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Volume 4, Issue 3 (9-2018)
jhehp 2018, 4(3): 138-143 |
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Moghanloo E, Ghorbani E, Beikverdi M S, Badameh P, Rezaei S, Piroozmand A, et al . The Netosis Formation of HL-60 Cell Differentiated to Neutrophil-Like Cells by LPS. jhehp 2018; 4 (3) :138-143
URL: http://jhehp.zums.ac.ir/article-1-165-en.html
URL: http://jhehp.zums.ac.ir/article-1-165-en.html
Ehsan Moghanloo1 
, Elham Ghorbani2 , Mohammad Sadegh Beikverdi3 , Parisa Badameh4 , Soheila Rezaei5 , Ahmad Piroozmand6 , Shahram Teimourian7 , Minoo Shahidi8 , Ahmad Khorshidi *1
, Elham Ghorbani2 , Mohammad Sadegh Beikverdi3 , Parisa Badameh4 , Soheila Rezaei5 , Ahmad Piroozmand6 , Shahram Teimourian7 , Minoo Shahidi8 , Ahmad Khorshidi *1
1- Department of Microbiology and Immunology, School of Medicine, Kashan University of Medical Sciences, Kashan, Iran.
2- Department of Genetics Campus2, Guilan University of Medical Science, Rasht, Iran.
3- Department of Biology Islamic Azad University Central Tehran Branch, Tehran, Iran.
4- Department of Microbiology, College of Basic Science and Biology,Varamin Pishva Branch, Islamic Azad University, Varamin, Iran.
5- Department of Molecular Genetic, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
6- Autoimmune Diseases Research Center, School of Medicine, Kashan University of Medical Sciences, Kashan, Iran.
7- Department of Medical Genetics, Iran University of Medical Sciences, Tehran, Iran.
8- Department of Hematology, School of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran.
2- Department of Genetics Campus2, Guilan University of Medical Science, Rasht, Iran.
3- Department of Biology Islamic Azad University Central Tehran Branch, Tehran, Iran.
4- Department of Microbiology, College of Basic Science and Biology,Varamin Pishva Branch, Islamic Azad University, Varamin, Iran.
5- Department of Molecular Genetic, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
6- Autoimmune Diseases Research Center, School of Medicine, Kashan University of Medical Sciences, Kashan, Iran.
7- Department of Medical Genetics, Iran University of Medical Sciences, Tehran, Iran.
8- Department of Hematology, School of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran.
Abstract: (9222 Views)
Background: Neutrophils are the most abundant white blood cells in humans. Recently, a novel strategy called the formation neutrophil extracellular traps (NETs) was described. NETs is a new strategy for pathogen response. This study focused on whether LPS induced NETs release in vitro in the HL60 cell line.
Methods: In this study, the HL60 cell line was used for culture and DMSO for induction and differentiation. Flow cytometry was used to evaluate CD11b in the differentiated cells, and the NBT assay was used to evaluate the functionality of the differentiated HL-60 cells. Neutrophil-like cells were incubated with LPS (200 ng/ml) for 45 min, followed by incubation for 25 min with 100 ng/ml Hoechst 33342. Trypan blue as vital staining was used for viability. The statistical significance of the difference between the control and treated groups was evaluated using a one-way ANOVA.
Results: Our results showed that 75% NETs was produced by HL-60 differentiated neutrophil cells exposed to 200 ng/ml LPS in 45 minues.
Conclusion: Consequently, the LPS-induced infection and lethality may occur through various mechanisms. Thus, understanding the molecular mechanisms regulating NET formation in LPS-induced neutrophil-like cells would support the development of new therapeutic methods.
Methods: In this study, the HL60 cell line was used for culture and DMSO for induction and differentiation. Flow cytometry was used to evaluate CD11b in the differentiated cells, and the NBT assay was used to evaluate the functionality of the differentiated HL-60 cells. Neutrophil-like cells were incubated with LPS (200 ng/ml) for 45 min, followed by incubation for 25 min with 100 ng/ml Hoechst 33342. Trypan blue as vital staining was used for viability. The statistical significance of the difference between the control and treated groups was evaluated using a one-way ANOVA.
