Detection of platelet vesicles by flow cytometry

Nolan, John P.; Jones, Jennifer C.

doi:10.1080/09537104.2017.1280602

Cited by 78 publications

(78 citation statements)

References 47 publications

(48 reference statements)

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“…AF4 separates EVs on the basis of hydrodynamic size and can identify and sort out vesicles ranging from few nanometers to an undefined level of micrometers [145]. Last but not least, flow cytometry (FC), commonly used for the analysis of cells, is being actively developed for EV analysis [146,147] and is adopted by an increasing number of research groups, mainly to study the larger EVs [148,149]. Direct FC analysis using tracers that stain the whole vital EV compartment, such as lipophilic carbocyanine dyes, combined with phalloidin, that selectively binds to F-actin, accurately discriminates EVs from artifacts [42][43][44]150].…”

Section: Methodological Approachesmentioning

confidence: 99%

Extracellular Vesicles as Signaling Mediators and Disease Biomarkers across Biological Barriers

Simeone

Bologna

Lanuti

et al. 2020

IJMS

138

117

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Extracellular vesicles act as shuttle vectors or signal transducers that can deliver specific biological information and have progressively emerged as key regulators of organized communities of cells within multicellular organisms in health and disease. Here, we survey the evolutionary origin, general characteristics, and biological significance of extracellular vesicles as mediators of intercellular signaling, discuss the various subtypes of extracellular vesicles thus far described and the principal methodological approaches to their study, and review the role of extracellular vesicles in tumorigenesis, immunity, non-synaptic neural communication, vascular-neural communication through the blood-brain barrier, renal pathophysiology, and embryo-fetal/maternal communication through the placenta.

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Section: Methodological Approachesmentioning

confidence: 99%

Extracellular Vesicles as Signaling Mediators and Disease Biomarkers across Biological Barriers

Simeone

Bologna

Lanuti

et al. 2020

IJMS

138

117

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“…The characterization of the plasma MPs presents technical challenges due to their submicron sizes (van der Pol et al 2010(van der Pol et al , 2013Nolan & Jones, 2017). The profile of diabetes-induced circulating MPs presented here is the result of our significant effort to optimize equipment and sample preparation procedures.…”

Section: Discussionmentioning

confidence: 99%

Increased circulating microparticles in streptozotocin‐induced diabetes propagate inflammation contributing to microvascular dysfunction

Feng

Stork

et al. 2019

The Journal of Physiology

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Key points Circulating microparticles (MPs) are elevated in many cardiovascular diseases and have been considered as biomarkers of disease prognosis; however, current knowledge of MP functions has been mainly derived from in vitro studies and their precise impact on vascular inflammation and disease progression remains obscure. Using a diabetic rat model, we identified a >130‐fold increase in MPs in plasma of diabetic rats compared to normal rats, the majority of which circulated as aggregates, expressing multiple cell markers and largely externalized phosphatidylserine; vascular images illustrate MP biogenesis and their manifestations in microvessels of diabetic rats. Using combined single microvessel perfusion and systemic cross‐transfusion approaches, we delineated how diabetic MPs propagate inflammation in the vasculature and transform normal microvessels into an inflammatory phenotype observed in the microvessels of diabetic rats. Our observations derived from animal studies resembling conditions in diabetic patients, providing a mechanistic insight into MP‐mediated pathogenesis of diabetes‐associated multi‐organ microvascular dysfunction. Abstract In various cardiovascular diseases, microparticles (MPs), the membrane‐derived vesicles released during cell activation, are markedly increased in the circulation. These MPs have been recognized to play diverse roles in the regulation of cellular functions. However, current knowledge of MP function has been largely derived from in vitro studies. The precise impact of disease‐induced MPs on vascular inflammation and disease progression remains obscure. In this study we investigated the biogenesis, profile and functional roles of circulating MPs using a streptozotocin‐induced diabetic rat model with well‐characterized microvascular functions. Our study revealed a >130‐fold increase in MPs in the plasma of diabetic rats compared to normal rats. The majority of these MPs originate from platelets, leukocytes and endothelial cells (ECs), and circulate as aggregates. Diabetic MPs show greater externalized phosphatidylserine (PS) than normal MPs. When diabetic plasma or isolated diabetic MPs were perfused into normal microvessels or systemically transfused into normal rats, MPs immediately adhered to endothelium and subsequently mediated leukocyte adhesion. These microvessels then exhibited augmented permeability responses to inflammatory mediators, replicating the microvascular manifestations observed in diabetic rats. These effects were abrogated when MPs were removed from diabetic plasma or when diabetic MPs were pre‐coated with a lipid‐binding protein, annexin V, suggesting externalized PS to be key in mediating MP interactions with endothelium and leukocytes. Our study demonstrated that the elevated MPs in diabetic plasma are actively involved in the propagation of vascular inflammation through their adhesive surfaces, providing mechanistic insight into the pathogenesis of multi‐organ vascular dysfunction that commonly occurs in diabetic patients.

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“…Over the past few years, significant improvements have been made regarding the sensitivity of FCM to detect single vesicles with a diameter of < 300 nm which have further established this methodology as the most promising tool for routine enumeration of EV subsets. 113,114 However, the lack of sensitivity of the current flow cytometers to characterize small MVs and to detect small amount of molecules at the MVs surface still impede this evaluation. Moreover, most FCM studies did not take into account the presence of other active PLs, such as PE or the presence of physiological inhibitors, regulating the procoagulant activity, such as TFPI.…”

Section: Methodological Considerationsmentioning

confidence: 99%

Microvesicles and Cancer Associated Thrombosis

Lacroix

Vallier

Bonifay

et al. 2019

Semin Thromb Hemost

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Microvesicles (MVs) are small membrane enclosed structures released into the extracellular space by virtually all cell types. Their composition varies according to the cell origin and the stimulus which caused their formation. They harbor functional molecules and participate in intercellular communication. Endothelium, inflammatory cells, and cancer cells produce procoagulant MVs which contribute to cancer-associated thrombosis (CAT) in animal models. The tissue factor (TF) conveyed by these MVs was shown to play a key role in different animal models of experimental CAT. Alternatively, other molecular mechanisms involving polyphosphates or phosphatidylethanolamine could also be involved. In clinical practice, an association between an increase in the number of TF-positive or the procoagulant activity of these MVs and the occurrence of CAT has indeed been demonstrated in pancreatic-biliary cancers, suggesting that they could behave as a biomarker predictive for CAT. However, to date, this association was not confirmed in other types of cancer. Potential causes explaining this limited associated between MVs and CAT are (1) the diversity of mechanisms associating MVs and different types of cancer; (2) a more complex role of MVs in hemostasis integrating their anticoagulant and fibrinolytic activity; and (3) the lack of sensitivity, reproducibility, and standardization of current methodologies permitting measurement of MVs. Each of these hypotheses constitutes an interesting exploration path for a future reassessment of the clinical interest of the MVs in CAT.

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Detection of platelet vesicles by flow cytometry

Cited by 78 publications

References 47 publications

Extracellular Vesicles as Signaling Mediators and Disease Biomarkers across Biological Barriers

Extracellular Vesicles as Signaling Mediators and Disease Biomarkers across Biological Barriers

Increased circulating microparticles in streptozotocin‐induced diabetes propagate inflammation contributing to microvascular dysfunction

Microvesicles and Cancer Associated Thrombosis

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