Activated signature of antiphospholipid syndrome neutrophils reveals potential therapeutic target

Knight, Jason S.; Meng, He; Coit, Patrick; Yalavarthi, Srilakshmi; Sule, Gautam; Gandhi, Alex A.; Grenn, Robert C.; Mazza, Levi F.; Ali, Ramadan A.; Renauer, Paul; Wren, Jonathan D.; Bockenstedt, Paula; Wang, Hui; Eitzman, Daniel T.; Sawalha, Amr H.

doi:10.1172/jci.insight.93897

Cited by 84 publications

(70 citation statements)

References 74 publications

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“…Alternatively, it is possible that neutrophils that have shed CD62L in vivo are strongly activated to the point that they have preferentially left circulation, thereby being unavailable for our analysis. Interestingly, we also detected increased surface expression of the APS autoantigen β 2 GPI on the surface of APS neutrophils by flow cytometry (Figure G), which is in line with our 2017 study demonstrating a 4.5‐fold increase in β 2 GPI gene expression in APS neutrophils . It should be noted that at least one other group has demonstrated similar β 2 GPI surface and expression phenotypes in circulating APS monocytes ().…”

Section: Discussionsupporting

confidence: 89%

“…( Figure 2G), which is in line with our 2017 study demonstrating a 4.5-fold increase in β 2 GPI gene expression in APS neutrophils (9). It should be noted that at least one other group has demonstrated similar β 2 GPI surface and expression phenotypes in circulating APS monocytes (34,35).…”

Section: Discussionsupporting

confidence: 88%

“…One possibility is that the patient neutrophils have had time to up‐regulate CD62L expression, thereby effectively compensating for shedding. In support of this idea, CD62L was up‐regulated at the gene level in our recent transcriptomic profiling of APS neutrophils . Alternatively, it is possible that neutrophils that have shed CD62L in vivo are strongly activated to the point that they have preferentially left circulation, thereby being unavailable for our analysis.…”

Section: Discussionmentioning

confidence: 56%

“…Indeed, dismantling NETs with deoxyribonuclease (7) and preventing NET release (NETosis) via activation of adenosine receptors (8) has proven effective in murine models of APS. In further support of neutrophil hyperactivity in APS, our group has demonstrated that the APS neutrophil transcriptome is characterized by the up-regulation of a number of meta-groups, including a cellular defense node that includes L-selectin and P-selectin glycoprotein I among other adhesion molecules (9).…”

Section: Introductionmentioning

confidence: 87%

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Increased Adhesive Potential of Antiphospholipid Syndrome Neutrophils Mediated by β2 Integrin Mac‐1

Sule

Kelley

Gockman

et al. 2019

Arthritis & Rheumatology

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Objective While the role of antiphospholipid antibodies in activating endothelial cells has been extensively studied, the impact of these antibodies on the adhesive potential of leukocytes has received less attention. This study was undertaken to investigate the extent to which antiphospholipid syndrome (APS) neutrophils adhere to resting endothelial cells under physiologic flow conditions and the surface molecules required for that adhesion. Methods Patients with primary APS (n = 43), patients with a history of venous thrombosis but negative test results for antiphospholipid antibodies (n = 11), and healthy controls (n = 38) were studied. Cells were introduced into a flow chamber and perfused across resting human umbilical vein endothelial cells (HUVECs). Surface adhesion molecules were quantified by flow cytometry. Neutrophil extracellular trap release (NETosis) was assessed in neutrophil‐HUVEC cocultures. Results Upon perfusion of anticoagulated blood through the flow chamber, APS neutrophils demonstrated increased adhesion as compared to control neutrophils under conditions representative of either venous (n = 8; P < 0.05) or arterial (n = 15; P < 0.0001) flow. At the same time, APS neutrophils were characterized by up‐regulation of CD64, CEACAM1, β2‐glycoprotein I, and activated Mac‐1 on their surface (n = 12–18; P < 0.05 for all markers). Exposing control neutrophils to APS plasma or APS IgG resulted in increased neutrophil adhesion (n = 10–11; P < 0.0001) and surface marker up‐regulation as compared to controls. A monoclonal antibody specific for activated Mac‐1 reduced the adhesion of APS neutrophils in the flow‐chamber assay (P < 0.01). The same monoclonal antibody reduced NETosis in neutrophil–HUVEC cocultures (P < 0.01). Conclusion APS neutrophils demonstrate increased adhesive potential, which is dependent upon the activated form of Mac‐1. In patients, this could lower the threshold for neutrophil–endothelium interactions, NETosis, and possibly thrombotic events.

