Stroud Leah J, Šlapeta Jan, Padula Matthew P, Druery Dylan, Tsiotsioras George, Coorssen Jens R, Stack Colin M
School of Science and Health, Western Sydney University, Campbelltown Campus, Penrith, NSW 2751, Australia.
Faculty of Veterinary Science, School of Life and Environmental Sciences, University of Sydney, NSW 2006, Australia.
Int J Parasitol. 2017 Mar;47(4):203-213. doi: 10.1016/j.ijpara.2016.11.004. Epub 2017 Jan 17.
Certain clinical isolates of Tritrichomonas foetus infect the urogenital tract of cattle while others infect the gastrointestinal tract of cats. Previous studies have identified subtle genetic differences between these isolates with the term "genotype" adopted to reflect host origin. The aim of this work was to seek evidence of host-specific adaptation and to clarify the relationship between T. foetus genotypes. To do this we characterised the proteomes of both genotypes using two-dimensional gel electrophoresis (2DE) coupled with LC-MS/MS. Our comparative analysis of the data revealed that both genotypes exhibited largely similar proteoform profiles; however differentiation was possible with 24 spots identified as having a four-fold or greater change. Deeper analysis using 2DE zymography and protease-specific fluorogenic substrates revealed marked differences in cysteine protease (CP) expression profiles between the two genotypes. These variances in CP activities could also account for the pathogenic and histopathological differences previously observed between T. foetus genotypes in cross-infection studies. Our findings highlight the importance of CPs as major determinants of parasite virulence and provide a foundation for future host-parasite interaction studies, with direct implications for the development of vaccines or drugs targeting T. foetus.
胎儿三毛滴虫的某些临床分离株感染牛的泌尿生殖道，而其他分离株感染猫的胃肠道。先前的研究已经确定了这些分离株之间的细微遗传差异，采用“基因型”一词来反映宿主来源。这项工作的目的是寻找宿主特异性适应的证据，并阐明胎儿三毛滴虫基因型之间的关系。为此，我们使用二维凝胶电泳（2DE）结合液相色谱-串联质谱（LC-MS/MS）对两种基因型的蛋白质组进行了表征。我们对数据的比较分析表明，两种基因型的蛋白质变体谱在很大程度上相似；然而，通过鉴定出24个斑点的变化达到四倍或更大，可以实现区分。使用2DE酶谱分析和蛋白酶特异性荧光底物进行的深入分析揭示了两种基因型之间半胱氨酸蛋白酶（CP）表达谱的显著差异。CP活性的这些差异也可以解释先前在交叉感染研究中观察到的胎儿三毛滴虫基因型之间的致病性和组织病理学差异。我们的研究结果突出了CP作为寄生虫毒力主要决定因素的重要性，并为未来的宿主-寄生虫相互作用研究奠定了基础，对开发针对胎儿三毛滴虫的疫苗或药物具有直接意义。
