豚鼠主动过敏试验无小鼠出现过敏性反应。结论细胞工厂工艺和10 L转瓶工艺制备的森林脑炎灭活疫苗均具有良好的免疫原性及安全性。

Immunogenicity and safety of inactivated tick-borne encephalitis vaccine prepared by two culture processes

Objective To evaluate the immunogenicity and safety of inactivated tick-borne encephalitis vaccine prepared by cell factory technique. Methods Tick-borne encephalitis virus(TBEV)was cultured in primary hamster kidney cells in10 L rolling bottle and cell factory respectively,and the virus liquid was harvested,inactivated,concentrated by ultrafiltration,purified by column chromatography and sterilized by filtration to prepare inactivated tick-borne encephalitis vaccine.The virus was observed for morphology by transmission electron microscopy and identified by SDS-PAGE. Mice were immunized with six batches of the vaccine prepared by two processes to evaluate the immunogenicity. The safety of vaccine was evaluated by irritation test in rabbits and active allergy test in guinea pigs. Results Round virus particles at sizes of about 50 nm were observed in six batches of bulks of vaccine prepared by two processes under transmission electron microscope,while SDS-PAGE showed protein bands with relative molecular masses of 50 000 and 40 000 ~ 45 000 respectively,which were consistent with those of TBEV. The immune protection indexes of vaccines prepared by the two processes were more than 1. 0 × 10~5,which met the requirements in Chinese Pharmacopoeia(Volume Ⅲ,2015 edition).No irritation in the injection site was observed in rabbit irritation test,and no allergic reaction in active allergic test in guinea pigs. Conclusion Both the inactivated tick-borne encephalitis vaccines prepared by cell factory technique and in10 L rolling bottle showed good immunogenicity and safety.