Neutrophil extracellular traps extrusion from neutrophils stably adhered to ICAM-1 by lipoteichoic acid stimulation_Journal of Biomedical Engineering

Authors：

HUANG Jiaqi ¹ , FANG Jinhua ² , WU Zhiwei ² , WU Jianhua ¹ ,  FANG Ying ¹ ,  LIN Jiangguo ²

1. School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, P. R. China;
2. Medical Research Institute, Guangdong Provincial People’s Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou 510080, P. R. China;

Corresponding author：

FANG Ying, Email: yfang@scut.edu.cn; LIN Jiangguo, Email: linjiangguo@gdph.org.cn

Keywords：

Neutrophil extracellular traps; Sepsis; Signaling pathway

DOI：

10.7507/1001-5515.202401062

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

The effect of neutrophil extracellular traps (NETs) on promoting intravascular microthrombi formation and exacerbating the severity of sepsis in patients has gained extensive attention. However, in sepsis, the mechanisms and key signaling molecules mediating NET formation during direct interactions of endothelial cells and neutrophils still need further explored. Herein, we utilized lipoteichoic acid (LTA), a component shared by Gram-positive bacteria, to induce NET extrusion from neutrophils firmly adhered to the glass slides coated with intercellular adhesion molecule-1(ICAM-1). We also used Sytox green to label NET-DNA and Flou-4 AM as the intracellular Ca²⁺ signaling indicator to observe the NET formation and fluctuation of Ca²⁺ signaling. Our results illustrated that LTA was able to induce NET release from neutrophils firmly attached to ICAM-1-coated glass slides, and the process was time-dependent. In addition, our study indicated that LTA-induced NET release by neutrophils stably adhered to ICAM-1 depended on Ca²⁺ signaling but not intracellular reactive oxygen species (ROS). This study reveals NET formation mediated by direct interactions between endothelial ICAM-1 and neutrophils under LTA stimulation and key signaling molecules involved, providing the theoretical basis for medicine development and clinical treatment for related diseases.

Citation： HUANG Jiaqi, FANG Jinhua, WU Zhiwei, WU Jianhua, FANG Ying, LIN Jiangguo. Neutrophil extracellular traps extrusion from neutrophils stably adhered to ICAM-1 by lipoteichoic acid stimulation. Journal of Biomedical Engineering, 2024, 41(2): 304-312. doi: 10.7507/1001-5515.202401062 Copy

