NETQUANT: Automated Quantification of Neutrophil Extracellular Traps

Mohanty, Tirthankar; Sørensen, Ole E.; Nordenfelt, Pontus

doi:10.3389/fimmu.2017.01999

Cited by 29 publications

(29 citation statements)

References 26 publications

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“…Desai et al have found that NET formation upon 2 h stimulation with PMA or crystals involves the RIPK1/ RIPK3/MLKL-dependent pathway of necroptosis [115,116] (st. 51). They therefore argue that NET formation that involves cell death is a passive process secondary to plasma membrane rupture induced by necroptosis or other forms of necrosis [117] (st. 53) This view is opposed by others who have seen RIP3/MLKL-independent extrusion of DNA choosing different experimental conditions [118] (st. 52) or who argue that the definition of NET formation includes a highly regulated and coordinated process that is different from both necroptosis and necrosis (st. 50,53). It also needs to be mentioned that two novel studies [119,120] have demonstrated that the rupture of the plasma membrane during ROS-dependent NET formation is mediated by gasdermin D, thus connecting NETs with pyroptosis [121,122].…”

Section: Pathways Of Net Formationmentioning

confidence: 99%

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To NET or not to NET:current opinions and state of the science regarding the formation of neutrophil extracellular traps

et al. 2019

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Since the discovery and definition of neutrophil extracellular traps (NETs) 14 years ago, numerous characteristics and physiological functions of NETs have been uncovered. Nowadays, the field continues to expand and novel mechanisms that orchestrate formation of NETs, their previously unknown properties, and novel implications in disease continue to emerge. The abundance of available data has also led to some confusion in the NET research community due to contradictory results and divergent scientific concepts, such as pro-and anti-inflammatory roles in pathologic conditions, demarcation from other forms of cell death, or the origin of the DNA that forms the NET scaffold. Here, we present prevailing concepts and state of the science in NET-related research and elaborate on open questions and areas of dispute.

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Section: Pathways Of Net Formationmentioning

confidence: 99%

“…2). The use of different methods of detection and quantification of NETs in vitro, in serum and in tissue [48][49][50][51][52][53][54][55][56] also impedes interpretation and/or comparison.…”

Section: Introductionmentioning

confidence: 99%

To NET or not to NET:current opinions and state of the science regarding the formation of neutrophil extracellular traps

et al. 2019

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“…Two additional conserved aspects include the composition and the stimuli capable of inducing NETs. Studies that have attempted to quantify ETs via software programming [166,167] defined NETs from human neutrophils as changes in DNA/chromatin staining pattern [168]. ETs have been described at around 153 nm in length as a bead-on-a-string model [169].…”

Section: Discussionmentioning

confidence: 99%

Extracellular Traps: An Ancient Weapon of Multiple Kingdoms

Neumann

Brogden²,

Köckritz-Blickwede

2020

Biology

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The discovery, in 2004, of extracellular traps released by neutrophils has extended our understanding of the mode of action of various innate immune cells. This fascinating discovery demonstrated the extracellular trapping and killing of various pathogens by neutrophils. During the last decade, evidence has accumulated showing that extracellular traps play a crucial role in the defence mechanisms of various cell types present in vertebrates, invertebrates, and plants. The aim of this review is to summarise the relevant literature on the evolutionary history of extracellular traps used as a weapon in various kingdoms of life. Biology 2020, 9, 34 2 of 23On the basis of findings from several studies mostly performed with human and murine neutrophils, the following three different pathways that lead to the formation of ETs by innate immune cells have been identified: (1) Release of nuclear DNA by ETosis, a suicidal cell death associated with the rupture of the nuclear membrane prior to cell lysis [14,15]; (2) vesicular release of nuclear DNA by viable cells [16,17]; and (3) release of mitochondrial DNA [18,19]. However, the exact molecular mechanisms leading to one or the other phenotype of ET formation has still not been entirely clarified. A group of renowned scientists and experts on NETs has recently published an opinionated review on the subject due to the abundance of available data that has also led to some confusion in the NET/ET research community because of contradictory results and divergent scientific concepts, for example, the molecular pathways of ET formation or the origin of the DNA that forms the ET scaffold [20]. There is a strong consensus about the composition of ETs among the findings that NETs contain a high amount of granule proteins, for example, cell-type-specific proteases and other antibacterial molecules that are associated with DNA-histone complexes. However, it is still unclear how different triggers or pathways have led to phenotypical differences about the source of DNA or viability of the ET-releasing cell.Comparison of ET phenotypical differences between host species in relation to evolutionary aspects, especially by comparing data from phylogenetic groups, would help to understand the pathways of ET formation in more detail.

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“…NET formation has been examined using correlative microscopy—combining TEM, SEM, immunofluorescence, and live‐cell imaging techniques . Such imaging approaches can be supported by automated image quantification software . A range of probe‐based methods are also available for quantifying NETosis .…”

Section: Apoptosis Autophagy and Netsmentioning

confidence: 99%

Nuclear cytometry and chromatin organization

2018

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The nuclear-targeting chemical probe, for the detection and quantification of DNA within cells, has been a mainstay of cytometry-from the colorimetric Feulgen stain to smart fluorescent agents with tuned functionality. The level of nuclear structure and function at which the probe aims to readout, or indeed at which a DNA-targeted drug acts, is shadowed by a wide range of detection modalities and analytical methods. These methods are invariably limited in terms of the resolution attainable versus the volume occupied by targeted chromatin structures. The scalar challenge arises from the need to understand the extent and different levels of compaction of genomic DNA and how such structures can be re-modeled, reported, or even perturbed by both probes and drugs. Nuclear cytometry can report on the complex levels of chromatin order, disorder, disassembly, and even active disruption by probes and drugs. Nuclear probes can report defining features of clinical and therapeutic interest as in NETosis and other cell death processes. New cytometric approaches continue to bridge the scalar challenges of analyzing chromatin organization. Advances in super-resolution microscopy address the resolution and depth of analysis issues in cellular systems. Typical of recent insights into chromatin organization enabled by exploiting a DNA interacting probe is ChromEM tomography (ChromEMT). ChromEMT uses the unique properties of the anthraquinone-based cytometric dye DRAQ5™ to reveal that local and global 3D chromatin structures effect differences in compaction. The focus of this review is nuclear and chromatin cytometry, with linked reference to DNA targeting probes and drugs as exemplified by the anthracenediones.

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NETQUANT: Automated Quantification of Neutrophil Extracellular Traps

Cited by 29 publications

References 26 publications

To NET or not to NET:current opinions and state of the science regarding the formation of neutrophil extracellular traps

To NET or not to NET:current opinions and state of the science regarding the formation of neutrophil extracellular traps

Extracellular Traps: An Ancient Weapon of Multiple Kingdoms

Nuclear cytometry and chromatin organization

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