Neutrophil extracellular traps in sepsis: trade-off between pros and cons

Abstract

Sepsis, a life-threatening syndrome driven by dysregulated immune responses to infection, presents significant global health challenges with high mortality rates. Neutrophil extracellular traps (NETs), composed of deoxyribonucleic acid and antimicrobial proteins, play a dual role in sepsis pathogenesis. While NETs trap pathogens and enhance immune responses via antimicrobial activity and immune cell activation, their overproduction exacerbates tissue damage, coagulopathy, and organ dysfunction. This review explores the mechanisms of NET formation, including suicidal, vital, and noncanonical NETosis, and their regulation through pattern recognition receptors, complement systems, and chemokine signaling. The interplay between NETs and immune cells-such as macrophages, T cells, and platelets-is highlighted, emphasizing NETs’ role in both pathogen clearance and inflammatory injury. Excessive NETs contribute to sepsis-associated coagulopathy by activating platelets and damaging endothelial cells, while histones and proteases within NETs mediate cytotoxicity. Emerging therapeutic strategies targeting NETs, such as deoxyribonuclease, PAD4 inhibitors, and anti-inflammatory agents, show promise in preclinical studies but face clinical challenges due to their dual roles and off-target effects. Balancing NETs’ protective and pathological functions remains critical for sepsis management. This review aims to provide a comprehensive understanding of NETs in sepsis, offering insights for future research and clinical applications.

Keywords

Neutrophils / Sepsis / NETs / Inflammation / Therapeutic strategies

References

[1] Singer M, Deutschman CS, Seymour CW, Shankar-Hari M, Annane D, Bauer M, et al. The third international consensus definitions for sepsis and septic shock (Sepsis-3). Jama 2016; 315: 801-10. https://doi.org/10.1001/jama.2016.0287.

[2] Martin-Loeches I, Singer M, Leone M. Sepsis: key insights, future directions, and immediate goals. A review and expert opinion. Intensive Care Med 2024; 50: 2043-9. https://doi.org/10.1007/s00134-024-07694-z.

[3] Wang Q, Deng J, Sun J, Zhang H, Liu D, Gao C, et al. PDGFR kinase inhibitor protects against septic death via regulation of BTLA. Sci China Life Sci 2022; 65: 1917-28. https://doi.org/10.1007/s11427-021-2136-y.

[4] Rudd KE, Johnson SC, Agesa KM, Shackelford KA, Tsoi D, Kievlan DR, et al. Global, regional, and national sepsis incidence and mortality, 1990-2017: analysis for the global burden of disease study. Lancet 2020; 395: 200-11. https://doi.org/10.1016/s0140-6736(19)32989-7.

[5] Meyer NJ, Prescott HC. Sepsis and septic shock. N Engl J Med 2024; 391: 2133-46. https://doi.org/10.1056/NEJMra2403213.

[6] Gotts JE, Matthay MA. Sepsis: pathophysiology and clinical management. BMJ 2016; 353: i1585. https://doi.org/10.1136/bmj.i1585.

[7] Yu HH, Yang YH, Chiang BL. Chronic granulomatous disease: a comprehensive review. Clin Rev Allergy Immunol 2021; 61: 101-13. https://doi.org/10.1007/s12016-020-08800-x.

[8] Mutua V, Gershwin LJ. A review of neutrophil extracellular traps (NETs) in disease: potential anti-NETs therapeutics. Clin Rev Allergy Immunol 2021; 61: 194-211. https://doi.org/10.1007/s12016-020-08804-7.

[9] Barbu EA, Mendelsohn L, Samsel L, Thein SL. Pro-inflammatory cytokines associate with NETosis during sickle cell vaso-occlusive crises. Cytokine 2020; 127: 154933. https://doi.org/10.1016/j.cyto.2019.154933.

[10] Messaoud-Nacer Y, Culerier E, Rose S, Maillet I, Rouxel N, Briault S, et al. STING agonist diABZI induces PANoptosis and DNA mediated acute respiratory distress syndrome (ARDS). Cell Death Dis 2022; 13: 269. https://doi.org/10.1038/s41419-022-04664-5.

[11] Basu S, Hodgson G, Katz M, Dunn AR. Evaluation of role of G-CSF in the production, survival, and release of neutrophils from bone marrow into circulation. Blood 2002; 100: 854-61. https://doi.org/10.1182/blood.v100.3.854.

[12] Summers C, Rankin SM, Condliffe AM, Singh N, Peters AM, Chilvers ER. Neutrophil kinetics in health and disease. Trends Immunol 2010; 31: 318-24. https://doi.org/10.1016/j.it.2010.05.006.

[13] Cowland JB, Borregaard N. Granulopoiesis and granules of human neutrophils. Immunol Rev 2016; 273: 11-28. https://doi.org/10.1111/imr.12440.

[14] Pillay J, den Braber I, Vrisekoop N, Kwast LM, de Boer RJ, Borghans JA, et al. In vivo labeling with 2H2O reveals a human neutrophil lifespan of 5.4 days. Blood 2010; 116: 625-7. https://doi.org/10.1182/blood-2010-01-259028.

[15] Adrover JM, Del Fresno C, Crainiciuc G, Cuartero MI, Casanova-Acebes M, Weiss LA, et al. A neutrophil timer coordinates immune defense and vascular protection. Immunity 2019; 50: 390-402.e10. https://doi.org/10.1016/j.immuni.2019.01.002.

[16] Zhang H, Wang Y, Qu M, Li W, Wu D, Cata JP, et al. Neutrophil, neutrophil extracellular traps and endothelial cell dysfunction in sepsis. Clin Transl Med 2023; 13: e1170. https://doi.org/10.1002/ctm2.1170.

