XI International Meeting on Paracoccidioidomycosis

XI International Meeting on Paracoccidioidomycosis

Resume:34-2

Oral / Poster

34-2	Proteomic study of Aspergillus fumigatus interaction with endothelial cells
Authors:	Nathália Curty de Andrade (UERJ - Universidade do Estado do Rio de Janeiro) ; Luis Felipe Clemente Velarde (UCM - Universidad Complutense de Madrid) ; Dolores Gutiérrez Blázquez (UCM - Universidad Complutense de Madrid) ; Concepcion Gil Garcia (UCM - Universidad Complutense de Madrid) ; Leila M. Lopes Bezerra (UERJ - Universidade do Estado do Rio de Janeiro)

Abstract

Aspergillus fumigatus is the main etiological agent of invasive aspergillosis, a life- threatening disease that affects patients with long-term neutropenia. The infection initiates by inhalation of airborne conidia which may reach the bronchioles and alveolar spaces. In immunocompromised patients conidia not eliminated by the innate immune system, germinate into angioinvasive hyphae. It is known that A. fumigatus hyphae but not conidia can activate endothelial cells. This study focuses on the infectome of human umbilical vein endothelial cells (HUVECs) upon interaction with A. fumigatus germelings. To achieve this goal a proteomic approach using iTRAQ (isobaric tags for relative and absolute quantitation) was delineate to analyze HUVEC proteins expressed upon interaction with this pathogen. HUVEC protein extracts of infected monolayers obtained upon 16 hour of interaction, were cleaved with trypsin and the resulting peptides labelled with iTRAQ 8-plex reagents. The labeled peptides were separated by isoelectric focusing using an Offgel system in 24 fractions. Each fraction was separated by reversed phase liquid chromatography (LC), in a nanoHPLC system and spotted in a MALDI plate by a robot for the MALDI TOF/TOF mass spectrometry (MS) analysis. The identification and quantification of the differentially expressed proteins was performed using ProteinPilot software. The relative amounts of each protein are being identified in comparison to control HUVECs. Using this approach it will be possible to accurately quantify over- and under expressed proteins in infected HUVEC monolayers. Also, protein-protein interactions and activated cellular routes can be inferred by bioinformatics analysis of this proteomic data. Financial support: CAPES-DGU, CNPq e Faperj.

Keyword: Aspergillus fumigatus, Células endoteliais, Interação fungo-hospedeiro

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