Quantification of NETs‐associated markers by flow cytometry and serum assays in patients with thrombosis and sepsis

Lee, Koon; Cavanaugh, Lauren; Leung, Halina; Feng, Yuping; Ahmadi, Zohra; Chong, Beng H.; Passam, Freda

doi:10.1111/ijlh.12800

Cited by 55 publications

(67 citation statements)

References 35 publications

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“…Indeed, patients with elevated H3Cit levels experienced a significantly higher cumulative incidence of VTE than patients with lower levels. The proportion of MPO/ H3Cit-positive neutrophilsa specific marker of NETosisdetected by flow cytometry, was also significantly higher in patients with DVT than in controls [67]. However, an association between NETosisspecific biomarkers and PTS has not been reported.…”

Section: Markers Of Netosismentioning

confidence: 94%

Inflammation in deep vein thrombosis: a therapeutic target?

et al. 2019

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Deep vein thrombosis is a common disease associated with a variety of complications including post-thrombotic syndrome as a late complication. It is now clear that in addition to classical deep vein thrombosis triggers such as blood flow disturbance, hypercoagulability, and vessel wall changes, inflammation has a key role in the pathophysiology of deep vein thrombosis, and there is a close relationship between inflammation and coagulation. As attested by changes in several plasma biomarkers, inflammation may have a significant role in the development of post-thrombotic syndrome. Here, we review the link between inflammation and deep vein thrombosis and thus the potential value of anti-inflammatory and/or anticoagulant drugs in the treatment of deep vein thrombosis and the prevention of postthrombotic syndrome.

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Section: Markers Of Netosismentioning

confidence: 94%

Inflammation in deep vein thrombosis: a therapeutic target?

et al. 2019

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“…To compensate for this shortcoming, lung tissue biopsies should be stained for NET detection in order to confirm the ex vivo data. In addition, cf DNA as well as granule and histone proteins are often quantified as "in vivo NET formation" 30 . Clearly, it is impossible to distinguish whether the released DNA/ protein complexes are owing to NET formation or neutrophil death 31,32 , the latter of which was reported to occur under in vivo conditions more than 50 years ago 33,34 .…”

Section: Nets and Eets In Bacterial Infectionsmentioning

confidence: 99%

“…Moreover, in elderly patients, levels of circulating extracellular DNA correlated with the extent of venous thromboembolism, inflammation as assessed by C-reactive protein and leukocytosis, and predicted mortality 133 . Recently, flow cytometric detection of MPO/citrullinated histone 3-positive neutrophils and serum dsDNA have been proposed for biomarker assessment 30 . However, as mentioned earlier, these biomarkers are not suitable for the definition of NET formation and can also not be used for mechanistic studies.…”

Section: Nets In Venous Thrombosis and Thromboembolismmentioning

confidence: 99%

In vivo evidence for extracellular DNA trap formation

et al. 2020

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Extracellular DNA trap formation is a cellular function of neutrophils, eosinophils, and basophils that facilitates the immobilization and killing of invading microorganisms in the extracellular milieu. To form extracellular traps, granulocytes release a scaffold consisting of mitochondrial DNA in association with granule proteins. As we understand more about the molecular mechanism for the formation of extracellular DNA traps, the in vivo function of this phenomenon under pathological conditions remains an enigma. In this article, we critically review the literature to summarize the evidence for extracellular DNA trap formation under in vivo conditions. Extracellular DNA traps have not only been detected in infectious diseases but also in chronic inflammatory diseases, as well as in cancer. While on the one hand, extracellular DNA traps clearly exhibit an important function in host defense, it appears that they can also contribute to the maintenance of inflammation and metastasis, suggesting that they may represent an interesting drug target for such pathological conditions. Facts •The demonstration of extracellular DNA traps in vivo requires sections of affected tissues, which are to be investigated with special staining techniques. These structures are seen in multiple inflammatory and cancer diseases.Is there a simple biomarker that reflects extracellular DNA trap formation?• What is the contribution of extracellular microbe killing compared to intracellular killing following phagocytosis?• The mechanism of DNA trap formation is unknown.

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“…The flow cytometry protocol was adapted from the protocol published by Lee et al [43]. In brief, 100 µL of whole blood were incubated with 10 µg immunoglobin G (IgG, 10% solution, Grifols, Barcelona, Spain) for 10 min in order to eliminate unspecific binding sites.…”

Section: Flow Cytometrymentioning

confidence: 99%

“…Second, the appropriate gating strategy to identify NET-releasing neutrophils is still under discussion [41,42]. In 2018, Lee et al published an optimized method of Gavillet et al's protocol, which aimed to quantify NET-releasing neutrophils by using whole blood probes without cell fixation [43]. Therefore, this method seems more feasible for implementation in an ICU.…”

Section: Introductionmentioning

confidence: 99%

Flow Cytometry-Based Quantification of Neutrophil Extracellular Traps Shows an Association with Hypercoagulation in Septic Shock and Hypocoagulation in Postsurgical Systemic Inflammation—A Proof-of-Concept Study

Schneck

Mallek

Schiederich

et al. 2020

JCM

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This proof-of-concept study aimed to evaluate a novel method of flow cytometry-based quantification of neutrophil extracellular traps (NETs) in septic shock patients and to identify possible interactions between the number of free-circulating NETs and alterations of the coagulatory system. Patients suffering from septic shock, a matched control group (CTRL), and patients suffering from systemic inflammation after cardiac (CABG) or major abdominal surgery (MAS) were enrolled in this prospective proof-of-concept study. Compared to the matched controls, free-circulating NETs were significantly elevated in septic shock and postsurgical patients (data are presented in median (IQR)); septic shock: (2.7 (1.9–3.9); CABG: 2.7 (2.1–3.7); MAS: 2.7 (2.1–3.9); CTRL: 1.6 (1–2); CTRL vs. septic shock: p = 0.001; CTRL vs. CABG: p < 0.001; CTRL vs. MAS: p < 0.001). NETs correlated positively with FIBTEM mean clot firmness (MCF) in septic shock patients (r = 0.37, p < 0.01) while they correlated negatively in surgical patients (CABG: r = −0.28, p < 0.01; MAS: r = −0.25, p = 0.03). Flow-cytometric quantification of NETs showed a significant increase in free-circulating NETs under inflammatory conditions. Furthermore, this study hints to an association of the number of NETs with hypercoagulation in septic shock patients and hypocoagulation in surgery-induced inflammation.

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Quantification of NETs‐associated markers by flow cytometry and serum assays in patients with thrombosis and sepsis

Cited by 55 publications

References 35 publications

Inflammation in deep vein thrombosis: a therapeutic target?

Inflammation in deep vein thrombosis: a therapeutic target?

In vivo evidence for extracellular DNA trap formation

Flow Cytometry-Based Quantification of Neutrophil Extracellular Traps Shows an Association with Hypercoagulation in Septic Shock and Hypocoagulation in Postsurgical Systemic Inflammation—A Proof-of-Concept Study

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