Enhancing Antiviral Immunity in the Gastrointestinal Epithelium: The Role of Fibroblast–Endothelium Interaction and Melatonin

The gastrointestinal (GI) tract is a major barrier against pathogens, including viruses. The antiviral responses in the GI epithelium have been broadly investigated, but data on the contribution of the stromal cells remain scarce. Melatonin, widely used to treat insomnia, has recently been proposed...

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Main Authors: Milda Šeškutė, Goda Laucaitytė, Rūta Inčiūraitė, Mantas Malinauskas, Lina Jankauskaitė
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Cells
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Online Access:https://www.mdpi.com/2073-4409/14/13/990
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author Milda Šeškutė
Goda Laucaitytė
Rūta Inčiūraitė
Mantas Malinauskas
Lina Jankauskaitė
author_facet Milda Šeškutė
Goda Laucaitytė
Rūta Inčiūraitė
Mantas Malinauskas
Lina Jankauskaitė
author_sort Milda Šeškutė
collection DOAJ
description The gastrointestinal (GI) tract is a major barrier against pathogens, including viruses. The antiviral responses in the GI epithelium have been broadly investigated, but data on the contribution of the stromal cells remain scarce. Melatonin, widely used to treat insomnia, has recently been proposed as an antiviral agent, yet its effect in the GI tract remains poorly understood. We compared the antiviral responses in Caco-2 monocultures and co-cultures with intestinal fibroblasts (HSIFs) and endothelial cells (HUVECs) after stimulation using Poly I:C. We evaluated the apoptosis, proliferation, key antiviral markers (IRF1, IRF3, IFNs, TBK1, STAT3), and mitochondrial and peroxisomal activation with and without melatonin. The Caco-2 cells cultured with the HSIFs and HUVECs demonstrated enhanced proliferation and reduced Poly I:C-induced apoptosis. The co-culture exhibited a more rapid IRF3-IFNλ1 response, higher TBK1 expression, and enhanced peroxisomal activation compared to these properties in the monoculture. Melatonin further reduced apoptosis and modulated organelle-specific antiviral signaling by suppressing peroxisomal activation and promoting mitochondrial activity. Reduced peroxisomal activation was associated with decreased TBK1, IRF3, and IFNλ1 levels and altered STAT3 signaling. These effects were more pronounced when melatonin was applied post-stimulation compared to that under prophylactic use. Fibroblast–endothelial interactions amplify the antiviral responses in the intestinal epithelial cells by activating the TBK1–IRF3–IFNλ1 axis. Melatonin modulates these responses, highlighting its therapeutic potential in viral GI infections.
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spelling doaj-art-987a039a3d2a4202aa9c517a1355ccb02025-08-20T03:50:17ZengMDPI AGCells2073-44092025-06-01141399010.3390/cells14130990Enhancing Antiviral Immunity in the Gastrointestinal Epithelium: The Role of Fibroblast–Endothelium Interaction and MelatoninMilda Šeškutė0Goda Laucaitytė1Rūta Inčiūraitė2Mantas Malinauskas3Lina Jankauskaitė4Department of Pediatrics, Lithuanian University of Health Sciences, 44307 Kaunas, LithuaniaDepartment of Pediatrics, Lithuanian University of Health Sciences, 44307 Kaunas, LithuaniaInstitute for Digestive Research, Lithuanian University of Health Sciences, 44307 Kaunas, LithuaniaInstitute of Physiology and Pharmacology, Lithuanian University of Health Sciences, 44307 Kaunas, LithuaniaDepartment of Pediatrics, Lithuanian University of Health Sciences, 44307 Kaunas, LithuaniaThe gastrointestinal (GI) tract is a major barrier against pathogens, including viruses. The antiviral responses in the GI epithelium have been broadly investigated, but data on the contribution of the stromal cells remain scarce. Melatonin, widely used to treat insomnia, has recently been proposed as an antiviral agent, yet its effect in the GI tract remains poorly understood. We compared the antiviral responses in Caco-2 monocultures and co-cultures with intestinal fibroblasts (HSIFs) and endothelial cells (HUVECs) after stimulation using Poly I:C. We evaluated the apoptosis, proliferation, key antiviral markers (IRF1, IRF3, IFNs, TBK1, STAT3), and mitochondrial and peroxisomal activation with and without melatonin. The Caco-2 cells cultured with the HSIFs and HUVECs demonstrated enhanced proliferation and reduced Poly I:C-induced apoptosis. The co-culture exhibited a more rapid IRF3-IFNλ1 response, higher TBK1 expression, and enhanced peroxisomal activation compared to these properties in the monoculture. Melatonin further reduced apoptosis and modulated organelle-specific antiviral signaling by suppressing peroxisomal activation and promoting mitochondrial activity. Reduced peroxisomal activation was associated with decreased TBK1, IRF3, and IFNλ1 levels and altered STAT3 signaling. These effects were more pronounced when melatonin was applied post-stimulation compared to that under prophylactic use. Fibroblast–endothelial interactions amplify the antiviral responses in the intestinal epithelial cells by activating the TBK1–IRF3–IFNλ1 axis. Melatonin modulates these responses, highlighting its therapeutic potential in viral GI infections.https://www.mdpi.com/2073-4409/14/13/990gastrointestinal co-culture modelantiviralmelatoninperoxisomesIFNλ1IRF3
spellingShingle Milda Šeškutė
Goda Laucaitytė
Rūta Inčiūraitė
Mantas Malinauskas
Lina Jankauskaitė
Enhancing Antiviral Immunity in the Gastrointestinal Epithelium: The Role of Fibroblast–Endothelium Interaction and Melatonin
Cells
gastrointestinal co-culture model
antiviral
melatonin
peroxisomes
IFNλ1
IRF3
title Enhancing Antiviral Immunity in the Gastrointestinal Epithelium: The Role of Fibroblast–Endothelium Interaction and Melatonin
title_full Enhancing Antiviral Immunity in the Gastrointestinal Epithelium: The Role of Fibroblast–Endothelium Interaction and Melatonin
title_fullStr Enhancing Antiviral Immunity in the Gastrointestinal Epithelium: The Role of Fibroblast–Endothelium Interaction and Melatonin
title_full_unstemmed Enhancing Antiviral Immunity in the Gastrointestinal Epithelium: The Role of Fibroblast–Endothelium Interaction and Melatonin
title_short Enhancing Antiviral Immunity in the Gastrointestinal Epithelium: The Role of Fibroblast–Endothelium Interaction and Melatonin
title_sort enhancing antiviral immunity in the gastrointestinal epithelium the role of fibroblast endothelium interaction and melatonin
topic gastrointestinal co-culture model
antiviral
melatonin
peroxisomes
IFNλ1
IRF3
url https://www.mdpi.com/2073-4409/14/13/990
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