Molecular Mechanisms of Trichomonas Infection

阴道毛滴虫是滴虫病的病原体，滴虫病是人类最广泛的非病毒性性传播疾病。感染包括缺氧氧的生态位、宿主细胞识别、铁离子的隔离、寄生虫与宿主组织接触后从鞭毛虫到变形虫的形态发生、对甲硝唑的抗性以及毛滴虫的厌氧线粒体氢酶体的能量代谢和蛋白质输入。目前关于滴虫感染的分子过程和机制的知识是有限的。特别是感染的非常早期阶段和宿主-寄生虫相互作用在很大程度上没有特征。在这里，我们提出了一个协调一致的努力，两个领先的欧洲实验室滴虫生物学工作，以揭示由阴道滴虫感染人类泌尿生殖道组织的分子过程。我们提案的前景问题包括i)寄生虫粘附的性质，ii)感染过程中核心碳和能量代谢的变化，以及iii)寄生虫与宿主组织接触时的转录组反应。我们的方法需要对早期感染期间的转录组进行深度测序，并结合与氢基因体输入和功能相关的单个组分的蛋白质组学和生化表征。为此，我们成功地将感染性阴道绦虫菌株T016与人阴道上皮细胞系MS74结合，建立了稳定的体外感染系统。该系统不仅帮助我们产生了丰富的关于感染过程的新颖初步数据来源，而且还为研究氢化基因体提供了新的方法，氢化基因体是甲硝唑治疗的靶点。

Trichomonas vaginalis is the causative agent of Trichomoniasis, the most widespread non-viral sexually transmitted disease in humans. Infection involves O2-poor niches, host cell recognition, Fe2+-sequestration, morphogenesis of the parasite from flagellate to amoeboid form upon contact with host tissue, resistance to metronidazole treatment, as well as energy metabolism in and protein import into hydrogenosomes, the anaerobic mitochondria of trichomonads. Current knowledge about the molecular processes and mechanisms of Trichomonas infection is limited. In particular the very early stages of infection and host-parasite interactions are largely uncharacterized. Here we propose a concerted effort of two leading European laboratories working on trichomonad biology to uncover the molecular processes underlying infection by Trichomonas vaginalis of human urogenital tract tissue. The questions in the foreground of our proposal include i) the nature of parasite adherence, ii) changes in core carbon and energy metabolism during infection, and iii) the parasites transcriptomic response upon contact with host tissue. Our approach entails deep-sequencing of transcriptomes during early infection, combined with proteomics and biochemical characterization of individual components associated with hydrogenosomal import and function. To this end we have successfully established a stable in vitro infection system by combining the infectious T. vaginalis strain T016 and the human vaginal epithelial cell line MS74. The system has helped us not only to generate a rich source of novel preliminary data regarding the infection process, but furthermore allows new approaches to study hydrogenosomes, which harbor the target of metronidazole treatment.