Toll-like Receptors and Cytokine Modulation by Goat Milk Extracellular Vesicles in a Model of Intestinal Inflammation
Extracellular vesicles (EVs) are nanometric spherical structures, enclosed in a lipid bilayer membrane and secreted by multiple cell types under specific physiologic and pathologic conditions. Their complex cargo modulates immune cells within an inflammatory microenvironment. Milk is one of the most...
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MDPI AG
2023-07-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/13/11096 |
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| author | Chiara Grazia De Ciucis Floriana Fruscione Livia De Paolis Samanta Mecocci Susanna Zinellu Lisa Guardone Giulia Franzoni Katia Cappelli Elisabetta Razzuoli |
| author_facet | Chiara Grazia De Ciucis Floriana Fruscione Livia De Paolis Samanta Mecocci Susanna Zinellu Lisa Guardone Giulia Franzoni Katia Cappelli Elisabetta Razzuoli |
| author_sort | Chiara Grazia De Ciucis |
| collection | DOAJ |
| description | Extracellular vesicles (EVs) are nanometric spherical structures, enclosed in a lipid bilayer membrane and secreted by multiple cell types under specific physiologic and pathologic conditions. Their complex cargo modulates immune cells within an inflammatory microenvironment. Milk is one of the most promising sources of EVs in terms of massive recovery, and milk extracellular vesicles (mEVs) have immunomodulatory and anti-inflammatory effects. The aim of this study was to characterize goat mEVs’ immunomodulating activities on Toll-like receptors (TLRs) and related immune genes, including cytokines, using a porcine intestinal epithelial cell line (IPEC-J2) after the establishment of a pro-inflammatory environment. IPEC-J2 was exposed for 2 h to pro-inflammatory stimuli as a model of inflammatory bowel disease (IBD), namely LPS for Crohn’s disease (CD) and H<sub>2</sub>O<sub>2</sub> for ulcerative colitis (UC); then, cells were treated with goat mEVs for 48 h. RT-qPCR and ELISA data showed that cell exposure to LPS or H<sub>2</sub>O<sub>2</sub> caused a pro-inflammatory response, with increased gene expression of <i>CXCL8</i>, <i>TNFA</i>, <i>NOS2</i> and the release of pro-inflammatory cytokines. In the LPS model, the treatment with mEVs after LPS determined the down-regulation of <i>NOS2</i>, <i>MMP9</i>, <i>TLR5</i>, <i>TGFB1</i>, <i>IFNB</i>, <i>IL18</i> and <i>IL12A</i> gene expressions, as well as lower release of IL-18 in culture supernatants. At the same time, we observed the increased expression of <i>TLR1</i>, <i>TLR2</i>, <i>TLR8</i> and <i>EBI3</i>. On the contrary, the treatment with mEVs after H<sub>2</sub>O<sub>2</sub> exposure, the model of UC, determined the increased expression of <i>MMP9</i> alongside the decrease in <i>TGFB1</i>, <i>TLR8</i> and <i>DEFB1</i>, with a lower release of IL-1Ra in culture supernatants. Overall, our data showed that a 48 h treatment with mEVs after a pro-inflammatory stimulus significantly modulated the expression of several TLRs and cytokines in swine intestinal cells, in association with a decreased inflammation. These results further highlight the immunomodulatory potential of these nanosized structures and suggest their potential application in vivo. |
| first_indexed | 2024-03-11T01:37:58Z |
| format | Article |
| id | doaj.art-3f2667af8260446ea60dc77fcef72038 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T01:37:58Z |
| publishDate | 2023-07-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-3f2667af8260446ea60dc77fcef720382023-11-18T16:48:04ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-07-0124131109610.3390/ijms241311096Toll-like Receptors and Cytokine Modulation by Goat Milk Extracellular Vesicles in a Model of Intestinal InflammationChiara Grazia De Ciucis0Floriana Fruscione1Livia De Paolis2Samanta Mecocci3Susanna Zinellu4Lisa Guardone5Giulia Franzoni6Katia Cappelli7Elisabetta Razzuoli8National Reference Center of Veterinary and Comparative Oncology (CEROVEC), Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d’Aosta, Piazza Borgo Pila 39-24, 16129 Genova, ItalyNational Reference Center of Veterinary and Comparative Oncology (CEROVEC), Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d’Aosta, Piazza Borgo Pila 39-24, 16129 Genova, ItalyNational Reference Center of Veterinary and Comparative Oncology (CEROVEC), Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d’Aosta, Piazza Borgo Pila 39-24, 16129 Genova, ItalyDepartment of Veterinary Medicine, University