Effect of duck Tembusu virus infection on exosomes of BHK-21 cells / 中国免疫学杂志

ABSTRACT

Objective:To explore the effect of duck Tembusu virus infection on secretion of exosomes in BHK-21 cells and the pathogenesis of the Tembusu virus.Methods:The exosomes were collected and purified from the culture supernatant of BHK-21 cells infected with duck Tembusu virus AH-F10 strain and the control BHK-21 cells by PEG precipitation method respectively.The purified exosomes were identified by electron microscopy,Western blot assay and mass spectrometry.Results: The classical exosome particle morphology was observed with hyperchromic cup-shaped vesicles and average particle size of 30-160 nm in diameter under transmission electron microscopy.The mean size of the exosome from the infected cells were bigger than the mean size of the exosome from the unin-fected.Western blot assay demonstrated that CD9 and CD63 were detected in purified exosomes as exosome marker molecula.A total of 106 proteins were identified by mass spectrometry assay,84 proteins of infected BHK-21 cells exosome,49 proteins of the uninfected, and the infected and the uninfected BHK-21 share 27 common proteins on exosomes.Conclusion:Duck Tembusu virus infection affect the exosome secretion of cells in connection to the particle size and protein molecular composition.This experiment can lay the foundation for further research of Tembusu virus infection and pathogenesis.