Quantitative detection of circulating endothelial cells in vasculitis: comparison of flow cytometry and immunomagnetic bead extraction

Clarke, L.; Shah, Vanita; Arrigoni, Francesca; Eleftheriou, D; Hong, Ying; Halcox, Julian; Klein, Nigel; Brogan, Paul A.

doi:10.1111/j.1538-7836.2008.02953.x

Cited by 36 publications

(47 citation statements)

References 24 publications

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“…We also measured CECs with immunomagnetic bead extraction as supporting evidence of endothelial injury due to active vasculitis, using the consensus protocol previously described by ourselves and others. 27,42 mAbs and Reagents…”

Section: Study Populationmentioning

confidence: 99%

Anti-Neutrophil Cytoplasmic Antibodies Stimulate Release of Neutrophil Microparticles

Hong

Eleftheriou

Hussain

et al. 2012

Journal of the American Society of Nephrology

138

151

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The mechanisms by which anti-neutrophil cytoplasmic antibodies (ANCAs) may contribute to the pathogenesis of ANCA-associated vasculitis are not well understood. In this study, both polyclonal ANCAs isolated from patients and chimeric proteinase 3-ANCA induced the release of neutrophil microparticles from primed neutrophils. These microparticles expressed a variety of markers, including the ANCA autoantigens proteinase 3 and myeloperoxidase. They bound endothelial cells via a CD18-mediated mechanism and induced an increase in endothelial intercellular adhesion molecule-1 expression, production of endothelial reactive oxygen species, and release of endothelial IL-6 and IL-8. Removal of the neutrophil microparticles by filtration or inhibition of reactive oxygen species production with antioxidants abolished microparticle-mediated endothelial activation. In addition, these microparticles promoted the generation of thrombin. In vivo, we detected more neutrophil microparticles in the plasma of children with ANCAassociated vasculitis compared with that in healthy controls or those with inactive vasculitis. Taken together, these results support a role for neutrophil microparticles in the pathogenesis of ANCA-associated vasculitis, potentially providing a target for future therapeutics.

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Section: Study Populationmentioning

confidence: 99%

Anti-Neutrophil Cytoplasmic Antibodies Stimulate Release of Neutrophil Microparticles

Hong

Eleftheriou

Hussain

et al. 2012

Journal of the American Society of Nephrology

138

151

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“…11,12,15 Importantly, CEC levels are predominantly affected by disease activity and endothelial injury and not treatment per se. 11,12,15 Our study now suggests that elevated CECs, and therefore persistence of endothelial injury, vary between children with single and recurrent AIS. CECs could therefore be an attractive candidate biomarker to detect those at risk of recurrent AIS, a finding that is consistent with the strong correlation between CECs and disease activity in systemic vasculitis.…”

mentioning

confidence: 99%

“…CECs were identified with immunomagnetic bead extraction based on an international consensus protocol. 11,24 CECs were defined as Ulex europaeus lectin (Sigma-Aldrich) bright cells .10 mm in size, with .5 magnetic beads attached (figure e-1A). 11,15,24 Cell-derived microparticles.…”

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confidence: 99%

Endothelial injury in childhood stroke with cerebral arteriopathy

et al. 2012

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Objective: Circulating endothelial cells (CECs) and microparticles (MPs) have been reported to reflect endothelial injury, cellular activation, and MP-mediated thrombin generation. We tested the hypothesis that these indices differ between children with cerebral arteriopathy and arterial ischemic stroke (AIS) recurrence, and those with a single event.Methods: This was a single-center cross-sectional study of 46 children with AIS and cerebral arteriopathy matched with pediatric controls. AIS recurrence was defined as new acute neurologic deficit with radiologic evidence of further cerebral infarction. CECs and MPs were identified with immunomagnetic bead extraction and flow cytometry, respectively. MP function as assessed by thrombin generation was determined using a fluorogenic assay.Results: Ten children had AIS recurrence while 36 had a single AIS event. CECs were raised in children with recurrent AIS, compared to those with no recurrence (p 5 0.0001), and in controls (p 5 0.0001). Total circulating annexin V1 MPs were significantly greater in children with recurrence than in those with no recurrence (p 5 0.0020). These MPs were of endothelial or platelet origin, and a subpopulation expressed tissue factor. Finally, MP-mediated thrombin generation was enhanced in children with recurrent AIS compared to those with no recurrence (p 5 0.0001), providing a link between inflammation, endothelial injury, and increased thrombotic tendency. GLOSSARY AIS 5 arterial ischemic stroke; AVM 5 arteriovenous malformation; CA 5 catheter arteriography; CEC 5 circulating endothelial cell; CI 5 confidence interval; EMP 5 endothelial microparticle; ETP 5 endogenous thrombin potential; FCA 5 focal cerebral arteriopathy; FITC 5 fluorescein isothiocyanate; FLAIR 5 fluid-attenuated inversion recovery; GOSH 5 Great Ormond Street Hospital; MP 5 microparticle; MPFP 5 microparticle-free plasma; MR 5 magnetic resonance; MRA 5 magnetic resonance angiography; OR 5 odds ratio; PACNS 5 primary angiitis of the CNS; PE 5 phycoerythrin; PPP 5 platelet-poor plasma; PSGL-1 5 P-selectin glycoprotein ligand 1; TF 5 tissue factor; TGA 5 thrombin generation assay; vWF 5 von Willebrand factor. Conclusion:Childhood arterial ischemic stroke (AIS) results in neurologic morbidity in over two-thirds of survivors and recurrence in up to 20%.

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“…Duda et al (17 ) proposed a cytometry protocol for phenotypic identification and enumeration of CECs and other cells in the blood, which has been adopted (6 ). However, in specific clinical setting, the agreement of the counts of CECs from the 2 techniques would be poor (3 ). Although flow cytometry is capable of high throughput and multiparametric analysis, capital and operational costs might prohibit its usage in resource-limited regions.…”

confidence: 99%

“…Reports have indicated that the amount of CECs can serve as an indicator of the health of blood vessels, and this finding has been leveraged to monitor cardiovascular events (1,2 ) and vascular disease (3,4 ). Nozaki et al (1 ) presented an assessment of endothelial dysfunction by plasma concentrations of endotheliumderived microparticles.…”

confidence: 99%

Detection of Circulating Endothelial Cells via a Microfluidic Disk

Chen¹,

Lee

Pan³

et al. 2011

Clinical Chemistry

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BACKGROUND:Circulating endothelial cells (CECs) in the blood are rare but have been shown to be associated with various diseases. With the ratio of CECs to peripheral blood mononuclear cells (PBMCs) less than 1 part per thousand, their separation from PBMCs and detection are challenging. We present a means of detecting CECs from PBMCs via an economical microfluidic disk with a model cell system [human umbilical vein endothelial cells (HUVECs) in PBMCs], along with demonstration of its efficacy clinically.

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Quantitative detection of circulating endothelial cells in vasculitis: comparison of flow cytometry and immunomagnetic bead extraction

Cited by 36 publications

References 24 publications

Anti-Neutrophil Cytoplasmic Antibodies Stimulate Release of Neutrophil Microparticles

Anti-Neutrophil Cytoplasmic Antibodies Stimulate Release of Neutrophil Microparticles

Endothelial injury in childhood stroke with cerebral arteriopathy

Detection of Circulating Endothelial Cells via a Microfluidic Disk

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