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首页> 外文期刊>Infection and immunity >Monoclonal antibodies with an expanded repertoire of specificities and potent neutralizing activity for Escherichia coli heat-labile enterotoxin.
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Monoclonal antibodies with an expanded repertoire of specificities and potent neutralizing activity for Escherichia coli heat-labile enterotoxin.

机译：具有扩展的全部库的特异性和对大肠杆菌热不稳定肠毒素的有效中和活性的单克隆抗体。

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Nine selected hybridoma cell lines that produced monoclonal antibodies against the heat-labile enterotoxin encoded by a plasmid from an Escherichia coli strain of human origin (LTh) were characterized. Hybridomas that produced anti-LTh antibodies with previously unrecognized specificities or reactivities were selected for cloning. Each monoclonal antibody was tested for isotype and for binding to LTh holotoxin, the A and B subunits derived from LTh (LTh-A and LTh-B), holotoxin encoded by a plasmid from an E. coli strain of porcine origin (LTp), and cholera enterotoxin (CT). Binding was also tested in Western blots with the following antigens: pentameric LTh-B, monomeric LTh-B, LTh-A, and the A1 and A2 fragments produced from LTh-A by treatment with trypsin. These monoclonal anti-LTh antibodies and selected anti-LTh and anti-CT monoclonal antibodies described previously were tested for neutralization of LTh, LTp, and CT. Five of the nine new monoclonal antibodies gave detectable cross-reactions with LTp and CT. Four reacted with determinants of LTh that were not present on CT; one of these four did not react with LTp and was specific for a unique epitope of LTh. Three antibodies were specific for LTh-B. All three reacted with pentameric LTh-B, but only one reacted in Western blots with monomeric LTh-B. Six antibodies were specific for LTh-A. Three reacted in Western blots with LTh-A and its A1 fragment; the other three did not react in Western blots. All nine of the new monoclonal antibodies neutralized LTh but not CT; the eight that cross-reacted with LTp in binding assays also neutralized LTp. Of four neutralizing anti-CT monoclonal antibodies that bound to LTh, none had significant neutralizing activity against LTh.
机译：表征了九种选择的杂交瘤细胞系，它们产生了抗热不稳定肠毒素的单克隆抗体，该单克隆抗体是由人类起源的大肠埃希氏菌菌株（LTh）的质粒编码的。选择产生具有先前无法识别的特异性或反应性的抗LTh抗体的杂交瘤进行克隆。测试了每种单克隆抗体的同种型以及与LTh全息毒素，源自LTh的A和B亚基（LTh-A和LTh-B）的结合，由猪源大肠杆菌菌株（LTp）的质粒编码的全毒素，和霍乱肠毒素（CT）。还用以下抗原在Western印迹中测试了结合：五聚体LTh-B，单体LTh-B，LTh-A以及通过胰蛋白酶处理从LTh-A产生的A1和A2片段。测试了这些单克隆抗LTh抗体以及先前所述的选定抗LTh和抗CT单克隆抗体对LTh，LTp和CT的中和作用。九种新的单克隆抗体中有五种与LTp和CT发生了可检测到的交叉反应。四个反应与CT中不存在的LTh决定簇反应;这四个中的一个不与LTp反应，并且对LTh的独特表位具有特异性。三种抗体对LTh-B具有特异性。所有三个都与五聚体LTh-B反应，但只有一个在Western blot中与单体LTh-B反应。六种抗体对LTh-A具有特异性。 3个在Western blot中与LTh-A及其A1片段反应;其他三个在Western blot中没有反应。所有九种新的单克隆抗体均中和了LTh，但未中和CT。在结合测定中与LTp交叉反应的8个也中和了LTp。在与LTh结合的四种中和性抗CT单克隆抗体中，没有一种对LTh具有明显的中和活性。

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