重组动物疱疹病毒活载体疫苗的构建与应用研究进展

Research Progress on Construction and Application of Recombinant Animal Herpesvirus Live Vector Vaccine

重组病毒活载体疫苗是通过基因工程方法构建的活载体疫苗,规避了传统疫苗的一些缺点,具有极大的优势和应用前景。目前被广泛用作重组病毒载体的动物疱疹病毒有火鸡疱疹病毒(Herpesvirus of turkey,HVT)、伪狂犬病病毒(Pseudorabies virus,PRV)、鸭瘟病毒(Duck enteritis virus,DEV)、鸡传染性喉气管炎病毒(Infectious laryngotracheitis virus,ILTV)等。对疱疹病毒基因组序列进行分析显示,其基因组大,复制非必需区域多,容纳外源基因的能力强。同时疱疹病毒具有宿主范围广泛、免疫持续期长、安全性较好及相关基因操作技术成熟等优点。笔者总结了当前构建重组动物疱疹病毒的主要方法,包括传统同源重组技术、细菌人工染色体(BAC)技术、Fosmid文库及CRISPR/Cas9基因编辑技术,简述了各个方法的基本原理及技术特点,比较了各个方法的优缺点,并分析了各个方法的最适运用条件,从而更准确地为研究工作提供理论及技术参考;对外源基因的表达方式进行分析,论证了不同启动子对外源基因表达的调控作用不同,同时对不同疱疹病毒的常用插入位点进行了研究成果的列举总结,并对当前重组病毒HVT、PRV、DEV及ILTV活载体疫苗的研究进展进行总结,对相关生物制品的注册应用信息进行了归纳,介绍了高选择率外源基因及高成功率插入位点,从而为相关生物制品的研制提供理论参考。

Recombinant virus live vector vaccines are live vector vaccines constructed through genetic engineering methods,avoiding some of the shortcomings of traditional vaccines,and have great advantages and application prospects.Animal Herpesviruses currently widely used as recombinant virus vectors include Herpesvirus of turkey(HVT),Pseudorabies virus(PRV),Duck enteritis virus(DEV),and avian Infectious laryngotracheitis virus(ILTV),etc.Analysis of the Herpesvirus genome sequence shows that its genome is large,there are many unnecessary regions for replication,and the ability to accommodate exogenous gene is strong.At the same time,the Herpesvirus has the advantages of wide host range,long duration of immunity,good safety and mature related gene manipulation technology.This article summarized the main methods of constructing recombinant animal Herpesviruses,including traditional homologous recombination technology,bacterial artificial chromosome technology(BAC),Fosmid library and CRISPR/Cas9 gene editing technology,and briefly described the basic principles and technical characteristics of each method,compared the advantages and disadvantages of each method,analyzed the optimal application conditions of each method,so as to provide more accurate theoretical and technical references for research work.The expression of exogenous genes were analyzed,and examples were given to demonstrate the different regulatory effects of different promoters on the expression of exogenous genes.At the same time,the common insertion sites of different Herpesviruses were listed and summarized.The research progress and the registration application information of related biological products of the current recombinant viruses HVT,PRV,DEV and ILTV live vector vaccines were summarized,and the high-selectivity exogenous genes and high-success rate insertion sites were induced,so as to provide a theoretical reference for the development of related biological products.