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Section: Discussionsupporting

confidence: 89%

Section: Discussionsupporting

confidence: 88%

Section: Discussionmentioning

confidence: 56%

Section: Introductionmentioning

confidence: 87%

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Increased Adhesive Potential of Antiphospholipid Syndrome Neutrophils Mediated by β2 Integrin Mac‐1

Sule

Kelley

Gockman

et al. 2019

Arthritis & Rheumatology

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“…We have previously demonstrated epigenetic aberrancies in SLE neutrophils, with robust demethylation in interferon-regulated genes [7]. More recently, we demonstrated a pro-inflammatory transcriptional signature in APS neutrophils, suggesting a role for neutrophils in the pathogenesis of APS [8]. The DNA methylation profile of APS remains to be characterized, however.…”

Section: Introductionmentioning

confidence: 99%

Genome-wide DNA methylation analysis in primary antiphospholipid syndrome neutrophils

Weeding

Coit

Yalavarthi

et al. 2018

Clinical Immunology

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Antiphospholipid syndrome (APS) is a systemic autoimmune disease characterized by thromboembolic events and pregnancy loss. We sought to characterize the DNA methylation profile of primary APS in comparison to healthy controls and individuals with SLE. In primary APS neutrophils compared to controls, 17 hypomethylated and 25 hypermethylated CpG sites were identified. Notable hypomethylated genes included ETS1, a genetic risk locus for SLE, and PTPN2, a genetic risk locus for other autoimmune diseases. Gene ontology analysis of hypomethylated genes revealed enrichment of genes involved in pregnancy. None of the differentially methylated sites in primary APS were differentially methylated in SLE neutrophils, and there was no demethylation of interferon signature genes in primary APS as is seen in SLE. Hypomethylation within a single probe in the IFI44L promoter (cg06872964) was able to distinguish SLE from primary APS with a sensitivity of 93.3% and specificity of 80.0% at a methylation fraction of 0.329.

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Anti–Neutrophil Extracellular Trap Antibodies and Impaired Neutrophil Extracellular Trap Degradation in Antiphospholipid Syndrome

Zuo

Yalavarthi

Gockman

et al. 2020

Arthritis & Rheumatology

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Objective The release of neutrophil extracellular traps (NETs) by hyperactive neutrophils has recently been recognized to play an important role in antiphospholipid syndrome (APS). This study was undertaken to evaluate autoantibodies targeting NETs in patients with primary APS, and to determine their potential functions and clinical associations. Methods We measured global anti‐NET activity in 76 patients with primary APS, 23 patients with systemic lupus erythematosus without antiphospholipid antibodies (aPL), 11 patients with a history of unprovoked venous thrombosis without aPL, and 44 healthy controls. The ability of APS sera to degrade NETs was also assessed. Results We found markedly elevated levels of anti‐NET IgG and IgM in patients with primary APS compared with healthy controls (for IgG, mean ± SD optical density 0.55 ± 0.34 versus 0.33 ± 0.17; for IgM, mean ± SD optical density 0.76 ± 0.51 versus 0.26 ± 0.23). This anti‐NET activity did not correlate with levels of traditional aPL and was relatively stable over time. Mechanistically, anti‐NET antibodies (especially of the IgG isotype) impaired the ability of patient sera to degrade NETs (r = 0.4, P = 0.003). Levels of anti‐NET IgM inversely correlated with complement C4 (r = 0.4, P = 0.019). Clinically, anti‐NET antibodies associated with certain APS clinical manifestations, and in particular recurrent venous thrombosis (odds ratio 4.3; P = 0.002). Interestingly, anti‐NET antibody levels also appeared to be associated with unprovoked venous thrombosis in the general population (for IgM, mean ± SD optical density 0.67 ± 0.34 versus 0.26 ± 0.23). Conclusion Our data indicate high levels of anti‐NET antibodies in patients with primary APS, which may impair NET clearance and activate the complement cascade. These findings may ultimately enable more effective risk stratification.

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Activated signature of antiphospholipid syndrome neutrophils reveals potential therapeutic target

Cited by 84 publications

References 74 publications

Increased Adhesive Potential of Antiphospholipid Syndrome Neutrophils Mediated by β2 Integrin Mac‐1

Increased Adhesive Potential of Antiphospholipid Syndrome Neutrophils Mediated by β2 Integrin Mac‐1

Genome-wide DNA methylation analysis in primary antiphospholipid syndrome neutrophils

Anti–Neutrophil Extracellular Trap Antibodies and Impaired Neutrophil Extracellular Trap Degradation in Antiphospholipid Syndrome

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