Evaluation of Immunomodulatory Effects of <i>Fusarium</i> Mycotoxins Using Bacterial Endotoxin-Stimulated Bovine Epithelial Cells and Macrophages in Co-Culture
Mycotoxins are secondary metabolites produced by a variety of fungi that contaminate animal food and feeds and are capable of inducing a wide range of toxicities. Predictive in vitro models represent valuable substitutes for animal experiments to assess the toxicity of mycotoxins. The complexities o...
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MDPI AG
2023-10-01
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| Series: | Genes |
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| Online Access: | https://www.mdpi.com/2073-4425/14/11/2014 |
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| author | Umesh K. Shandilya Ankita Sharma Ran Xu Maria Malane M. Muniz Niel A. Karrow |
| author_facet | Umesh K. Shandilya Ankita Sharma Ran Xu Maria Malane M. Muniz Niel A. Karrow |
| author_sort | Umesh K. Shandilya |
| collection | DOAJ |
| description | Mycotoxins are secondary metabolites produced by a variety of fungi that contaminate animal food and feeds and are capable of inducing a wide range of toxicities. Predictive in vitro models represent valuable substitutes for animal experiments to assess the toxicity of mycotoxins. The complexities of the interactions between epithelial and innate immune cells, vital for upholding barrier integrity and averting infections, remain inadequately understood. In the current study, a co-culture model of bovine epithelial cells (MAC-T) and macrophages (BoMac) was used to investigate the impact of exposure to <i>Fusarium</i> mycotoxins, namely deoxynivalenol (DON), zearalenone (ZEN), enniatin B (ENB), and beauvericin (BEA), on the inflammatory response elicited by the bacterial lipopolysaccharide (LPS) endotoxin. The MAC-T cells and BoMac were seeded on the apical side of a Transwell membrane and in the lower chamber, respectively, and mycotoxin exposure on the apical side of the membrane was carried out with the different mycotoxins (LC20; concentrations that elicited 20% cytotoxicity) for 48 h followed by an LPS immunity challenge for 24 h. The culture supernatants were collected from the basolateral compartment and these samples were submitted for cytokine/chemokine multiplex analysis. RNA-Seq analysis was performed using total RNA extracted from the MAC-T cells to acquire a more detailed insight into their cellular functions. The multiplex analysis indicated that IFN-γ, IL-1α, IL-8, and MCP-1 were significantly induced post-DON treatment when compared to control cells, and levels of IL-1α and IL-8 were enhanced significantly in all mycotoxin-treated groups post-LPS challenge. Analysis of the sequencing results showed that there were 341, 357, and 318 differentially expressed MAC-T cell genes that were up-regulated in the DON, ENB, and BEA groups, respectively. Gene ontology and pathway analysis revealed that these DEGs were significantly enriched in various biological processes and pathways related to inflammation, apoptosis signaling, and Wnt signaling. These results provide a comprehensive analysis of the co-culture cytokine/chemokine production and MAC-T cells’ gene expression profiles elicited by <i>Fusarium</i> mycotoxins, which further contributes to the understanding of early endotoxemia post-mycotoxin exposure. |
| first_indexed | 2024-03-09T16:48:09Z |
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| institution | Directory Open Access Journal |
| issn | 2073-4425 |
| language | English |
| last_indexed | 2024-03-09T16:48:09Z |
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| spelling | doaj.art-ac4af9b520114482b2d701c94bb0078f2023-11-24T14:43:43ZengMDPI AGGenes2073-44252023-10-011411201410.3390/genes14112014Evaluation of Immunomodulatory Effects of <i>Fusarium</i> Mycotoxins Using Bacterial Endotoxin-Stimulated Bovine Epithelial Cells and Macrophages in Co-CultureUmesh K. Shandilya0Ankita Sharma1Ran Xu2Maria Malane M. Muniz3Niel A. Karrow4Department of Animal Biosciences, University of Guelph, 50 Stone Rd. E., Guelph, ON N1G2W1, CanadaDepartment of Animal Biosciences, University of Guelph, 50 Stone Rd. E., Guelph, ON N1G2W1, CanadaDepartment of Animal Biosciences, University