Biomolecular Corona Remodelling of <i>Aspergillus fumigatus</i> Extracellular Vesicles Shapes Immune Recognition and Macrophage Activation Programs

ABSTRACT Extracellular vesicles (EVs) released by Aspergillus fumigatus play key roles in fungal–host communication. Similar to synthetic nanoparticles, fungal EVs acquire a biomolecular corona composed of a tightly bound “hard” corona layer of fungal proteins and a dynamic “soft” layer of host‐derived molecules adsorbed upon contact with biological fluids. Although the corona is thought to modulate EV identity and immunological outcomes, its role in fungal EV–immune cell interactions remains poorly defined. Here, we investigated how corona removal and reconstitution alter the biophysical properties and immunomodulatory function of A. fumigatus EVs. Corona removal reduced EV‐associated protein content and impaired EV‐macrophage interaction, whereas serum exposure promoted secondary corona formation and markedly increased EV association with AMJ2‐C11 macrophages. Native and serum‐coated EVs induced robust secretion of both pro‐inflammatory (IL‐1β, IL‐6, TNF‐α, IFN‐γ) and anti‐inflammatory (IL‐4, IL‐10) cytokines, whereas stripped EVs elicited a markedly attenuated response. At the transcriptional level, serum‐coated EVs upregulated genes involved in innate recognition and signalling pathways ( Tlr2 , Tlr4 , Dectin1 , DC‐Sign and Nf‐κb) as well as macrophage activation programs ( Nos2 , Arg1 , Chi3l3 , Retnla and Tgfb1 ). These findings establish the biomolecular corona as a key determinant of fungal EV immunomodulatory activity. Beyond advancing our understanding of fungal EV biology, this work highlights corona engineering as a potential strategy to reprogram EV–host interactions, with implications for antifungal therapeutic development.