EpCAM-dependent extracellular vesicles from intestinal epithelial cells maintain intestinal tract immune balance

Abstract How the intestinal tract develops a tolerance to foreign antigens is still largely unknown. Here we report that extracellular vesicles (EVs) with TGF-β1-dependent immunosuppressive activity are produced by intestinal epithelial cells (IECs) under physiological conditions. Transfer of these EVs into inflammatory bowel disease (IBD) mice induced by dextran sulfate sodium salt decreases IBD severity by inducing regulatory T cells and immunosuppressive dendritic cells. In contrast, decreased endogenous EV production promotes IBD development. IECs produce EVs with increased levels of TGF-β1 upon IBD development in an ERK-dependent manner. Furthermore, these EVs tend to localize in the intestinal tract associated with epithelial cell adhesion molecule (EpCAM). Knockdown of EpCAM in vivo increases the severity of murine IBD, and the protective effect of EVs from IECs with decreased EpCAM on murine IBD is blunted. Therefore, our study indicates that EVs from IECs participate in maintaining the intestinal tract immune balance.

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Erratum: Corrigendum: EpCAM-dependent extracellular vesicles from intestinal epithelial cells maintain intestinal tract immune balance

Nature Communications ◽

10.1038/ncomms16006 ◽

2017 ◽

Vol 8 (1) ◽

Cited By ~ 2

Author(s):

Lingling Jiang ◽

Yingying Shen ◽

Danfeng Guo ◽

Diya Yang ◽

Jiajun Liu ◽

...

Keyword(s):

Epithelial Cells ◽

Extracellular Vesicles ◽

Intestinal Tract ◽

Intestinal Epithelial Cells ◽

Intestinal Epithelial ◽

Immune Balance

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Author Correction: EpCAM-dependent extracellular vesicles from intestinal epithelial cells maintain intestinal tract immune balance

Nature Communications ◽

10.1038/s41467-020-17430-y ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Lingling Jiang ◽

Yingying Shen ◽

Danfeng Guo ◽

Diya Yang ◽

Jiajun Liu ◽

...

Keyword(s):

Epithelial Cells ◽

Extracellular Vesicles ◽

Intestinal Tract ◽

Intestinal Epithelial Cells ◽

Intestinal Epithelial ◽

Immune Balance

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Comparison of gene expression and activation of transcription factors in organoid-derived monolayer intestinal epithelial cells and organoids

Bioscience Biotechnology and Biochemistry ◽

10.1093/bbb/zbab136 ◽

2021 ◽

Author(s):

Yu Takahashi ◽

Yu Inoue ◽

Keitaro Kuze ◽

Shintaro Sato ◽

Makoto Shimizu ◽

...

Keyword(s):

Gene Expression ◽

Epithelial Cells ◽

Gene Expression Regulation ◽

Intestinal Epithelial Cells ◽

Three Dimensional ◽

Culture Conditions ◽

Dependent Manner ◽

Intestinal Epithelial

Abstract Intestinal organoids better represent in vivo intestinal properties than conventionally used established cell lines in vitro. However, they are maintained in three-dimensional culture conditions that may be accompanied by handling complexities. We characterized the properties of human organoid-derived two-dimensionally cultured intestinal epithelial cells (IECs) compared with those of their parental organoids. We found that the expression of several intestinal markers and functional genes were indistinguishable between monolayer IECs and organoids. We further confirmed that their specific ligands equally activate intestinal ligand-activated transcriptional regulators in a dose-dependent manner. The results suggest that culture conditions do not significantly influence the fundamental properties of monolayer IECs originating from organoids, at least from the perspective of gene expression regulation. This will enable their use as novel biological tools to investigate the physiological functions of the human intestine.

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Endocytosis of commensal antigens by intestinal epithelial cells regulates mucosal T cell homeostasis

Science ◽

10.1126/science.aat4042 ◽

2019 ◽

Vol 363 (6431) ◽

pp. eaat4042 ◽

Cited By ~ 43

Author(s):

Mark S. Ladinsky ◽

Leandro P. Araujo ◽

Xiao Zhang ◽

John Veltri ◽

Marta Galan-Diez ◽

...

Keyword(s):

T Cell ◽

Epithelial Cells ◽

Intestinal Epithelial Cells ◽

Commensal Bacteria ◽

Dependent Manner ◽

Intestinal Epithelial ◽

T Helper 17 ◽

Associated Proteins ◽

Gut Microbe

Commensal bacteria influence host physiology, without invading host tissues. We show that proteins from segmented filamentous bacteria (SFB) are transferred into intestinal epithelial cells (IECs) through adhesion-directed endocytosis that is distinct from the clathrin-dependent endocytosis of invasive pathogens. This process transfers microbial cell wall–associated proteins, including an antigen that stimulates mucosal T helper 17 (TH17) cell differentiation, into the cytosol of IECs in a cell division control protein 42 homolog (CDC42)–dependent manner. Removal of CDC42 activity in vivo led to disruption of endocytosis induced by SFB and decreased epithelial antigen acquisition, with consequent loss of mucosal TH17 cells. Our findings demonstrate direct communication between a resident gut microbe and the host and show that under physiological conditions, IECs acquire antigens from commensal bacteria for generation of T cell responses to the resident microbiota.

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Extracellular Vesicles: A New Nano Tool for the Treatment of Inflammatory Bowel Diseases

Current Nanoscience ◽

10.2174/1573413715666190411141126 ◽

2019 ◽

Vol 15 (6) ◽

pp. 589-595

Author(s):

Nitin Tandra ◽

Peipei Wu ◽

Xinyuan Hu ◽

Fei Mao ◽

Wenrong Xu ◽

...