1.	Salomao R, Ferreira B L, Salomao M C, et al. Sepsis: Evolving concepts and challenges. Braz J Med Biol Res, 2019, 52(4): e8595.
2.	Srzic I, Nesek Adam V, Tunjic Pejak D. Sepsis definition: What’s new in the treatment guidelines. Acta Clin Croat, 2022, 61(Suppl 1): 67-72.
3.	Iba T, Helms J, Connors J M, et al. The pathophysiology, diagnosis, and management of sepsis-associated disseminated intravascular coagulation. J Intensive Care, 2023, 11(1): 24.
4.	Martin G S, Mannino D M, Eaton S, et al. The epidemiology of sepsis in the United States from 1979 through 2000. N Engl J Med, 2003, 348(16): 1546-1554.
5.	Thome S, Begandt D, Pick R, et al. Intracellular beta2 integrin (CD11/CD18) interacting partners in neutrophil trafficking. Eur J Clin Invest, 2018, 48(Suppl 2): e12966.
6.	Chen Z, Zhang H, Qu M, et al. Review: The emerging role of neutrophil extracellular traps in sepsis and sepsis-associated thrombosis. Front Cell Infect Microbiol, 2021, 11: 653228.
7.	Hidalgo A, Libby P, Soehnlein O, et al. Neutrophil extracellular traps: From physiology to pathology. Cardiovasc Res, 2022, 118(13): 2737-2753.
8.	Pires R H, Felix S B, Delcea M. The architecture of neutrophil extracellular traps investigated by atomic force microscopy. Nanoscale, 2016, 8(29): 14193-14202.
9.	Scozzi D, Liao F, Krupnick A S, et al. The role of neutrophil extracellular traps in acute lung injury. Front Immunol, 2022, 13: 953195.
10.	Morales-Primo A U, Becker I, Zamora-Chimal J. Neutrophil extracellular trap-associated molecules: A review on their immunophysiological and inflammatory roles. Int Rev Immunol, 2022, 41(2): 253-274.
11.	Thakur M, Junho C V C, Bernhard S M, et al. NETs-induced thrombosis impacts on cardiovascular and chronic kidney disease. Circ Res, 2023, 132(8): 933-949.
12.	Mazetto B M, Hounkpe B W, da Silva Saraiva S, et al. Association between neutrophil extracellular traps (NETs) and thrombosis in antiphospholipid syndrome. Thromb Res, 2022, 214: 132-137.
13.	Kaltenmeier C, Yazdani H O, Handu S, et al. The role of neutrophils as a driver in hepatic ischemia-reperfusion injury and cancer growth. Front Immunol, 2022, 13: 887565.
14.	Ling S, Xu J W. NETosis as a pathogenic factor for heart failure. Oxid Med Cell Longev, 2021, 2021: 6687096.
15.	Wang Y, Li Y, Chen Z, et al. GSDMD-dependent neutrophil extracellular traps promote macrophage-to-myofibroblast transition and renal fibrosis in obstructive nephropathy. Cell Death Dis, 2022, 13(8): 693.
16.	Petrelli A, Popp S K, Fukuda R, et al. The contribution of neutrophils and NETs to the development of type 1 diabetes. Front Immunol, 2022, 13: 930553.
17.	Mutua V, Gershwin L J. A review of neutrophil extracellular traps (NETs) in disease: Potential anti-nets therapeutics. Clin Rev Allergy Immunol, 2021, 61(2): 194-211.
18.	Lin H, Liu J, Li N, et al. NETosis promotes chronic inflammation and fibrosis in systemic lupus erythematosus and COVID-19. Clin Immunol, 2023, 254: 109687.
19.	Bissenova S, Ellis D, Mathieu C, et al. Neutrophils in autoimmunity: When the hero becomes the villain. Clin Exp Immunol, 2022, 210(2): 128-140.
20.	Yang X, Ma Y, Chen X, et al. Mechanisms of neutrophil extracellular trap in chronic inflammation of endothelium in atherosclerosis. Life Sci, 2023, 328: 121867.
21.	Kenny E F, Herzig A, Kruger R, et al. Diverse stimuli engage different neutrophil extracellular trap pathways. Elife, 2017, 6: e24437.
22.	Sakurai K, Miyashita T, Okazaki M, et al. Role for neutrophil extracellular traps (NETs) and platelet aggregation in early sepsis-induced hepatic dysfunction. In Vivo, 2017, 31(6): 1051-1058.
23.	洪天添, 刘望, 黄嘉祺, 等. LPS刺激稳定黏附于ICAM-1上的中性粒细胞形成胞外诱捕网依赖于整合素Mac-1和细胞骨架蛋白. 生物医学工程学杂志, 2021, 38(5): 903-910.
24.	Lotz S, Aga E, Wilde I, et al. Highly purified lipoteichoic acid activates neutrophil granulocytes and delays their spontaneous apoptosis via CD14 and TLR2. J Leukoc Biol, 2004, 75(3): 467-477.
25.	Hann J, Bueb J L, Tolle F, et al. Calcium signaling and regulation of neutrophil functions: Still a long way to go. J Leukoc Biol, 2020, 107(2): 285-297.
26.	de Bont C M, Koopman W J H, Boelens W C, et al. Stimulus-dependent chromatin dynamics, citrullination, calcium signalling and ROS production during NET formation. Biochim Biophys Acta Mol Cell Res, 2018, 1865(11 Pt A): 1621-1629.
27.	Rochael N C, Guimaraes-Costa A B, Nascimento M T, et al. Classical ROS-dependent and early/rapid ROS-independent release of neutrophil extracellular traps triggered by Leishmania parasites. Sci Rep, 2015, 5: 18302.
28.	Mazzoleni V, Zimmermann K, Smirnova A, et al. Staphylococcus aureus Panton-Valentine Leukocidin triggers an alternative NETosis process targeting mitochondria. FASEB J, 2021, 35(2): e21167.
29.	Schonrich G, Raftery M J, Samstag Y. Devilishly radical network in COVID-19: Oxidative stress, neutrophil extracellular traps (NETs), and T cell suppression. Adv Biol Regul, 2020, 77: 100741.
30.	Venkataranganayaka Abhilasha K, Kedihithlu Marathe G. Bacterial lipoproteins in sepsis. Immunobiology, 2021, 226(5): 152128.
31.	Ginsburg I. Role of lipoteichoic acid in infection and inflammation. Lancet Infect Dis, 2002, 2(3): 171-179.
32.	Tan C, Aziz M, Wang P. The vitals of NETs. J Leukoc Biol, 2021, 110(4): 797-808.
33.	Lee S K, Goh S Y, Wong Y Q, et al. Response of neutrophils to extracellular haemoglobin and LTA in human blood system. EBioMedicine, 2015, 2(3): 225-233.
34.	Pilsczek F H, Salina D, Poon K K, et al. A novel mechanism of rapid nuclear neutrophil extracellular trap formation in response to Staphylococcus aureus. J Immunol, 2010, 185(12): 7413-7425.

