Research Center for Hydatid Disease in Iran, School of Medicine, Kerman University of Medical Sciences, Kerman, 7616914115, Iran.
Department of Parasitology, School of Medicine, Shahrekord University of Medical Sciences, Shahrekord, Iran.
Parasit Vectors. 2020 Apr 10;13(1):190. doi: 10.1186/s13071-020-04052-y.
Cystic echinococcosis, caused by the cestode Echinococcus granulosus, is a neglected tropical disease with remarkable morbidity in humans and a problem of worldwide economic importance in livestock industry. Understanding the molecular basis of the parasite growth and development is essential for the disease diagnosis, management and control. The tetraspanin (TSP) family of proteins are transmembrane proteins with a role in many physiological processes of eukaryotic organisms. TSPs present in the tegumental surface of platyhelminths play pivotal roles in host-parasite interaction. However, little is known about the role of TSPs in growth and development in the Platyhelminthes. To understand the role of TSP1 in the growth and development of E. granulosus we investigated the effect of EgTSP1-specific long dsRNA in different in vitro stages of the parasite.
Different stages of E. granulosus, protoscoleces and strobilated worms, were cultivated In vitro in di-phasic media. Using long dsRNA and two delivery methods, i.e. electroporation and electro-soaking, EgTSP1 silencing was performed with an EgTSP1-specific dsRNA. The TSP1 expression profile was assessed as well as the biological and ultrastructural properties of the parasites.
After three days of dsRNA treatment, EgTSP1 expression was significantly reduced in both stages of E. granulosus as compared to irrelevant/unrelated dsRNA and untreated controls. Silencing expression of EgTSP1 in different stages of E. granulosus resulted in reduced viability and body contractions, inhibition of protoscoleces evagination and distinctive tegumental changes. Ultrastructural morphology of the strobilated worms treated with EgTSP1-specific dsRNA was indicative of the microtriches impairments and vacuolated tegument compared to the control helminths.
Results of the present study suggest that EgTSP1 plays important structural roles in tegument configuration in E. granulosus. EgTSP1 is proved to be a potential target for the development of vaccines and RNAi-based drugs.
由带绦虫细粒棘球蚴引起的包虫病是一种被忽视的热带病，在人类中发病率很高，在畜牧业中也是一个具有全球经济重要性的问题。了解寄生虫生长和发育的分子基础对于疾病诊断、管理和控制至关重要。四跨膜蛋白（TSP）家族的蛋白是具有真核生物许多生理过程作用的跨膜蛋白。在扁形动物的表皮表面存在的 TSP 在宿主-寄生虫相互作用中起着关键作用。然而，对于 TSP 在扁形动物生长和发育中的作用知之甚少。为了了解 TSP1 在细粒棘球蚴生长和发育中的作用，我们研究了 EgTSP1 特异性长 dsRNA 在寄生虫不同体外阶段的影响。
用两相培养基在体外培养不同阶段的细粒棘球蚴，原头蚴和有钩绦虫。用 EgTSP1 特异性 dsRNA 通过电穿孔和电浸泡两种方法进行 EgTSP1 沉默。评估 TSP1 的表达谱以及寄生虫的生物学和超微结构特性。
与无关/dsRNA 和未处理对照相比，用 EgTSP1 特异性 dsRNA 处理三天后，两种阶段的细粒棘球蚴的 EgTSP1 表达均显著降低。在不同阶段的细粒棘球蚴中沉默 EgTSP1 的表达导致活力和身体收缩降低，原头蚴出芽抑制和独特的表皮变化。用 EgTSP1 特异性 dsRNA 处理的有钩绦虫的超微结构形态表明微纤毛损伤和空泡化的表皮与对照寄生虫相比。
本研究结果表明，EgTSP1 在细粒棘球蚴表皮结构中起重要结构作用。EgTSP1 被证明是疫苗和基于 RNAi 的药物开发的潜在靶标。
