<i>Fasciola hepatica</i> Soluble Antigen (<i>Fh</i>Ag)-Induced NETs Under Hypoxic Conditions Exert Cytotoxic Effects on Hepatic Cells In Vitro
<i>Fasciola hepatica</i> is a parasitic trematode that causes fasciolosis in sheep, provoking a decrease in their reproductive capacity, weight gain, meat and milk production, and wool quality. In the pathogenesis of <i>F. hepatica</i>, the penetration and migration of parasi...
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2024-11-01
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| author | Tamara Muñoz-Caro Pamela Quiroz Cristina Abarca Marcela Gómez-Ceruti Pablo Alarcón Stefanie Teuber Max Navarro Anja Taubert Carlos Hermosilla Rafael A. Burgos |
| author_facet | Tamara Muñoz-Caro Pamela Quiroz Cristina Abarca Marcela Gómez-Ceruti Pablo Alarcón Stefanie Teuber Max Navarro Anja Taubert Carlos Hermosilla Rafael A. Burgos |
| author_sort | Tamara Muñoz-Caro |
| collection | DOAJ |
| description | <i>Fasciola hepatica</i> is a parasitic trematode that causes fasciolosis in sheep, provoking a decrease in their reproductive capacity, weight gain, meat and milk production, and wool quality. In the pathogenesis of <i>F. hepatica</i>, the penetration and migration of parasitic stages through the liver provoke intense inflammatory immune responses and tissue damage. The aim of this study was to investigate the cytotoxic effects of <i>Fascila hepatica</i>-induced ovine NETs in exposed hepatocytes in vitro, and to analyze whether <i>F. hepatica</i> antigens (<i>Fh</i>Ag) trigger the release of ovine NETs under hypoxic conditions as well as the roles of matrix metalloproteinase-9 (MMP-9) and CD11b in this cellular process in vitro. Here, isolated ovine PMNs were co-cultured with <i>Fh</i>Ag under hypoxia (5% O<sub>2</sub>) and NETs were visualized via immunofluorescence analyses, confirming their classical characteristics. The quantification of NETs in response to <i>Fh</i>Ag in hypoxic conditions significantly enhanced the formation of anchored and cell-free NETs (<i>p</i> < 0.01), and NADPH oxidase (NOX) inhibitor diphenylene iodonium (DPI) significantly reduced their production (<i>p</i> < 0.05). Furthermore, the cytotoxic effect of NETs on hepatic cells was determined by using a live/dead-staining with Sytox Orange, thereby demonstrating that <i>Fh</i>Ag-induced NETs are cytotoxic for hepatic cells (<i>p</i> = 0.001). We additionally analyzed PMN supernatants to determine the enzymatic activity of MMP-9, observing that <i>Fh</i>Ag exposure enhances MMP-9 release in ovine PMNs (<i>p</i> < 0.05) but not in bovine PMNs. Interestingly, by using flow cytometric analysis, we determined that the exposure of PMNs to <i>Fh</i>Ag does not increase the CD11b surface expression of ovine PMNs. This could be an effect of the activation of other surface receptors or transcription factors involved in <i>F. hepatica</i>-induced NETosis. Consequently, we hypothesize that <i>F. hepatica</i>-induced NETs play a role in the pathogenesis of fasciolosis, contributing to liver tissue damage if released in an uncontrolled manner. |
| format | Article |
| id | doaj-art-4d9cf4a3541941bf83947ae55d8beff2 |
| institution | Kabale University |
| issn | 2076-2615 |
| language | English |
| publishDate | 2024-11-01 |
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| spelling | doaj-art-4d9cf4a3541941bf83947ae55d8beff22024-12-13T16:21:17ZengMDPI AGAnimals2076-26152024-11-011423345610.3390/ani14233456<i>Fasciola hepatica</i> Soluble Antigen (<i>Fh</i>Ag)-Induced NETs Under Hypoxic Conditions Exert Cytotoxic Effects on Hepatic Cells In VitroTamara Muñoz-Caro0Pamela Quiroz1Cristina Abarca2Marcela Gómez-Ceruti3Pablo Alarcón4Stefanie Teuber5Max Navarro6Anja Taubert7Carlos Hermosilla8Rafael A. Burgos9Escuela de Medicina Veterinaria, Facultad de Medicina Veterinaria y Recursos Naturales, Universidad Santo Tomás, Talca 3460000, ChileEscuela de Medicina Veterinaria, Facultad de Medicina Veterinaria y Recursos Naturales, Universidad Santo Tomás, Talca 3460000, ChileEscuela de Medicina Veterinaria, Facultad de Medicina Veterinaria y Recursos Naturales, Universidad Santo Tomás, Talca 3460000, ChileCentro de Investigación de Ovinos Para El Secano OVISNOVA, Facultad de Medicina Veterinaria y Recursos Naturales, Universidad Santo Tomás, Talca 3460000, ChileLaboratory of Inflammation Pharmacology, Institute of Pharmacology and Morphophysiology, Faculty of Veterinary Sciences, Universidad Austral de Chile, Valdivia 5090000, ChileLaboratory of Inflammation