F81 extracellular vesicles mediate upregulation of antiviral cytokine expression by IFN-ω
This study aims to investigate the role of extracellular vesicles(EVs)from feline kidney cells(F81)in mediating the regulation of antiviral cytokines through feline omega interferon(IFN-ω),providing a theoretical foundation for research on EV-mediated interferon antiviral mechanisms.IFN-ω-stimulated IFN-ω-EVs secreted by F81 cells were extracted and characterized for their marker proteins,morphology,and size using Western blotting,transmission electron microscopy,and nanoscale particle technology,respectively.F81 cells were incubated with IFN-ω-EVs containing IFN-ω,and fluorescence colocalization was analyzed using a laser confocal microscope to determine whether IFN-ω-EVs enter F81 cells.The expression levels of ISG15,ISG56,and IFN-ω cytokines were detected by RT-qPCR to investigate the antiviral effects of IFN-ω-EVs against VSV,in comparison with IFN-ω and a blank control.IFN-ω-EVs were primarily identified as cup-shaped vesicular structures carrying specific marker proteins CD63 and CD81,with particle sizes ranging between 50-100 nm and 250-600 nm.Fluorescence colocalization analysis revealed that IFN-ω-EVs could enter F81 cells.RT-qPCR results indicated that IFN-ω-EVs,like IFN-ω,significantly upregulated the expression levels of antiviral cytokines such as ISG15,ISG56,and IFN-ω.Importantly,the anti-VSV activity of IFN-ω-EVs post-VSV infection was significantly higher than that of IFN-ω alone(P＜0.05).IFN-ω-EVs extracted from F81 cells,can mediate the entry of IFN-ω by F81 cells,significantly upregulate the expression of antiviral cytokines such as ISG15,ISG56,and IFN-ω,and inhibit the inhibition of VSV virus proli-feration.
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