Cleaved N-terminal histone tails distinguish between NADPH oxidase (NOX)-dependent and NOX-independent pathways of neutrophil extracellular trap formation
| Autor: | Jelle Gerretsen, Sebastian Boeltz, Peter Pickkers, Martin Herrmann, Cansu Yanginlar, Johan van der Vlag, Luis E. Muñoz, Elmar Pieterse, Nils Rother, Luuk B. Hilbrands |
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| Rok vydání: | 2018 |
| Předmět: |
inorganic chemicals
0301 basic medicine Immunology lnfectious Diseases and Global Health Radboud Institute for Molecular Life Sciences [Radboudumc 4] Enzyme-Linked Immunosorbent Assay Extracellular Traps General Biochemistry Genetics and Molecular Biology Histones 03 medical and health sciences chemistry.chemical_compound All institutes and research themes of the Radboud University Medical Center 0302 clinical medicine Rheumatology Western blot In vivo Rheumatic Diseases Sepsis medicine Humans Immunology and Allergy NADPH oxidase biology medicine.diagnostic_test business.industry Antibodies Monoclonal NADPH Oxidases Neutrophil extracellular traps respiratory system Molecular biology Renal disorders Radboud Institute for Molecular Life Sciences [Radboudumc 11] 030104 developmental biology Histone chemistry 030220 oncology & carcinogenesis Neutrophil elastase Myeloperoxidase cardiovascular system biology.protein Renal disorders Radboud Institute for Health Sciences [Radboudumc 11] business Nicotinamide adenine dinucleotide phosphate |
| Zdroj: | Annals of the Rheumatic Diseases, 77, 1790-1798 Annals of the Rheumatic Diseases, 77, 12, pp. 1790-1798 |
| ISSN: | 0003-4967 |
| Popis: | ObjectivesNeutrophil extracellular traps (NETs) act in various rheumatic diseases. Although NET formation was originally described as a nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX)-dependent pathway, it appears that there are also NOX-independent pathways of NET release. Currently, no tools are available that can discriminate between both NET-forming pathways. We aimed to develop a serological method allowing the discrimination between NETs generated through NOX-dependent or NOX-independent pathways.MethodsHistones from in vitro generated NOX-dependent and NOX-independent NETs were characterised with a panel of lupus-derived antibodies against N-terminal histone tails using immunofluorescence microscopy, western blot and ELISA. NETs in patients with NET-associated diseases, that is, rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), psoriatic arthritis (PsA) and sepsis, were characterised in sandwich ELISAs employing antibodies against myeloperoxidase (MPO) and N-terminal histone tails as detecting and capturing antibodies, respectively. Functional responses of endothelial cells to NOX-dependent and NOX-independent NETs were assessed as well.ResultsNeutrophil elastase cleaves the N-terminal tails of core histones during NOX-dependent, but not during NOX-independent NET formation. Consequently, the detection of MPO–histone complexes with antibodies against N-terminal histone tails allows discrimination between NETs formed through a NOX-dependent or NOX-independent manner. Characterisation of in vivo circulating NETs revealed the presence of NOX-independent NETs in RA, SLE and sepsis, but NOX-dependent NETs in PsA. NOX-independent NETs displayed an increased capacity to activate endothelial cells when compared with NOX-dependent NETs.ConclusionsThese results indicate heterogeneity in NET-forming pathways in vivo and highlight the need for disease-specific strategies to prevent NET-mediated pathology. |
| Databáze: | OpenAIRE |
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