Estudo proteômico da interação do Aspergillus fumigatus com células endoteliais da veia umbilical humana (HUVECs)

• Ano de defesa: 2012
• Autor(a) principal: Andrade, Nathália Curty
• Orientador(a): Bezerra, Leila Maria Lopes
• Banca de defesa: Ferreira, Prescilla Emy Nagao , Cunha, Marcel Menezes Lyra da , Baruffi, Marcelo Dias , Barboza, Luciana Pizzatti
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade do Estado do Rio de Janeiro
• Programa de Pós-Graduação: Programa de Pós-Graduação em Biociências
• Departamento: Centro Biomédico::Instituto de Biologia Roberto Alcantara Gomes
• País: BR
• Palavras-chave em Português: Aspergillus fumigatus; Δugm1; HUVEC; Galectina-1; Anexina A2; Células endoteliais da veia umbilical humana; Galectina 1; Anexina 2; Aspergilose pulmonar invasiva
• Palavras-chave em Inglês: Aspergillus fumigates; Δugm1; HUVEC; Galectin-1; Annexin A2
• Área do conhecimento CNPq: CNPQ::CIENCIAS BIOLOGICAS::MICROBIOLOGIA::BIOLOGIA E FISIOLOGIA DOS MICROORGANISMOS::MICOLOGIA
• Link de acesso: http://www.bdtd.uerj.br/handle/1/16167

Aspergillus fumigatus is the main etiological agent of invasive aspergillosis, the main opportunistic fungal infection of Hematologial Unity s patients, especially those with long-term neutropenia. Upon filamentation, this angioinvasive fungus can activate and damage the human umbilical vein endothelial cells (HUVEC), which in response switch to a pro-thrombotic phenotype. HUVEC activation is mediated by TNF-α once cell-cell contact occurs. This activation is characterized by the expression of pro-inflammatory molecules such cytokines, chemokines and adhesion molecules. Recently, our group performed the comparison of HUVEC activation upon interaction with a wild type and the UGM1 mutant strains of A. fumigatus. The Δugm1 strain, which presents an increased production of the cell wall galactosaminogalactan, showed a hyper adherent phenotype and an increased capability to cause endothelial cell stimulation and activation, when compared with the wild type strain. The receptors involved in the pathogen-host interaction or the signaling pathways after endothelial activation by A. fumigatus remain unknown. Thus, the aim of this study was to investigate the differentially expressed proteins in HUVECs upon interaction with A. fumigatus, using the 2D-DIGE proteomic approach. Briefly, HUVECs were challenged with germlings of A. fumigatus wild type Af293 and Δugm1 strains and then submitted to protein extraction. The total HUVEC protein extracts were labeled with different CyDyes and fractionated by 2D electrophoresis. Quantitative analysis to determine the differences in protein abundance amongst interacted cells vs. control endothelial cells was performed using the software DeCyder. Five differentially expressed proteins were identified by MS/MS including galectin-1 and annexin A2, both overexpressed after the interaction. These two proteins are described elsewhere to be associated with host-pathogen interaction. Besides, galectin-1 showed an ~25% increase after interaction with the Δugm1 strain and it is plausible that this particular protein could be a putative receptor for galactose-containing polymers of the A. fumigatus cell wall and annexin A2 could be involved in signalizing pathways upon interaction. However, other experimental evidences, under development, are necessary to confirm this hypothesis.