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首页> 外文期刊>Parasitology >Human recombinant antibodies against Trypanosoma cruzi ribosomal P2β protein
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Human recombinant antibodies against Trypanosoma cruzi ribosomal P2β protein

机译：抗克氏锥虫核糖体P2β蛋白的人重组抗体

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Patients with chronic Chagas' Heart Disease (cChHD) develop an antibody response that is suspected to be involved in the cardiac pathogenesis. The response against Trypanosoma cruzi ribosomal P proteins is of particular interest, as these antibodies can cross-react with host cardiac receptors causing electrophysiological alterations. To better understand the humoral anti-P response we constructed a single-chain variable fragment library derived from a cChHD patient. The variable heavy and light regions were amplified from bone-marrow RNA and subcloned into the vector pComb3X. The phage library was subsequently panned against T. cruzi ribosomal P2β protein (TcP2β). We obtained 3 different human recombinant antibodies that specifically reacted with TcP2β in ELISA and Western blots. Two of them reacted with the C-terminal region of TcP2β, peptide R13, as the recombinant autoanti-P antibodies from Systemic Lupus Erythematosus (SLE) patients. Interestingly, the third one was specific for TcP2β but did not recognize R13, confirming the specific nature of the anti-P response in Chagas disease. Neither sequence nor VH usage similarities between Chagas and SLE anti-P autoantibodies were observed. Herein, the first human mAbs against TcP2β have been obtained and characterized showing that the humoral anti-P response is directed against the parasite and does not include an autoimmune component.
机译：患有慢性恰加斯氏心脏病（cChHD）的患者产生抗体应答，该抗体应答被怀疑与心脏发病机制有关。对克鲁斯锥虫核糖体P蛋白的反应特别受关注，因为这些抗体可与宿主心脏受体发生交叉反应，从而引起电生理改变。为了更好地理解体液抗P反应，我们构建了一个来自cChHD患者的单链可变片段文库。从骨髓RNA扩增出可变的重和轻区，并将其亚克隆到载体pComb3X中。噬菌体文库随后针对克鲁斯氏锥虫核糖体P2β蛋白（TcP2β）进行淘选。我们获得了3种不同的人类重组抗体，它们在ELISA和Western印迹中与TcP2β特异性反应。它们中的两个与TcP2β的C末端区域（肽R13）反应，作为系统性红斑狼疮（SLE）患者的重组自身抗P抗体。有趣的是，第三个是特异于TcP2β的，但不识别R13，证实了Chagas病中抗P反应的特异性。 Chagas和SLE抗P自身抗体之间没有序列或VH使用相似性。在本文中，已经获得了针对TcP2β的第一批人mAb，其特征在于表明体液抗P应答是针对寄生虫的，并且不包括自身免疫成分。

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