[17] Liew PX, Kubes P. The neutrophil’s role during health and disease. Physiol Rev 2019; 99: 1223-48. https://doi.org/10.1152/physrev.00012.2018.

[18] Silva CMS, Wanderley CWS, Veras FP, Sonego F, Nascimento DC, Gonçalves AV, et al. Gasdermin D inhibition prevents multiple organ dysfunction during sepsis by blocking NET formation. Blood 2021; 138: 2702-13. https://doi.org/10.1182/blood.2021011525.

[19] Ma Y, Zhao Y, Zhang X. Factors affecting neutrophil functions during sepsis: human microbiome and epigenetics. J Leukoc Biol 2024; 116: 672-88. https://doi.org/10.1093/jleuko/qiae107.

[20] Sun BW, Wang YE, Yang YX. Neutrophil and burn sepsis. Chin J Burns Wounds 2024; 40: 618-24. https://doi.org/10.3760/cma.j.cn501225-20240329-00109.

[21] de Bont CM, Boelens WC, Pruijn GJM. NETosis, complement, and coagulation: a triangular relationship. Cell Mol Immunol 2018; 16: 19-27. https://doi.org/10.1038/s41423-018-0024-0.

[22] Zhang D, Guo J, Shi C, Wang Y, Zhang Y, Zhang X, et al. MPO-DNA complexes and cf-DNA in patients with sepsis and their clinical value. Biomedicines 2024; 12: 2190. https://doi.org/10.3390/biomedicines12102190.

[23] Herster F, Bittner Z, Archer NK, Dickhofer S, Eisel D, Eigenbrod T, et al. Neutrophil extracellular trap-associated RNA and LL37 enable self-amplifying inflammation in psoriasis. Nat Commun 2020; 11: 105. https://doi.org/10.1038/s41467-019-13756-4.

[24] Brinkmann V, Reichard U, Goosmann C, Fauler B, Uhlemann Y, Weiss DS, et al. Neutrophil extracellular traps kill bacteria. Science 2004; 303: 1532-5. https://doi.org/10.1126/science.1092385.

[25] Hidalgo A, Libby P, Soehnlein O, Aramburu IV, Papayannopoulos V, Silvestre-Roig C. Neutrophil extracellular traps: from physiology to pathology. Cardiovasc Res 2022; 118: 2737-53. https://doi.org/10.1093/cvr/cvab329.

[26] Li X, Xiao S, Filipczak N, Yalamarty SSK, Shang H, Zhang J, et al. Role and therapeutic targeting strategies of neutrophil extracellular traps in inflammation. Int J Nanomedicine 2023; 18: 5265-87. https://doi.org/10.2147/ijn.S418259.

[27] Iba T, Levy JH. Inflammation and thrombosis: roles of neutrophils, platelets and endothelial cells and their interactions in thrombus formation during sepsis. J Thromb Haemost 2018; 16: 231-41. https://doi.org/10.1111/jth.13911.

[28] Dehghani T, Panitch A. Endothelial cells, neutrophils and platelets: getting to the bottom of an inflammatory triangle. Open Biol 2020; 10: 200161. https://doi.org/10.1098/rsob.200161.

[29] Wang Y, Liu Y. Neutrophil-induced liver injury and interactions between neutrophils and liver sinusoidal endothelial cells. Inflammation 2021; 44: 1246-62. https://doi.org/10.1007/s10753-021-01442-x.

[30] Castanheira FVS, Kubes P. Neutrophils and NETs in modulating acute and chronic inflammation. Blood 2019; 133: 2178-85. https://doi.org/10.1182/blood-2018-11-844530.

[31] Papayannopoulos V. Neutrophil extracellular traps in immunity and disease. Nat Rev Immunol 2018; 18: 134-47. https://doi.org/10.1038/nri.2017.105.

[32] Zhang R, Sun C, Han Y, Huang L, Sheng H, Wang J, et al. Neutrophil autophagy and NETosis in COVID-19: perspectives. Autophagy 2023; 19: 758-67. https://doi.org/10.1080/15548627.2022.2099206.

[33] Klopf J, Brostjan C, Eilenberg W, Neumayer C. Neutrophil extracellular traps and their implications in cardiovascular and inflammatory disease. Int J Mol Sci 2021; 22: 559. https://doi.org/10.3390/ijms22020559.

[34] Najmeh S, Cools-Lartigue J, Giannias B, Spicer J, Ferri LE. Simplified human neutrophil extracellular traps (NETs) isolation and handling. J Vis Exp 2015; 98: 52687. https://doi.org/10.3791/52687.

[35] Fuchs TA, Abed U, Goosmann C, Hurwitz R, Schulze I, Wahn V, et al. Novel cell death program leads to neutrophil extracellular traps. J Cell Biol 2007; 176: 231-41. https://doi.org/10.1083/jcb.200606027.

[36] Nagaoka I, Tamura H, Reich J. Therapeutic potential of cathelicidin peptide LL-37, an antimicrobial agent, in a murine sepsis model. Int J Mol Sci 2020; 21: 5973. https://doi.org/10.3390/ijms21175973.

[37] Violi F, Bartimoccia S, Cangemi R, Nocella C, D’Amico A, Oliva A, et al. Neutrophil cathepsin G and thrombosis in COVID-19. Circ Res 2024; 135: 350-2. https://doi.org/10.1161/circresaha.124.324649.

[38] Burgener SS, Leborgne NGF, Snipas SJ, Salvesen GS, Bird PI, Benarafa C. Cathepsin G inhibition by Serpinb1 and Serpinb6 prevents programmed necrosis in neutrophils and monocytes and reduces GSDMD-driven inflammation. Cell Rep 2019; 27: 3646-3656.e5. https://doi.org/10.1016/j.celrep.2019.05.065.

