Effect of Adhesion and Substrate Elasticity on Neutrophil Extracellular Trap Formation
Neutrophils are the most abundant type of white blood cells. Upon stimulation, they are able to decondense and release their chromatin as neutrophil extracellular traps (NETs). This process (NETosis) is part of immune defense mechanisms but also plays an important role in many chronic and inflammato...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2019-10-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fimmu.2019.02320/full |
| _version_ | 1818269818606845952 |
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| author | Luise Erpenbeck Antonia Luise Gruhn Galina Kudryasheva Gökhan Günay Gökhan Günay Daniel Meyer Julia Busse Elsa Neubert Elsa Neubert Michael P. Schön Michael P. Schön Florian Rehfeldt Sebastian Kruss |
| author_facet | Luise Erpenbeck Antonia Luise Gruhn Galina Kudryasheva Gökhan Günay Gökhan Günay Daniel Meyer Julia Busse Elsa Neubert Elsa Neubert Michael P. Schön Michael P. Schön Florian Rehfeldt Sebastian Kruss |
| author_sort | Luise Erpenbeck |
| collection | DOAJ |
| description | Neutrophils are the most abundant type of white blood cells. Upon stimulation, they are able to decondense and release their chromatin as neutrophil extracellular traps (NETs). This process (NETosis) is part of immune defense mechanisms but also plays an important role in many chronic and inflammatory diseases such as atherosclerosis, rheumatoid arthritis, diabetes, and cancer. For this reason, much effort has been invested into understanding biochemical signaling pathways in NETosis. However, the impact of the mechanical micro-environment and adhesion on NETosis is not well-understood. Here, we studied how adhesion and especially substrate elasticity affect NETosis. We employed polyacrylamide (PAA) gels with distinctly defined elasticities (Young's modulus E) within the physiologically relevant range from 1 to 128 kPa and coated the gels with integrin ligands (collagen I, fibrinogen). Neutrophils were cultured on these substrates and stimulated with potent inducers of NETosis: phorbol 12-myristate 13-acetate (PMA) and lipopolysaccharide (LPS). Interestingly, PMA-induced NETosis was neither affected by substrate elasticity nor by different integrin ligands. In contrast, for LPS stimulation, NETosis rates increased with increasing substrate elasticity (E > 20 kPa). LPS-induced NETosis increased with increasing cell contact area, while PMA-induced NETosis did not require adhesion at all. Furthermore, inhibition of phosphatidylinositide 3 kinase (PI3K), which is involved in adhesion signaling, completely abolished LPS-induced NETosis but only slightly decreased PMA-induced NETosis. In summary, we show that LPS-induced NETosis depends on adhesion and substrate elasticity while PMA-induced NETosis is completely independent of adhesion. |
| first_indexed | 2024-12-12T21:00:26Z |
| format | Article |
| id | doaj.art-632562c8a15a4a23800bbfea26ac1128 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-12T21:00:26Z |
| publishDate | 2019-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-632562c8a15a4a23800bbfea26ac11282022-12-22T00:12:10ZengFrontiers Media S.A.Frontiers in Immunology1664-32242019-10-011010.3389/fimmu.2019.02320473479Effect of Adhesion and Substrate Elasticity on Neutrophil Extracellular Trap FormationLuise Erpenbeck0Antonia Luise Gruhn1Galina Kudryasheva2Gökhan Günay3Gökhan Günay4Daniel Meyer5Julia Busse6Elsa Neubert7Elsa Neubert8Michael P. Schön9Michael P. Schön10Florian Rehfeldt11Sebastian Kruss12Department of Dermatology, Venereology and Allergology, University Medical Center, Göttingen University, Göttingen, GermanyDepartment of Dermatology, Venereology and Allergology, University Medical Center, Göttingen University, Göttingen, GermanyThird Institute of Physics–Biophysics, Göttingen University, Göttingen, GermanyDepartment of Dermatology, Venereology and Allergology, University Medical Center, Göttingen University, Göttingen, GermanyDepartment of Chemistry, Institute of Physical Chemistry, Göttingen University, Göttingen, GermanyDepartment