of Guelph, 50 Stone Rd. E., Guelph, ON N1G2W1, CanadaDepartment of Animal Biosciences, University of Guelph, 50 Stone Rd. E., Guelph, ON N1G2W1, CanadaDepartment of Animal Biosciences, University of Guelph, 50 Stone Rd. E., Guelph, ON N1G2W1, CanadaMycotoxins are secondary metabolites produced by a variety of fungi that contaminate animal food and feeds and are capable of inducing a wide range of toxicities. Predictive in vitro models represent valuable substitutes for animal experiments to assess the toxicity of mycotoxins. The complexities of the interactions between epithelial and innate immune cells, vital for upholding barrier integrity and averting infections, remain inadequately understood. In the current study, a co-culture model of bovine epithelial cells (MAC-T) and macrophages (BoMac) was used to investigate the impact of exposure to <i>Fusarium</i> mycotoxins, namely deoxynivalenol (DON), zearalenone (ZEN), enniatin B (ENB), and beauvericin (BEA), on the inflammatory response elicited by the bacterial lipopolysaccharide (LPS) endotoxin. The MAC-T cells and BoMac were seeded on the apical side of a Transwell membrane and in the lower chamber, respectively, and mycotoxin exposure on the apical side of the membrane was carried out with the different mycotoxins (LC20; concentrations that elicited 20% cytotoxicity) for 48 h followed by an LPS immunity challenge for 24 h. The culture supernatants were collected from the basolateral compartment and these samples were submitted for cytokine/chemokine multiplex analysis. RNA-Seq analysis was performed using total RNA extracted from the MAC-T cells to acquire a more detailed insight into their cellular functions. The multiplex analysis indicated that IFN-γ, IL-1α, IL-8, and MCP-1 were significantly induced post-DON treatment when compared to control cells, and levels of IL-1α and IL-8 were enhanced significantly in all mycotoxin-treated groups post-LPS challenge. Analysis of the sequencing results showed that there were 341, 357, and 318 differentially expressed MAC-T cell genes that were up-regulated in the DON, ENB, and BEA groups, respectively. Gene ontology and pathway analysis revealed that these DEGs were significantly enriched in various biological processes and pathways related to inflammation, apoptosis signaling, and Wnt signaling. These results provide a comprehensive analysis of the co-culture cytokine/chemokine production and MAC-T cells’ gene expression profiles elicited by <i>Fusarium</i> mycotoxins, which further contributes to the understanding of early endotoxemia post-mycotoxin exposure.https://www.mdpi.com/2073-4425/14/11/2014co-culturecytokinesinflammatory responsemacrophagesmycotoxin |
| spellingShingle | Umesh K. Shandilya Ankita Sharma Ran Xu Maria Malane M. Muniz Niel A. Karrow Evaluation of Immunomodulatory Effects of <i>Fusarium</i> Mycotoxins Using Bacterial Endotoxin-Stimulated Bovine Epithelial Cells and Macrophages in Co-Culture Genes co-culture cytokines inflammatory response macrophages mycotoxin |
| title | Evaluation of Immunomodulatory Effects of <i>Fusarium</i> Mycotoxins Using Bacterial Endotoxin-Stimulated Bovine Epithelial Cells and Macrophages in Co-Culture |
| title_full | Evaluation of Immunomodulatory Effects of <i>Fusarium</i> Mycotoxins Using Bacterial Endotoxin-Stimulated Bovine Epithelial Cells and Macrophages in Co-Culture |
| title_fullStr | Evaluation of Immunomodulatory Effects of <i>Fusarium</i> Mycotoxins Using Bacterial Endotoxin-Stimulated Bovine Epithelial Cells and Macrophages in Co-Culture |
| title_full_unstemmed | Evaluation of Immunomodulatory Effects of <i>Fusarium</i> Mycotoxins Using Bacterial Endotoxin-Stimulated Bovine Epithelial Cells and Macrophages in Co-Culture |
| title_short | Evaluation of Immunomodulatory Effects of <i>Fusarium</i> Mycotoxins Using Bacterial Endotoxin-Stimulated Bovine Epithelial Cells and Macrophages in Co-Culture |
| title_sort | evaluation of immunomodulatory effects of i fusarium i mycotoxins using bacterial endotoxin stimulated bovine epithelial cells and macrophages in co culture |
| topic | co-culture cytokines inflammatory response macrophages mycotoxin |
| url | https://www.mdpi.com/2073-4425/14/11/2014 |
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