[39] Rodrigues DAS, Prestes EB, Gama AMS, Silva LS, Pinheiro AAS, Ribeiro JMC, et al. CXCR4 and MIF are required for neutrophil extracellular trap release triggered by plasmodium-infected erythrocytes. PLoS Pathog 2020; 16: e1008230. https://doi.org/10.1371/journal.ppat.1008230.

[40] Papayannopoulos V, Metzler KD, Hakkim A, Zychlinsky A. Neutrophil elastase and myeloperoxidase regulate the formation of neutrophil extracellular traps. J Cell Biol 2010; 191: 677-91. https://doi.org/10.1083/jcb.201006052.

[41] Lee KH, Kronbichler A, Park DD, Park Y, Moon H, Kim H, et al. Neutrophil extracellular traps (NETs) in autoimmune diseases: a comprehensive review. Autoimmun Rev 2017; 16: 1160-73. https://doi.org/10.1016/j.autrev.2017.09.012.

[42] Wigerblad G, Kaplan MJ.Neutrophil extracellular traps in systemic autoimmune and autoinflammatory diseases. Nat Rev Immunol 2023; 23: 274-88. https://doi.org/10.1038/s41577-022-00787-0.

[43] Huang J, Hong W, Wan M, Zheng L. Molecular mechanisms and therapeutic target of NETosis in diseases. MedComm 2020; 3: e162. https://doi.org/10.1002/mco2.162.

[44] Mou Z, Chen Y, Hu J, Hu Y, Zou L, Chen X, et al. Icaritin inhibits the progression of urothelial cancer by suppressing PADI2-mediated neutrophil infiltration and neutrophil extracellular trap formation. Acta Pharm Sin B 2024; 14: 3916-30. https://doi.org/10.1016/j.apsb.2024.06.029.

[45] Burgener SS, Schroder K. Neutrophil extracellular traps in host defense. Cold Spring Harb Perspect Biol 2020; 12: a037028. https://doi.org/10.1101/cshperspect.a037028.

[46] Zou S, Han X, Luo S, Tan Q, Huang H, Yao Z, et al. Bay-117082 treats sepsis by inhibiting neutrophil extracellular traps (NETs) formation through down-regulating NLRP3/N-GSDMD. Int Immunopharmacol 2024; 141: 112805. https://doi.org/10.1016/j.intimp.2024.112805.

[47] Liu X, Arfman T, Wichapong K, Reutelingsperger CPM, Voorberg J, Nicolaes GAF. PAD4 takes charge during neutrophil activation: impact of PAD4 mediated NET formation on immune-mediated disease. J Thromb Haemost 2021; 19: 1607-17. https://doi.org/10.1111/jth.15313.

[48] Thirugnanasambandham I, Radhakrishnan A, Kuppusamy G, Kumar Singh S, Dua K. Peptidylarginine deiminase-4: medico-formulative strategy towards management of rheumatoid arthritis. Biochem Pharmacol 2022; 200: 115040. https://doi.org/10.1016/j.bcp.2022.115040.

[49] Singhal A, Kumar S. Neutrophil and remnant clearance in immunity and inflammation. Immunology 2022; 165: 22-43. https://doi.org/10.1111/imm.13423.

[50] Chen KW, Monteleone M, Boucher D, Sollberger G, Ramnath D, Condon ND, et al. Noncanonical inflammasome signaling elicits gasdermin D-dependent neutrophil extracellular traps. Sci Immunol 2018; 3: eaar6676. https://doi.org/10.1126/sciimmunol.aar6676.

[51] Zhou L, Li Y, You J, Wu C, Zuo L, Chen Y, et al. Salmonella spvC gene suppresses macrophage/neutrophil antibacterial defense mediated by gasdermin D. Inflamm Res 2024; 73: 19-33. https://doi.org/10.1007/s00011-023-01818-9.

[52] Kawai T, Ikegawa M, Ori D, Akira S. Decoding toll-like receptors: recent insights and perspectives in innate immunity. Immunity 2024; 57: 649-73. https://doi.org/10.1016/j.immuni.2024.03.004.

[53] Futosi K, Fodor S, Mócsai A. Neutrophil cell surface receptors and their intracellular signal transduction pathways. Int Immunopharmacol 2013; 17: 638-50. https://doi.org/10.1016/j.intimp.2013.06.034.

[54] Tong M, Smith AH, Abrahams VM. Activated neutrophils propagate fetal membrane inflammation and weakening through ERK and neutrophil extracellular trap-induced TLR-9 signaling. J Immunol 2021; 206: 1039-45. https://doi.org/10.4049/jimmunol.2001268.

[55] Breda LCD, Breda CNS, de Almeida JRF, Paulo LNM, Jannuzzi GP, Menezes IG, et al. Fonsecaeapedrosoi conidia and hyphae activate neutrophils distinctly: requirement of TLR-2 and TLR-4 in neutrophil effector functions. Front Immunol 2020; 11: 540064. https://doi.org/10.3389/fimmu.2020.540064.

[56] Chen Z, Xiao G, Ao J. Resveratrol attenuates rheumatoid arthritis induce neutrophil extracellular traps via TLR-4 mediated inflammation in C57BL/6 mice. Physiol Res 2024; 73: 91-104. https://doi.org/10.33549/physiolres.935172.

[57] Whitmore LC, Hook JS, Philiph AR, Hilkin BM, Bing X, Ahn C, et al. A common genetic variant in TLR1 enhances human neutrophil priming and impacts length of intensive care stay in pediatric sepsis. J Immunol 2016; 196: 1376-86. https://doi.org/10.4049/jimmunol.1500856.

