甲型H7N9流感裂解疫苗制备及免疫保护效果的评价  

Preparation and immunoprotective efficacy evaluation of recombinant H7N9 influenza vaccines

作　　者：陈锐[1,2] 段跃强[2] 赵忠鹏[2] 张良艳[2] 邢丽[2] 刘娜[3] 张立 周娅[1] 王希良[2] 杨鹏辉[2] 

机构地区：[1]宁夏医科大学基础医学院病原生物学和免疫学教研室,银川750004 [2]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071 [3]解放军第302医院医疗部,北京100039 [4]总参警卫局卫生保健处,北京100071

出　　处：《免疫学杂志》2014年第7期608-611,共4页Immunological Journal

基　　金：国家"863"项目(SS2012AA020905)

摘　　要：目的构建、拯救重配甲型H7N9流感病毒疫苗候选株并制备甲型H7N9流感裂解疫苗,动物实验评价甲型H7N9流感裂解疫苗的免疫原性及免疫保护性效果。方法采用反向遗传学技术将A/Anhui/1/2013(H7N9)疫苗株的HA、NA基因和A/Puerto Rico/8/34(PR8)毒株的PB2、PB1、PA、NP、M、NS基因进行重配,转染细胞后筛选拯救甲型H7N9流感病毒疫苗候选株,制备rgPR8-H7N9流感裂解疫苗抗原。腹腔注射免疫Balb/c小鼠,检测血清IgG、IgG1、IgG2a、HI效价,进一步用野生株攻毒,评价rgPR8-H7N9流感裂解疫苗的免疫保护效果。结果成功拯救甲型H7N9流感病毒疫苗候选株rgPR8-H7N9。制备的重配甲型H7N9流感裂解疫苗对小鼠产生较高的HI抗体效价。IgG1/IgG2a亚型检测结果表明小鼠体内以诱导体液免疫为主。攻毒实验显示甲型H7N9流感裂解疫苗能够有效降低肺部的病毒载量,肺组织病变显著减轻、体质量下降后趋于稳定,疫苗剂量达到15μg即可全部存活。A/Anhui/1/2013(H7N9)野生株流感病毒攻毒,甲型H7N9流感裂解疫苗能够达到保护小鼠效果。结论成功拯救重配甲型H7N9流感病毒疫苗候选株rgPR8-H7N9,制备的甲型H7N9流感裂解疫苗具有较好的免疫原性及免疫保护性,为H7N9流感裂解疫苗的研发及进入临床研究提供了实验依据。On March 31, 20 13, the Centers for Disease Control and Prevention of China first reported 3 laboratory confirmed cases of human infection with influenza A subtype H7N9 virus, a new avian influenza virus （AIV） in the eastern China. As of January 19, 2014, a total of more than 200 cases of laboratory confirmed infection have been reported. Most reported cases have severe respiratory illness. Development a safe and effective H7N9 vaccine is urgently needed. In this study, we synthesized the haemagglutinin （HA） and neuraminidase （NA） gene sequence of A/Anhui/1/2013, and others from A/Puerto Rico/8/34（PR8） to recombine a H7N9 influenza vaccine rgPR8-H7N9. Then a formalin inactivated split vaccine was prepared from this virus and used to immunize mice. The immunogenicity of the formalin inactivated split vaccine was evaluated by measuring IgG, IgG1, IgG2a, and HI titers. Furthermore, the vaccines in 15 μg or more doses completely protected mice from subsequently lethal wildtype H7N9 virus challenge. Our study suggested that the success of recombinant A stubtye H7N9 influenza vaccine make it a promising candidate for further evaluation in human.

关 键 词：反向遗传学 H7N9流感病毒 裂解疫苗 免疫保护性 

分 类 号：R392[医药卫生—免疫学]