H2减毒株甲型病毒性肝炎灭活疫苗制备及免疫剂量分析
Preparation and immunizing dose analysis of inactivated hepatitis A vaccine using attenuated H2 strain
李宏森 1平玲 2王正鑫 1蒋厚菲 1侯丁琳 1张艺榕 1王灵犀 1杨净思2
作者信息
- 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 昆明 650118
- 2. 中国医学科学院北京协和医学院医学生物学研究所,云南 昆明 650118;昆明市新发突发高致病性病原体疫苗研发和产业化企业科技创新中心,云南 昆明 650118
- 折叠
摘要
目的 利用甲型肝炎(甲肝)病毒(HAV)H2减毒株制备甲肝灭活疫苗,分析免疫剂量,为甲肝灭活疫苗的研发与生产提供参考.方法 采用HAV H2减毒株感染人胚肺二倍体细胞(KMB17)增殖病毒,收获含病毒的细胞,经提取、纯化和灭活后,制备成疫苗原液和抗原含量为1 280 EU/mL的试验疫苗;根据《中华人民共和国药典》三部(2020年版)对疫苗的各项指标进行检定.110只小鼠随机分为11组,包括试验疫苗、参比疫苗各5个剂量组(80、160、320、640和1 280 EU/剂)和佐剂对照组,腹腔注射免疫2次,检测血清HAV抗体,分析抗体几何平均滴度(GMT)和抗体阳转率,判断免疫剂量.结果 病毒收获液的抗原含量和病毒滴度分别为5 120 EU/mL和8.33 lgCCID50/mL,杂蛋白去除率为98.05%,抗原回收率为66.25%;试验疫苗各项指标检定结果均符合《中华人民共和国药典》三部(2020年版)的要求.试验疫苗和参比疫苗各剂量组二次免疫后小鼠血清HAV抗体GMT较初次免疫平均增加2倍以上;无论初次免疫还是二次免疫,试验疫苗160 EU/剂及以上剂量组小鼠血清HAV抗体GMT(log2)高于80 EU/剂组(均P<0.05),但160 EU/剂及以上剂量组之间差异无统计学意义(均P＞0.05);二次免疫后,试验疫苗160 EU/剂及以上组HAV抗体阳转率均为100.00%.结论 本研究制备的H2减毒株甲肝灭活疫苗在小鼠体内具有良好的免疫原性,可作为甲肝灭活疫苗的候选疫苗,初步分析其免疫剂量为320 EU/剂(儿童)和640 EU/剂(成人).
Abstract
Objective To prepare an inactivated hepatitis A vaccine using a attenuated strain of hepatitis A virus(HAV)H2 and to analyze its immunizing dose,so as to provide the reference for development and production of inactivated hepatitis A vaccines.Methods Human embryonic lung diploid cells(KMB17)were infected with attenuated HAV H2 strain to proliferate the virus,then the cells containing viruses were harvested,extracted and purified.The obtained vi-rus concentrate was prepared into vaccine bulk and test vaccines with 1 280 EU/mL antigen content.Vaccine testing was carried out according to the inactivated hepatitis A vaccine standards specified in the Part Ⅲ of the Pharmacopoe-ia of the People's Republic of China(2020 edition).A total of 110 mice were randomly divided into 11 groups,includ-ing 5 dose groups(80,160,320,640 and 1 280 EU/dose)of the test vaccine and the reference vaccine,as well as the adjuvant control group.Mice were immunized twice by intraperitoneal injection,their serum HAV antibodies were de-tected,and the geometric mean titer(GMT)and positive conversion rate of antibodies were analyzed to evaluate the im-munising dose of the vaccine.Results The antigen content and viral titer of the virus harvest solution were 5 120 EU/mL and 8.33 lgCCID50/mL,respectively.The removal rate of foreign protein reached 98.05%and the recovery rate of anti-gen was 66.25%.The test vaccine met the requirements of Part Ⅲ of the Pharmacopoeia of the People's Republic of China(2020 edition).The GMTs of HAV antibodies in the test vaccine and the reference vaccine dose groups after the second immunization were more than twice higher than those after the first immunization.Regardless of primary immuni-zation or secondary immunization,the GMTs(log2)of HAV antibodies in the test vaccine groups with doses of 160 EU/dose and above were higher than those in the 80 EU/dose group(all P<0.05),while there was no statistically signifi-cant differences between the dose groups of 160 EU/dose and above(all P>0.05).The antibody positive conversion rate of 160 EU/dose and above of the test vaccine was 100.00%after the secondary immunization.Conclusions The inacti-vated hepatitis A vaccine of attenuated H2 strain tested in this study demonstrates strong immunogenicity in mice,sug-gesting its potential as a candidate vaccine.The preliminary analysis indicates an immunizing dose of 320 EU/dose for children and 640 EU/dose for adults.
关键词
H2减毒株/甲肝灭活疫苗/免疫原性/免疫剂量Key words
attenuated H2 strain/inactivated hepatitis A vaccine/immunogenicity/immunizing dose引用本文复制引用
基金项目
云南省科技重大专项(202002AA100009)
云南省科技重大专项(202302AA310003)
出版年
2024
NETL
NSTL
维普
万方数据