title: Infection-induced NETosis is a dynamic process involving neutrophil multitasking in vivo.
abstract: Neutrophil extracellular traps (NETs) are released as neutrophils die in vitro in a process requiring hours, leaving a temporal gap that invasive microbes may exploit. Neutrophils capable of migration and phagocytosis while undergoing NETosis have not been documented. During Gram-positive skin infections, we directly visualized live polymorphonuclear cells (PMNs) in vivo rapidly releasing NETs, which prevented systemic bacterial dissemination. NETosis occurred during crawling, thereby casting large areas of NETs. NET-releasing PMNs developed diffuse decondensed nuclei, ultimately becoming devoid of DNA. Cells with abnormal nuclei showed unusual crawling behavior highlighted by erratic pseudopods and hyperpolarization consistent with the nucleus being a fulcrum for crawling. A requirement for both Toll-like receptor 2 and complement-mediated opsonization tightly regulated NET release. Additionally, live human PMNs injected into mouse skin developed decondensed nuclei and formed NETS in vivo, and intact anuclear neutrophils were abundant in Gram-positive human abscesses. Therefore early in infection NETosis involves neutrophils that do not undergo lysis and retain the ability to multitask.
authors.affiliation: Department of Critical Care Medicine, University of Calgary, Calgary, Alberta, Canada. 2The Calvin, Phoebe and Joan Snyder Institute for Chronic Diseases, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Bryan G
authors.initials: BG
authors.lastname: Yipp
authors.forename: Björn
authors.initials: B
authors.lastname: Petri
authors.forename: Davide
authors.initials: D
authors.lastname: Salina
authors.forename: Craig N
authors.initials: CN
authors.lastname: Jenne
authors.forename: Brittney N V
authors.initials: BN
authors.lastname: Scott
authors.forename: Lori D
authors.initials: LD
authors.lastname: Zbytnuik
authors.forename: Keir
authors.initials: K
authors.lastname: Pittman
authors.forename: Muhammad
authors.initials: M
authors.lastname: Asaduzzaman
authors.forename: Kaiyu
authors.initials: K
authors.lastname: Wu
authors.forename: H Christopher
authors.initials: HC
authors.lastname: Meijndert
authors.forename: Stephen E
authors.initials: SE
authors.lastname: Malawista
authors.forename: Anne
authors.initials: A
authors.lastname: de Boisfleury Chevance
authors.forename: Kunyan
authors.initials: K
authors.lastname: Zhang
authors.forename: John
authors.initials: J
authors.lastname: Conly
authors.forename: Paul
authors.initials: P
authors.lastname: Kubes
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D005110:Extracellular Space / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D009068:Movement / Q000502:physiology*; D009504:Neutrophils / Q000276:immunology* / Q000378:metabolism / Q000502:physiology; D009895:Opsonin Proteins / Q000378:metabolism; D017192:Skin Diseases, Bacterial / Q000276:immunology* / Q000378:metabolism; D051195:Toll-Like Receptor 2 / Q000378:metabolism
chemical_list: D009895:Opsonin Proteins; D051195:Toll-Like Receptor 2; D049452:Green Fluorescent Proteins