Paracoccidioides brasiliensis induces neutrophil extracellular traps in vitro
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1
Universidade Estadual Paulista "Julio de Mesquita Filho" - UNESP, Departamento de Microbiologia e Imunologia, Brazil
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2
Universidade Estadual Paulista "Julio de Mesquita Filho" - UNESP, Departamento de Patologia, Brazil
Paracoccidioides brasiliensis is a dimorphic fungus that is the causative agent of paracoccidioidomycosis, a chronic, subacute or acute mycosis, with visceral and cutaneous involvement. Recently, studies have focused on the role of neutrophils that are involved in primary response to the fungus. In addition to their ability to eliminate pathogens by phagocytosis and antimicrobial secretions, it has been shown recently that neutrophils can trap and kill microorganisms by release of extracellular structures composed by DNA and antimicrobial proteins, called neutrophil extracellular traps (NETs).The aim of this study was to demonstrate whether P. brasiliensis induces NETs release by human neutrophils activated or not with cytokines that promote fungicidal activity of these cells. Human neutrophils were isolated from peripheral blood of healthy donors and incubated with P. brasiliensis for 60, 90, 120 and 150 minutes. Some of these cultures were pré-treated with DNAse (to degrade NETs) or PMA (positive control of NETs release). Samples analyzed by scanning electron microscopy and photomicrographs demonstrated that P. brasiliensis induces NETs 60, 90, 120 and 150 minutes after interaction with neutrophils. DNAse treatment degraded these structures, whereas PMA induced their formation. Moreover, by using confocal imunofluorescence microscopy the following NET constituents were identified in activated cells with TNF-α, IFN-γ and GM-CSF: DNA (stained with DAPI, elastase (stained with FITC) and histones (stained with TEXAS RED). Thus, we demonstrated that Paracoccidioides brasiliensis is able to induce NETs.
Acknowledgements
FAPESP 2010/17405-5; FAPESP 2010/18957-1
Keywords:
Paracoccidioides brasiliensis,
neutrophil extracellular traps,
confocal imunofluorescence microscopy,
Scanning electron microscopy,
innate immunity
Conference:
15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013.
Presentation Type:
Abstract
Topic:
Host-pathogen interactions
Citation:
Bachiega
TF,
Fernandes
RK,
Rodrigues
DR,
Balderramas
HA,
Dias-Melicio
LA and
Soares
ÂM
(2013). Paracoccidioides brasiliensis induces neutrophil extracellular traps in vitro.
Front. Immunol.
Conference Abstract:
15th International Congress of Immunology (ICI).
doi: 10.3389/conf.fimmu.2013.02.00502
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Received:
29 Apr 2013;
Published Online:
22 Aug 2013.
*
Correspondence:
Miss. Tatiana F Bachiega, Universidade Estadual Paulista "Julio de Mesquita Filho" - UNESP, Departamento de Microbiologia e Imunologia, Botucatu, São Paulo, 18618970, Brazil, tatybachiega@hotmail.com
Prof. Ângela M Soares, Universidade Estadual Paulista "Julio de Mesquita Filho" - UNESP, Departamento de Microbiologia e Imunologia, Botucatu, São Paulo, 18618970, Brazil, acsoares@ibb.unesp.br