1. Salomao R, Ferreira B L, Salomao M C, et al. Sepsis: Evolving concepts and challenges. Braz J Med Biol Res, 2019, 52(4): e8595.
2. Srzic I, Nesek Adam V, Tunjic Pejak D. Sepsis definition: What’s new in the treatment guidelines. Acta Clin Croat, 2022, 61(Suppl 1): 67-72.
3. Iba T, Helms J, Connors J M, et al. The pathophysiology, diagnosis, and management of sepsis-associated disseminated intravascular coagulation. J Intensive Care, 2023, 11(1): 24.
4. Martin G S, Mannino D M, Eaton S, et al. The epidemiology of sepsis in the United States from 1979 through 2000. N Engl J Med, 2003, 348(16): 1546-1554.
5. Thome S, Begandt D, Pick R, et al. Intracellular beta2 integrin (CD11/CD18) interacting partners in neutrophil trafficking. Eur J Clin Invest, 2018, 48(Suppl 2): e12966.
6. Chen Z, Zhang H, Qu M, et al. Review: The emerging role of neutrophil extracellular traps in sepsis and sepsis-associated thrombosis. Front Cell Infect Microbiol, 2021, 11: 653228.
7. Hidalgo A, Libby P, Soehnlein O, et al. Neutrophil extracellular traps: From physiology to pathology. Cardiovasc Res, 2022, 118(13): 2737-2753.
8. Pires R H, Felix S B, Delcea M. The architecture of neutrophil extracellular traps investigated by atomic force microscopy. Nanoscale, 2016, 8(29): 14193-14202.
9. Scozzi D, Liao F, Krupnick A S, et al. The role of neutrophil extracellular traps in acute lung injury. Front Immunol, 2022, 13: 953195.
10. Morales-Primo A U, Becker I, Zamora-Chimal J. Neutrophil extracellular trap-associated molecules: A review on their immunophysiological and inflammatory roles. Int Rev Immunol, 2022, 41(2): 253-274.
11. Thakur M, Junho C V C, Bernhard S M, et al. NETs-induced thrombosis impacts on cardiovascular and chronic kidney disease. Circ Res, 2023, 132(8): 933-949.
12. Mazetto B M, Hounkpe B W, da Silva Saraiva S, et al. Association between neutrophil extracellular traps (NETs) and thrombosis in antiphospholipid syndrome. Thromb Res, 2022, 214: 132-137.
13. Kaltenmeier C, Yazdani H O, Handu S, et al. The role of neutrophils as a driver in hepatic ischemia-reperfusion injury and cancer growth. Front Immunol, 2022, 13: 887565.
14. Ling S, Xu J W. NETosis as a pathogenic factor for heart failure. Oxid Med Cell Longev, 2021, 2021: 6687096.
15. Wang Y, Li Y, Chen Z, et al. GSDMD-dependent neutrophil extracellular traps promote macrophage-to-myofibroblast transition and renal fibrosis in obstructive nephropathy. Cell Death Dis, 2022, 13(8): 693.
16. Petrelli A, Popp S K, Fukuda R, et al. The contribution of neutrophils and NETs to the development of type 1 diabetes. Front Immunol, 2022, 13: 930553.
17. Mutua V, Gershwin L J. A review of neutrophil extracellular traps (NETs) in disease: Potential anti-nets therapeutics. Clin Rev Allergy Immunol, 2021, 61(2): 194-211.
18. Lin H, Liu J, Li N, et al. NETosis promotes chronic inflammation and fibrosis in systemic lupus erythematosus and COVID-19. Clin Immunol, 2023, 254: 109687.
