Exosomes are cell-derived vesicles that are secreted by many eukaryotic cells.It has recently attracted attention as vehicles of intercellular communication.Virus-infected cells release exosomes,which contain viral proteins,RNA,and pathogenic molecules.However,the role of exosomes in virus infection process remains unclear and needs to be further investigated.In this study,we aimed to evaluate the effects of exosomes on rabies virus infection.OptiPrepTM density gradient centrifugation was used to isolate exosomes from rabies virus-infected cell culture supernatants.A rabies virus G protein enzyme-linked immunosorbent assay and acetylcholinesterase activity assays were performed to verify the centrifugation fractions.Exosomes were then characterized using transmission electron microscopy and Western blotting.Our results showed that rabies virus infection increased the release of exosomes.Treatment with GW4869 and si-Rab27a,two exosomal secretion inhibitors,inhibited exosome release.Furthermore,the inhibitors reduced the levels of extracellular and intracellular viral RNA.These data indicated that exosomes may participate in the viral infection process.Moreover,our results establish a basis for future research into the roles of exosomes in rabies virus infection and as potential targets for developing new antiviral strategies.