Barbour A G, Tessier S L, Todd W J
Infect Immun. 1983 Aug;41(2):795-804. doi: 10.1128/iai.41.2.795-804.1983.
Ixodid tick-associated spirochetes have been implicated as the etiological agents of Lyme disease. We raised a murine monoclonal antibody (H5332) against a spirochete, strain B31, isolated from Ixodes dammini ticks. In indirect immunofluorescence assays and western blot analyses, H5332 reacted with whole cells or isolated components of not only strain B31 but also spirochetes isolated from Ixodes ricinus ticks, a field mouse, a raccoon, and patients with Lyme disease. In contrast, H5332 did not bind to representative borreliae, treponemes, and leptospires. Using indirect immunofluorescence assays and immune electron microscopy, we found the H5332 determinant to be diffusely distributed over the surface of prefixed spirochetes but to be aggregated in patches when the organisms were incubated with H5332 and a second ligand before fixation. Radioimmunoprecipitation and western blot studies revealed the H5332 determinant to be either on or tightly associated with an abundant outer membrane protein with an apparent subunit molecular weight of 31,000.
硬蜱相关螺旋体被认为是莱姆病的病原体。我们制备了一种针对从达敏硬蜱分离出的螺旋体菌株B31的鼠单克隆抗体(H5332)。在间接免疫荧光试验和蛋白质印迹分析中,H5332不仅与菌株B31的全细胞或分离成分发生反应,还与从蓖麻硬蜱、一只田鼠、一只浣熊以及莱姆病患者分离出的螺旋体发生反应。相比之下,H5332不与代表性的疏螺旋体、密螺旋体和钩端螺旋体结合。通过间接免疫荧光试验和免疫电子显微镜,我们发现H5332决定簇在固定前的螺旋体表面呈弥散分布,但当生物体在固定前与H5332和第二种配体一起孵育时,会聚集形成斑块。放射免疫沉淀和蛋白质印迹研究表明,H5332决定簇位于一种明显亚基分子量为31000的丰富外膜蛋白上或与之紧密相关。