Traditional and emerging Fusarium mycotoxins disrupt homeostasis of bovine mammary cells by altering cell permeability and innate immune function

Xu, Ran; Shandilya, Umesh K.; Yiannikouris, Alexandros; Karrow, Niel A.

doi:10.1016/j.aninu.2022.10.007

Cited by 5 publications

(4 citation statements)

References 121 publications

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“…These findings suggest that the damage caused by DON exposure to the barrier and tight junction of the epithelial monolayer may facilitate the diffusion of LPS through the epithelial layer to macrophages in the lower compartment of the co-culture. In support of this hypothesis, our previous research reported that DON, ENB, and BEA differentially disrupted the paracellular permeability of MAC-T cells [34]. The maintenance of intact barrier functions, as well as innate and adaptive immune responses, is crucial for effectively eliminating infectious pathogens and toxic metabolites.…”

Section: Discussionmentioning

confidence: 70%

Evaluation of Immunomodulatory Effects of Fusarium Mycotoxins Using Bacterial Endotoxin-Stimulated Bovine Epithelial Cells and Macrophages in Co-Culture

Shandilya,

Sharma,

et al. 2023

Genes

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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 Fusarium 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 Fusarium mycotoxins, which further contributes to the understanding of early endotoxemia post-mycotoxin exposure.

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Section: Discussionmentioning

confidence: 70%

Evaluation of Immunomodulatory Effects of Fusarium Mycotoxins Using Bacterial Endotoxin-Stimulated Bovine Epithelial Cells and Macrophages in Co-Culture

Shandilya,

Sharma,

et al. 2023

Genes

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“…It was also anticipated to provide sufficient contact time for adsorption to reach equilibrium based on previous studies [ 24 , 53 , 54 ]. Our group previously reported that DON, BEA, OTA, and CIT decreased cell viability of MAC-T cells [ 41 , 55 ]. Coupled with an adsorption isotherm model-based approach, a MAC-T cell-based bioassay was used as an in vitro model of bovine mammary epithelium to investigate the cytotoxic effects of residual mycotoxin concentrations present in the supernatant and their impact on cell viability with or without the inclusion of three yeast cell wall-based mycotoxin treatments tested individually.…”

Section: Discussionmentioning

confidence: 99%

“…A volume of 1 mL supernatant subsamples of DON (2 and 4 μg/mL), BEA (20 and 40 μg/mL), OTA (20 and 30 μg/mL), and CIT (50 and 68 μg/mL) preserved from the GIT incubation procedure ( Section 2.2 ) were lyophilized using a freeze dryer (Harvest Right, Canada). The concentrations that were selected had the highest two cytotoxic concentrations reported in our previously published studies [ 41 , 55 ].…”

Section: Methodsmentioning

confidence: 99%

“…5.6.2. Cytotoxicity Assay MAC-T cells were seeded at a density of 2  10 4 cells per well in 96-well microplates [41,55]. A volume of 1 mL supernatant subsamples of DON (2 and 4 g/mL), BEA (20 and 40 g/mL), OTA (20 and 30 g/mL), and CIT (50 and 68 g/mL) preserved from the GIT incubation procedure (Section 2.2) were lyophilized using a freeze dryer (Harvest Right, Canada).…”

Section: Cell Culturementioning

confidence: 99%

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Comparative Assessment of Different Yeast Cell Wall-Based Mycotoxin Adsorbents Using a Model- and Bioassay-Based In Vitro Approach

Yiannikouris

Shandilya

et al. 2023

Toxins

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Frequently reported occurrences of deoxynivalenol (DON), beauvericin (BEA), and, to a lesser extent, ochratoxin A (OTA) and citrinin (CIT) in ruminant feed or feedstuff could represent a significant concern regarding feed safety, animal health, and productivity. Inclusion of yeast cell wall-based mycotoxin adsorbents in animal feeds has been a common strategy to mitigate adverse effects of mycotoxins. In the present study, an in vitro approach combining adsorption isotherm models and bioassays was designed to assess the efficacy of yeast cell wall (YCW), yeast cell wall extract (YCWE), and a postbiotic yeast cell wall-based blend (PYCW) products at the inclusion rate of 0.5% (w/v) (ratio of adsorbent mass to buffer solution volume). The Hill’s adsorption isotherm model was found to best describe the adsorption processes of DON, BEA, and CIT. Calculated binding potential for YCW and YCWE using the Hill’s model exhibited the same ranking for mycotoxin adsorption, indicating that BEA had the highest adsorption rate, followed by DON and CIT, which was the least adsorbed. PYCW had the highest binding potential for BEA compared with YCW and YCWE. In contrast, the Freundlich isotherm model presented a good fit for OTA adsorption by all adsorbents and CIT adsorption by PYCW. Results indicated that YCW was the most efficacious for sequestering OTA, whereas YCWE was the least efficacious. PYCW showed greater efficacy at adsorbing OTA than CIT. All adsorbents exhibited high adsorption efficacy for BEA, with an overall percentage average of bound mycotoxin exceeding 60%, whereas moderate efficacies for the other mycotoxins were observed (up to 37%). Differences in adsorbent efficacy of each adsorbent significantly varied according to experimental concentrations tested for each given mycotoxin (p < 0.05). The cell viability results from the bioassay using a bovine mammary epithelial cell line (MAC-T) indicated that all tested adsorbents could potentially mitigate mycotoxin-related damage to bovine mammary epithelium. Results from our studies suggested that all tested adsorbents had the capacity to adsorb selected mycotoxins in vitro, which could support their use to mitigate their effects in vivo.

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Involvement of pro-inflammatory mediators and cell cycle disruption in neuronal cells induced by gliotoxin and ochratoxin A after individual and combined exposure

Penalva-Olcina,

Juan,

Fernández-Franzón

et al. 2024

Toxicology Letters

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Traditional and emerging Fusarium mycotoxins disrupt homeostasis of bovine mammary cells by altering cell permeability and innate immune function

Cited by 5 publications

References 121 publications

Evaluation of Immunomodulatory Effects of Fusarium Mycotoxins Using Bacterial Endotoxin-Stimulated Bovine Epithelial Cells and Macrophages in Co-Culture

Evaluation of Immunomodulatory Effects of Fusarium Mycotoxins Using Bacterial Endotoxin-Stimulated Bovine Epithelial Cells and Macrophages in Co-Culture

Comparative Assessment of Different Yeast Cell Wall-Based Mycotoxin Adsorbents Using a Model- and Bioassay-Based In Vitro Approach

Involvement of pro-inflammatory mediators and cell cycle disruption in neuronal cells induced by gliotoxin and ochratoxin A after individual and combined exposure

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