28 December 2021: Review Articles
A Review of the Impact of Neutrophils and Neutrophil Extracellular Traps (NETs) on the Development of Aortic Aneurysms in Animal and Human Studies
Milena Michalska ABCDEFG* , Tadeusz Grochowiecki AEFG , Tomasz Jakimowicz DEF , Sławomir Nazarewski DDOI: 10.12659/MSM.935134
Med Sci Monit 2021; 27:e935134
Background
Use of Animal Models in AA
Pathogenesis of Aortic Aneurysm and Neutrophilic Role
Inflammation
Atherosclerosis and Thrombosis
NETosis in Vascular Pathologies
Process of Visualization and Measurement of NETs
NETs in AAA
NETs in Other Diseases that Can Be Risk Factors of AAA
Conclusions
References
Table 2 Summary of published studies that include influence on neutrophil extracellular traps in aortic aneurysm.
| No | Author | Year | Model | Population | Control group |
|---|---|---|---|---|---|
| 1. | Sandrine Delbosc et al []64 | 2011 | |||
| 2. | Humin Yan et al []67 | 2016 | |||
| 3. | Akshaya K. Meher et al []47 | 2018 | |||
| 4. | Michael Spinosa et al []62 | 2018 | |||
| 5. | Silvia D. Visonà et al []72 | 2018 | – | ||
| 6. | Patrick Haider et al []73 | 2020 | – | ||
| 7. | Wolf Eilenberg et al []74 | 2021 | |||
| 8. | Ming Wei et al []75 | 2021 | – | ||
| 9. | Akrivi Chrysantho-poulou et al []77 | 2021 | |||
| 1. | Cancer, infection, and any immune-mediated disease | H1, Cit-H4, elastase | Immunofluorescence | NETs are abundant in AAA ILTsThe enlarged concentration of cf-DNA in plasma and conditioned medium between AAA patients and healthy volunteersIncreased circulating MPO-DNA complexes in AAAChronic -bacteremia prompt neutrophil recruitment in experimental AAA | |
| Cit-H4 | Western blot | ||||
| cf-DNA | Quant- it Picogreen, dsDNA Reagent, Invitrogen | ||||
| MPO, MPO-DNA complexes, Anti-P.gingivalis Ig | ELISA | ||||
| MMP-9 | Gelatin zymography | ||||
| Bacterial endotoxin released by ILT | Limulus Amebocyte Lysate (LAL) chromogenic endpoint assay) | ||||
| DNA extraction and bacterial DNA amplification | QIamp DNA Blood Midi Kit (Qiagen) PCR | ||||
| Neutrophil isolation | Immunofluorescence | ||||
| 2. | – | DNA-CRAMP complexes | Generation of DNA-CRAMP complexes | In elastase-induced murine aortic tissues NETs are observed, mainly in the adventitiaNeutrophil proteases and NETs play an important role in the development of AAA | |
| Elastin degradation,macrophage, CD3+ T cells, DCs | Immunohistochemistry | AAA is triggered by the DNA-CRAMP complexINF I and DCs promote the inflammatory responses of AAA | |||
| IFN I, Histone H2B, MPO, CRAMP | Immunofluorescence | ||||
| DCs | Flow cytometric analysis | ||||
| Gelatinase | Zymography | ||||
| NET stimulation (by LPS or C5a) | Fluorescence microscope | ||||
| Cytokine analysis | Cytometric bead arrays | ||||
| 3. | – | NE | Immunofluorescence | NETosis is reduced by blocking IL-1β, in human AAA NETs coexist with IL-1β | |
| Cit-H3 | Western blot | ||||
| Ly-6B.2 (neutrophil indicator) Alfa-smooth muscle alkaline phosphatase | Immunohistochemistry | Human neutrophils can produce ceramide by IL-1β stimulation, decreased ceramide synthesis attenuates NETosisThe initial phase of murine experimental AAA includes development of NETsFor AAA mice development, IL-1β expression in neutrophils is necessary | |||
| cf-DNA | Quant-iT PicoGreen dsDNA Reagent ThermoFisher, Scientific | ||||
