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Characterization of Aspergillus fumigatus Extracellular Vesicles and Their Effects on Macrophages and Neutrophils
Jéssica Amanda Marques Souza1, Ludmila de Matos Baltazar2, Virgínia Mendes Carregal3
1Centro de Pesquisa e Desenvolvimento de Fármacos, Departamento de Genética, Ecologia e Evolução, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
Abstract:
Extracellular vesicles (EVs) has been considered an alternative process for intercellular communication. EVs release by filamentous fungi and the role of vesicular secretion during fungus-host cells interaction remain unknown. Here, we identified the secretion of EVs from the pathogenic filamentous fungus, Aspergillus fumigatus. Analysis of the structure of EVs demonstrated that A. fumigatus produces round shaped bilayer structures ranging from 100 to 200 nm size, containing ergosterol and a myriad of proteins involved in REDOX, cell wall remodeling and metabolic functions of the fungus. We demonstrated that macrophages can phagocytose A. fumigatus EVs. Phagocytic cells, stimulated with EVs, increased fungal clearance after A. fumigatus conidia challenge. EVs were also able to induce the production of TNF-α and CCL2 by macrophages and a synergistic effect was observed in the production of these mediators when the cells were challenged with the conidia. In bone marrow-derived neutrophils (BMDN) treated with EVs, there was enhancement of the production of TNF-α and IL-1β in response to conidia. Together, our results demonstrate, for the first time, that A. fumigatus produces EVs containing a diverse set of proteins involved in fungal physiology and virulence. Moreover, EVs are biologically active and stimulate production of inflammatory mediators and fungal clearance.
Insights
Filamentous fungi like Aspergillus fumigatus release extracellular vesicles (EVs) that modulate immune responses. These EVs enhance fungal clearance and stimulate inflammatory mediators in host cells.
Area of Science:
- Mycology
- Immunology
- Cell Biology
Background:
- Extracellular vesicles (EVs) mediate intercellular communication.
- The role of EVs in fungal-host interactions, particularly from filamentous fungi, is largely unknown.
- Aspergillus fumigatus is a pathogenic fungus with significant clinical implications.
Purpose of the Study:
- To identify and characterize EVs secreted by Aspergillus fumigatus.
- To investigate the biological activity and immune-modulating effects of A. fumigatus EVs on host phagocytic cells.
Main Methods:
- Isolation and structural analysis of EVs from A. fumigatus.
- Assessment of EV uptake by macrophages.
- Measurement of inflammatory mediator production (TNF-α, CCL2, IL-1β) by host cells stimulated with EVs and/or A. fumigatus conidia.
- Evaluation of fungal clearance by phagocytic cells.
Main Results:
- A. fumigatus secretes EVs (100-200 nm) containing fungal proteins involved in REDOX, cell wall remodeling, and metabolism.
- Macrophages phagocytose A. fumigatus EVs, leading to enhanced fungal clearance.
- EVs induce TNF-α and CCL2 production in macrophages and TNF-α and IL-1β in neutrophils, with synergistic effects when combined with fungal conidia.
Conclusions:
- Aspergillus fumigatus produces biologically active EVs containing virulence-associated proteins.
- These EVs play a role in modulating host immune responses, including enhancing fungal clearance and stimulating inflammation.
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