Análise metabôlica do paracoccidioides brasiliensis durante a infecçâo em macrófagos

Abstract

Several fungal species have been emerging as human pathogens and have gained special attention in public health. Among these, Paracoccidioidesbrasiliensis presents as the causative agent of the systemic mycose called Paracoccidioimycose (PCM). It is a dimorphic pathogen which presents two forms, yeast at 37°C and mycelium at 25°C. During infection Paracoccidioides needs a variety of elements to survive within the host. The metallic micronutrients are essential elements required for the cell homeostasis in virtually all organisms Metals have a great participation in countless metabolic processes, such as DNA metabolism, regulation of protein synthesis, virulence, DNA synthesis, amino acid metabolism, the post transcriptional mechanism and others. In this work it was performed a quantitative metallomic analysis of P. brasiliensis during infection of bone marrow derived macrophages (BMDM) and J774 immortalized lineage. Also it was evaluated the importance of iron availability in fungus-macrophage interaction. Fungal cells recovered from macrophages, treated with iron citrate, revealed higher survival rate compared with non-treated ones. Using Inductively Coupled Plasma Mass Spectrometry (ICP-MS), we detected a significant change in magnesium, manganese and zinc levels in yeast cells recovered from macrophages. Additionally, a clear decrease in cooper and iron amounts was also seen. It was also detected a significant increase in the levels of calcium and cobalt in infected J774 macrophages when compared with non infected ones. These results indicate that the limitation of Mg, Mn, Zn, Cu and Fe might be a mechanism of macrophages to hinder the fungal survival during infection. It was observed an increase of calcium and cobalt amounts in infected macrophages indicating that those micronutrients might be important to an efficient immune response against P. brasiliensis. These findings suggest that other metals, beyond iron and cooper, play crucial functions in the P. brasiliensis-macrophage interaction and their homeostasis could be targets for further studies and treatments.