| Neutrophil culture and NETs induce, CerS6 | Flow cytometry | ||||
| Ceramide isolation | Liquid chromatography-mass spectrometry | ||||
| 4. | – | RvD1IL-1beta | RvD1 ELISA | In the Ang II mouse model, RvD1 reduced AAAs and decreased Cit-H3 | |
| CitH3 | Western blot | Treatment RvD1 displays lower stain for markers of NETosis | |||
| NE | Immunohistochemistry | ||||
| MMP-2MMP-9 | Gelatin zymography | In the treatment group RvD1, IL-1β is decreased, whereas IL-10 is increased | |||
| 5. | Chronic aortitis and periaortitis, purely traumatic dissections, incomplete sampling | α-smooth muscle alpha-actin (SMA) MPOCit-H3α-CD3, α-CD68 (macrophages identificator)CD31 and CD34 (EC identificator) | Immunohistochemistry | The presence of NEU and NETs were increased significantly in subacute infiltering dissections but decreased in early organized and late scarring groupsThere were high numbers of MPO neutrophils in acute dissection but the Cit-H3 NETS levels were relatively little | |
| Dissection age (4 phases) | Histological | ||||
| 6. | – | Cit-H4MPOCD68Ly6GPolarization of macrophages | Immunofluorescence | NETs develop voluntarily in human blood clotDifferently polarized macrophages can degrade NETs by DNA uptake by individuals’ mechanismsNETs were found in all vessels aortic wall; most NET were found in the abluminal section and then in the luminal part | |
| DNA uptake from blood clots by macrophages | Flow cytometry | ||||
| Degradation of NETs in vitro | Fluorescence | DNase1L3 can be produced by generated macrophages | |||
| Protein quantification (DNase) | ELISA and flow cytometry | Blockage of macropinocytosis cause enlarge NET burden and decreased thrombus degradation | |||
| DNase 1L1 | Immunofluorescence | Reginal concertation of macrophages is inversely connected with NETs in human AAA | |||
| Apoptosis assessment | Flow cytometry | ||||
| DNA extraction | PCR and electrophoresis | ||||
| 7. | Recent tumors ( | Cit-H3MPOcf-DNA-histone complexesIL-1beta | ELISA | The level of Cit-H3 in plasma can be a biomarker of AAA enlargementAfter repairing AAA level of Cit-H3 is decreased | |
| Ly6GCit-H4 | Immunofluorescence | Blocking the citrullination in histone can prevent the development of AAA in an Ang II-induced model | |||
| NGALelastase | Mass spectrometers (Luminex MagPix) | ||||
| 8. | MPO | Immunohistochemistry | In vitro and in vivo NET formation is enhanced by Ang IIIn vitro PAD4 inhibition (by YW3-56) reduced Ang II-induced NET formationPAD4 inhibition prevent AAA rapture and decline VSMC apoptosisBy p38/JNK pathway NET enhance VSMC apoptosis | ||
| Cit-H3Ly6GNE | Immunofluorescence | ||||
| dsDNAPAD4 | Quant-iT PicoGreen dsDNA Assay Kit Thermo Fisher, Scientific | ||||
| Depletion of VSMC | Annexin V/PI apoptosis detection kit (556547, BD biosciencesFlow cytometry | ||||
| Neutrophil isolation | Immunofluorescence | ||||
| p38p-JNK | Western blot | ||||
| 9. | Age | MPONECit-H4 | Immunofluorescence | The development of NETs is induced by Ang II in a ROS/autophagy-dependent mechanismIn fibrotic renal and aneurysmal aortic tissue are noticeable NETs from NEU with tissue factor expression | |
| Cit-H3 | ELISA | ||||
| TF | TF activity assay | ||||
| MPO/DNA complexes | ELISA | ||||
| RNA isolation | PCR | ||||
| Neutrophil isolation and viability | Flow cytometry | ||||