1型登革病毒感染病人血清中非结构蛋白1B细胞线性表位鉴定  

Identification of B-cell linear epitopes of nonstructural protein 1 from dengue virus type 1 infected patients

作　　者：林亚英 丁细霞[1] 温坤[1] 余楠[1] 任瑞文 陈月 LIN Ya-ying;DING Xi-xia;WEN Kun;YU Nan;REN Rui-wen;CHEN Yue(Department of Laboratory Medicine Zhujiang Hospital,Southern Medical University,Guangzhou 510282,China;Center for Disease Prevention and Control of the Southern Theatre Command of the PLA,Guangdong Arbovirus Diseases Emergency Technology Research Center,Guangzhou 510507,China)

机构地区：[1]南方医科大学珠江医院检验医学部,广州510282 [2]南部战区疾病预防控制中心,广东省虫媒病毒性传染疾病应急技术研究中心,广州510507

出　　处：《中国人兽共患病学报》2023年第7期639-644,共6页Chinese Journal of Zoonoses

基　　金：广东省基础与应用基础研究基金项目(No.2020A1515011171,2022A1515011132);全军实验动物专项(No.SYDW[2018]04)。

摘　　要：目的鉴定1型登革病毒感染病人血清中1型登革病毒非结构蛋白1(nonstructural Protein 1 of Dengue virus type 1,DENV-1 NS1)B细胞线性表位。方法以重组DENV-1 NS1为抗原,免疫印迹法筛选NS1阳性病人血清。一组重叠DENV-1 NS1的重叠多肽同这些NS1阳性血清反应鉴定NS1蛋白线性B细胞表位。采用梯度稀释的1-4型重组DENVNS1竞争抑制阳性多肽同病人血清反应鉴定其反应特异性和交叉反应性。结果12份DENV-1核酸阳性病人中筛选得到5份抗DENV-1 NS1阳性病人血清。重叠多肽法、竞争抑制法和生物信息学方法筛选出3条阳性NS1多肽(氨基酸残基序列第1-15,131-145,271-285),其中aa1-15在已报道的DENV-1分离株高度保守,提示这个区域可能存在DENV-1血清型特异性表位。aa131-145和aa271-285对应的蛋白序列比对分析表明以上区域中的133-FI/LIDGP-138和271-GKLEL/M/IDF-277在已报道的4种血清型DENV分离株中高度保守,提示这2个区域存在4种血清型DENV共同表位。结论发现3个NS1上B细胞线性表位,其中aa131-145和aa271-285为首次报道,结果有助于疫苗和诊断试剂的研发。Dengue virus nonstructural protein 1(DENV NS1),the major nonstructural protein thatinduces a protective antibody response,has potential in vaccinedevelopment.However,epitopes of DENV NS1,particularly B-cell linear epitopes in infected patients,have not been comprehensively mapped.We identified B-cell linear epitopes of DENV-1 NS1 in DENV-1 infected patients byusing serum from anti-NS1-positive patients.The serum was subjected to western blot analysiswith recombinant DENV-1 NS1 protein as an antigen.Epitope recognition analysis was performed with a set of 15-mer sequential overlapping peptides spanning the entire NS1 protein sequence of DENV-1.Among the 12 DENV-1 nucleic acid positive patients,five anti-DENV-1 NS1-positive sera were identified.Most of these five sera reacted with three regions of NS1:amino acid residues(aa)1-15,131-145,and 271-285.The serotype specificity and epitope cross-reactivity were further characterized by testing their reactivity with each virus serotype in competitivebinding assayswitha range of dilutions of 1-4 type recombinant DENV NS1.NS1 sequences in different flavivirus family members were compared by alignment of NS1 residues 1-15 from DENV-1,-2,-3,-4,WNV,YFV,and JEV.The alignment revealed that the amino acid sequence of peptide 1(aa1-15)was completely conserved among DENV-1 serotypes but differed among the flaviviruses,thus suggesting that the NS1 region was a DENV-1 serotype specific epitope.We also identified two group-specific B-cell epitopes that were highly conserved among isolates of the four DENV serotypes.Alignment of group-specific epitopes(aa131-145 and 271-285)with NS1 from DENV-1,-2,-3,and-4 isolates revealed that the amino acid sequences 133-FI/LIDGP-138 and 271-GKLEL/M/IDF-277 were similar among the serotypes,thus suggesting high conservation of these NS1 regions among the four DENV serotypes.Three linear B-cell epitopes on NS1 may aid in the development of vaccines and diagnostic reagents.

关 键 词：登革病毒 非结构蛋白1 重叠多肽 B细胞表位 

分 类 号：R373.3[医药卫生—病原生物学]