School of Basic Medicine, Ningxia Medical University, Yinchuan, China.
Department of Molecular Biology, Shanghai Centre for Clinical Laboratory, Shanghai, China.
Front Cell Infect Microbiol. 2022 Mar 10;12:805010. doi: 10.3389/fcimb.2022.805010. eCollection 2022.
The globally distributed cystic echinococcosis (CE) is caused by the larval stage of (), a cosmopolitan and zoonotic disease with potentially life-threatening complications in humans. The emerging roles for extracellular vesicles (EVs) in parasitic infection include transferring proteins and modifying host cell gene expression to modulate host immune responses. Few studies focused on the host-derived EVs and its protein profiles. We focused on the EVs from mouse infected with at different stages. ExoQuick kit was used for isolating EVs from mouse plasma and ExoEasy Maxi kit was used for isolating protoscolex culture supernatant (PCS) and hydatid cyst fluid (HCF). Firstly, EVs were characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and immunoblot. Secondly, the proteins of plasma EVs were identified using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The resulting LC-MS/MS data were processed using Maxquant search engine (v 1.5.2.8). Tandem mass spectra were researched against the mice and proteins database in the NCBI. The differentially expressed proteins are performed by proteomic label-free quantitative analysis and bioinformatics. Thirdly, experiment, the results of co-culture of plasma EVs and spleen mononuclear cells showed that 7W-EVs can increase the relative abundance of regulatory T (Treg) cells and IL-10. We further verified that EVs can be internalized by CD4 and CD8 T cells, B cells, and myeloid-derived suppressor cells (MDSC). These results implied host-derived EVs are multidirectional immune modulators. The findings can contribute to a better understanding of the role of host-derived EVs which are the optimal vehicle to transfer important cargo into host immune system. In addition, we have found several important proteins associated with and identified in infected mouse plasma at different stages. Furthermore, our study further highlighted the proteomics and immunological function of EVs from mouse infected with protoscoleces at different infection stages. We have laid a solid foundation for the role of EVs in cystic echinococcosis in the future research and supplemented a unique dataset for this
全球分布的包虫病（CE）是由幼虫阶段的()引起的，这是一种世界性的人畜共患病，在人类中可能导致危及生命的并发症。细胞外囊泡（EVs）在寄生虫感染中的新作用包括传递蛋白质和修饰宿主细胞基因表达，以调节宿主免疫反应。很少有研究关注宿主来源的 EVs 及其蛋白谱。我们专注于在不同阶段感染的小鼠的 EVs。使用 ExoQuick 试剂盒从小鼠血浆中分离 EVs，使用 ExoEasy Maxi 试剂盒从原头节培养上清液（PCS）和包虫囊液（HCF）中分离 EVs。首先，通过透射电子显微镜（TEM）、纳米颗粒跟踪分析（NTA）和免疫印迹法对 EVs 进行表征。其次，使用液相色谱-串联质谱（LC-MS/MS）鉴定血浆 EVs 的蛋白质。使用 Maxquant 搜索引擎（v1.5.2.8）处理所得 LC-MS/MS 数据。串联质谱针对 NCBI 中的小鼠和()蛋白质数据库进行研究。通过蛋白质组学无标记定量分析和生物信息学对差异表达蛋白进行分析。第三，在共培养血浆 EVs 和脾单核细胞的实验中，结果表明 7W-EVs 可以增加调节性 T（Treg）细胞和 IL-10 的相对丰度。我们进一步验证了 EVs 可以被 CD4 和 CD8 T 细胞、B 细胞和髓源性抑制细胞（MDSC）内化。这些结果表明宿主来源的 EVs 是多向免疫调节剂。这些发现有助于更好地理解宿主来源的 EVs 的作用，宿主来源的 EVs 是将重要货物转移到宿主免疫系统的最佳载体。此外，我们发现了一些与感染不同阶段的小鼠血浆中相关的重要蛋白质。此外，我们的研究进一步强调了不同感染阶段感染原头节的小鼠的 EVs 的蛋白质组学和免疫功能。为今后的研究奠定了坚实的基础，为 EVs 在包虫病中的作用补充了独特的数据集。