[58] Sugawara I, Yamada H, Li C, Mizuno S, Takeuchi O, Akira S. Mycobacterial infection in TLR2 and TLR6 knockout mice. Microbiol Immunol 2003; 47: 327-36. https://doi.org/10.1111/j.1348-0421.2003.tb03404.x.

[59] Liu R, Zhu G, Sun Y, Li M, Hu Z, Cao P, et al. Neutrophil infiltration associated genes on the prognosis and tumor immune microenvironment of lung adenocarcinoma. Front Immunol 2023; 14: 1304529. https://doi.org/10.3389/fimmu.2023.1304529.

[60] Cambier LC, Heinen MP, Bagut ET, Antoine NA, Mignon BR. Overexpression of TLR-2 and TLR-4 mRNA in feline polymorphonuclear neutrophils exposed to Microsporum canis. Vet Dermatol 2016; 27: 78-81e22. https://doi.org/10.1111/vde.12295.

[61] Zhu Y, Han Q, Wang L, Wang B, Chen J, Cai B, et al. Jinhua Qinggan granules attenuates acute lung injury by promotion of neutrophil apoptosis and inhibition of TLR4/MyD88/NF-κB pathway. J Ethnopharmacol 2023; 301: 115763. https://doi.org/10.1016/j.jep.2022.115763.

[62] Kim YC, Lee SE, Kim SK, Jang HD, Hwang I, Jin S, et al. Toll-like receptor mediated inflammation requires FASN-dependent MYD88 palmitoylation. Nat Chem Biol 2019; 15: 907-16. https://doi.org/10.1038/s41589-019-0344-0.

[63] Kirsebom FCM, Kausar F, Nuriev R, Makris S, Johansson C. Neutrophil recruitment and activation are differentially dependent on MyD88/TRIF and MAVS signaling during RSV infection. Mucosal Immunol 2019; 12: 1244-55. https://doi.org/10.1038/s41385-019-0190-0.

[64] Li W, Hsiao HM, Higashikubo R, Saunders BT, Bharat A, Goldstein DR, et al. Heart-resident CCR2(+) macrophages promote neutrophil extravasation through TLR9/MyD88/CXCL5 signaling. JCI Insight 2016; 1: e87315. https://doi.org/10.1172/jci.insight.87315.

[65] Lu J, Zhong X, Guo C, Tang L, Yu N, Peng C, et al. TLR7-MyD88-DC-CXCL16 axis results neutrophil activation to elicit inflammatory response in pustular psoriasis. Cell Death Dis 2023; 14: 315. https://doi.org/10.1038/s41419-023-05815-y.

[66] Bugl S, Wirths S, Radsak MP, Schild H, Stein P, André MC, et al. Steady-state neutrophil homeostasis is dependent on TLR4/TRIF signaling. Blood 2013; 121: 723-33. https://doi.org/10.1182/blood-2012-05-429589.

[67] Muendlein HI, Connolly WM, Cameron J, Jetton D, Magri Z, Smirnova I, et al. Neutrophils and macrophages drive TNF-induced lethality via TRIF/CD14-mediated responses. Sci Immunol 2022; 7: eadd0665. https://doi.org/10.1126/sciimmunol.add0665.

[68] Prince LR, Whyte MK, Sabroe I, Parker LC. The role of TLRs in neutrophil activation. Curr Opin Pharmacol 2011; 11: 397-403. https://doi.org/10.1016/j.coph.2011.06.007.

[69] Li K, Underhill DM. C-type lectin receptors in phagocytosis. Curr Top Microbiol Immunol 2020; 429: 1-18. https://doi.org/10.1007/82_2020_198.

[70] Sung PS, Peng YC, Yang SP, Chiu CH, Hsieh SL. CLEC5A is critical in Pseudomonas aeruginosa-induced NET formation and acute lung injury. JCI Insight 2022; 7: e156613. https://doi.org/10.1172/jci.insight.156613.

[71] Malamud M, Whitehead L, McIntosh A, Colella F, Roelofs AJ, Kusakabe T, et al. Recognition and control of neutrophil extracellular trap formation by MICL. Nature 2024; 633: 442-50. https://doi.org/10.1038/s41586-024-07820-3.

[72] Branzk N, Lubojemska A, Hardison SE, Wang Q, Gutierrez MG, Brown GD, et al. Neutrophils sense microbe size and selectively release neutrophil extracellular traps in response to large pathogens. Nat Immunol 2014; 15: 1017-25. https://doi.org/10.1038/ni.2987.

[73] Alyami HM, Finoti LS, Teixeira HS, Aljefri A, Kinane DF, Benakanakere MR. Role of NOD1/NOD2 receptors in fusobacterium nucleatum mediated NETosis. Microb Pathog 2019; 131: 53-64. https://doi.org/10.1016/j.micpath.2019.03.036.

[74] Scozzi D, Gelman AE. Avoid being trapped by your liver: ischemia-reperfusion injury in liver transplant triggers S1P-mediated NETosis. J Clin Invest 2023; 133: e167012. https://doi.org/10.1172/jci167012.

[75] Clark HL, Abbondante S, Minns MS, Greenberg EN, Sun Y, Pearlman E. Protein deiminase 4 and CR3 regulate aspergillus fumigatus and β-glucan-induced neutrophil extracellular trap formation, but hyphal killing is dependent only on CR3. Front Immunol 2018; 9: 1182. https://doi.org/10.3389/fimmu.2018.01182.

[76] Raftery MJ, Lalwani P, Krautkrämer E, Peters T, Scharffetter-Kochanek K, Krüger R, et al. β2 integrin mediates hantavirus-induced release of neutrophil extracellular traps. J Exp Med 2014; 211: 1485-97. https://doi.org/10.1084/jem.20131092.

