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  • Early Endosome Antigen 1 (EEA1) decreases in macrophages infected with Paracoccidioides brasiliensis.

Early Endosome Antigen 1 (EEA1) decreases in macrophages infected with Paracoccidioides brasiliensis.

Medical mycology (2013-04-10)
Aline Raquel Voltan, Janaina De Cassia Orlandi Sardi, Christiane Pienna Soares, Marcelo Pelajo Machado, Ana Marisa Fusco Almeida, Maria José Soares Mendes-Giannini
ABSTRACT

Paracoccidioidomycosis (PCM) is a chronic granulomatous disease caused by the dimorphic fungus Paracoccidioides brasiliensis, endemic in Latin America. P. brasiliensis has been observed in epithelial cells in vivo and in vitro, as well as within the macrophages. The identification of the mechanism by which it survives within the host cell is fertile ground for the discovery of its pathogenesis since this organism has the ability to induce its own endocytosis in epithelial cells and most likely in macrophages. The study of the expression of endocytic proteins pathway and co-localization of microorganisms enable detection of the mechanism by which microorganisms survive within the host cell. The aim of this study was to evaluate the expression of the endocytic protein EEA1 (early endosome antigen 1) in macrophages infected with P. brasiliensis. For detection of EEA1, three different techniques were employed: immunofluorescence, real-time polymerase chain reaction (PCR) and immunoblotting. In the present study, decreased expression of EEA1 as well as the rearrangement of the actin was observed when the fungus was internalized, confirming that the input mechanism of the fungus in macrophages occurs through phagocytosis.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Mouse IgG (Fab specific)–Peroxidase antibody produced in goat, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Anti-EEA1 antibody, Mouse monoclonal, clone EEA1-N19, purified from hybridoma cell culture