Results: Our results showed that 75% NETs was produced by HL-60 differentiated neutrophil cells exposed to 200 ng/ml LPS in 45 minues.
Conclusion: Consequently, the LPS-induced infection and lethality may occur through various mechanisms. Thus, understanding the molecular mechanisms regulating NET formation in LPS-induced neutrophil-like cells would support the development of new therapeutic methods.
Type of Study: Original Article |
Subject:
Public Health
Received: 2018/08/11 | Accepted: 2018/09/8 | Published: 2018/09/21
Received: 2018/08/11 | Accepted: 2018/09/8 | Published: 2018/09/21
References
1. Nathan C. Neutrophils and Immunity: Challenges and Opportunities. Nat Rev Immunol. 2006; 6(3): 173-82. [Crossref]
2. Le Cabec V, Calafat J, Borregaard N. Sorting of the Specific Granule Protein, NGAL, During Granulocytic Maturation of HL-60 Cells. Blood. 1997; 89(6): 2113-21.
3. Itakura A. Dynamic Regulation of Neutrophil Activation, Migration and Death. Oregon Health Sci Univ OHSU Digital Common. 2013.
4. 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(2): 231-41. [Crossref]
5. Brinkmann V, Reichard U, Goosmann C, Fauler B, Uhlemann Y, Weiss DS, et al. Neutrophil Extracellular Traps Kill Bacteria. Sci. 2004; 303(5663): 1532-5. [Crossref]
6. Shin SH, Lee HR, Jeong J, Kim JH, Moon BG, Sohn KY, et al. PLAG Ameliorates LPS-induced ALI by Attenuation of Neutrophil Infiltration into Alveolar via a Prompt Resolution of TLR4 Signaling. Am Assoc Immnol. 2018; 200(1).
7. Urban CF, Ermert D, Schmid M, Abu-Abed U, Goosmann C, Nacken W, et al. Neutrophil Extracellular Traps Contain Calprotectin, a Cytosolic Protein Complex Involved in Host Defense Against Candida albicans. PLoS Pathog. 2009; 5(10): e1000639.
8. Megens RT, Vijayan S, Lievens D, Doering Y, van Zandvoort MA, Grommes J, et al. Presence of Luminal Neutrophil Extracellular Traps in Atherosclerosis. Thromb Haemost. 2012; 107(3): 597-8. [Crossref]
9. Brill A, Fuchs T, Savchenko A, Thomas G, Martinod K, De Meyer S, et al. Neutrophil Extracellular Traps Promote Deep Vein Thrombosis in Mice. J Thromb Haemost. 2012; 10(1): 136-44. [Crossref]
10. Cools-Lartigue J, Spicer J, McDonald B, Gowing S, Chow S, Giannias B, et al. Neutrophil Extracellular Traps Sequester Circulating Tumor Cells and Promote Metastasis. J Clin Investig. 2013; 123(8): 3446-58. [Crossref]
11. Dubois AV, Gauthier A, Bréa D, Varaigne F, Diot P, Gauthier F, et al. Influence of DNA on the Activities and Inhibition of Neutrophil Serine Proteases in Cystic Fibrosis Sputum. Am J Respir Cell Mol Biol. 2012; 47(1): 80-6. [Crossref]
12. Teimourian S, Moghanloo E. Role of PTEN in Neutrophil Extracellular Trap Formation. Mol Immunol. 2015; 66(2): 319-24. [Crossref]
13. Shimomura Y, Suga M, Kuriyama N, Nakamura T, Sakai T, Kato Y, et al. Recombinant Human Thrombomodulin Inhibits Neutrophil Extracellular Trap formation in vitro. J Intensive Care. 2016; 4: 48. [Crossref]
14. Amulic B, Sollberger G, Zychlinsky A. Neutrophil Extracellular Trap Formation is Independent of de Novo Gene Expression. PLoS One. 2016; 11(6): e0157454.
