基于恒定链载体不同抗原表位串联嵌合体免疫原性分析  

Influence of series multiepitopes on immune enhancement mediated by invariant chain functional segments

作　　者：熊冉 艾珊珊 萧晟[1] XIONG Ran;AI Shan-shan;XIAO Sheng(Wuhu Institute of Technology,Wuhu,Anhui 241003,China)

机构地区：[1]芜湖职业技术学院

出　　处：《中国兽医学报》2019年第8期1460-1465,共6页Chinese Journal of Veterinary Science

基　　金：安徽省高校科学研究基金资助项目(KJ2018A0693);芜湖职业技术学院校级科研基金资助项目(Wzyzr201711);安徽省特色(品牌)专业动物防疫与检疫专业基金资助项目(2016tszy087)

摘　　要：为研究多抗原表位串联在恒定链(inviraint chain,Ii)功能片段载体增强免疫作用中的特性,自行设计一系列引物,克隆新城疫病毒HN和鸡传染性法氏囊病病毒VP2基因片段,用连接序列将其串联,并进一步连接小鼠Ii功能片段Cyt/TM,构建HN/VP2、Cyt/TM/VP2、Cyt/TM/HN、Cyt/TM/HN/VP2和Cyt/TM/VP2/HN共5个嵌合体,定向插入pET-32a原核表达载体,转化大肠杆菌诱导表达并纯化融合蛋白,分别免疫小鼠,经间接ELISA法测定血清抗体效价。结果显示,Ii功能片段可增强小鼠分泌特异性抗体,无论是Ii功能片段连接单一抗原表位,还是Ii功能片段连接多抗原表位串联,均比单用抗原表位免疫组提高抗体效价3倍以上,而多抗原表位串联不同试验组之间差异不明显。结果表明,Ii功能片段(Cyt/TM)具有增强免疫的作用,其连接的抗原表位串联不影响其免疫原性。To study the charactrism of the order in series multiepitopes linked with Ii(invariant chain) functional segments in enhancing immune response.A series of primers were designed.The genetic segments of NDV(Newcastle disease ivrus)-HN and IBDV(infectious bursal disease virus)-VP2 were cloned respectively and then the both segments were linked with the primers in different orders.Farther these segments were linked with Ii functinal segments(Cyt/TM) to construct five chimers,HN/VP2,Cyt/TM/VP2,Cyt/TM/HN,Cyt/TM/HN/VP2 and Cyt/TM/VP2/HN.These sequences were inserted into the prokaryotic expression vector pET-32 a or pGEX-4 T-1 and transformed into E.coli Rosetta(DE3) to induce the expression of the recombinant proteins.Finally mice were immunized with the purified proteins,and the specific antibody titers were detected with ELISA respectively.Ii functional segments,which were linked with single epitope as well as with series mutiepitopes,enhanced production of specific antibody.The antibody titers were three times higher than the control groups immunized with single epitopes.The antibody titers between different order of multiepitopes appeared no significant differences.The Ii functional segments showed evident immune enchancement effect,while the order of series multiepitopes in chimers had no influence on the immunogenicity.

关 键 词：恒定链载体 串联基因 免疫原性 新城疫病毒 鸡传染性法氏囊病病毒 HN VP2 

分 类 号：S852.4[农业科学—基础兽医学]