Heterogeneity in Neutrophil Extracellular Traps from Healthy Human Subjects
Neutrophil extracellular traps (NETs), a key component of early defense against microbial infection, are also associated with tissue injury. NET composition has been reported to vary with some disease states, but the composition and variability of NETs across many healthy subjects provide a critical...
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| Format: | Article |
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MDPI AG
2023-12-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/25/1/525 |
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| author | Margaret S. Collins Michelle A. Imbrogno Elizabeth J. Kopras James A. Howard Nanhua Zhang Elizabeth L. Kramer Kristin M. Hudock |
| author_facet | Margaret S. Collins Michelle A. Imbrogno Elizabeth J. Kopras James A. Howard Nanhua Zhang Elizabeth L. Kramer Kristin M. Hudock |
| author_sort | Margaret S. Collins |
| collection | DOAJ |
| description | Neutrophil extracellular traps (NETs), a key component of early defense against microbial infection, are also associated with tissue injury. NET composition has been reported to vary with some disease states, but the composition and variability of NETs across many healthy subjects provide a critical comparison that has not been well investigated. We evaluated NETs from twelve healthy subjects of varying ages isolated from multiple blood draws over a three-and-one-half-year period to delineate the variability in extracellular DNA, protein, enzymatic activities, and susceptibility to protease inhibitors. We calculated correlations for NET constituents and loss of human bronchial epithelial barrier integrity, measured by transepithelial electrical resistance, after NET exposure. We found that although there was some variability within the same subject over time, the mean NET total DNA, dsDNA, protein, LDH, neutrophil elastase (NE), and proteinase 3 (PR3) in isolated NETs were consistent across subjects. NET serine protease activity varied considerably within the same donor from day to day. The mean NET cathepsin G and MPO were significantly different across donors. IL-8 > IL-1RA > G-CSF were the most abundant cytokines in NETs. There was no significant difference in the mean concentration or variability of IL-8, IL-1RA, G-CSF, IL-1α, IL-1β, or TNF-α in different subjects’ NETs. NET DNA concentration was correlated with increased NET neutrophil elastase activity and higher NET IL-1RA concentrations. The mean reduction in protease activity by protease inhibitors was significantly different across donors. NET DNA concentration correlated best with reductions in the barrier integrity of human bronchial epithelia. Defining NET concentration by DNA content correlates with other NET components and reductions in NET-driven epithelial barrier dysfunction, suggesting DNA is a reasonable surrogate measurement for these complex structures in healthy subjects. |
| first_indexed | 2024-03-08T15:04:32Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-08T15:04:32Z |
| publishDate | 2023-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-2ea3adffb516462cae7561ac3696b6002024-01-10T14:59:50ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-12-0125152510.3390/ijms25010525Heterogeneity in Neutrophil Extracellular Traps from Healthy Human SubjectsMargaret S. Collins0Michelle A. Imbrogno1Elizabeth J. Kopras2James A. Howard3Nanhua Zhang4Elizabeth L. Kramer5Kristin M. Hudock6Division of Pulmonary, Critical Care & Sleep Medicine, Department of Medicine, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USADivision of Pulmonary, Critical Care & Sleep Medicine, Department of Medicine, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USADivision of Pulmonary, Critical Care & Sleep Medicine, Department of Medicine, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USADepartment of Pharmacology & Systems Physiology, University of Cincinnati, Cincinnati, OH 45267, USADepartment of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH 45229, USADepartment of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH 45229, USADivision of Pulmonary, Critical Care & Sleep Medicine, Department of Medicine, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USANeutrophil extracellular traps (NETs), a key component of early defense against microbial infection, are also associated with tissue injury. NET composition has been reported to vary with some disease states, but the composition and variability of NETs across many healthy subjects provide a critical comparison that has not been well investigated. We evaluated NETs from twelve healthy subjects of varying ages isolated from multiple blood draws over a three-and-one-half-year period to delineate the variability in extracellular DNA, protein, enzymatic activities, and susceptibility to protease inhibitors. We calculated correlations for NET constituents and loss of human bronchial epithelial barrier integrity, measured by transepithelial electrical resistance, after NET exposure. We found that although there was some variability within the same subject over time, the mean NET total DNA, dsDNA, protein, LDH, neutrophil elastase (NE), and proteinase 3 (PR3) in isolated NETs were consistent across subjects. NET serine protease activity varied considerably within the same donor from day to day. The mean NET cathepsin G and MPO were significantly different across donors. IL-8 > IL-1RA > G-CSF were the most abundant cytokines in NETs. There was no significant difference in the mean concentration or variability of IL-8, IL-1RA, G-CSF, IL-1α, IL-1β, or TNF-α in different subjects’ NETs. NET DNA concentration was correlated with increased NET neutrophil elastase activity and higher NET IL-1RA concentrations. The mean reduction in protease activity by protease inhibitors was significantly different across donors. NET DNA concentration correlated best with reductions in the barrier integrity of human bronchial epithelia. Defining NET concentration by DNA content correlates with other NET components and reductions in NET-driven epithelial barrier dysfunction, suggesting DNA is a reasonable surrogate measurement for these complex structures in healthy subjects.https://www.mdpi.com/1422-0067/25/1/525NET heterogeneityneutrophil extracellular trapsDNAneutrophil elastase |
| spellingShingle | Margaret S. Collins Michelle A. Imbrogno Elizabeth J. Kopras James A. Howard Nanhua Zhang Elizabeth L. Kramer Kristin M. Hudock Heterogeneity in Neutrophil Extracellular Traps from Healthy Human Subjects International Journal of Molecular Sciences NET heterogeneity neutrophil extracellular traps DNA neutrophil elastase |
| title | Heterogeneity in Neutrophil Extracellular Traps from Healthy Human Subjects |
| title_full | Heterogeneity in Neutrophil Extracellular Traps from Healthy Human Subjects |
| title_fullStr | Heterogeneity in Neutrophil Extracellular Traps from Healthy Human Subjects |
| title_full_unstemmed | Heterogeneity in Neutrophil Extracellular Traps from Healthy Human Subjects |
| title_short | Heterogeneity in Neutrophil Extracellular Traps from Healthy Human Subjects |
| title_sort | heterogeneity in neutrophil extracellular traps from healthy human subjects |
| topic | NET heterogeneity neutrophil extracellular traps DNA neutrophil elastase |
| url | https://www.mdpi.com/1422-0067/25/1/525 |
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