Abstract Objective: To isolate and identify exosomes from Toxoplasma gondii RH strain tachyzoites.Methods: The supernatant of T. gondii tachyzoites co-cultured with SW480 cells was collected. After centrifugating and filtrating, exosomes were isolated from supernatant by exosome isolation kit based on polymer precipitation technology. The morphological characteristics were observed by transmission electron microscopy (TEM). The size of exosomes was determined by nanoparticle tracking analysis (NTA). Western blotting was used to detect the expression of CD63 and HSP70 protein, which were considered as specific-classic molecules of exosomes, as well as surface antigen 1 (SAG1), rhoptry protein 16 (ROP16), ROP18 and dense granule protein 1 (GRA1) protein related to T. gondii. Results: The vesicles, which were separated from co-culture supernatant, were round with a complete membrane structure, 30-150 nm in diameter. It not only expressed CD63 and HSP70 protein, but also expressed SAG1, ROP16, ROP18 and GRA1 protein. Conclusion: Exosomes of T. gondii were obtained from the supernatant of co-culture media.
[Key words]Toxoplasma gondii; exosomes; isolation; identification; co-culture; nanoparticle tracking analysis
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