Accumulating evidence clarifies that intestinal barrier function, for example, by the mucus layer, antimicrobial peptides, immune systems, and epithelial tight junctions, plays crucial roles in maintaining our health. We reported previously that yogurt fermented with Lactobacillus delbrueckii subsp. bulgaricus 2038 and Streptococcus thermophilus 1131 induced the gene expression of the regenerating family member 3 (REG3) family, which encodes antimicrobial peptides in the small intestine, although it was unclear how the yogurt activated the intestinal cells related to it. Here, we evaluated the cytokine production from the intestinal immune cells stimulated by these strains in vitro and in vivo to elucidate the mechanism for the induction of the REG3 family by the yogurt. The results showed that stimulation by both strains induced interleukin (IL)-23 production from bone marrow-derived dendritic cells (DCs) and IL-22 production from small intestinal lamina propria (LP) cells. In addition, oral administration of these strains to mice increased IL-23p19⁺ LPDCs and IL-22⁺ type 3 innate lymphoid cells and induced the expression of Reg3g in small intestinal tissue. Moreover, we showed that the activities for the induction of IL-23 by DCs were strain dependent on L. bulgaricus and S. thermophilus and that S. thermophilus 1131, which is the predominant species in the yogurt, exhibited relatively higher activity compared to the other strains of S. thermophilus. Our findings suggested that these yogurt starter strains, L. bulgaricus 2038 and S. thermophilus 1131, have the potential to maintain and improve intestinal barrier function by stimulating immune cells in the LP.

Lactic acid bacteria (LAB) are commonly used probiotics that improve human health in various aspects. We previously reported that yogurt starter strains, Lactobacillus delbrueckii subsp. bulgaricus 2038 and Streptococcus thermophilus 1131, potentially enhance the intestinal epithelial barrier function by inducing the expression of antimicrobial peptides in the small intestine. However, their effects on physical barrier functions remain unknown. In this study, we found that both strains ameliorated the decreased trans-epithelial resistance and the increased permeability of fluorescein isothiocyanate-dextran induced by tumor necrosis factor (TNF)-α and interferon (IFN)-γ in Caco-2 cells. We also demonstrated that LAB prevented a decrease in the expression and disassembly of tight junctions (TJs) induced by TNF-α and IFN-γ. To assess the repair activity of TJs, a calcium switch assay was performed. Both strains were found to promote the reassembly of TJs, and their activity was canceled by the inhibitor of AMP-activated protein kinase (AMPK). Moreover, these strains showed increased AMPK phosphorylation. These observations suggest that the strains ameliorated physical barrier dysfunction via the activation of AMPK. The activities preventing barrier destruction induced by TNF-α and IFN-γ were strain-dependent. Several strains containing L. bulgaricus 2038 and S. thermophilus 1131 significantly suppressed the barrier impairment, and L. bulgaricus 2038 showed the strongest activity among them. Our findings suggest that the intake of L. bulgaricus 2038 and S. thermophilus 1131 is a potential strategy for the prevention and repair of leaky gut.

We investigated the effects of Lactobacillus paragasseri OLL2716 on gastrointestinal symptoms in healthy adults with gastric complaints. In this randomized, double-blind, placebo-controlled trial, 174 healthy Japanese adults were randomly assigned to an OLL2716 or placebo group, and each group consumed 85 g of yogurt containing L. paragasseri OLL2716 or placebo yogurt daily for 12 weeks. The primary endpoint was the change in gastric symptoms from baseline as per the participants’ questionnaires at 6 and 12 weeks. The secondary endpoints were changes from baseline in the short-form Nepean Dyspepsia Index (SF-NDI), the Gastrointestinal Symptom Rating Scale (GSRS), and the Council on Nutrition Appetite Questionnaire-Japanese (CNAQ-J) scores at 6 and 12 weeks. The primary endpoint data showed that the changes in “epigastric pain” at 6 and 12 weeks were significantly decreased in the OLL2716 group compared with those in the placebo group. Additionally, the changes in “epigastric pain syndrome-like symptoms” were significantly decreased in the OLL2716 group compared with those in the placebo group at 6 weeks. The SF-NDI items that improved at 6 weeks were “irritable, tense, or frustrated”, “enjoyment of eating or drinking”, and “tension”, which are sub-scales related to mental stress. The items “Over-all” in the GSRS and “feeling hungry” in the CNAQ-J significantly improved in the OLL2716 group compared with the placebo group at 12 weeks. The results suggest that regular intake of L. paragasseri OLL2716 may improve both gastric discomfort and mental stress in healthy adults with gastric complaints, such as postprandial fullness or early satiety.

