Neutrophil Extracellular Traps Identification in Tegumentary Lesions of Patients with Paracoccidioidomycosis and Different Patterns of NETs Generation In Vitro.
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.
Department of Pathology, UNESP - São Paulo State University, Botucatu Medical School, Botucatu, São Paulo, Brazil.
Amanda Manoel
AM
Della Coletta
Department of Microbiology and Immunology, UNESP-São Paulo State University, Biosciences Institute, Botucatu, São Paulo, Brazil.
Tatiana Fernanda
TF
Bachiega
Department of Pathology, UNESP - São Paulo State University, Botucatu Medical School, Botucatu, São Paulo, Brazil.
Juliana Carvalho
JC
de Quaglia e Silva
Department of Microbiology and Immunology, UNESP-São Paulo State University, Biosciences Institute, Botucatu, São Paulo, Brazil.
Ângela Maria Victoriano de Campos
ÂM
Soares
Department of Pathology, UNESP - São Paulo State University, Botucatu Medical School, Botucatu, São Paulo, Brazil.
Julio
J
De Faveri
Department of Dermatology and Radiotherapy, UNESP - São Paulo State University, Botucatu Medical School, Botucatu, São Paulo, Brazil.
Silvio Alencar
SA
Marques
Department of Pathology, UNESP - São Paulo State University, Botucatu Medical School, Botucatu, São Paulo, Brazil.
Mariângela Esther Alencar
ME
Marques
Department of Chemistry, UNESP-São Paulo State University, School of Sciences, Bauru, São Paulo, Brazil.
Valdecir Farias
VF
Ximenes
Department of Pathology, UNESP - São Paulo State University, Botucatu Medical School, Botucatu, São Paulo, Brazil.
Luciane Alarcão
LA
Dias-Melicio
D000368:Aged; D001938:Brazil; D065206:Extracellular Traps / Q000378:metabolism*; D006801:Humans; D008297:Male; D018613:Microscopy, Confocal; D008875:Middle Aged; D009504:Neutrophils / Q000276:immunology*; D010228:Paracoccidioides / Q000276:immunology*; D010229:Paracoccidioidomycosis / Q000276:immunology / Q000473:pathology*; D011446:Prospective Studies; D013057:Spectrum Analysis