Low-density granulocytes: functionally distinct, immature neutrophils in rheumatoid arthritis with altered properties and defective TNF signalling

Journal of Leukocyte Biology - Tập 101 Số 2 - Trang 599-611 - 2017
Helen L. Wright1, Fatima Makki1, Robert J. Moots2, Steven W. Edwards1
1Institute of Integrative Biology, University of Liverpool, Liverpool, United Kingdom
2Institute of Ageing and Chronic Disease, University Hospital Aintree, University of Liverpool , Liverpool , United Kingdom

Tóm tắt

Abstract Our aim was to determine whether rheumatoid arthritis (RA) low-density granulocytes (LDGs) are functionally different from RA neutrophils. LDGs from 32 RA patients were characterized using flow cytometry and quantitative PCR (qPCR). RNA sequencing (RNA-Seq) was carried out on paired RA LDGs and neutrophils (n = 4) and validated using qPCR. Functional assays included chemotaxis, phagocytosis, reactive oxygen species (ROS) production, cell-cycle analysis, apoptosis, neutrophil extracellular trap (NET)osis, and measurement of cytokine production (n ≥ 5 paired RA LDGs/neutrophils). RA LDGs had a substantially altered transcriptome, expressing >5000 genes at significantly different levels compared with RA neutrophils, including elevated levels of transcripts for granule proteins [including elastase and myeloperoxidase (MPO)] and cell-cycle genes [including cyclin-dependent kinase (CDK)2, CDK4, and CDK6]. Approximately 1% of RA LDGs stained positive for the G2/S phase of the cell cycle. RA LDGs had a significantly lower constitutive rate of apoptosis compared with RA neutrophils and did not respond to TNF-α in culture. Expression of transcripts for cytokines and cytokine receptors was lower in RA LDGs. NET formation was lower in LDGs in response to PMA compared with RA neutrophils. Chemotaxis and phagocytosis was lower in RA LDGs compared with neutrophils. RA LDGs produced significantly lower amounts of ROS in response to fMLP following priming with TNF-α. Expression of TNFR1 and -2 mRNA and protein was significantly lower in LDGs. We conclude that RA LDGS are functionally different from RA neutrophils, representing an immature neutrophil population within peripheral blood. Their enhanced survival properties and decreased TNF signaling are likely to have important consequences for disease pathology and response to therapy.

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Tài liệu tham khảo

Wright, 2010, Neutrophil function in inflammation and inflammatory diseases, Rheumatology (Oxford), 49, 1618, 10.1093/rheumatology/keq045

Wright, 2014, The multifactorial role of neutrophils in rheumatoid arthritis, Nat. Rev. Rheumatol., 10, 593, 10.1038/nrrheum.2014.80

Mantovani, 2011, Neutrophils in the activation and regulation of innate and adaptive immunity, Nat. Rev. Immunol., 11, 519, 10.1038/nri3024

Wright, 2015, Interferon gene expression signature in rheumatoid arthritis neutrophils correlates with a good response to TNFi therapy, Rheumatology (Oxford), 54, 188, 10.1093/rheumatology/keu299

Wright, 2011, Changes in expression of membrane TNF, NF-kappaB activation and neutrophil apoptosis during active and resolved inflammation, Ann. Rheum. Dis., 70, 537, 10.1136/ard.2010.138065

Cross, 2003, Synovial fluid neutrophils transcribe and express class II major histocompatibility complex molecules in rheumatoid arthritis, Arthritis Rheum., 48, 2796, 10.1002/art.11253

Cross, 2004, Secretion of oncostatin M by neutrophils in rheumatoid arthritis, Arthritis Rheum., 50, 1430, 10.1002/art.20166

Hacbarth, 1986, Low density neutrophils in patients with systemic lupus erythematosus, rheumatoid arthritis, and acute rheumatic fever, Arthritis Rheum., 29, 1334, 10.1002/art.1780291105

Denny, 2010, A distinct subset of proinflammatory neutrophils isolated from patients with systemic lupus erythematosus induces vascular damage and synthesizes type I IFNs, J. Immunol., 184, 3284, 10.4049/jimmunol.0902199

Villanueva, 2011, Netting neutrophils induce endothelial damage, infiltrate tissues, and expose immunostimulatory molecules in systemic lupus erythematosus, J. Immunol., 187, 538, 10.4049/jimmunol.1100450

Martinelli, 2004, Induction of genes mediating interferon-dependent extracellular trap formation during neutrophil differentiation, J. Biol. Chem., 279, 44123, 10.1074/jbc.M405883200

Theilgaard-Mönch, 2005, The transcriptional program of terminal granulocytic differentiation, Blood, 105, 1785, 10.1182/blood-2004-08-3346

Rørvig, 2013, Proteome profiling of human neutrophil granule subsets, secretory vesicles, and cell membrane: correlation with transcriptome profiling of neutrophil precursors, J. Leukoc. Biol., 94, 711, 10.1189/jlb.1212619