Human induced pluripotent stem cell-derived small intestinal epithelial cell (hiPSC-SIEC) monolayers are useful in vitro models for evaluating the gut mucosal barrier; however, their reactivity to cytokines, which are closely related to the regulation of mucosal barrier function, remains unclear. Interleukin (IL)-22 is a cytokine that contributes to regulate the mucosal barrier in the intestinal epithelia. Using microarray and Gene Set Enrichment Analysis, we found that hiPSC-SIEC monolayers activate the immune response and enhance the mucosal barrier in response to IL-22. Moreover, hiPSC-SIEC monolayers induced the gene expression of antimicrobials including the regenerating islet-derived protein (REG) 3 family. Furthermore, IL-22 stimulation upregulated Mucin 2 secretion and gene expression of an enzyme that modifies sugar chains, suggesting alteration of the state of the mucus layer of hiPSC-SIEC monolayers. To evaluate its physiological significance, we measured the protective activity agaist Salmonella enterica subsp. enterica infection in hiPSC-SIEC monolayers and found that pre-stimulation with IL-22 reduced the number of viable intracellular bacteria. Collectively, these results suggest that hiPSC-SIEC monolayers enhance the mucosal barrier and inhibit infection by pathogenic bacteria in response to IL-22, as previously reported. These results can contribute to the further application of hiPSC-SIECs in evaluating mucosal barriers.

The plastic is broken into nanoscale particles (nanoplastics) that are harmful to the human body. Nanoplastics orally ingested are internalized into various cells, causing adverse effects such as oxidative stress and apoptosis; however, methods for preventing nanoplastic internalization are lacking. By elucidating this method, it is possible to make a significant contribution to human health. Then, following previous reports that Lactobacillus delbrueckii subsp. bulgaricus 2038 and Streptococcus thermophilus 1131 ameliorated barrier dysfunction in the small intestine, we examined their effects on the internalization of polystyrene nanoplastics (PSNPs) in a differentiated Caco-2 monolayer by flow cytometry and immunofluorescent staining. Both yogurt starter strains significantly suppressed fluorescently labeled PSNP internalization by Caco-2 cells, even when the strains were non-viable. Moreover, both strains significantly suppressed the transcellular permeability of PSNPs, suggesting reduced PSNP accumulation in blood vessels and various tissues. Finally, the suppression of PSNP internalization was strain dependent, and both strains were the most potent strains among multiple strains from the same species. This study highlights the potential of L. bulgaricus 2038 and S. thermophilus 1131 for ameliorating the adverse health effects of PSNP intake.

Aims: This study aimed to elucidate in vitro how orally administered Streptococcus thermophilus 1131 and Lactobacillus delbrueckii subsp. bulgaricus 2038, stimulated immune cells in the gut lamina propria and thereby enhanced the intestinal barrier function, as reported in our previous study. Methods and Results: We prepared an M cell model by coculturing small intestinal epithelial cells derived from human induced pluripotent stem cells with Raji cells. In this model, we demonstrated that both strains were incorporated into M cells and translocated to the basolateral side via transcytosis. Evaluation of the immune response using peripheral blood mononuclear cells showed that S. thermophilus 1131, translocated via M cells, induced the secretion of several cytokines, including IL-22 and IL-23. Conclusion: We showed that both strains were transcytosed by M cells in a human gut model. The results suggested that oral administration of both strains induced immune responses by translocating from the gut lumen to the lamina propria via M cells.