Pharmacology, Institute of Pharmacology and Morphophysiology, Faculty of Veterinary Sciences, Universidad Austral de Chile, Valdivia 5090000, ChileInstitute of Veterinary Clinical Science, Faculty of Veterinary Sciences, Universidad Austral de Chile, Valdivia 5090000, ChileInstitute of Parasitology, Faculty of Veterinary Medicine, Justus Liebig University Giessen, 35392 Giessen, GermanyInstitute of Parasitology, Faculty of Veterinary Medicine, Justus Liebig University Giessen, 35392 Giessen, GermanyLaboratory of Inflammation Pharmacology, Institute of Pharmacology and Morphophysiology, Faculty of Veterinary Sciences, Universidad Austral de Chile, Valdivia 5090000, Chile<i>Fasciola hepatica</i> is a parasitic trematode that causes fasciolosis in sheep, provoking a decrease in their reproductive capacity, weight gain, meat and milk production, and wool quality. In the pathogenesis of <i>F. hepatica</i>, the penetration and migration of parasitic stages through the liver provoke intense inflammatory immune responses and tissue damage. The aim of this study was to investigate the cytotoxic effects of <i>Fascila hepatica</i>-induced ovine NETs in exposed hepatocytes in vitro, and to analyze whether <i>F. hepatica</i> antigens (<i>Fh</i>Ag) trigger the release of ovine NETs under hypoxic conditions as well as the roles of matrix metalloproteinase-9 (MMP-9) and CD11b in this cellular process in vitro. Here, isolated ovine PMNs were co-cultured with <i>Fh</i>Ag under hypoxia (5% O<sub>2</sub>) and NETs were visualized via immunofluorescence analyses, confirming their classical characteristics. The quantification of NETs in response to <i>Fh</i>Ag in hypoxic conditions significantly enhanced the formation of anchored and cell-free NETs (<i>p</i> < 0.01), and NADPH oxidase (NOX) inhibitor diphenylene iodonium (DPI) significantly reduced their production (<i>p</i> < 0.05). Furthermore, the cytotoxic effect of NETs on hepatic cells was determined by using a live/dead-staining with Sytox Orange, thereby demonstrating that <i>Fh</i>Ag-induced NETs are cytotoxic for hepatic cells (<i>p</i> = 0.001). We additionally analyzed PMN supernatants to determine the enzymatic activity of MMP-9, observing that <i>Fh</i>Ag exposure enhances MMP-9 release in ovine PMNs (<i>p</i> < 0.05) but not in bovine PMNs. Interestingly, by using flow cytometric analysis, we determined that the exposure of PMNs to <i>Fh</i>Ag does not increase the CD11b surface expression of ovine PMNs. This could be an effect of the activation of other surface receptors or transcription factors involved in <i>F. hepatica</i>-induced NETosis. Consequently, we hypothesize that <i>F. hepatica</i>-induced NETs play a role in the pathogenesis of fasciolosis, contributing to liver tissue damage if released in an uncontrolled manner.https://www.mdpi.com/2076-2615/14/23/3456<i>Fasciola hepatica</i>neutrophil extracellular trapsNETosisROShypoxiacytotoxicity |
| spellingShingle | Tamara Muñoz-Caro Pamela Quiroz Cristina Abarca Marcela Gómez-Ceruti Pablo Alarcón Stefanie Teuber Max Navarro Anja Taubert Carlos Hermosilla Rafael A. Burgos <i>Fasciola hepatica</i> Soluble Antigen (<i>Fh</i>Ag)-Induced NETs Under Hypoxic Conditions Exert Cytotoxic Effects on Hepatic Cells In Vitro Animals <i>Fasciola hepatica</i> neutrophil extracellular traps NETosis ROS hypoxia cytotoxicity |
| title | <i>Fasciola hepatica</i> Soluble Antigen (<i>Fh</i>Ag)-Induced NETs Under Hypoxic Conditions Exert Cytotoxic Effects on Hepatic Cells In Vitro |
| title_full | <i>Fasciola hepatica</i> Soluble Antigen (<i>Fh</i>Ag)-Induced NETs Under Hypoxic Conditions Exert Cytotoxic Effects on Hepatic Cells In Vitro |
| title_fullStr | <i>Fasciola hepatica</i> Soluble Antigen (<i>Fh</i>Ag)-Induced NETs Under Hypoxic Conditions Exert Cytotoxic Effects on Hepatic Cells In Vitro |
| title_full_unstemmed | <i>Fasciola hepatica</i> Soluble Antigen (<i>Fh</i>Ag)-Induced NETs Under Hypoxic Conditions Exert Cytotoxic Effects on Hepatic Cells In Vitro |
| title_short | <i>Fasciola hepatica</i> Soluble Antigen (<i>Fh</i>Ag)-Induced NETs Under Hypoxic Conditions Exert Cytotoxic Effects on Hepatic Cells In Vitro |
| title_sort | i fasciola hepatica i soluble antigen i fh i ag induced nets under hypoxic conditions exert cytotoxic effects on hepatic cells in vitro |
| topic | <i>Fasciola hepatica</i> neutrophil extracellular traps NETosis ROS hypoxia cytotoxicity |
| url | https://www.mdpi.com/2076-2615/14/23/3456 |
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