CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES >

2020 , Vol. 38 >Issue 6: 695 - 701

DOI: https://doi.org/10.12140/j.issn.1000-7423.2020.06.004

ORIGINAL ARTICLES

Proteomic analysis of human proteins in extracellular vesicles secreted by protoscoleces of Echinococcus granulosus

  • Chun-li SHI ,
  • Hui YANG ,
  • Wen PAN ,
  • Xin ZHANG ,
  • Xiao-ting ZHU ,
  • Jia-qing ZHAO
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  • 1 Basic Medical College of Ningxia Medical University, Yinchuan 750004, China
    2 Science and Technology Center of Ningxia Medical University, Yinchuan 750004, China
    3 Institute of Medical Sciences of Ningxia Hui Autonomous Region, Yinchuan 750004, China
    4 Clinical Medical College of Ningxia Medical University, Yinchuan 750004, China

Received date: 2020-08-07

  Online published: 2021-01-12

Supported by

Ningxia Hui Autonomous Region Key Research Program(2020BEG03014);Ningxia Natural Science Found Project(2019AAC03266);Ningxia Natural Science Found Project(2020AAC03426)

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Abstract

Objective To identify the protein of human source in extracellular vesicles (EVs) secreted by protoscoleces of Echinococcus granulosus by proteomic analysis. Methods Liver intact cysts were collected from cystic echinococcosis patients, and the protoscoleces were separated under aseptic conditions and then cultured in 30% exosome-free fetal bovine serum in vitro, with medium replacement every three days. The supernatant was collected, from which EVs were isolated by a simple and rapid extracellular vesicle separation kit. A transmission electron microscope was employed to observe and identify the morphology of EVs, and the concentration and size of EVs were evaluated by nanoparticle tracking analysis. The EVs underwent Liquid chromatography-mass spectrometry analysis for identification and characterization of their protein content. The secondary mass spectrometry data were used to retrieve and annotated with Maxquant (v1.5.2.8). The enrichment analysis of tissues, disease and pathway for the human source proteins in EVs were were carried out using online tool David. Results The EVs secreted by protoscoleces in vitro were cup-shaped bilayer membranous vesicles with a diameter less than 200 nm. The proteomic analysis found a total of 20 317 peptides, and identified 1 461 proteins, including 328 human source proteins. Exosome marker proteins, including filament protein, 14-3-3 protein, heat shock protein 90, heat shock protein 70 and pyruvate kinase, are highly enriched in EVs secreted by protoscolex. The top 3 tissues in enrichment extent were plasma, Cajal-Retzius cells and liver. The disease enrichment found involving infection, immunity and cancer. The Bio Carta pathways analysis indicated top 3 enriched pathways, which were the alternative complement pathway, lectin complement pathway and complement pathway; the top 3 of KEGG pathways enrichment were adhesion plaque, extracellular matrix receptor-related action, and proteasome. Conclusion The proteins of human source in EVs secreted by protoscoleces cultured in vitro are related to human plasma and liver, indicating that the protoscoleces exert substance exchange with human via seretion of EVs.

Key words： Echinococcus granulosus; Protosoleces; Extracellular vesicle; Immune escape; Proteomics

Cite this article

Chun-li SHI , Hui YANG , Wen PAN , Xin ZHANG , Xiao-ting ZHU , Jia-qing ZHAO . Proteomic analysis of human proteins in extracellular vesicles secreted by protoscoleces of Echinococcus granulosus[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2020 , 38(6) : 695 -701 . DOI: 10.12140/j.issn.1000-7423.2020.06.004

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