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Monoclonal Antibodies to Immunodominant Epitope of Tropheryma whipplei

机译:Whipplei鞭毛螺旋体免疫抗原决定簇的单克隆抗体

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Recent isolation of Tropheryma whipplei (formerly Trophyrema whippelii), the agent of Whipple’s disease, from the cardiac valve of a patient with Whipple’s disease endocarditis now allows the detection of reactive epitopes that could be used in a serological assay. In order to propose an enzyme-linked immunosorbent assay (ELISA) that uses recombinant T. whipplei antigen, we first determined by Western blotting of human, mouse, and rabbit antisera that the common immunodominant epitope is an 84-kDa protein. We then produced 13 monoclonal antibodies (MAbs) against T. whipplei, 12 of which recognize this immunodominant epitope. These MAbs did not react with phylogenetically closely related bacteria or bacteria previously shown to be cross-reactive with T. whipplei, but they did react with two other strains of T. whipplei isolated, one from an ocular sample and the other from a duodenal biopsy specimen. By confocal microscopy, the MAbs allowed detection of T. whipplei within infected fibroblasts. The identification of the 84-kDa antigen with our MAbs will make it possible to develop a diagnostic antigen for use in a diagnostic ELISA for Whipple’s disease.
机译:最近从患有Whipple病性心内膜炎的患者的心脏瓣膜中分离出Whipple病的病原体-Tropheryma whipplei (原称​​ Trophyrema whippelii ),现在可以检测出反应性抗原决定簇,可用于血清学检测。为了提出一种使用重组 T的酶联免疫吸附测定(ELISA)。首先通过人,小鼠和兔抗血清的Western印迹法确定了whipplei抗原,常见的免疫优势表位是84 kDa蛋白。然后,我们产生了13种针对 T的单克隆抗体(MAb)。 whipplei ,其中12个可以识别这种免疫优势表位。这些单克隆抗体不会与系统发育上密切相关的细菌或先前显示与 T交叉反应的细菌发生反应。 whipplei ,但它们确实与另外两个 T菌株发生了反应。 Whipplei 分离,一个来自眼样本,另一个来自十二指肠活检样本。通过共聚焦显微镜检查,单克隆抗体可检测出 T。被感染的成纤维细胞中的whipplei 。用我们的单克隆抗体鉴定84 kDa抗原将有可能开发出一种用于Whipple病诊断ELISA的诊断抗原。

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