Identification and molecular characterization of exosome-like vesicles derived from the Taenia asiatica adult worm

Taenia asiatica is an important food-borne parasite that poses a threat to food-safety and animal husbandry hygine, yet little is known about its specific infection and immune escape mechanisms. Exosome-like vesicles have recently emerged as a regulator in the interactions between parasites and hosts, providing a new direction for research on infection of T. asiatica. In this experiment, exosome-like vesicles were collected from the excretory/secretory products of cultured T. asiatica and isolated by differential centrifugation. The purified vesicles, ranging from 30 to 150 nm in size, were identified as exosome-like vesicles by transmission electron microscope and Nanoparticle tracking analysis. Proteomics analysis identified 455 proteins in the exosome-like vesicles. Of these proteins, enzymes involved in metabolic processes were identified, including glyceraldehyde 3 phosphate dehydrogenase, fructose-1, 6-bisphosphate aldolase, cytosolic malate dehydrogenase, and enolase. The two most abundant proteins from proteomic analysis, 14-3-3 and enolase, were shown to be present in the exosome-like vesicles by immunogold labeling. High-throughput RNA sequencing yielded twenty known miRNAs present in exosome-like vesicle sRNA libraries. Nine of the miRNAs, including six known miRNAs (tas-miR-71, tas-miR-1, tas-miR-7, tas-miR-9, tas-miR-10, and tas-let-7) and three newly discovered miRNAs (tas-m0022-3p, tas-m0816-3p, tas-m0082-5p), were confirmed by RT-qPCR as present in T. asiatica adult worm extracts and secreted exosome-like vesicles in T. asiatica. Additionally, we demonstrated that exosome-like vesicles experimentally labeled with PKH67 were internalized by LoVo cells in vitro. These findings provide new insights into the interaction between tapeworms and hosts mediated by exosome-like vesicles.