重组肠道病毒71型疫苗的免疫原性和保护效果研究  

Evaluation of immunogenicity and protective efficacy of the recombinant EV-A71 vaccine

作　　者：顾美荣[1] 高帆[2] 张改梅[1] 肖霞 李国顺[1] 陈磊[1] 郭林[1] 肖海峰 刘建凯[1] 毛群颖[2] 郑海发[1] GU Mei-rong;GAO Fan;ZHANG Gai-mei;XIAO Xia;LI Guo-shun;CHEN Lei;GUO Lin;XIAO Hai-feng;LIU Jian-kai;MAO Qun-ying;ZHENG Hai-fa(Center for Research and Development,Beijing Minhai Biotechnology Co.,Ltd,Beijing 102600,China)

机构地区：[1]北京民海生物科技有限公司北京市新型联合疫苗工程技术研究中心,北京102600 [2]中国食品药品检定研究院卫生部生物技术产品检定方法及其标准化重点实验室,北京102629

出　　处：《中国病毒病杂志》2018年第6期451-455,共5页Chinese Journal of Viral Diseases

摘　　要：目的评价重组肠道病毒71型(EV-A71)疫苗的免疫原性和保护效果.方法将重组EV-A71疫苗按0、14d程序免疫BALB/c小鼠,采用微量细胞病变法测定一针免疫、二针免疫14d后的EV-A71中和抗体效价,应用酶联免疫吸附法测定二针免疫14d后小鼠脾单个核细胞分泌的细胞因子水平.乳鼠颅内攻击EV-A71病毒,同时将疫苗免疫后小鼠血清通过腹腔注射乳鼠,计算乳鼠生存率及半数动物保护性抗体水平.结果重组EV-A71疫苗一针免疫小鼠后诱导产生的EV-A71中和抗体阳转率即达100%,二针免疫小鼠后诱导产生的EV-A71中和抗体效价显著高于一针免疫(P=0.000),疫苗可诱导小鼠产生Th1和Th2类细胞因子应答.乳鼠颅内攻击20μl原倍EV-A71/M6病毒,同时将疫苗免疫小鼠血清160、40、10、2.5、0.625和0.15625U分别经腹腔注射乳鼠,连续观察22d,乳鼠生存率分别为100%、100%、70%、20%、0和0,对照组乳鼠于第8天全部死亡,采用Reed-Muench法计算平均半数动物保护中和抗体效价(ED50)为5.59U.结论重组EV-A71疫苗可诱导小鼠产生良好的体液免疫和细胞免疫应答,并可保护乳鼠免受EV-A71病毒感染.Objective To evaluate the immunogenicity and protective efficacy of the recombinant enterovirus 71(EV-A71) vaccine.Methods BALB/c mice were immunized with recombinant EV-A71vaccines at 0-day and 14-day.Cytopathic effect (CPE) was assayed to determine the EV-A71neutralizing antibody (NtAb) on the 14dafter first and second vaccination.Enzyme-linked immunosorbent assay (ELISA) was employed to de-tect the cytokines secretion of mouse spleen mononuclear cells (MNCs) on the 14dafter second vaccina-tion.The sucking mice were immunized intraperitoneally (i.p.) with diluted antiserum from immunized mice and challenged with EV-A71intracranially.The survival rates of the post-challenged newborn mice were ob-served for 22days and the 50%animal protective antibody level was evaluated.Results After first vaccina-tion,the seroconversion rates of EV-A71NtAb were 100%.After second vaccination,the EV-A71NtAb titers all increased significantly (P =0.000) .Th1and Th2cytokine secretion could be induced by the recombinant vaccines in BALB/c mice.After the challenge with 20 μ l EV-A71/M6virus intracranially,no survival in con-trol group at 8days post-challenge ; while at day 22post-challenge,the survival rates were 100%,100%,70%,20%,0and 0in mice immunized with diluted antiserum of NtAb titers at 160,40,10,2.5,0.625and 0.156 25U,respectively.The average animal protective NtAb titer was 5.59Ucalculated by Reed-Muench method.Conclusions Recombinant EV-A71vaccines showed effective humoral and cellular immune respon-ses in BALB/c mice and provided effective protections against lethal infection of EV-A71in the sucking mice.

关 键 词：手足口病 肠道病毒71型 疫苗 免疫原性 体液免疫 细胞免疫 保护效果 

分 类 号：R373.2[医药卫生—病原生物学]