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Paracoccidioidomycosis (PCM) is a systemic mycosis, endemic in most Latin American countries, especially in Brazil. It is caused by the thermo-dimorphic fungus of the genus  Paracoccidioides  ( Paracoccidioides brasiliensis  and  Paracoccidioides lutzii ). Innate immune response plays a crucial role in host defense against fungal infections, and neutrophils (PMNs) are able to combat microorganisms with three different mechanisms: phagocytosis, secretion of granular proteins, which have antimicrobial properties, and the most recent described mechanism called NETosis. This new process is characterized by the release of net-like structures called Neutrophil Extracellular Traps (NETs), which is composed of nuclear (decondensed DNA and histones) and granular material such as elastase. Several microorganisms have the ability of inducing NETs formation, including gram-positive and gram-negative bacteria, viruses and some fungi. We proposed to identify NETs in tegumentary lesions of patients with PCM and to analyze the interaction between two strains of  P .  brasiliensis  and human PMNs by NETs formation  in vitro . In this context, the presence of NETs  in vivo  was evidenced in tegumentary lesions of patients with PCM by confocal spectrum analyzer. Furthermore, we showed that the high virulent  P .  brasiliensis  strain 18 (Pb18) and the lower virulent strain Pb265 are able to induce different patterns of NETs formation  in vitro . The quantification of extracellular DNA corroborates the idea of the ability of  P .  brasiliensis  in inducing NETs release. In conclusion, our data show for the first time the identification of NETs in lesions of patients with PCM and demonstrate distinct patterns of NETs in cultures challenged with fungi  in vitro . The presence of NETs components both  in vivo  and  in vitro  open new possibilities for the detailed investigation of immunity in PCM.

plos neglected tropical diseases

Neutrophil Extracellular Traps Identification in Tegumentary Lesions of Patients with Paracoccidioidomycosis and Different Patterns of NETs Generation In Vitro