水痘-带状疱疹病毒高效价中和抗体的制备及应用  

Preparation and application of high titer neutralizing antibody against varicella-herpes zoster virus

作　　者：邵宏秋 魏巍 黄晓娜 李景新 魏国良 张芸畅 修华谦 刘婷婷 王宇迪 孙博[5] 卢井才 石亮 SHAO Hongqiu;WEI Wei;HUANG Xiaona;LI Jingxin;WEI Guoliang;ZHANG Yunchang;XIU Huaqian;LIU Tingting;WANG Yudi;SUN Bo;LU Jingcai;SHI Liang(College of Life Science,Jilin Agricultural University,Changchun 130018,Jilin Province,China;不详)

机构地区：[1]吉林农业大学生命科学学院,吉林长春130018 [2]长春百克生物科技股份公司,吉林长春130013 [3]长春生物制品研究所有限责任公司,吉林长春130062 [4]吉林省药品监督管理局长春检查分局,吉林长春130000 [5]吉林大学生命科学学院,吉林长春130000

出　　处：《中国生物制品学杂志》2023年第12期1430-1433,1441,共5页Chinese Journal of Biologicals

基　　金：吉林省科技发展计划(20210204173YY)。

摘　　要：目的制备水痘-带状疱疹病毒(varicella-herpes zoster virus,VZV)高效价特异性免疫血清,用于水痘减毒活疫苗及带状疱疹减毒活疫苗质量控制。方法将高纯度重组gE糖蛋白分别与弗氏佐剂、氢氧化铝佐剂及MF59佐剂组合,免疫雄性家兔,每组2只,免疫后第56天,每只最大量采血(心脏或颈动脉)制备血清,与VZV混合进行中和反应后,接种至长满单层MRC-5株人二倍体细胞的6孔板中,孵育7 d,计数空斑数,检测免疫血清的中和效价和中和病毒能力;选择高中和效价和高中和病毒能力血清进行水痘减毒活疫苗及带状疱疹减毒活疫苗VZV(Oka株)工作种子批毒种鉴别试验和外源病毒因子检查。结果重组gE糖蛋白的多种组合免疫家兔后制备的免疫血清均具有中和活性,其中重组gE糖蛋白与弗氏佐剂组合免疫后制备的血清中和效价最高,为1:512,中和病毒能力可达240000 PFU/mL;制备的免疫血清用于VZV(Oka株)工作种子批毒种鉴别试验和外源病毒因子检查,结果均符合要求。结论重组gE糖蛋白可用于VZV高效价中和抗体制备,制备的高效价中和抗体适用于水痘减毒活疫苗及带状疱疹减毒活疫苗质量控制。Objective To prepare high titer specific immune serum of varicella-herpes zoster virus(VZV)for the quality control of live attenuated varicella vaccine and live attenuated herpes zoster vaccine.MethodsMale rabbits were immunized with high purity recombinant gE glycoprotein combined with Freund's adjuvant,aluminum hydroxide adjuvant or MF59 adjuvant,2 rabbits in each group.On the 56th day after immunization,the maximum blood samples(heart or carotid artery)were collected from each rabbit to prepare serum,which was mixed with VZV for neutralization reaction,and then inoculated into a 6-well plate full of monolayer of MRC-5 human diploid cells.After incubation for 7 d,the number of plaques was counted and the neutralizing titer and virus neutralizing ability of immune serumwere determined.The serum with high neutralizing titer and virus neutralizing ability was selected for the identification test of live attenuated varicella vaccine and live attenuated herpes zoster vaccine VZV(Oka strain)working seed lot and the detection of exogenous virus factors.ResultsThe immune sera prepared by immunizing rabbits with various combinations of recombinant gE glycoprotein all showed neutralizing activity,among which the serum prepared by the combination of recombinant gE glycoprotein and Freund's adjuvant had the highest neutralizing titer of 1∶512 and the virus neutralizing ability of 240000 PFU/mL;The prepared immune serum was usedfor the identification test of VZV(Oka strain)working seed lot and the detection of exogenous virus factors,of which all the results were in line with the requirements.Conclusion The recombinantgE glycoprotein could be used for the preparation of high titer neutralizing antibody against VZV,and the prepared high titer neutralizing antibody is suitable for thequality control of live attenuated varicella vaccine and live attenuated herpes zoster vaccine.

关 键 词：水痘-带状疱疹病毒 重组gE糖蛋白 水痘减毒活疫苗 带状疱疹减毒活疫苗 特异性血清 中和抗体 

分 类 号：R373.9[医药卫生—病原生物学]