19. Bissenova S, Ellis D, Mathieu C, et al. Neutrophils in autoimmunity: When the hero becomes the villain. Clin Exp Immunol, 2022, 210(2): 128-140.
20. Yang X, Ma Y, Chen X, et al. Mechanisms of neutrophil extracellular trap in chronic inflammation of endothelium in atherosclerosis. Life Sci, 2023, 328: 121867.
21. Kenny E F, Herzig A, Kruger R, et al. Diverse stimuli engage different neutrophil extracellular trap pathways. Elife, 2017, 6: e24437.
22. Sakurai K, Miyashita T, Okazaki M, et al. Role for neutrophil extracellular traps (NETs) and platelet aggregation in early sepsis-induced hepatic dysfunction. In Vivo, 2017, 31(6): 1051-1058.
23. 洪天添, 刘望, 黄嘉祺, 等. LPS刺激稳定黏附于ICAM-1上的中性粒细胞形成胞外诱捕网依赖于整合素Mac-1和细胞骨架蛋白. 生物医学工程学杂志, 2021, 38(5): 903-910.
24. Lotz S, Aga E, Wilde I, et al. Highly purified lipoteichoic acid activates neutrophil granulocytes and delays their spontaneous apoptosis via CD14 and TLR2. J Leukoc Biol, 2004, 75(3): 467-477.
25. Hann J, Bueb J L, Tolle F, et al. Calcium signaling and regulation of neutrophil functions: Still a long way to go. J Leukoc Biol, 2020, 107(2): 285-297.
26. de Bont C M, Koopman W J H, Boelens W C, et al. Stimulus-dependent chromatin dynamics, citrullination, calcium signalling and ROS production during NET formation. Biochim Biophys Acta Mol Cell Res, 2018, 1865(11 Pt A): 1621-1629.
27. Rochael N C, Guimaraes-Costa A B, Nascimento M T, et al. Classical ROS-dependent and early/rapid ROS-independent release of neutrophil extracellular traps triggered by Leishmania parasites. Sci Rep, 2015, 5: 18302.
28. Mazzoleni V, Zimmermann K, Smirnova A, et al. Staphylococcus aureus Panton-Valentine Leukocidin triggers an alternative NETosis process targeting mitochondria. FASEB J, 2021, 35(2): e21167.
29. Schonrich G, Raftery M J, Samstag Y. Devilishly radical network in COVID-19: Oxidative stress, neutrophil extracellular traps (NETs), and T cell suppression. Adv Biol Regul, 2020, 77: 100741.
30. Venkataranganayaka Abhilasha K, Kedihithlu Marathe G. Bacterial lipoproteins in sepsis. Immunobiology, 2021, 226(5): 152128.
31. Ginsburg I. Role of lipoteichoic acid in infection and inflammation. Lancet Infect Dis, 2002, 2(3): 171-179.
32. Tan C, Aziz M, Wang P. The vitals of NETs. J Leukoc Biol, 2021, 110(4): 797-808.
33. Lee S K, Goh S Y, Wong Y Q, et al. Response of neutrophils to extracellular haemoglobin and LTA in human blood system. EBioMedicine, 2015, 2(3): 225-233.
34. Pilsczek F H, Salina D, Poon K K, et al. A novel mechanism of rapid nuclear neutrophil extracellular trap formation in response to Staphylococcus aureus. J Immunol, 2010, 185(12): 7413-7425.

Journal of Biomedical Engineering

Neutrophil extracellular traps extrusion from neutrophils stably adhered to ICAM-1 by lipoteichoic acid stimulation

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content