Dynamic NETosis is Carried Out by Live Neutrophils in Human and Mouse Bacterial Abscesses and During Severe Gram-Positive Infection

Neutrophil extracellular traps (NETs) are released, as neutrophils die in vitro, in a process requiring hours, leaving a temporal gap for invasive microbes to exploit. Functional neutrophils undergoing NETosis have not been documented. During Gram-positive skin infections, we directly visualized live PMN in vivo rapidly releasing NETs, which prevented bacterial Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms Corresponding author: Dr. Paul Kubes. HRIC 4AA16, 3330 Hospital Drive NW, Calgary, AB, Canada T2N 4N1. tel 403-220-8558, fax 403-270-7516. [email protected]. 9These authors contributed equally. The authors declare no conflict-of-interest or competing financial interests. Author Contributions B.G.Y. and B.P. designed the overall study, performed experiments, analyzed the data and wrote the manuscript, D.S. designed, performed and analyzed the TEM experiments, C.N.J. designed, performed and analyzed the immunofluorescence microscopy experiments, B.N.V.S performed intravital imaging, L.D.Z performed intravital imaging, Xenogen and dissemination experiments and analyzed the data, K.P. performed human neutrophil experiments, M.A. performed in vitro NETs assays, K.W. developed the transgenic bacteria, H.C.M helped develop the imaging analysis protocols, S.E.M and A.B.C designed and performed the human cytoplast experiments, K.Z. designed and supervised the development of the transgenic bacteria and provided expert advice on microbiology, J.C. designed the clinical experiments, supervised the acquisition of patient samples and clinical histories and provided expert advice on infectious diseases and microbiology, P.K. provided overall supervision, analyzed the data and wrote the manuscript. HHS Public Access Author manuscript Nat Med. Author manuscript; available in PMC 2015 August 07. Published in final edited form as: Nat Med. 2012 September ; 18(9): 1386–1393. doi:10.1038/nm.2847. A uhor M anscript