Cargando…
Enhanced neutrophil extracellular trap generation in rheumatoid arthritis: analysis of underlying signal transduction pathways and potential diagnostic utility
INTRODUCTION: Neutrophil extracellular traps (NETs) have recently been implicated in a number of autoimmune conditions, including rheumatoid arthritis (RA). We examined the underlying signaling pathways triggering enhanced NETosis in RA and ascertained whether the products of NETosis had diagnostic...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2014
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4229860/ https://www.ncbi.nlm.nih.gov/pubmed/24928093 http://dx.doi.org/10.1186/ar4579 |
_version_ | 1782344182005760000 |
---|---|
author | Sur Chowdhury, Chanchal Giaglis, Stavros Walker, Ulrich A Buser, Andreas Hahn, Sinuhe Hasler, Paul |
author_facet | Sur Chowdhury, Chanchal Giaglis, Stavros Walker, Ulrich A Buser, Andreas Hahn, Sinuhe Hasler, Paul |
author_sort | Sur Chowdhury, Chanchal |
collection | PubMed |
description | INTRODUCTION: Neutrophil extracellular traps (NETs) have recently been implicated in a number of autoimmune conditions, including rheumatoid arthritis (RA). We examined the underlying signaling pathways triggering enhanced NETosis in RA and ascertained whether the products of NETosis had diagnostic implications or usefulness. METHODS: Neutrophils were isolated from RA patients with active disease and from controls. Spontaneous NET formation from RA and control neutrophils was assessed in vitro with microscopy and enzyme-linked immunosorbent assay (ELISA) for NETosis-derived products. The analysis of the signal-transduction cascade included reactive oxygen species (ROS) production, myeloperoxidase (MPO), neutrophil elastase (NE), peptidyl arginine deiminase 4 (PAD4), and citrullinated histone 3 (citH3). NET formation was studied in response to serum and synovial fluid and immunoglobulin G (IgG) depleted and reconstituted serum. Serum was analyzed for NETosis-derived products, for which receiver operator characteristic (ROC) curves were calculated. RESULTS: Neutrophils from RA cases exhibited increased spontaneous NET formation in vitro, associated with elevated ROS production, enhanced NE and MPO expression, nuclear translocation of PAD4, PAD4-mediated citrullination of H3, and altered nuclear morphology. NET formation in both anti-citrullinated peptide antibody (ACPA)-positive and -negative RA was abolished by IgG depletion, but restored only with ACPA-positive IgG. NETosis-derived products in RA serum demonstrated diagnostic potential, the ROC area under the curve for cell-free nucleosomes being >97%, with a sensitivity of 91% and a specificity of 92%. No significant difference was observed between ACPA-positive and -negative cases. CONCLUSIONS: Signaling elements associated with the extrusion of NETs are significantly enhanced to promote NETosis in RA compared with healthy controls. NETosis depended on the presence of ACPA in ACPA-positive RA serum. The quantitation of NETosis-derived products, such as cell-free nucleosomes in serum, may be a useful complementary tool to discriminate between healthy controls and RA cases. |
format | Online Article Text |
id | pubmed-4229860 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-42298602014-11-14 Enhanced neutrophil extracellular trap generation in rheumatoid arthritis: analysis of underlying signal transduction pathways and potential diagnostic utility Sur Chowdhury, Chanchal Giaglis, Stavros Walker, Ulrich A Buser, Andreas Hahn, Sinuhe Hasler, Paul Arthritis Res Ther Research Article INTRODUCTION: Neutrophil extracellular traps (NETs) have recently been implicated in a number of autoimmune conditions, including rheumatoid arthritis (RA). We examined the underlying signaling pathways triggering enhanced NETosis in RA and ascertained whether the products of NETosis had diagnostic implications or usefulness. METHODS: Neutrophils were isolated from RA patients with active disease and from controls. Spontaneous NET formation from RA and control neutrophils was assessed in vitro with microscopy and enzyme-linked immunosorbent assay (ELISA) for NETosis-derived products. The analysis of the signal-transduction