Extracellular vesicles from virulent P. brasiliensis induce TLR4 and dectin-1 expression in innate cells and promote enhanced Th1/Th17 response

ABSTRACT

Extracellular vesicles (EVs) are membrane-enclosed nanoparticles that transport several biomolecules and are involved in important mechanisms and functions related to the pathophysiology of fungal diseases. EVs from Paracoccidioides brasiliensis, the main causative agent of Paracoccidioidomycosis (PCM), modulate the immune response of macrophages. In this study, we assessed the EVs proteome from a virulent P. brasiliensis isolated from granulomatous lesions and compared their immunomodulatory ability with EVs isolated from the fungus before the animal passage (control EVs) when challenging macrophages and dendritic cells (DCs). Proteome showed that virulent EVs have a higher abundance of virulence factors such as GP43, protein 14-3-3, GAPDH, as well as virulence factors never described in PCM, such as aspartyl aminopeptidase and a SidJ analogue compared with control EVs. Virulent extracellular vesicles induced higher expression of TLR4 and Dectin-1 than control EVs in macrophages and dendritic cells (DCs). In opposition, a lower TLR2 expression was induced by virulent EVs. Additionally, virulent EVs induced lower expression of CD80, CD86 and TNF-α, but promoted a higher expression of IL-6 and IL-10, suggesting that EVs isolated from virulent P. brasiliensis-yeast promote a milder DCs and macrophage maturation. Herein, we showed that EVs from virulent fungi stimulated a higher frequency of Th1/Tc1, Th17, and Treg cells, which gives new insights into fungal extracellular vesicles. Taken together, our results suggest that P. brasiliensis utilizes its EVs as virulence bags that manipulate the immune system in its favour, creating a milder immune response and helping with fungal evasion from the immune system.

KEYWORDS:

• Paracoccidioides brasiliensis
• extracellular vesicles, proteome
• granuloma
• Paracoccidioidomycosis
• dendritic cells
• macrophages

Acknowledgements

We want to thank the Mass Spectrometry Facility (MAS) of the Brazilian Biosciences National Laboratory (LNBio), part of the Brazilian Centre for Research in Energy and Materials (CNPEM), a private non-profit organization under the supervision of the Brazilian Ministry for Science, Technology, and Innovations (MCTI) for their assistance.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data Availability statement

Proteome data are available via ProteomeXchange with identifier PXD046549. Any other data are available upon request.

Supplemental data

Supplemental data for this article can be accessed online at https://doi.org/10.1080/21505594.2024.2329573

Authors contribution statement

BMB: Conceptualization, Data curation, Formal Analysis, Investigation, Methodology, Project administration, Validation, Visualization, Writing – original draft, Writing – review & editing. MGS: Data curation, Formal Analysis, Investigation, Visualization. NYP: Formal Analysis, Investigation, Methodology, Visualization, Writing – review & editing. VLK: Formal Analysis, Investigation, Methodology, Visualization, Writing – review & editing. GT: Investigation, Methodology, Writing – review & editing. FA: Conceptualization, Methodology, Writing – original draft, Writing – review & editing; FVL: Conceptualization, Funding acquisition, Investigation, Methodology, Project administration, Resources, Supervision, Writing – original draft, Writing – review & editing.

Additional information

Funding

We thank the Fundação de Amparo à Pesquisa do Estado de São Paulo [FAPESP] for funding this work, under the grant numbers 2018-14762-3 [FVL], 2021/09962-6 [BMB], 2022/10654-7 [MGS], and 2021/06794-5 [FA].