Fibroblasts Impact Goblet Cell Responses to Lactic Acid Bacteria After Exposure to Inflammatory Cytokines and Mucus Disruptors

Abstract

Scope Mucus produced by goblet cells contributes to gut barrier function. Lactic acid bacteria (LAB) have been shown to impact mucus production. It is not completely known whether mucus production is influenced by the abundantly present fibroblasts in the intestine. Methods and results The influence of fibroblasts on mucus‐related genes including mucin‐2 (MUC2), trefoil factor 3 (TFF3), resistin‐like molecule β (RETNLB), carbohydrate sulfotransferase 5 (CHST5), and galactose‐3‐O‐sulfotransferase 2 (GAL3ST2) is examined after co‐culture of LS174T‐goblet cells and CCD‐18Co colonic fibroblasts in the presence and absence of LAB‐strains known to impact mucus function. This is also tested after exposure to TNF‐α, IL‐13, or the mucin synthesis inhibitor tunicamycin (Tm). Effects of fibroblasts are treatment duration‐ and bacterial species‐dependent under homeostatic conditions. During TNF‐α challenge, fibroblasts reverse Lactobacillus (L.) rhamnosus CCFM237‐elicited declined TFF3 expression. After IL‐13 exposure, L. rhamnosus CCFM237 and L. fermentum CCFM787 attenuate enhanced TFF3 and RETNLB expression, respectively, only in the presence of fibroblasts. LAB has no effects on Tm‐induced decreased expression of goblet cell‐related genes regardless of the presence of fibroblasts. Conclusion It is demonstrated that goblet cell–fibroblast crosstalk impacts mucus synthesis and influences the effects of LAB on goblet cell‐related genes. Effects are LAB‐species and stressor dependent.

Human Milk Oligosaccharides Differently Modulate Goblet Cells Under Homeostatic, Proinflammatory Conditions and ER Stress

“…The reason for choosing this cell type and not other commonly used epithelial cell lines such as Caco2 is that LS174T cells are highly secretory and exhibit more of a clear intestinal mucus secreting goblet‐cell phenotype. For that reason it is widely used in goblet cell function studies . Non‐transformed mucin‐producing intestine epithelial cells could be a better choice but are to the best of our knowledge not available yet.…”

“…For that reason it is widely used in goblet cell function studies. [38,40,43] Nontransformed mucin-producing intestine epithelial cells could be a better choice but are to the best of our knowledge not available yet. LS174T is also favored because it expresses all the tested genes under homeostatic condition, proinflammatory condition, and ER-stress, [38] which allows studying of the effects of individual hMOs under pro-inflammatory or other diseased conditions.…”

Distribution of intraepithelial lymphocytes, mast cells, and goblet cells in the intestine of alpaca

“…It is well known that goblet cells in the intestine mainly secrete mucins, which, as an important part of the intestinal mucus layer and intestinal epithelial cells, can not only lubricate and protect the intestinal mucosal epithelium Ren et al, 2019), but are also involved in intercellular signalling transduction and the regulation of intestinal mucosa‐associated immune factors (Castagnoli et al, 2021), which can be divided into the secreted mucins and the membrane‐bound mucins according to their existing forms (Etzold & Juge, 2014). It has been shown that bacterial products and their inflammatory factors can induce the formation of sulphonyltransferase, thus making MUC2 sulphate (Ren et al, 2019).…”

“…It is well known that goblet cells in the intestine mainly secrete mucins, which, as an important part of the intestinal mucus layer and intestinal epithelial cells, can not only lubricate and protect the intestinal mucosal epithelium Ren et al, 2019), but are also involved in intercellular signalling transduction and the regulation of intestinal mucosa‐associated immune factors (Castagnoli et al, 2021), which can be divided into the secreted mucins and the membrane‐bound mucins according to their existing forms (Etzold & Juge, 2014). It has been shown that bacterial products and their inflammatory factors can induce the formation of sulphonyltransferase, thus making MUC2 sulphate (Ren et al, 2019). However, when animals are infected with specific pathogens, the goblet cells hyperplasia is often accompanied with alterations in mucin terminal sugar residues, which increases sialylated MUC2 (Grondin et al, 2020).…”

“…It has been shown that bacterial products and their inflammatory factors can induce the formation of sulphonyltransferase, thus making MUC2 sulphate (Ren et al, 2019). However, when animals are infected with specific pathogens, the goblet cells hyperplasia is often accompanied with alterations in mucin terminal sugar residues, which increases sialylated MUC2 (Grondin et al, 2020).…”