[77] Skendros P, Mitsios A, Chrysanthopoulou A, Mastellos DC, Metallidis S, Rafailidis P, et al. Complement and tissue factor-enriched neutrophil extracellular traps are key drivers in COVID-19 immunothrombosis. J Clin Invest 2020; 130: 6151-7. https://doi.org/10.1172/jci141374.

[78] Mongirdienė A, Liuizė A, Kašauskas A. Novel knowledge about molecular mechanisms of heparin-induced thrombocytopenia type II and treatment targets. Int J Mol Sci 2023; 24. https://doi.org/10.3390/ijms24098217.

[79] Teijeira Á, Garasa S, Gato M, Alfaro C, Migueliz I, Cirella A, et al. CXCR1 and CXCR2 chemokine receptor agonists produced by tumors induce neutrophil extracellular traps that interfere with immune cytotoxicity. Immunity 2020; 52: 856-871.e8. https://doi.org/10.1016/j.immuni.2020.03.001.

[80] Veras FP, Pontelli MC, Silva CM, Toller-Kawahisa JE, de Lima M, Nascimento DC, et al. SARS-CoV-2-triggered neutrophil extracellular traps mediate COVID-19 pathology. J Exp Med 2020; 217:e20201129. https://doi.org/10.1084/jem.20201129.

[81] Hann J, Bueb JL, Tolle F, Bréchard S. Calcium signaling and regulation of neutrophil functions: still a long way to go. J Leukoc Biol 2020; 107: 285-97. https://doi.org/10.1002/jlb.3ru0719-241r.

[82] Cao C, Yu P, Chu C, Wang Z, Xu W, Cheng F, et al. Magnesium hydride attenuates intestinal barrier injury during hemorrhage shock by regulating neutrophil extracellular trap formation via the ROS/MAPK/PAD4 pathway. Int Immunopharmacol 2024; 130: 111688. https://doi.org/10.1016/j.intimp.2024.111688.

[83] Poli V, Pui-Yan Ma V, Di Gioia M, Broggi A, Benamar M, Chen Q, et al. Zinc-dependent histone deacetylases drive neutrophil extracellular trap formation and potentiate local and systemic inflammation. iScience 2021; 24: 103256. https://doi.org/10.1016/j.isci.2021.103256.

[84] Goggs R, Jeffery U, LeVine DN, Li RHL. Neutrophil-extracellular traps, cell-free DNA, and immunothrombosis in companion animals: a review. Vet Pathol 2020; 57: 6-23. https://doi.org/10.1177/0300985819861721.

[85] Al-Kuraishy HM, Al-Gareeb AI, Al-Hussaniy HA, Al-Harcan NAH, Alexiou A, Batiha GE. Neutrophil extracellular traps (NETs) and Covid-19: a new frontiers for therapeutic modality. Int Immunopharmacol 2022; 104: 108516. https://doi.org/10.1016/j.intimp.2021.108516.

[86] Jenne CN, Wong CH, Zemp FJ, McDonald B, Rahman MM, Forsyth PA, et al. Neutrophils recruited to sites of infection protect from virus challenge by releasing neutrophil extracellular traps. Cell Host Microbe 2013; 13: 169-80. https://doi.org/10.1016/j.chom.2013.01.005.

[87] Silva JC, Rodrigues NC, Thompson-Souza GA, Muniz VS, Neves JS, Figueiredo RT. Mac-1 triggers neutrophil DNA extracellular trap formation to aspergillus fumigatus independently of PAD4 histone citrullination. J Leukoc Biol 2020; 107: 69-83. https://doi.org/10.1002/jlb.4a0119-009rr.

[88] Shirakawa K, Kobayashi E, Ichihara G, Kitakata H, Katsumata Y, Sugai K, et al. H(2) inhibits the formation of neutrophil extracellular traps. JACC Basic Transl Sci 2022; 7: 146-61. https://doi.org/10.1016/j.jacbts.2021.11.005.

[89] Thiam HR, Wong SL, Wagner DD, Waterman CM. Cellular mechanisms of NETosis. Annu Rev Cell Dev Biol 2020; 36: 191-218. https://doi.org/10.1146/annurev-cellbio-020520-111016.

[90] Singel KL, Segal BH. NOX2-dependent regulation of inflammation. Clin Sci (Lond) 2016; 130: 479-90. https://doi.org/10.1042/cs20150660.

[91] Cao Z, Huang D, Tang C, Zeng M, Hu X. PFKL, a novel regulatory node for NOX2-dependent oxidative burst and NETosis. J Zhejiang Univ Sci B 2022; 23: 607-12. https://doi.org/10.1631/jzus.B2101029.

[92] Dömer D, Walther T, Möller S, Behnen M, Laskay T. Neutrophil extracellular traps activate proinflammatory functions of human neutrophils. Front Immunol 2021; 12: 636954. https://doi.org/10.3389/fimmu.2021.636954.

[93] Murata K, Murao A, Tan C, Wang P, Aziz M. B-1a cells scavenge NETs to attenuate sepsis. J Leukoc Biol 2024; 116: 632-43. https://doi.org/10.1093/jleuko/qiae066.

[94] Karmakar U, Vermeren S. Crosstalk between B cells and neutrophils in rheumatoid arthritis. Immunology 2021; 164: 689-700. https://doi.org/10.1111/imm.13412.

[95] Gestermann N, Di Domizio J, Lande R, Demaria O, Frasca L, Feldmeyer L, et al. Netting neutrophils activate autoreactive B cells in lupus. J Immunol 2018; 200: 3364-71. https://doi.org/10.4049/jimmunol.1700778.

[96] Lande R, Pietraforte I, Mennella A, Palazzo R, Spinelli FR, Giannakakis K, et al. Complementary effects of carbamylated and citrullinated LL37 in autoimmunity and inflammation in systemic lupus erythematosus. Int J Mol Sci 2021; 22: 1650. https://doi.org/10.3390/ijms22041650.

