RT Journal Article
SR Electronic
T1 Proteomic, biomechanical and functional analyses define neutrophil heterogeneity in systemic lupus erythematosus
JF Annals of the Rheumatic Diseases
JO Ann Rheum Dis
FD BMJ Publishing Group Ltd and European League Against Rheumatism
SP 209
OP 218
DO 10.1136/annrheumdis-2020-218338
VO 80
IS 2
A1 Kathleen R Bashant
A1 Angel M Aponte
A1 Davide Randazzo
A1 Paniz Rezvan Sangsari
A1 Alexander JT Wood
A1 Jack A Bibby
A1 Erin E West
A1 Arlette Vassallo
A1 Zerai G Manna
A1 Martin P Playford
A1 Natasha Jordan
A1 Sarfaraz Hasni
A1 Marjan Gucek
A1 Claudia Kemper
A1 Andrew Conway Morris
A1 Nicole Y Morgan
A1 Nicole Toepfner
A1 Jochen Guck
A1 Nehal N Mehta
A1 Edwin R Chilvers
A1 Charlotte Summers
A1 Mariana J Kaplan
YR 2021
UL http://ard.bmj.com/content/80/2/209.abstract
AB Objectives Low-density granulocytes (LDGs) are a distinct subset of proinflammatory and vasculopathic neutrophils expanded in systemic lupus erythematosus (SLE). Neutrophil trafficking and immune function are intimately linked to cellular biophysical properties. This study used proteomic, biomechanical and functional analyses to further define neutrophil heterogeneity in the context of SLE.Methods Proteomic/phosphoproteomic analyses were performed in healthy control (HC) normal density neutrophils (NDNs), SLE NDNs and autologous SLE LDGs. The biophysical properties of these neutrophil subsets were analysed by real-time deformability cytometry and lattice light-sheet microscopy. A two-dimensional endothelial flow system and a three-dimensional microfluidic microvasculature mimetic (MMM) were used to decouple the contributions of cell surface mediators and biophysical properties to neutrophil trafficking, respectively.Results Proteomic and phosphoproteomic differences were detected between HC and SLE neutrophils and between SLE NDNs and LDGs. Increased abundance of type 1 interferon-regulated proteins and differential phosphorylation of proteins associated with cytoskeletal organisation were identified in SLE LDGs relative to SLE NDNs. The cell surface of SLE LDGs was rougher than in SLE and HC NDNs, suggesting membrane perturbances. While SLE LDGs did not display increased binding to endothelial cells in the two-dimensional assay, they were increasingly retained/trapped in the narrow channels of the lung MMM.Conclusions Modulation of the neutrophil proteome and distinct changes in biophysical properties are observed alongside differences in neutrophil trafficking. SLE LDGs may be increasingly retained in microvasculature networks, which has important pathogenic implications in the context of lupus organ damage and small vessel vasculopathy.