cascade included reactive oxygen species (ROS) production, myeloperoxidase (MPO), neutrophil elastase (NE), peptidyl arginine deiminase 4 (PAD4), and citrullinated histone 3 (citH3). NET formation was studied in response to serum and synovial fluid and immunoglobulin G (IgG) depleted and reconstituted serum. Serum was analyzed for NETosis-derived products, for which receiver operator characteristic (ROC) curves were calculated. RESULTS: Neutrophils from RA cases exhibited increased spontaneous NET formation in vitro, associated with elevated ROS production, enhanced NE and MPO expression, nuclear translocation of PAD4, PAD4-mediated citrullination of H3, and altered nuclear morphology. NET formation in both anti-citrullinated peptide antibody (ACPA)-positive and -negative RA was abolished by IgG depletion, but restored only with ACPA-positive IgG. NETosis-derived products in RA serum demonstrated diagnostic potential, the ROC area under the curve for cell-free nucleosomes being >97%, with a sensitivity of 91% and a specificity of 92%. No significant difference was observed between ACPA-positive and -negative cases. CONCLUSIONS: Signaling elements associated with the extrusion of NETs are significantly enhanced to promote NETosis in RA compared with healthy controls. NETosis depended on the presence of ACPA in ACPA-positive RA serum. The quantitation of NETosis-derived products, such as cell-free nucleosomes in serum, may be a useful complementary tool to discriminate between healthy controls and RA cases. BioMed Central 2014 2014-06-13 /pmc/articles/PMC4229860/ /pubmed/24928093 http://dx.doi.org/10.1186/ar4579 Text en Copyright © 2014 Sur Chowdhury et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/4.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Article Sur Chowdhury, Chanchal Giaglis, Stavros Walker, Ulrich A Buser, Andreas Hahn, Sinuhe Hasler, Paul Enhanced neutrophil extracellular trap generation in rheumatoid arthritis: analysis of underlying signal transduction pathways and potential diagnostic utility |
title | Enhanced neutrophil extracellular trap generation in rheumatoid arthritis: analysis of underlying signal transduction pathways and potential diagnostic utility |
title_full | Enhanced neutrophil extracellular trap generation in rheumatoid arthritis: analysis of underlying signal transduction pathways and potential diagnostic utility |
title_fullStr | Enhanced neutrophil extracellular trap generation in rheumatoid arthritis: analysis of underlying signal transduction pathways and potential diagnostic utility |
title_full_unstemmed | Enhanced neutrophil extracellular trap generation in rheumatoid arthritis: analysis of underlying signal transduction pathways and potential diagnostic utility |
title_short | Enhanced neutrophil extracellular trap generation in rheumatoid arthritis: analysis of underlying signal transduction pathways and potential diagnostic utility |
title_sort | enhanced neutrophil extracellular trap generation in rheumatoid arthritis: analysis of underlying signal transduction pathways and potential diagnostic utility |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4229860/ https://www.ncbi.nlm.nih.gov/pubmed/24928093 http://dx.doi.org/10.1186/ar4579 |
work_keys_str_mv | AT surchowdhurychanchal enhancedneutrophilextracellulartrapgenerationinrheumatoidarthritisanalysisofunderlyingsignaltransductionpathwaysandpotentialdiagnosticutility AT giaglisstavros enhancedneutrophilextracellulartrapgenerationinrheumatoidarthritisanalysisofunderlyingsignaltransductionpathwaysandpotentialdiagnosticutility AT walkerulricha enhancedneutrophilextracellulartrapgenerationinrheumatoidarthritisanalysisofunderlyingsignaltransductionpathwaysandpotentialdiagnosticutility AT buserandreas enhancedneutrophilextracellulartrapgenerationinrheumatoidarthritisanalysisofunderlyingsignaltransductionpathwaysandpotentialdiagnosticutility AT hahnsinuhe enhancedneutrophilextracellulartrapgenerationinrheumatoidarthritisanalysisofunderlyingsignaltransductionpathwaysandpotentialdiagnosticutility AT haslerpaul enhancedneutrophilextracellulartrapgenerationinrheumatoidarthritisanalysisofunderlyingsignaltransductionpathwaysandpotentialdiagnosticutility |