[97] Fang H, Shao S, Xue K, Yuan X, Qiao P, Zhang J, et al. Neutrophil extracellular traps contribute to immune dysregulation in bullous pemphigoid via inducing B-cell differentiation and antibody production. FASEB J 2021; 35: e21746. https://doi.org/10.1096/fj.202100145R.

[98] Tillack K, Breiden P, Martin R, Sospedra M. T lymphocyte priming by neutrophil extracellular traps links innate and adaptive immune responses. J Immunol 2012; 188: 3150-9. https://doi.org/10.4049/jimmunol.1103414.

[99] Roy M, Chakraborty S, Kumar Srivastava S, Kaushik S, Jyoti A, Kumar SV. Entamoeba histolytica induced NETosis and the dual role of NETs in amoebiasis. Int Immunopharmacol 2023; 118: 110100. https://doi.org/10.1016/j.intimp.2023.110100.

[100] Torres-Ruiz J, Alcalá-Carmona B, Alejandre-Aguilar R, Gómez-Martín D. Inflammatory myopathies and beyond: the dual role of neutrophils in muscle damage and regeneration. Front Immunol 2023; 14: 1113214. https://doi.org/10.3389/fimmu.2023.1113214.

[101] Rahman S, Sagar D, Hanna RN, Lightfoot YL, Mistry P, Smith CK, et al. Low-density granulocytes activate T cells and demonstrate a non-suppressive role in systemic lupus erythematosus. Ann Rheum Dis 2019; 78: 957-66. https://doi.org/10.1136/annrheumdis-2018-214620.

[102] Miranda FJB, Rocha BC, Pereira MCA, Pereira LMN, de Souza EHM, Marino AP, et al. Toxoplasma gondii-induced neutrophil extracellular traps amplify the innate and adaptive response. mBio 2021; 12: e0130721. https://doi.org/10.1128/mBio.01307-21.

[103] Wilson AS, Randall KL, Pettitt JA, Ellyard JI, Blumenthal A, Enders A, et al. Neutrophil extracellular traps and their histones promote Th17 cell differentiation directly via TLR2. Nat Commun 2022; 13: 528. https://doi.org/10.1038/s41467-022-28172-4.

[104] Wang H, Zhang H, Wang Y, Brown ZJ, Xia Y, Huang Z, et al. Regulatory T-cell and neutrophil extracellular trap interaction contributes to carcinogenesis in non-alcoholic steatohepatitis. J Hepatol 2021; 75: 1271-83. https://doi.org/10.1016/j.jhep.2021.07.032.

[105] Shi Y, Wu D, Wang Y, Shao Y, Zeng F, Zhou D, et al. Treg and neutrophil extracellular trap interaction contributes to the development of immunosuppression in sepsis. JCI Insight 2024; 9: e180132. https://doi.org/10.1172/jci.insight.180132.

[106] Grégoire M, Uhel F, Lesouhaitier M, Gacouin A, Guirriec M, Mourcin F, et al. Impaired efferocytosis and neutrophil extracellular trap clearance by macrophages in ARDS. Eur Respir J 2018; 52: 1702590. https://doi.org/10.1183/13993003.02590-2017.

[107] Hou FF, Mi JH, Wang Q, Tao YL, Guo SB, Ran GH, et al. Macrophage polarization in sepsis: emerging role and clinical application prospect. Int Immunopharmacol 2025; 144: 113715. https://doi.org/10.1016/j.intimp.2024.113715.

[108] Guimarães-Costa AB, Rochael NC, Oliveira F, Echevarria-Lima J, Saraiva EM. Neutrophil extracellular traps reprogram IL-4/GM-CSF-induced monocyte differentiation to anti-inflammatory macrophages. Front Immunol 2017; 8: 523. https://doi.org/10.3389/fimmu.2017.00523.

[109] Monteith AJ, Miller JM, Maxwell CN, Chazin WJ, Skaar EP. Neutrophil extracellular traps enhance macrophage killing of bacterial pathogens. Sci Adv 2021; 7: eabj2101. https://doi.org/10.1126/sciadv.abj2101.

[110] Sutton JAF, Carnell OT, Lafage L, Gray J, Biboy J, Gibson JF, et al. Staphylococcus aureus cell wall structure and dynamics during host-pathogen interaction. PLoS Pathog 2021; 17: e1009468. https://doi.org/10.1371/journal.ppat.1009468.

[111] Chen L, Zhao Y, Lai D, Zhang P, Yang Y, Li Y, et al. Neutrophil extracellular traps promote macrophage pyroptosis in sepsis. Cell Death Dis 2018; 9: 597. https://doi.org/10.1038/s41419-018-0538-5.

[112] Zhu L, Zheng Q, Liu X, Ding H, Ma M, Bao J, et al. HMGB1 lactylation drives neutrophil extracellular trap formation in lactate-induced acute kidney injury. Front Immunol 2024; 15: 1475543. https://doi.org/10.3389/fimmu.2024.1475543.

[113] Rigamonti A, Villar J, Segura E. Monocyte differentiation within tissues: a renewed outlook. Trends Immunol 2023; 44: 999-1013. https://doi.org/10.1016/j.it.2023.10.005.

[114] Goudot C, Coillard A, Villani AC, Gueguen P, Cros A, Sarkizova S, et al. Aryl hydrocarbon receptor controls monocyte differentiation into dendritic cells versus macrophages. Immunity 2017; 47: 582-596.e6. https://doi.org/10.1016/j.immuni.2017.08.016.

