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首页> 外文期刊>Infection and immunity >Generation and Characterization of Human Monoclonal scFv Antibodies against Helicobacter pylori Antigens
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Generation and Characterization of Human Monoclonal scFv Antibodies against Helicobacter pylori Antigens

机译：抗幽门螺杆菌抗原的人单克隆scFv抗体的产生与表征

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Infection with Helicobacter pylori is chronic despite a vigorous cellular and humoral immune response and causes severe pathology in some patients. In this study, phage display was used as a new approach in order to investigate the role of the host's humoral immune response in the pathogenesis of H. pylori gastritis. Human monoclonal single-chain Fv (scFv) antibody fragments against H. pylori cell lysate and the H. pylori urease were isolated from an immune phage display library, constructed from peripheral blood lymphocytes of an H. pylori-infected patient. After affinity selection, 23% of the clones tested showed binding activity against a lysate of the H. pylori Sydney strain in enzyme-linked immunosorbent assay (ELISA) and 9% bound the H. pylori urease. Further characterization by PCR-fingerprint analysis and sequencing revealed that two closely related H. pylori binders and one antiurease scFv could be isolated. The selected scFvs were highly specific as analyzed by ELISA and immunoblots using various bacterial lysates and recombinant proteins. Analysis of the humoral immune response following H. pylori infection using human monoclonal antibodies might contribute to a better understanding of the pathogenesis of the disease. Moreover, using immune phage display libraries, it might be possible for relevant epitopes of H. pylori antigens to be determined, which might be of use for vaccine development.
机译：尽管有强烈的细胞和体液免疫反应，幽门螺杆菌仍是慢性感染，并在某些患者中引起严重的病理。在这项研究中，噬菌体展示被用作一种新方法，以研究宿主的体液免疫应答在 H发病机理中的作用。幽门螺杆菌针对 H的人单克隆单链Fv（scFv）抗体片段。幽门螺杆菌细胞裂解液和 H。从免疫噬菌体展示文库中分离出幽门螺杆菌脲酶，该文库由 H的外周血淋巴细胞构建。幽门螺杆菌感染的患者亲和力选择后，测试的克隆中有23％表现出针对 H裂解物的结合活性。酶联免疫吸附试验（ELISA）中的pylori Sydney菌株，其中9％与 H结合。幽门螺杆菌脲酶通过PCR指纹分析和测序进一步表征，发现两个密切相关的 H。可以分离出幽门螺杆菌结合剂和一种抗脲酶scFv。如通过ELISA和使用各种细菌裂解物和重组蛋白进行的免疫印迹分析，所选择的scFv具有高度特异性。 H后的体液免疫反应分析。使用人类单克隆抗体感染幽门螺杆菌可能有助于更好地了解该病的发病机理。而且，使用免疫噬菌体展示文库，可能存在 H的相关表位。待确定的幽门螺杆菌抗原，可能用于疫苗开发。

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