Grayson, 2015, Neutrophil-related gene expression and low-density granulocytes associated with disease activity and response to treatment in antineutrophil cytoplasmic antibody-associated vasculitis, Arthritis Rheumatol., 67, 1922, 10.1002/art.39153

Cloke, 2012, Characterization of a novel population of low-density granulocytes associated with disease severity in HIV-1 infection, PLoS One, 7, e48939, 10.1371/journal.pone.0048939

Midgley, 2016, Increased expression of low density granulocytes in juvenile-onset systemic lupus erythematosus patients correlates with disease activity, Lupus, 25, 407, 10.1177/0961203315608959

Fu, 2014, Neutrophil-like low-density granulocytes are elevated in patients with moderate to severe persistent asthma, Ann. Allergy Asthma Immunol., 113, 635, 10.1016/j.anai.2014.08.024

Arnett, 1988, The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis, Arthritis Rheum., 31, 315, 10.1002/art.1780310302

Zhang, 2005, Selection of reference genes for gene expression studies in human neutrophils by real-time PCR, BMC Mol. Biol., 6, 4, 10.1186/1471-2199-6-4

Trapnell, 2009, TopHat: discovering splice junctions with RNA-Seq, Bioinformatics, 25, 1105, 10.1093/bioinformatics/btp120

Trapnell, 2012, Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and Cufflinks, Nat. Protoc., 7, 562, 10.1038/nprot.2012.016

Ramsköld, 2009, An abundance of ubiquitously expressed genes revealed by tissue transcriptome sequence data, PLOS Comput. Biol., 5, e1000598, 10.1371/journal.pcbi.1000598

Wright, 2013, RNA-seq reveals activation of both common and cytokine-specific pathways following neutrophil priming, PLoS One, 8, e58598, 10.1371/journal.pone.0058598

Thomas, 2015, Whose gene is it anyway? The effect of preparation purity on neutrophil transcriptome studies, PLoS One, 10, e0138982, 10.1371/journal.pone.0138982

Saeed, 2006, TM4 microarray software suite, Methods Enzymol., 411, 134, 10.1016/S0076-6879(06)11009-5

Borregaard, 1997, Granules of the human neutrophilic polymorphonuclear leukocyte, Blood, 89, 3503, 10.1182/blood.V89.10.3503

Klausen, 2004, End-stage differentiation of neutrophil granulocytes in vivo is accompanied by up-regulation of p27kip1 and down-regulation of CDK2, CDK4, and CDK6, J. Leukoc. Biol., 75, 569, 10.1189/jlb.1003474

Chiewchengchol, 2015, Differential changes in gene expression in human neutrophils following TNF-α stimulation: Up-regulation of anti-apoptotic proteins and down-regulation of proteins involved in death receptor signaling, Immun. Inflamm. Dis., 4, 35, 10.1002/iid3.90

Tamarozzi, 2014, A lack of confirmation with alternative assays questions the validity of IL-17A expression in human neutrophils using immunohistochemistry, Immunol. Lett., 162, 194, 10.1016/j.imlet.2014.10.025

Huang, 2015, Impaired phagocytosis of apoptotic cell material in serologically active clinically quiescent patients with systemic lupus erythematosis, Int. J. Rheum. Dis.

Ballantine, 2015, Increased soluble phagocytic receptors sMer, sTyro3 and sAxl and reduced phagocytosis in juvenile-onset systemic lupus erythematosus, Pediatr. Rheumatol. Online J., 13, 10, 10.1186/s12969-015-0007-y

Courtney, 1999, Increased apoptotic peripheral blood neutrophils in systemic lupus erythematosus: relations with disease activity, antibodies to double stranded DNA, and neutropenia, Ann. Rheum. Dis., 58, 309, 10.1136/ard.58.5.309

Midgley, 2009, The role of neutrophil apoptosis in juvenile-onset systemic lupus erythematosus, Arthritis Rheum., 60, 2390, 10.1002/art.24634

Khandpur, 2013, NETs are a source of citrullinated autoantigens and stimulate inflammatory responses in rheumatoid arthritis, Sci. Transl. Med., 5, 178ra40, 10.1126/scitranslmed.3005580

Pelletier, 2010, Evidence for a cross-talk between human neutrophils and Th17 cells, Blood, 115, 335, 10.1182/blood-2009-04-216085

Dal-Secco, 2015, A dynamic spectrum of monocytes arising from the in situ reprogramming of CCR2+ monocytes at a site of sterile injury, J. Exp. Med., 212, 447, 10.1084/jem.20141539

Strauss-Ayali, 2007, Monocyte subpopulations and their differentiation patterns during infection, J. Leukoc. Biol., 82, 244, 10.1189/jlb.0307191

Elghetany, 2002, Surface antigen changes during normal neutrophilic development: a critical review, Blood Cells Mol. Dis., 28, 260, 10.1006/bcmd.2002.0513

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Journal of Leukocyte Biology

Tập 101 Số 2

599-611

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