Rheumatoid arthritis synovial fluid neutrophils drive inflammation through production of chemokines, reactive oxygen species and neutrophil extracellular traps



Wright, Helen ORCID: 0000-0003-0442-3134, Lyon, Max, Chapman, Elinor ORCID: 0000-0002-4398-1705, Moots, Robert and Edwards, Steven ORCID: 0000-0002-7074-0552
(2020) Rheumatoid arthritis synovial fluid neutrophils drive inflammation through production of chemokines, reactive oxygen species and neutrophil extracellular traps. Unpublished. 2020.07.16.20155291-.

[thumbnail of SF-preprint-manuscript.pdf] Text
SF-preprint-manuscript.pdf - Submitted version

Download (11MB) | Preview

Abstract

<h4>ABSTRACT</h4> Rheumatoid arthritis (RA) is a chronic inflammatory disorder affecting synovial joints. Neutrophils are believed to play an important role in both the initiation and progression of RA, and large numbers of activated neutrophils are found within both synovial fluid (SF) and synovial tissue from RA joints. In this study we analysed paired blood and SF neutrophils from patients with severe, active RA (DAS28> 5.1, n=3) using RNA-seq. 772 genes were significantly different between blood and SF neutrophils. IPA analysis predicted that SF neutrophils had increased expression of chemokines and ROS production, delayed apoptosis, and activation of signalling cascades regulating the production of NETs. This activated phenotype was confirmed experimentally by incubating healthy control neutrophils in cell-free RA SF, which was able to delay apoptosis and induce ROS production in both unprimed and TNF α primed neutrophils (p< 0.05). RA SF significantly increased neutrophil migration through 3mM transwell chambers (p< 0.05) and also increased production of NETs by healthy control neutrophils, including exposure of myeloperoxidase (MPO) and citrullinated histone-H3-positive DNA NETs. IPA analysis predicted NET production was mediated by signalling networks including AKT, RAF1, SRC and NF- κ B. Our results expand the understanding of the molecular changes that take place in the neutrophil transcriptome during migration into inflamed joints in RA, and the altered phenotype in RA SF neutrophils. Specifically, RA SF neutrophils lose their migratory properties, residing within the joint to generate signals that promote joint damage, as well as inflammation via recruitment and activation of both innate and adaptive immune cells. We propose that this activated SF neutrophil phenotype contributes to the chronic inflammation and progressive damage to cartilage and bone observed in patients with RA.

Item Type: Article
Uncontrolled Keywords: 31 Biological Sciences, 32 Biomedical and Clinical Sciences, 3202 Clinical Sciences, Arthritis, Rheumatoid Arthritis, Women's Health, Clinical Research, Autoimmune Disease, 1 Underpinning research, 2 Aetiology, 1.1 Normal biological development and functioning, 2.1 Biological and endogenous factors, Inflammatory and immune system
Depositing User: Symplectic Admin
Date Deposited: 20 Jul 2020 07:39
Last Modified: 20 Jun 2024 16:33
DOI: 10.1101/2020.07.16.20155291
Related URLs:
URI: https://livrepository.liverpool.ac.uk/id/eprint/3094568