Stryiński Robert, Fiedorowicz Ewa, Mateos Jesús, Andronowska Aneta, Łopieńska-Biernat Elżbieta, Carrera Mónica
Department of Biochemistry, Faculty of Biology and Biotechnology, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland.
Department of Food Technology, Institute of Marine Research, Spanish National Research Council, Vigo, Spain.
Front Immunol. 2025 Feb 3;16:1509984. doi: 10.3389/fimmu.2025.1509984. eCollection 2025.
Anisakis simplex sensu stricto (s. s.) is one of the most widespread parasitic nematodes of marine organisms, with humans as accidental hosts. While many studies have explored nematode biology and host interactions, the role of extracellular vesicles (EVs) as signaling molecules in parasitic nematodes is less understood.
Therefore, the proteins present in the EVs of A. simplex (s. s.) (Anis-EVs) were identified. In addition, a cross-talk proteomic approach was used to identify differentially regulated proteins (DRPs) in the proteome of the human intestinal epithelial cell line (Caco-2) co-cultured with L3 larvae of A. simplex (s. s.) or directly exposed to two concentrations (low or high) of Anis-EVs. In addition, DRPs were identified in the proteome of A. simplex (s. s.) larvae affected by co-culture with Caco-2. To achieve this goal, the shotgun proteomics method based on isobaric mass labeling (via tandem mass tags; TMT) was used with a combination of nano high-performance liquid chromatography (nLC) coupled with an LTQ-Orbitrap Elite mass spectrometer. In addition, ELISA assays were used to demonstrate if Caco-2 respond to A. simplex (s. s.) larvae and Anis-EVs with significant changes in selected cytokines secretion.
The results of this study indicate the anti-inflammatory character of Anis-EVs in relation to Caco-2. At the same time, direct treatment with Anis-EVs resulted in more significant changes in the Caco-2 proteome than co-culture with L3 larvae.
The results obtained should lead to a better understanding of the molecular mechanisms underlying the development of A. simplex (s. s.) infection in humans and will complement the existing knowledge on the role of EVs in host-parasite communication.
简单异尖线虫(狭义)是海洋生物中分布最广泛的寄生线虫之一,人类为偶然宿主。虽然许多研究探讨了线虫生物学及其与宿主的相互作用,但细胞外囊泡(EVs)作为寄生线虫信号分子的作用尚不清楚。
因此,对简单异尖线虫(狭义)的细胞外囊泡(Anis-EVs)中的蛋白质进行了鉴定。此外,采用相互作用蛋白质组学方法,鉴定与简单异尖线虫(狭义)L3幼虫共培养或直接暴露于两种浓度(低或高)Anis-EVs的人肠上皮细胞系(Caco-2)蛋白质组中的差异调节蛋白(DRPs)。此外,还鉴定了与Caco-2共培养影响的简单异尖线虫(狭义)幼虫蛋白质组中的DRPs。为实现这一目标,使用了基于等压质量标记(通过串联质量标签;TMT)的鸟枪法蛋白质组学方法,并结合了纳升高效液相色谱(nLC)与LTQ-Orbitrap Elite质谱仪。此外,采用酶联免疫吸附测定(ELISA)来证明Caco-2是否通过选定细胞因子分泌的显著变化对简单异尖线虫(狭义)幼虫和Anis-EVs作出反应。
本研究结果表明Anis-EVs对Caco-2具有抗炎特性。同时,与L3幼虫共培养相比,直接用Anis-EVs处理导致Caco-2蛋白质组发生更显著的变化。
所获得的结果将有助于更好地理解人类感染简单异尖线虫(狭义)的分子机制,并将补充关于细胞外囊泡在宿主-寄生虫通讯中作用的现有知识。