[115] Villar J, Cros A, De Juan A, Alaoui L, Bonte PE, Lau CM, et al. ETV3 and ETV6 enable monocyte differentiation into dendritic cells by repressing macrophage fate commitment. Nat Immunol 2023; 24: 84-95. https://doi.org/10.1038/s41590-022-01374-0.

[116] Barrientos L, Bignon A, Gueguen C, de Chaisemartin L, Gorges R, Sandré C, et al. Neutrophil extracellular traps downregulate lipopolysaccharide-induced activation of monocyte-derived dendritic cells. J Immunol 2014; 193: 5689-98. https://doi.org/10.4049/jimmunol.1400586.

[117] Tateosian NL, Reiteri RM, Amiano NO, Costa MJ, Villalonga X, Guerrieri D, et al. Neutrophil elastase treated dendritic cells promote the generation of CD4(+)FOXP3(+) regulatory T cells in vitro. Cell Immunol 2011; 269: 128-34. https://doi.org/10.1016/j.cellimm.2011.03.013.

[118] Kotov DI, Lee OV, Fattinger SA, Langner CA, Guillen JV, Peters JM, et al. Early cellular mechanisms of type I interferon-driven susceptibility to tuberculosis. Cell 2023; 186: 5536-5553.e22. https://doi.org/10.1016/j.cell.2023.11.002.

[119] Zheng LY, Duan Y, He PY, Wu MY, Wei ST, Du XH, et al. Dysregulated dendritic cells in sepsis: functional impairment and regulated cell death. Cell Mol Biol Lett 2024; 29: 81. https://doi.org/10.1186/s11658-024-00602-9.

[120] Osawa I, Goto T, Kudo D, Hayakawa M, Yamakawa K, Kushimoto S, et al. Targeted therapy using polymyxin B hemadsorption in patients with sepsis: a post-hoc analysis of the JSEPTIC-DIC study and the EUPHRATES trial. Crit Care 2023; 27: 245. https://doi.org/10.1186/s13054-023-04533-3.

[121] Zhu L, Dong H, Li L, Liu X. The mechanisms of sepsis induced coagulation dysfunction and its treatment. J Inflamm Res 2025; 18: 1479-95. https://doi.org/10.2147/jir.S504184.

[122] Qiu P, Liu Y, Zhang J. Review: the role and mechanisms of macrophage autophagy in sepsis. Inflammation 2019; 42: 6-19. https://doi.org/10.1007/s10753-018-0890-8.

[123] Cox D, Kerrigan SW, Watson SP. Platelets and the innate immune system: mechanisms of bacterial-induced platelet activation. J Thromb Haemost 2011; 9: 1097-107. https://doi.org/10.1111/j.1538-7836.2011.04264.x.

[124] Mammadova-Bach E, Ollivier V, Loyau S, Schaff M, Dumont B, Favier R, et al. Platelet glycoprotein VI binds to polymerized fibrin and promotes thrombin generation. Blood 2015; 126: 683-91. https://doi.org/10.1182/blood-2015-02-629717.

[125] Willemsen JF, Wenskus J, Lenz M, Rhode H, Trochimiuk M, Appl B, et al. DNases improve effectiveness of antibiotic treatment in murine polymicrobial sepsis. Front Immunol 2023; 14: 1254838. https://doi.org/10.3389/fimmu.2023.1254838.

[126] McDonald B, Davis RP, Kim SJ, Tse M, Esmon CT, Kolaczkowska E, et al. Platelets and neutrophil extracellular traps collaborate to promote intravascular coagulation during sepsis in mice. Blood 2017; 129: 1357-67. https://doi.org/10.1182/blood-2016-09-741298.

[127] Denorme F, Portier I, Rustad JL, Cody MJ, de Araujo CV, Hoki C, et al. Neutrophil extracellular traps regulate ischemic stroke brain injury. J Clin Invest 2022; 132: e154225. https://doi.org/10.1172/jci154225.

[128] He L, Liu R, Yue H, Zhang X, Pan X, Sun Y, et al. Interaction between neutrophil extracellular traps and cardiomyocytes contributes to atrial fibrillation progression. Signal Transduct Target Ther 2023; 8: 279. https://doi.org/10.1038/s41392-023-01497-2.

[129] Boufenzer A, Carrasco K, Jolly L, Brustolin B, Di-Pillo E, Derive M, et al. Potentiation of NETs release is novel characteristic of TREM-1 activation and the pharmacological inhibition of TREM-1 could prevent from the deleterious consequences of NETs release in sepsis. Cell Mol Immunol 2021; 18: 452-60. https://doi.org/10.1038/s41423-020-00591-7.

[130] Carmona-Rivera C, Zhao W, Yalavarthi S, Kaplan MJ. Neutrophil extracellular traps induce endothelial dysfunction in systemic lupus erythematosus through the activation of matrix metalloproteinase-2. Ann Rheum Dis 2015; 74: 1417-24. https://doi.org/10.1136/annrheumdis-2013-204837.

[131] Kolaczkowska E, Jenne CN, Surewaard BG, Thanabalasuriar A, Lee WY, Sanz MJ, et al. Molecular mechanisms of NET formation and degradation revealed by intravital imaging in the liver vasculature. Nat Commun 2015; 6: 6673. https://doi.org/10.1038/ncomms7673.

[132] Abrams ST, Zhang N, Manson J, Liu T, Dart C, Baluwa F, et al. Circulating histones are mediators of trauma-associated lung injury. Am J Respir Crit Care Med 2013; 187: 160-9. https://doi.org/10.1164/rccm.201206-1037OC.

[133] Wu G, Pan B, Shi H, Yi Y, Zheng X, Ma H, et al. Neutrophils’ dual role in cancer: from tumor progression to immunotherapeutic potential. Int Immunopharmacol 2024; 140: 112788. https://doi.org/10.1016/j.intimp.2024.112788.

