IBERS, Aberystwyth University, Aberystwyth, UK.
Centre for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands.
J Extracell Vesicles. 2015 Oct 5;4:28665. doi: 10.3402/jev.v4.28665. eCollection 2015.
Penetration of skin, migration through tissues and establishment of long-lived intravascular partners require Schistosoma parasites to successfully manipulate definitive host defences. While previous studies of larval schistosomula have postulated a function for excreted/secreted (E/S) products in initiating these host-modulatory events, the role of extracellular vesicles (EVs) has yet to be considered. Here, using preparatory ultracentrifugation as well as methodologies to globally analyse both proteins and small non-coding RNAs (sncRNAs), we conducted the first characterization of Schistosoma mansoni schistosomula EVs and their potential host-regulatory cargos.
Transmission electron microscopy analysis of EVs isolated from schistosomula in vitro cultures revealed the presence of numerous, 30-100 nm sized exosome-like vesicles. Proteomic analysis of these vesicles revealed a core set of 109 proteins, including homologs to those previously found enriched in other eukaryotic EVs, as well as hypothetical proteins of high abundance and currently unknown function. Characterization of E/S sncRNAs found within and outside of schistosomula EVs additionally identified the presence of potential gene-regulatory miRNAs (35 known and 170 potentially novel miRNAs) and tRNA-derived small RNAs (tsRNAs; nineteen 5' tsRNAs and fourteen 3' tsRNAs).
The identification of S. mansoni EVs and the combinatorial protein/sncRNA characterization of their cargo signifies that an important new participant in the complex biology underpinning schistosome/host interactions has now been discovered. Further work defining the role of these schistosomula EVs and the function/stability of intra- and extra-vesicular sncRNA components presents tremendous opportunities for developing novel schistosomiasis diagnostics or interventions.
为了成功地操纵终宿主的防御机制,血吸虫寄生虫需要穿透皮肤、迁移穿过组织并建立长期存在的血管内伙伴。虽然先前对幼虫毛蚴的研究假设分泌/排泄 (E/S) 产物在启动这些宿主调节事件中具有功能,但细胞外囊泡 (EV) 的作用尚未得到考虑。在这里,我们使用预备超速离心以及全面分析蛋白质和小非编码 RNA (sncRNA) 的方法,首次对曼氏血吸虫毛蚴 EV 及其潜在的宿主调节 cargos 进行了表征。
对体外培养的毛蚴 EV 进行透射电子显微镜分析,发现存在大量 30-100nm 大小的类外泌体囊泡。对这些囊泡进行的蛋白质组学分析揭示了一组核心 109 种蛋白质,包括与其他真核 EV 中富集的蛋白质同源的蛋白质,以及高丰度和目前未知功能的假想蛋白质。对毛蚴内外的 E/S sncRNA 的特征分析还鉴定了潜在的基因调节 miRNA(35 种已知和 170 种潜在新 miRNA)和 tRNA 衍生的小 RNA(tsRNA;19 种 5' tsRNA 和 14 种 3' tsRNA)。
曼氏血吸虫 EV 的鉴定以及对其 cargo 的组合蛋白质/sncRNA 特征表明,一种在血吸虫/宿主相互作用的复杂生物学中非常重要的新参与者现已被发现。进一步定义这些毛蚴 EV 的作用以及囊内外 sncRNA 成分的功能/稳定性,为开发新的血吸虫病诊断或干预方法提供了巨大的机会。