of Chemistry, Institute of Physical Chemistry, Göttingen University, Göttingen, GermanyDepartment of Dermatology, Venereology and Allergology, University Medical Center, Göttingen University, Göttingen, GermanyDepartment of Dermatology, Venereology and Allergology, University Medical Center, Göttingen University, Göttingen, GermanyDepartment of Chemistry, Institute of Physical Chemistry, Göttingen University, Göttingen, GermanyDepartment of Dermatology, Venereology and Allergology, University Medical Center, Göttingen University, Göttingen, GermanyLower Saxony Institute of Occupational Dermatology, Göttingen, GermanyThird Institute of Physics–Biophysics, Göttingen University, Göttingen, GermanyDepartment of Chemistry, Institute of Physical Chemistry, Göttingen University, Göttingen, GermanyNeutrophils are the most abundant type of white blood cells. Upon stimulation, they are able to decondense and release their chromatin as neutrophil extracellular traps (NETs). This process (NETosis) is part of immune defense mechanisms but also plays an important role in many chronic and inflammatory diseases such as atherosclerosis, rheumatoid arthritis, diabetes, and cancer. For this reason, much effort has been invested into understanding biochemical signaling pathways in NETosis. However, the impact of the mechanical micro-environment and adhesion on NETosis is not well-understood. Here, we studied how adhesion and especially substrate elasticity affect NETosis. We employed polyacrylamide (PAA) gels with distinctly defined elasticities (Young's modulus E) within the physiologically relevant range from 1 to 128 kPa and coated the gels with integrin ligands (collagen I, fibrinogen). Neutrophils were cultured on these substrates and stimulated with potent inducers of NETosis: phorbol 12-myristate 13-acetate (PMA) and lipopolysaccharide (LPS). Interestingly, PMA-induced NETosis was neither affected by substrate elasticity nor by different integrin ligands. In contrast, for LPS stimulation, NETosis rates increased with increasing substrate elasticity (E > 20 kPa). LPS-induced NETosis increased with increasing cell contact area, while PMA-induced NETosis did not require adhesion at all. Furthermore, inhibition of phosphatidylinositide 3 kinase (PI3K), which is involved in adhesion signaling, completely abolished LPS-induced NETosis but only slightly decreased PMA-induced NETosis. In summary, we show that LPS-induced NETosis depends on adhesion and substrate elasticity while PMA-induced NETosis is completely independent of adhesion.https://www.frontiersin.org/article/10.3389/fimmu.2019.02320/fullneutrophil extracellular traps (NET)substrate elasticitystiffness and its variationsinflammationimmunomodulationadhesion |
| spellingShingle | Luise Erpenbeck Antonia Luise Gruhn Galina Kudryasheva Gökhan Günay Gökhan Günay Daniel Meyer Julia Busse Elsa Neubert Elsa Neubert Michael P. Schön Michael P. Schön Florian Rehfeldt Sebastian Kruss Effect of Adhesion and Substrate Elasticity on Neutrophil Extracellular Trap Formation Frontiers in Immunology neutrophil extracellular traps (NET) substrate elasticity stiffness and its variations inflammation immunomodulation adhesion |
| title | Effect of Adhesion and Substrate Elasticity on Neutrophil Extracellular Trap Formation |
| title_full | Effect of Adhesion and Substrate Elasticity on Neutrophil Extracellular Trap Formation |
| title_fullStr | Effect of Adhesion and Substrate Elasticity on Neutrophil Extracellular Trap Formation |
| title_full_unstemmed | Effect of Adhesion and Substrate Elasticity on Neutrophil Extracellular Trap Formation |
| title_short | Effect of Adhesion and Substrate Elasticity on Neutrophil Extracellular Trap Formation |
| title_sort | effect of adhesion and substrate elasticity on neutrophil extracellular trap formation |
| topic | neutrophil extracellular traps (NET) substrate elasticity stiffness and its variations inflammation immunomodulation adhesion |
| url | https://www.frontiersin.org/article/10.3389/fimmu.2019.02320/full |
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