of Perugia, 06123 Perugia, ItalyDepartment of Animal Health, Istituto Zooprofilattico Sperimentale della Sardegna, 07100 Sassari, ItalyNational Reference Center of Veterinary and Comparative Oncology (CEROVEC), Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d’Aosta, Piazza Borgo Pila 39-24, 16129 Genova, ItalyDepartment of Animal Health, Istituto Zooprofilattico Sperimentale della Sardegna, 07100 Sassari, ItalyDepartment of Veterinary Medicine, University of Perugia, 06123 Perugia, ItalyNational Reference Center of Veterinary and Comparative Oncology (CEROVEC), Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d’Aosta, Piazza Borgo Pila 39-24, 16129 Genova, ItalyExtracellular vesicles (EVs) are nanometric spherical structures, enclosed in a lipid bilayer membrane and secreted by multiple cell types under specific physiologic and pathologic conditions. Their complex cargo modulates immune cells within an inflammatory microenvironment. Milk is one of the most promising sources of EVs in terms of massive recovery, and milk extracellular vesicles (mEVs) have immunomodulatory and anti-inflammatory effects. The aim of this study was to characterize goat mEVs’ immunomodulating activities on Toll-like receptors (TLRs) and related immune genes, including cytokines, using a porcine intestinal epithelial cell line (IPEC-J2) after the establishment of a pro-inflammatory environment. IPEC-J2 was exposed for 2 h to pro-inflammatory stimuli as a model of inflammatory bowel disease (IBD), namely LPS for Crohn’s disease (CD) and H<sub>2</sub>O<sub>2</sub> for ulcerative colitis (UC); then, cells were treated with goat mEVs for 48 h. RT-qPCR and ELISA data showed that cell exposure to LPS or H<sub>2</sub>O<sub>2</sub> caused a pro-inflammatory response, with increased gene expression of <i>CXCL8</i>, <i>TNFA</i>, <i>NOS2</i> and the release of pro-inflammatory cytokines. In the LPS model, the treatment with mEVs after LPS determined the down-regulation of <i>NOS2</i>, <i>MMP9</i>, <i>TLR5</i>, <i>TGFB1</i>, <i>IFNB</i>, <i>IL18</i> and <i>IL12A</i> gene expressions, as well as lower release of IL-18 in culture supernatants. At the same time, we observed the increased expression of <i>TLR1</i>, <i>TLR2</i>, <i>TLR8</i> and <i>EBI3</i>. On the contrary, the treatment with mEVs after H<sub>2</sub>O<sub>2</sub> exposure, the model of UC, determined the increased expression of <i>MMP9</i> alongside the decrease in <i>TGFB1</i>, <i>TLR8</i> and <i>DEFB1</i>, with a lower release of IL-1Ra in culture supernatants. Overall, our data showed that a 48 h treatment with mEVs after a pro-inflammatory stimulus significantly modulated the expression of several TLRs and cytokines in swine intestinal cells, in association with a decreased inflammation. These results further highlight the immunomodulatory potential of these nanosized structures and suggest their potential application in vivo.https://www.mdpi.com/1422-0067/24/13/11096goat EVsRT-qPCRIPEC-J2Toll-like receptormodel of intestinal inflammationanti-inflammatory |
| spellingShingle | Chiara Grazia De Ciucis Floriana Fruscione Livia De Paolis Samanta Mecocci Susanna Zinellu Lisa Guardone Giulia Franzoni Katia Cappelli Elisabetta Razzuoli Toll-like Receptors and Cytokine Modulation by Goat Milk Extracellular Vesicles in a Model of Intestinal Inflammation International Journal of Molecular Sciences goat EVs RT-qPCR IPEC-J2 Toll-like receptor model of intestinal inflammation anti-inflammatory |
| title | Toll-like Receptors and Cytokine Modulation by Goat Milk Extracellular Vesicles in a Model of Intestinal Inflammation |
| title_full | Toll-like Receptors and Cytokine Modulation by Goat Milk Extracellular Vesicles in a Model of Intestinal Inflammation |
| title_fullStr | Toll-like Receptors and Cytokine Modulation by Goat Milk Extracellular Vesicles in a Model of Intestinal Inflammation |
| title_full_unstemmed | Toll-like Receptors and Cytokine Modulation by Goat Milk Extracellular Vesicles in a Model of Intestinal Inflammation |
| title_short | Toll-like Receptors and Cytokine Modulation by Goat Milk Extracellular Vesicles in a Model of Intestinal Inflammation |
| title_sort | toll like receptors and cytokine modulation by goat milk extracellular vesicles in a model of intestinal inflammation |
| topic | goat EVs RT-qPCR IPEC-J2 Toll-like receptor model of intestinal inflammation anti-inflammatory |
| url | https://www.mdpi.com/1422-0067/24/13/11096 |
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