Fungal extracellular vesicles (EVs) mediate intra- and interspecies communication and are critical in host–fungus interaction, modulating inflammation and immune responses. In this study, we evaluated thein vitropro- and anti-inflammatory properties ofAspergillus fumigatusEVs over innate leukocytes.A. fumigatusEVs induced a partial proinflammatory response by macrophages, characterized by increased tumor necrosis factor-alpha production, and increased gene expression of induced nitric oxide synthase and adhesion molecules. EVs induce neither NETosis in human neutrophils nor cytokine secretion by peripheral mononuclear cells. However, prior inoculation ofA. fumigatusEVs inGalleria mellonellalarvae resulted in increased survival after the fungal challenge. Taken together, these findings show thatA. fumigatusEVs play a role in protection against fungal infection, although they induce a partial pro-inflammatory response.