Mouse neutrophil extracellular traps in microbial infections.
Neutrophil extracellular traps (NETs) play an important role in innate immunity to microbial infections. NETs have been described in several species, but the molecular details of NET formation and their role in infection has not been addressed, partly because we lack optimal experimental models. Here we describe tools to investigate NET formation in neutrophils isolated from mice. Upon in vitro stimulation of wild-type mouse neutrophils with PMA, we analyzed 3 important steps in the process of NET formation: reactive oxygen species (ROS) production, NET cell death and NET release. As expected, neutrophils from NADPH oxidase-deficient mice failed to produce ROS and did not die nor release NETs upon stimulation. We found that neutrophils from several mouse strains produced NETs with different efficiency and that NET formation correlated with the amount of ROS produced. Activation with Candida albicans also resulted in ROS production and NET cell death. The hyphal form of this fungus induced NETs more effectively than the yeast form. With this work, we provide tools to study in vitro NET assembly in the mouse system.
Ermert
David
D
Department for Cellular Microbiology, Max Planck Institute for Infection Biology, Berlin, Germany.
Urban
Constantin F
CF
Laube
Britta
B
Goosmann
Christian
C
Zychlinsky
Arturo
A
Brinkmann
Volker
V
D000818:Animals; D002176:Candida albicans / Q000276:immunology / Q000472:pathogenicity; D002177:Candidiasis / Q000276:immunology* / Q000382:microbiology; D016923:Cell Death; D022082:Cellular Structures / Q000378:metabolism / Q000648:ultrastructure*; D004247:DNA / Q000378:metabolism; D004195:Disease Models, Animal; D007113:Immunity, Innate; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D018345:Mice, Knockout; D008855:Microscopy, Electron, Scanning; D019255:NADPH Oxidases / Q000235:genetics; D018375:Neutrophil Activation / Q000276:immunology*; D009504:Neutrophils / Q000166:cytology / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure; D011506:Proteins / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D013045:Species Specificity; D013755:Tetradecanoylphorbol Acetate / Q000276:immunology / Q000494:pharmacology
D011506:Proteins; D017382:Reactive Oxygen Species; D004247:DNA; D019255:NADPH Oxidases; D013755:Tetradecanoylphorbol Acetate