一种新型1型单纯疱疹病毒mRNA疫苗的制备  

Developing a Novel Vaccine Against Herpes Simplex Virus Type 1 by mRNA Technology

作　　者：刘彪 蔡林康 苏红霞 汪洋 刘滨磊 LIU Biao;CAI Linkang;SU Hongxia;WANG Yang;LIU Binlei(School of Bioengineering and Food,Hubei Univ.of Tech.,Wuhan 430068,China;Wuhan Binhui Biotechnology Co.,Ltd,Wuhan 430073,China)

机构地区：[1]湖北工业大学生物工程与食品学院,湖北武汉430068 [2]武汉滨会生物科技股份有限公司,湖北武汉430073

出　　处：《湖北工业大学学报》2024年第5期96-99,114,共5页Journal of Hubei University of Technology

摘　　要：制备用于防治1型单纯疱疹病毒感染的新型mRNA疫苗,并检测其免疫效果。利用同源重组技术构建HSV1糖蛋白gD胞外域(HSV1-gD^(Ectodomain))重组质粒,经PCR反应扩增HSV1-gD^(Ectodomain)基因模板,体外转录反应得到HSV1-gD^(Ectodomain)mRNA,利用LNP包封HSV1-gD^(Ectodomain)mRNA得到mRNA-LNP疫苗复合物,小鼠大腿肌肉接种mRNA-LNP疫苗复合物。ELISA法检测小鼠血清HSV1-gD抗体效价;TCID50法检测HSV1-gD血清中和效价。结果显示:经HSV1-gD^(Ectodomain)mRNA疫苗接种后,小鼠血清抗体滴度达到1∶12800,中和实验表明抗体可保护Vero细胞不受HSV1病毒感染。成功制备一种新型HSV1 mRNA疫苗,用其接种小鼠可诱导产生特异性免疫应答,并具有良好的保护作用。To develop a novel mRNA vaccine that can be used to prevent and test the herpes simplex virus infection type 1,the coding sequence for HSV1 glycoprotein gD extracellular domain was cloned into the pUC57 vector to obtain a recombinant plasmid by homologous recombination.The HSV1-gD^(Ectodomain) gene template was amplified by PCR reaction,and HSV1-gD^(Ectodomain) mRNA was obtained by in vitro transcription reaction.The LNP was used to encapsulate HSV1-gD^(Ectodomain) mRNA to obtain mRNA-LNP complex,and the mRNA-LNP complex was injected intramuscularly into the thighs of mice.HSV1-gD antibody titer was determined by ELISA assay.The neutralization antibody titer of HSV1-gD was detected by TCID50 method.The results showed HSV1-gD antibody titer of the immunized mice reached to 1:12800,and the neutralization test showed Vero cells were protected from HSV1 virus infection(green fluorescent GFP and plaques did not appear in the experimental group).All in all,The HSV1 vaccine prepared by mRNA technology can induce specific immune response and has good protective effect.

关 键 词：HSV-1 gD胞外域基因 mRNA疫苗 特异性抗体 

分 类 号：R392[医药卫生—免疫学]