Crouch M L, Benchimol M, Alderete J F
Department of Microbiology, The University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78284-7758, USA.
Microb Pathog. 2001 Sep;31(3):131-44. doi: 10.1006/mpat.2001.0455.
We have reported that Trichomonas vaginalis, the causative agent of the most common, non-viral sexually transmitted disease, bound to cover slips coated with fibronectin (FN) (Crouch & Alderete, Microbiol 1999 145: 2835-43). In this study, we extend that observation by showing that FN binding is specific, and we present data on the requirements of FN binding by T. vaginalis. Immunofluorescence and immuno-gold labelling readily detected FN throughout the trichomonal surface. Parasites bound to(125)I-labelled FN in a time- and concentration-dependent fashion. In the absence of protease inhibitor, iodinated FN was released from the trichomonad surface. Unlabelled FN specifically competed for binding in a concentration-dependent fashion with the(125)I-labelled FN. Interestingly, the amount of FN bound by T. vaginalis organisms was dependent on iron. High-iron-grown trichomonads acquired lower numbers of molecules but with 10-fold higher affinity than low-iron-grown organisms. Further, we show that for iron-replete organisms, calcium (Ca(2+)) at physiological levels increased amounts of bound FN. The increase in binding was rapid, occurring within 5 min of Ca(2+)addition, and required de novo protein synthesis. Co-incubation of live parasites with Ca(2+)in the presence of FN was necessary to increase the amount of FN bound. Treatment of trichomonads with okadaic acid, but not other phosphatase inhibitors, resulted in a 50% decrease in binding of FN, regardless of the presence of Ca(2+), suggesting a role for phosphatase in FN association. These results indicate that depending on the iron status of T. vaginalis organisms in vivo, Ca(2+)may influence trichomonad recognition and binding to FN during host parasitism.
我们已经报道过,最常见的非病毒性传播疾病的病原体阴道毛滴虫能够结合到包被有纤连蛋白(FN)的盖玻片上(克劳奇和阿尔德雷特,《微生物学》1999年,第145卷:2835 - 43页)。在本研究中,我们通过证明FN结合具有特异性来扩展这一观察结果,并给出阴道毛滴虫结合FN的相关需求数据。免疫荧光和免疫金标记很容易在整个滴虫表面检测到FN。寄生虫以时间和浓度依赖的方式结合到(125)I标记的FN上。在没有蛋白酶抑制剂的情况下,碘化FN从滴虫表面释放出来。未标记的FN以浓度依赖的方式与(125)I标记的FN特异性竞争结合。有趣的是,阴道毛滴虫结合的FN量取决于铁。高铁培养的滴虫获得的分子数量较少,但亲和力比低铁培养的生物体高10倍。此外,我们表明对于铁充足的生物体,生理水平的钙(Ca(2 +))会增加结合的FN量。结合的增加很快,在添加Ca(2 +)后5分钟内就会发生,并且需要从头合成蛋白质。在有FN存在的情况下,将活寄生虫与Ca(2 +)共同孵育对于增加结合的FN量是必要的。用冈田酸处理滴虫,但不是其他磷酸酶抑制剂,无论是否存在Ca(2 +),都会导致FN结合减少50%,这表明磷酸酶在FN结合中起作用。这些结果表明,根据体内阴道毛滴虫生物体的铁状态,Ca(2 +)可能在宿主寄生期间影响滴虫对FN的识别和结合。