Mimotopes selected with a neutralizing antibody against urease B from Helicobacter pylori induce enzyme inhibitory antibodies in mice upon vaccination.
Mimotopes selected with a neutralizing antibody against urease B from Helicobacter pylori induce enzyme inhibitory antibodies in mice upon vaccination.
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用针对幽门螺杆菌脲酶 B 的中和抗体选择的模拟表位在接种疫苗后在小鼠中诱导酶抑制抗体
DOI：
10.1186/1472-6750-10-84
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发表时间：
2010-11-30
影响因子：
3.5
通讯作者：
Li M
中科院分区：
文献类型：
--
作者：
Li Y;Ning Y;Wang Y;Peng D;Jiang Y;Zhang L;Long M;Luo J;Li M
背景：尿素酶B是幽门螺杆菌定植于胃粘膜所必需的重要毒力因子。抑制尿素酶B酶活性的鼠单抗将作为疫苗用于预防和治疗幽门螺杆菌感染。在这里，我们生产了针对尿素酶B的小鼠单抗，它可以中和该酶的活性。利用噬菌体展示文库对其表位进行了定位，并对所选模拟表位进行了体内免疫原性研究。DQ141576)，重组pGEX-4T-1/UreeB在大肠杆菌中以92 kDa的形式与谷胱甘肽-S转移酶(GST)融合表达。以尿素酶B-GST为免疫原，采用杂交瘤技术制备了5株单抗U001-U005。只有U001能抑制尿素酶B的活性。通过噬菌体展示文库的免疫筛选，发现了两个不同的尿素酶B蛋白模拟表位：来自ph.D.12-库的EXXXHDM和来自ph.D.C7C的EXXXHSM，它们在347-353aa与尿素酶B蛋白相匹配。经筛选的噬菌体克隆诱导的抗血清可识别尿素酶B蛋白并抑制其酶活性，表明噬菌体表位诱导的免疫反应具有抗原特异性。结论：噬菌体展示模拟表位可与小鼠免疫系统结合并触发体液反应。尿素酶B模拟表位为幽门螺杆菌感染的诊断和治疗疫苗的研制提供了一种新的、有希望的途径。
Background:Urease B is an important virulence factor that is required for Helicobacter pylori to colonise the gastric mucosa. Mouse monoclonal antibodies (mAbs) that inhibit urease B enzymatic activity will be useful as vaccines for the prevention and treatment of H. pylori infection. Here, we produced murine mAbs against urease B that neutralize the enzyme's activity. We mapped their epitopes by phage display libraries and investigated the immunogenicity of the selected mimotopes in vivo.Results:The urease B gene was obtained (GenBank accession No. DQ141576) and the recombinant pGEX-4T-1/UreaseB protein was expressed in Escherichia coli as a 92-kDa recombinant fusion protein with glutathione-S-transferase (GST). Five mAbs U001-U005 were produced by a hybridoma-based technique with urease B-GST as an immunogen. Only U001 could inhibit urease B enzymatic activity. Immunoscreening via phage display libraries revealed two different mimotopes of urease B protein; EXXXHDM from ph.D.12-library and EXXXHSM from ph.D.C7C that matched the urease B proteins at 347-353 aa. The antiserum induced by selected phage clones clearly recognised the urease B protein and inhibited its enzymatic activity, which indicated that the phagotope-induced immune responses were antigen specific.Conclusions:The present work demonstrated that phage-displayed mimotopes were accessible to the mouse immune system and triggered a humoral response. The urease B mimotope could provide a novel and promising approach for the development of a vaccine for the diagnosis and treatment of H. pylori infection.
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影响因子：
46.9
作者：
De Berardinis, P;Sartorius, R;Guardiola, J
通讯作者：
Guardiola, J
影响因子：
46.9
作者：
Agadjanyan, M;Luo, P;KieberEmmons, T
通讯作者：
KieberEmmons, T
DOI：
10.2165/00063030-200721030-00002
发表时间：
2007
期刊：
BioDrugs : clinical immunotherapeutics, biopharmaceuticals and gene therapy
影响因子：
--
作者：
Gershoni JM;Roitburd-Berman A;Siman-Tov DD;Tarnovitski Freund N;Weiss Y
通讯作者：
Weiss Y
影响因子：
3.7
作者：
Bastien, N;Trudel, M;Simard, C
通讯作者：
Simard, C
影响因子：
3.1
作者：
Larralde, Osmany G.;Martinez, Raiza;Perez, Ela M.
通讯作者：
Perez, Ela M.