PXD050398-1

PXD050398 is an original dataset announced via ProteomeXchange.

Title	Genetic-based reporter systems for labelling and differentiation of host-derived extracellular vesicles reveal their association with a human-pathogenic fungus
Description	Extracellular vesicles (EVs) have gained attention as a mechanism facilitating intercellular as well as interkingdom communication during host-microbe interactions. Recently we showed that upon infection, host polymorphonuclear leukocytes produce antifungal EVs targeting the clinically important fungal pathogen Aspergillus fumigatus; however, the small size of EVs complicates their tracking and functional analysis. We employed a more tractable, reporter-based system to label host alveolar epithelial cell-derived EVs and enable their visualization and tracking during in vitro A. fumigatus interaction. Proteome analysis of the cargo of these EVs confirmed the EV marker proteins, tetraspanins CD63, CD9, and CD81, and revealed limited antifungal components, which was corroborated by mito-GFP fungal killing assays. Fusion of the EV marker proteins with a nanoluciferase (NLuc) and a green fluorescent protein (GFP) facilitated their relative quantification by luminescence and visualization by fluorescence signal. The use of an NLuc fused with a GFP is advantageous as it allows quantification and visualisation of EVs simultaneously without additional external manipulation. Using this system, visualisation of EVs was possible using confocal laser scanning microscopy and revealed the ability of host cell-derived EVs to associate with A. fumigatus hyphae. In conclusion, we have created a series of tools to better define the complex interplay of host-derived EVs with microbial pathogens.
HostingRepository	PRIDE
AnnounceDate	2024-11-13
AnnouncementXML	Submission_2024-11-13_00:11:06.476.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Thomas Krüger
SpeciesList	scientific name: Aspergillus fumigatus CEA17; NCBI TaxID: 1266464; scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive Plus

Revision	Datetime	Status	ChangeLog Entry
0	2024-03-06 04:01:50	ID requested
⏵ 1	2024-11-13 00:11:06	announced

10.1093/FEMSML/UQAE022;

submitter keyword: green fluorescent protein, Aspergillus fumigatus, A549 epithelial cells, CD9,Extracellular vesicles, CD81, CD63, Nanoluciferase

Axel A. Brakhage
contact affiliation	Molecular and Applied Microbiology, Leibniz Institute for Natural Product Research and Infection Biology - Hans Knöll Institute (Leibniz-HKI)
contact email	axel.brakhage@leibniz-hki.de
lab head
Thomas Krüger
contact affiliation	Leibniz Institute for Natural Product Research and Infection Biology - Hans Knöll Institute
contact email	thomas.krueger@leibniz-hki.de
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2024/11/PXD050398

PRIDE project URI

• PRIDE
  1. PXD050398
     1. Label: PRIDE project
     2. Name: Genetic-based reporter systems for labelling and differentiation of host-derived extracellular vesicles reveal their association with a human-pathogenic fungus