乙型脑炎Vero细胞灭活疫苗毒株的遗传稳定性  

Genetic stability of a Vero-cell-derived, inactivated Japanese encephalitis vaccine （P3 strain）

作　　者：张海燕[1] 曹晗[1] 王俊荣[1] 张名[1] 梁疆莉[1] 马艳[1] 顾琴[1] 杨卉娟[1] 孙明波[1] 

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所云南省重大传染病疫苗研发重点实验室,昆明650118

出　　处：《中华实验和临床病毒学杂志》2014年第3期191-193,共3页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的 研究流行性乙型脑炎Vero细胞灭活疫苗毒种（P3毒株）在生产过程中的遗传稳定性,为疫苗的安全性和免疫原性评价提供依据.方法 检测P3毒株鼠脑传代一代毒株、主种子、工作种子、疫苗及疫苗续传5代后病毒E蛋白基因核苷酸及氨基酸序列,并与GenBank中乙脑病毒P3株（AF036919）进行比对分析,同时比较主种子、工作种子、疫苗及疫苗续传5代后毒株的病毒滴度、抗原含量及效价.结果 以上5代次病毒的E蛋白基因核苷酸及蛋白质序列完全相同,同源性均为100％,与GenBank中乙脑病毒P3株（AF036919）比较显示：核苷酸序列中第E9、E10、E324、E330、E1223、E1338基因位点存在差异,同源性为99.73％,其中只有E1223突变引起相应氨基酸aE408突变（L→S）,同源性为99.80％,但该位点为非毒力相关位点,而其他位点均为沉默突变.4个代次疫苗毒株病毒滴度均≥8.0 lgLD50/ml,且各代次抗原含量及效价较为相似.结论 在生产乙型脑炎灭活疫苗（P3株）过程中建立的以Vero细胞为基质的乙脑毒种库具有良好的遗传稳定性.Objective To investigate genetic stability of P3 strain of Vero cell derived inactivated Japanese encephalitis vaccine.Methods The nucleotide and amino acid sequences of E protein of Japanese encephalitis virus （JEV） P3 strain at different culture period including the mouse brain one passage seed,master seed,working seed the vaccine lot in addition to its 5 passages lot were determined,while the E gene and protein sequences were compared with JEV wide stain （AF036919） from the GenBank.Furthermore,the master seed,working seed,vaccine lot and its 5 passages lot of P3 stain were determined for virus titer,antigen concentration and the vaccine potency.Results The E gene and protein sequences of the above 5 passages of vaccine strain prepared for JEV vaccine showed no difference with homologies of 100%.When the 5 passages of vaccine stain compared with those JEV wide stain （AF036919）,the gene sequence at E9,E10,E324,E330,E1223,E1338 showed difference with homologies of 99.73%.No silent mutation were investigated except the amino acid mutation at aE408 （L→S） but was no virulence-associated sites with homologies of 99.80%.The virus titers of the master seed,working seed,vaccine lot and its 5 passages lot of P3 stain were higher than 8.0 lgLD50/ml,while the antigen concentrations and the vaccine potency showed no difference.Conclusion The virus seed bank of P3 strain for Vero cell derived inactivated Japanese encephalitis vaccine showed high genetic stable.

关 键 词：脑炎病毒 日本 疫苗 灭活 遗传学 生物化学 蛋白质E 

分 类 号：R512.32[医药卫生—内科学]