基于黏附素HpaA B细胞表位的多抗原肽疫苗的免疫原性鉴定  

Preparing a multi-antigen peptide vaccine based on B-cell epitope of Helicobacter pylori adhesin HpaA and determining its immunogenicity

作　　者：孙艳 李瑞芳 格桑卓玛 冯明旨 张骏 SUN Yan;LI Ruifang;KELSANG Drolma;FENG Mingzhi;ZHANG Jun(Department of Gastroenterology,Zhejiang Provincial People's Hospital(Affiliated People's Hospital of Hangzhou Medical College),Hangzhou 310014,China)

机构地区：[1]浙江省人民医院(杭州医学院附属人民医院)消化内科,杭州310014 [2]浙江中医药大学附属第一医院(浙江省中医院)消化内科

出　　处：《浙江医学》2024年第15期1573-1579,I0003,共8页Zhejiang Medical Journal

基　　金：浙江省医药卫生重大科技计划项目(WKJ-ZJ-2018);浙江省中医药科技计划项目(2023ZL266)。

摘　　要：目的制备以幽门螺杆菌(Hp)黏附素HpaA B细胞表位为基础的多抗原肽(MAP)疫苗,并分析其免疫原性。方法在GenBank数据库检索Hp黏附素HpaA的氨基酸及核苷酸序列。采用表位分析软件预测HpaA的B细胞表位,以此合成8分支MAP并免疫白毛黑眼兔,体外试验测定MAP与兔抗Hp多克隆抗体的亲和力,体内试验检测免疫血清抗体效价和抗体的特异性。结果HpaA的优势B细胞表位可能位于其氨基酸序列的第130~142和第212~219区段,并分别以上述B细胞表位合成8分支MAP1和MAP2。MAP1和MAP2均能在体外与兔抗Hp多克隆抗体结合,且MAP1具有较高的亲和力。仅MAP1能在体内诱导免疫应答,产生高滴度特异性抗体。结论HpaA氨基酸序列的第130~142区段为其优势B细胞表位,基于此合成的MAP具有较强的免疫原性。Objective To prepare a multiple antigenic polypetide(MAP)vaccine based on the B-cell epitope of Helicobacter pylori(Hp)adhesin HpaA and to determine its immunogenicity.Methods Amino acid and nucleotide sequences of HpaA were obtained from GenBank.Epitope analysis softwares were utilized for HpaA B-cell epitope prediction.An 8-branch MAP was synthesized and used to immunize white-haired black eye(WHBE)rabbits.The affinity of MAP and rabbit anti-Hp polyclonal antibodies in vitro was determined,and the serum antibody titer and antibody specificity in vivo were detected.Results The dominant B cell epitopes of HpaA were located in the 130-142 and 212-219 amino acid segments,the 8-branched MAP1 and MAP2 were synthesized according to above B cell epitopes.Both MAP1 and MAP2 showed binding ability to rabbit anti-Hp polyclonal antibodies in vitro,while MAP1 exhibited higher affinity.Only MAP1 induced immune response and produced high-titer specific antibody in vivo.Conclusion The dominant B-cell epitope is located in 130-142 amino acid segment of HpaA,the corresponding MAP was synthesized and the vaccine was prepared in the study,which demonstrates strong immunogenicity.

关 键 词：幽门螺杆菌 黏附素HpaA B细胞表位 多抗原肽 免疫原性 

分 类 号：R392-33[医药卫生—免疫学]