CAJ | 학술논문

目的筛选安全有效新型EV71候选疫苗,为将来EV71疫苗开发应用奠定基础.方法以重组蛋白VP1为疫苗设计靶点,不同候选疫苗包括灭活疫苗、DNA疫苗、VP1蛋白疫苗在0、2周、4周分别按照不同剂量和肌内注射途径免疫BALB/c雌性小鼠,在0、2周、4周、6周、8周、10周、16周分别采集小鼠尾静脉血,16周处死小鼠,收集小鼠脾脏细胞,检测小鼠体液免疫和细胞免疫功能,筛选评价候选疫苗的疗效和安全性.结果灭活病毒疫苗、VP1 DNA质粒疫苗、VP1蛋白疫苗免疫小鼠2周后IgG抗体滴度即开始升高,4周后明显升高,8周后达高峰,至少维持16周以上;IgG亚类以IgG2a和IgG1为主.三种疫苗能诱导以γ-IFN和IL-4为主的细胞免疫反应.灭活疫苗疗效优于其他候选疫苗,未发现疫苗相关不良反应.结论灭活病毒疫苗、VP1 DNA质粒疫苗、VP1蛋白疫苗均能诱导持久的特异性细胞免疫和中和抗体反应,以灭活病毒疫苗疗效较好,需要将来攻毒实验可进一步验证其免疫力.
목적 사선안전유효신형EV71후선역묘,위장래EV71역묘개발응용전정기출.방법 이중조단백VP1위역묘설계파점,불동후선역묘포괄멸활역묘、DNA역묘、VP1단백역묘재0、2주、4주분별안조불동제량화기내주사도경면역BALB/c자성소서,재0、2주、4주、6주、8주、10주、16주분별채집소서미정맥혈,16주처사소서,수집소서비장세포,검측소서체액면역화세포면역공능,사선평개후선역묘적료효화안전성.결과 멸활병독역묘、VP1 DNA질립역묘、VP1단백역묘면역소서2주후IgG항체적도즉개시승고,4주후명현승고,8주후체고봉,지소유지16주이상;IgG아류이IgG2a화IgG1위주.삼충역묘능유도이γ-IFN화IL-4위주적세포면역반응.멸활역묘료효우우기타후선역묘,미발현역묘상관불량반응.결론 멸활병독역묘、VP1 DNA질립역묘、VP1단백역묘균능유도지구적특이성세포면역화중화항체반응,이멸활병독역묘료효교호,수요장래공독실험가진일보험증기면역력.
Objective To Screen for safe and effective vaccine candidates for EV71,provide a theoretical basis for development of EV71 vaccines in the future.Methods VP1 gene of enterovirus was used to design a target for development of EV71 vaccines.Different vaccine candidates,including inactivated EV71 vaccines,VP1 protein vaccine,DNA vaccines of different doses,were used to challenge female BALB/c mice by intramuscular injection at baseline (0),2 weeks,4 weeks,and caudal vein blood was collected at 0,2,4,6,8,10,and 16 weeks,and BALB/c mice were sacrificed and the spleen cells were collected for detection of both humoral immunity and cellular immunity to evaluate the efficacy and safety of the vaccine candidates.Results IgG antibody titers were increased at 2 weeks,remarkably increased at 4 weeks,reached a peak at 8 weeks,at least sustained for 16 weeks during the whole observation period,subtypes of IgG1 and IgG2a were the major component.The three vaccines could induce cellular immunity characterized by EV71 specific γ-IFN and IL-4 production.Our results indicated that inactivated EV71 vaccine was superior to the other vaccine candidates.Conclusions Inactivated EV71 vaccines,VP1 protein vaccine,DNA vaccines can induce both strong and sustainable humoral and cellular immunities in challenged mice,and the inactivated EV71 vaccine is superior to the other vaccine candidates,which needs to be proved their immunity by challenge assay in the future.