[134] Demkow U.Molecular mechanisms of neutrophil extracellular trap (NETs) degradation. Int J Mol Sci 2023; 24. https://doi.org/10.3390/ijms24054896.

[135] Yang LY, Luo Q, Lu L, Zhu WW, Sun HT, Wei R, et al. Increased neutrophil extracellular traps promote metastasis potential of hepatocellular carcinoma via provoking tumorous inflammatory response. J Hematol Oncol 2020; 13: 3. https://doi.org/10.1186/s13045-019-0836-0.

[136] Xu C, Ye Z, Jiang W, Wang S, Zhang H. Cyclosporine a alleviates colitis by inhibiting the formation of neutrophil extracellular traps via the regulating pentose phosphate pathway. Mol Med 2023; 29: 169. https://doi.org/10.1186/s10020-023-00758-8.

[137] Li Y, Li H, Wang Y, Guo J, Zhang D. Potential biomarkers for early diagnosis, evaluation, and prognosis of sepsis-induced coagulopathy. Clin Appl Thromb Hemost 2023; 29: 10760296231195089. https://doi.org/10.1177/10760296231195089.

[138] Healy LD, Puy C, Fernández JA, Mitrugno A, Keshari RS, Taku NA, et al. Activated protein C inhibits neutrophil extracellular trap formation in vitro and activation in vivo. J Biol Chem 2017; 292: 8616-29. https://doi.org/10.1074/jbc.M116.768309.

[139] Zhao J, Zhen N, Zhou Q, Lou J, Cui W, Zhang G, et al. NETs promote inflammatory injury by activating cGAS-STING pathway in acute lung injury. Int J Mol Sci 2023; 24: 5125. https://doi.org/10.3390/ijms24065125.

[140] Fonseca Z, Díaz-Godínez C, Mora N, Alemán OR, Uribe-Querol E, Carrero JC, et al. Entamoeba histolytica induce signaling via Raf/MEK/ERK for neutrophil extracellular trap (NET) formation. Front Cell Infect Microbiol 2018; 8: 226. https://doi.org/10.3389/fcimb.2018.00226.

[141] Mai SH, Khan M, Dwivedi DJ, Ross CA, Zhou J, Gould TJ, et al. Delayed but not early treatment with DNase reduces organ damage and improves outcome in a murine model of sepsis. Shock 2015; 44: 166-72. https://doi.org/10.1097/shk.0000000000000396.

[142] Zalghout S, Martinod K. Therapeutic potential of DNases in immunothrombosis: promising succor or uncertain future? J Thromb Haemost 2025; 23: 760-78. https://doi.org/10.1016/j.jtha.2024.11.028.

[143] Martinod K, Fuchs TA, Zitomersky NL, Wong SL, Demers M, Gallant M, et al. PAD4-deficiency does not affect bacteremia in polymicrobial sepsis and ameliorates endotoxemic shock. Blood 2015; 125: 1948-56. https://doi.org/10.1182/blood-2014-07-587709.

[144] Liu X, Li T, Chen H, Yuan L, Ao H. Role and intervention of PAD4 in NETs in acute respiratory distress syndrome. Respir Res 2024; 25: 63. https://doi.org/10.1186/s12931-024-02676-7.

[145] He W, Xi Q, Cui H, Zhang P, Huang R, Wang T, et al. Liang-Ge decoction ameliorates coagulation dysfunction in cecal ligation and puncture-induced sepsis model rats through inhibiting PAD4-dependent neutrophil extracellular trap formation. Evid Based Complement Alternat Med 2023; 2023: 5042953. https://doi.org/10.1155/2023/5042953.

[146] Wildhagen KC, García de Frutos P, Reutelingsperger CP, Schrijver R, Aresté C, Ortega-Gómez A, et al. Nonanticoagulant heparin prevents histone-mediated cytotoxicity in vitro and improves survival in sepsis. Blood 2014; 123: 1098-101. https://doi.org/10.1182/blood-2013-07-514984.

[147] Mansour A, Bachelot-Loza C, Nesseler N, Gaussem P, Gouin-Thibault I. P2Y(12) Inhibition beyond thrombosis: effects on inflammation. Int J Mol Sci 2020; 21: 1391. https://doi.org/10.3390/ijms21041391.

[148] Vardon-Bounes F, Ruiz S, Gratacap MP, Garcia C, Payrastre B, Minville V. Platelets are critical key players in sepsis. Int J Mol Sci 2019; 20: 3494. https://doi.org/10.3390/ijms20143494.

[149] A feedback loop between platelets and NETs amplifies inflammation in severe sepsis. Nat Cardiovasc Res 2022; 1: 698-9. https://doi.org/10.1038/s44161-022-00110-z.

[150] Kato Y, Nishida O, Kuriyama N, Nakamura T, Kawaji T, Onouchi T, et al. Effects of thrombomodulin in reducing lethality and suppressing neutrophil extracellular trap formation in the lungs and liver in a lipopolysaccharide-induced murine septic shock model. Int J Mol Sci 2021; 22: 4933. https://doi.org/10.3390/ijms22094933.

[151] Su F, Moreau A, Savi M, Salvagno M, Annoni F, Zhao L, et al. Circulating nucleosomes as a novel biomarker for sepsis: a scoping review. Biomedicines 2024; 12: 1385. https://doi.org/10.3390/biomedicines12071385.

[152] Garcia B, Su F, Dewachter L, Wang Y, Li N, Remmelink M, et al. Neutralization of extracellular histones by sodium-B-O-methyl cellobioside sulfate in septic shock. Crit Care 2023; 27: 458. https://doi.org/10.1186/s13054-023-04741-x.