Histone H3 clipping is a novel signature of human neutrophil extracellular traps

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Abstract

Neutrophils are critical to host defence, executing diverse strategies to perform their antimicrobial and regulatory functions. One tactic is the production of neutrophil extracellular traps (NETs). In response to certain stimuli neutrophils decondense their lobulated nucleus and release chromatin into the extracellular space through a process called NETosis. However, NETosis, and the subsequent degradation of NETs, can become dysregulated. NETs are proposed to play a role in infectious as well as many non-infection related diseases including cancer, thrombosis, autoimmunity and neurological disease. Consequently, there is a need to develop specific tools for the study of these structures in disease contexts. In this study, we identified a NET-specific histone H3 cleavage event and harnessed this to develop a cleavage site-specific antibody for the detection of human NETs. By microscopy, this antibody distinguishes NETs from chromatin in purified and mixed cell samples. It also detects NETs in tissue sections. We propose this antibody as a new tool to detect and quantify NETs.

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Data generated or analysed during this study are included in the manuscript. Source data files have been provided.

Article and author information

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Dorothea Ogmore Tilley

Department of Cellular Microbiology, Max Planck Institute for Infection Biology, Berlin, Germany

Competing interests
Dorothea Ogmore Tilley, has made a patent application for this antibody hybridoma cell line and sequence and its use in the detection of NETs outside of research purposes. No. EP 21 159 757.0.

"This ORCID iD identifies the author of this article:" 0000-0003-3254-6991
Ulrike Abuabed

Microscopy Core Facility, Max Planck Institute for Infection Biology, Berlin, Germany

Competing interests
No competing interests declared.
Ursula Zimny Arndt

Protein Analysis Core Facility, Max Planck Institute for Infection Biology, Berlin, Germany

Competing interests
No competing interests declared.
Monika Schmid

Protein Analysis Core Facility, Max Planck Institute for Infection Biology, Berlin, Germany

Competing interests
No competing interests declared.
Stefan Florian

Institut für Pathologie, Charité - Universitätsmedizin Berlin, Berlin, Germany

Competing interests
No competing interests declared.
Peter R Jungblut

Protein Analysis Core Facility, Max Planck Institute for Infection Biology, Berlin, Germany

Competing interests
No competing interests declared.
Volker Brinkmann

Microscopy Core Facility, Max Planck Institute for Infection Biology, Berlin, Germany

Competing interests
No competing interests declared.
Alf Herzig

Department of Cellular Microbiology, Max Planck Institute for Infection Biology, Berlin, Germany

Competing interests
Alf Herzig, has made a patent application for this antibody hybridoma cell line and sequence and its use in the detection of NETs outside of research purposes. No. EP 21 159 757.0.
Arturo Zychlinsky

Department of Cellular Microbiology, Max Planck Institute for Infection Biology, Berlin, Germany

For correspondence
zychlinsky@mpiib-berlin.mpg.de

Competing interests
Arturo Zychlinsky, has made a patent application for this antibody hybridoma cell line and sequence and its use in the detection of NETs outside of research purposes. No. EP 21 159 757.0.

"This ORCID iD identifies the author of this article:" 0000-0001-6018-193X

Funding

Max Planck Institute for Infection Biology

Arturo Zychlinsky

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Human subjects: Samples were collected from healthy donors who had provided informed consent according to the Declaration of Helsinki. Ethical approval was provided by the ethics committee of Charité-Universitätsmedizin Berlin and blood was donated anonymously at Charité Hospital Berlin. For histological tissue samples, tissue was obtained from historical archives and used in an anonymised way after approval through the Charité Ethics Committee (Project EA4/124/19, July 24, 2019). Informed consent from patients for use of biomaterials for research was obtained as part of the institutional treatment contract at Charité.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.