第2.3.2.1分支H5亚型禽流感变异株候选疫苗株的构建及免疫效力试验

蒋文明，李金平，侯广宇，王素春，袁万哲，孙文博，彭程，陈继明; Jiang Wenming，Li Jinping，Hou Guangyu，Wang Suchun，Yuan Wanzhe，Sun Wenbo，Peng Cheng，Chen Jiming

首页 > 过刊浏览>2017年第34卷第9期 >79-82. DOI:10.3969/j.issn.1005-944X.2017.09.022

第2.3.2.1分支H5亚型禽流感变异株候选疫苗株的构建及免疫效力试验
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                        10.3969/j.issn.1005-944X.2017.09.022
                    
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中图分类号:S852.65
基金项目:科技部科技基础性工作专项（2012FY111000）；山东省博士基金（BS2013SW022）

Generation and Protective Efficacy of Candidate Vaccine Virus against Clade2.3.2.1 H5 Avian Influenza Variant

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摘要:

第2.3.2.1分支H5N1亚型禽流感病毒（AIV）抗原性目前已发生显著变异。为应对变异株可能引发的流行，利用反向遗传技术，以鸡胚适应毒A/Puerto Rico/8/34（PR8）的内部基因为骨架，以H5N1亚型AIV A/duck/JS/ZJ/2016（H5N1）（ZJ，第2.3.2.1d分支）、A/Chicken/SD/YT/2015（H5N1）（YT，第2.3.2.1e分支）的基因组为模板，扩增其HA及NA基因，并对HA基因进行修饰，去除裂解位点处的多个碱性氨基酸，使其获得低致病性AIV的分子特征，成功构建了第2.3.2.1分支H5N1亚型AIV疫苗候选株rZJ和rYT。抗原性分析显示，rZJ和rYT与Re-6疫苗株之间的抗原性已有显著差异。免疫效力试验表明，rZJ和rYT重组灭活疫苗免疫21 d的平均HI抗体效价分别达到8.8 log2和8.9 log2，说明重组疫苗具备良好的免疫原性。免疫攻毒保护试验表明，rZJ和rYT重组灭活疫苗可以提供SPF鸡抵抗同源H5N1病毒100%的保护，而针对第2.3.2.1分支的Re-6疫苗仅能提供大约40%和30%的保护。利用反向遗传技术构建的rZJ和rYT重组候选疫苗株为该分支病毒的防控提供了技术支持。

Abstract:

The significant antigenic changes of clade 2.3.2.1 H5 subtype of Avian influenza virus（AIV） have occurred. To control the potential pandemic threat of variations to poultry，in this study，two reassortant viruses，named rZJ and rYT，were generated using reverse genetics. The viruses contained modified HA and NA genes of epidemic strains A/duck/JS/ZJ/2016（H5N1）and A/Chicken/SD/YT/2015（H5N1）respectively，with the internal genes derived from A/Puerto Rico/8/34（PR8）. The reassortant candidate vaccine viruses was attenuated by removal of the multi-basic amino acid motif in the HA gene by mutation and deletion. Antigenicity analysis showed that there was significant difference between rZJ and rYT，compared with Re-6 vaccine strain. Immune efficacy tests showed that the HI antibody titers could reach 8.8 log2 and 8.9 log2，respectively，after 21 days from immunization of rZJ and rYT inactivated vaccines，showing a good immunogenicity. Immune challenge tests showed that rZJ and rYT recombinant inactivated vaccines could provide SPF chickens with 100% of the protection against homologous H5N1 viruses，and Re-6 vaccine against clade 2.3.2.1 viruses could only provide about 40% and 30% of the protection，respectively. And rZJ and rYT recombinant vaccine candidate strains using reverse genetics technology provided technical support for prevention and control of the viruses of clade 2.3.2.1.