15. Yost CC, Cody MJ, Harris ES, Thornton NL, McInturff AM, Martinez ML, et al. Impaired Neutrophil Extracellular Trap (NET) Formation: a Novel Innate Immune Deficiency of Human Neonates. Blood. 2009; 113(25): 6419-27. [Crossref]
16. Remijsen Q, Kuijpers T, Wirawan E, Lippens S, Vandenabeele P, Berghe TV. Dying for a Cause: NETosis, Mechanisms Behind an Antimicrobial Cell Death Modality. Cell Death Differ. 2011; 18(4): 581-8. [Crossref]
17. Liu S, Su X, Pan P, Zhang L, Hu Y, Tan H, et al. Neutrophil Extracellular Traps are Indirectly Triggered by Lipopolysaccharide and Contribute to Acute Lung Injury. Sci Rep. 2016; 6: 37252. [Crossref]
18. Onouchi T, Shiogama K, Matsui T, Mizutani Y, Sakurai K, Inada K, et al. Visualization of Neutrophil Extracellular Traps and Fibrin Meshwork in Human Fibrinopurulent Inflammatory Lesions: II. Ultrastructural Study. Acta Histochem Cytochem. 2016; 49(4): 117-23. [Crossref]
19. Cheng T. Hematopoietic Differentiation of Human Pluripotent Stem Cells. Springer; 2015.
20. Morishima T, Watanabe Ki, Niwa A, Fujino H, Matsubara H, Adachi S, et al. Neutrophil Differentiation from Human‐Induced Pluripotent Stem Cells. J Cell Physiol. 2011; 226(5): 1283-91. [Crossref]
21. Teimourian S, Moghanloo E. Thwarting PTEN Expression by siRNA Augments HL-60 Cell Differentiation to Neutrophil-Like Cells by DMSO and ATRA. DNA Cell Biol. 2016; 35(10): 591-8. [Crossref]
22. Bateman J, Parida S, Nash G. Neutrophil Integrin Assay for Clinical Studies. Cell Biochem Funct. 1993; 11(2): 87-91. [Crossref]
23. Nathan DG, Baehner RL, Weaver DK. Failure of Nitro Blue Tetrazolium Reduction in the Phagocytic Vacuoles of Leukocytes in Chronic Granulomatous Disease. J Clin Invest. 1969; 48(10): 1895-904. [Crossref]
24. Segal AW. Nitroblue-Tetrazolium Tests. Lancet. 1974; 2(7891): 1248-52. [Crossref]
25. Zheng X, Wang Y, Liu B, Liu C, Liu D, Zhu J, et al. Bmi-1-shRNA Inhibits the Proliferation of Lung Adenocarcinoma Cells by Blocking the G1/S Phase Through Decreasing Cyclin D1 and Increasing p21/p27 Levels. Nucleic Acid Ther. 2014; 24(3): 210-6. [Crossref]
26. McInturff AM, Cody MJ, Elliott EA, Glenn JW, Rowley JW, Rondina MT, et al. Mammalian Target of Rapamycin Regulates Neutrophil Extracellular Trap Formation via Induction of Hypoxia-Inducible Factor 1 alpha. Blood. 2012; 120(15): 3118-25. [Crossref]
27. Clark SR, Ma AC, Tavener SA, McDonald B, Goodarzi Z, Kelly MM, et al. Platelet TLR4 Activates Neutrophil Extracellular Traps to Ensnare Bacteria in Septic Blood. Nat Med. 2007; 13(4): 463-9. [Crossref]
28. Shimomura Y, Suga M, Kuriyama N, Nakamura T, Sakai T, Kato Y, et al. Recombinant Human Thrombomodulin Inhibits Neutrophil Extracellular Trap Formation in vitro. J Intensive Care. 2016; 4(1): 48. [Crossref]
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