Keyword(s):

Extracellular Vesicles ◽

Intestinal Tract ◽

Intestinal Epithelial Cells ◽

Therapeutic Strategies ◽

Intestinal Stem Cells ◽

Inflammatory Disorder ◽

Intestinal Epithelial ◽

Bowel Diseases ◽

Inflammatory Bowel

The intestinal tract is a complex and important physiological and immunological organ. Intestinal tract homeostasis requires a series of coordinated interactions involving gut microbiota, the crypt intestinal stem cells (ISC) and the surrounding niche, including the intestinal epithelial cells, endothelial cells, dendritic cells, and macrophages. The destruction of intestinal homeostasis leads to autoimmune diseases, such as inflammatory bowel disease (IBD). IBD is a non-specific, and remittent- relapsing inflammatory disorder of the gastrointestinal tract. There is no effective method to keep patients in remission for a long term. It has been reported that extracellular vesicles (EVs) exert immune activation and immunosuppressive effects in the pathogenesis of IBD. In order to explore new therapeutic strategies for IBD, in this review, we summarize the observations on the immune properties and functions of EVs in intestinal mucosal immunity.

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Endocytic pathways of luminal antigens intersect MHC class I and class II pathways in multivesicular late endosomes – Antigen trafficking in intestinal epithelial cells studied in vivo in Crohn's disease patients

Zeitschrift für Gastroenterologie ◽

10.1055/s-2006-950626 ◽

2006 ◽

Vol 44 (08) ◽

Author(s):

G Hundorfean ◽

S Strobel ◽

KP Zimmer ◽

A Gebert ◽

D Ludwig ◽

...

Keyword(s):

Epithelial Cells ◽

Mhc Class I ◽

Intestinal Epithelial Cells ◽

Class Ii ◽

Class I ◽

Intestinal Epithelial ◽

Late Endosomes ◽

Antigen Trafficking ◽

Endocytic Pathways

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Positive cross talk between protein kinase D and β-catenin in intestinal epithelial cells: impact on β-catenin nuclear localization and phosphorylation at Ser552

AJP Cell Physiology ◽

10.1152/ajpcell.00302.2015 ◽

2016 ◽

Vol 310 (7) ◽

pp. C542-C557 ◽

Cited By ~ 9

Author(s):

Jia Wang ◽

Liang Han ◽

James Sinnett-Smith ◽

Li-Li Han ◽

Jan V. Stevens ◽

...

Keyword(s):

Protein Kinase ◽

Epithelial Cells ◽

Transgenic Mice ◽

Nuclear Localization ◽

Cross Talk ◽

Intestinal Epithelial Cells ◽

Dependent Manner ◽

Intestinal Epithelial ◽

Ang Ii ◽

Potent Inducer

Given the fundamental role of β-catenin signaling in intestinal epithelial cell proliferation and the growth-promoting function of protein kinase D1 (PKD1) in these cells, we hypothesized that PKDs mediate cross talk with β-catenin signaling. The results presented here provide several lines of evidence supporting this hypothesis. We found that stimulation of intestinal epithelial IEC-18 cells with the G protein-coupled receptor (GPCR) agonist angiotensin II (ANG II), a potent inducer of PKD activation, promoted endogenous β-catenin nuclear localization in a time-dependent manner. A significant increase was evident within 1 h of ANG II stimulation ( P < 0.01), peaked at 4 h ( P < 0.001), and declined afterwards. GPCR stimulation also induced a marked increase in β-catenin-regulated genes and phosphorylation at Ser552 in intestinal epithelial cells. Exposure to preferential inhibitors of the PKD family (CRT006610 or kb NB 142-70) or knockdown of the isoforms of the PKD family prevented the increase in β-catenin nuclear localization and phosphorylation at Ser552 in response to ANG II. GPCR stimulation also induced the formation of a complex between PKD1 and β-catenin, as shown by coimmunoprecipitation that depended on PKD1 catalytic activation, as it was abrogated by cell treatment with PKD family inhibitors. Using transgenic mice that express elevated PKD1 protein in the intestinal epithelium, we detected a marked increase in the localization of β-catenin in the nucleus of crypt epithelial cells in the ileum of PKD1 transgenic mice, compared with nontransgenic littermates. Collectively, our results identify a novel cross talk between PKD and β-catenin in intestinal epithelial cells, both in vitro and in vivo.

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Expression of lysophosphatidic acid receptor 5 is necessary for the regulation of intestinal Na+/H+ exchanger 3 by lysophosphatidic acid in vivo

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00130.2018 ◽

2018 ◽

Vol 315 (4) ◽

pp. G433-G442 ◽

Cited By ~ 3

Author(s):

Kayte A. Jenkin ◽

Peijian He ◽

C. Chris Yun

Keyword(s):

Epithelial Cells ◽

Lysophosphatidic Acid ◽

Direct Evidence ◽

Intestinal Epithelial Cells ◽

Regulatory Role ◽

Intestinal Epithelial ◽

Fluid Absorption ◽

Wild Type

Lysophosphatidic acid (LPA) is a bioactive lipid molecule, which regulates a broad range of pathophysiological processes. Recent studies have demonstrated that LPA modulates electrolyte flux in the intestine, and its potential as an antidiarrheal agent has been suggested. Of six LPA receptors, LPA5 is highly expressed in the intestine. Recent studies by our group have demonstrated activation of Na+/H+ exchanger 3 (NHE3) by LPA5. However, much of what has been elucidated was achieved using colonic cell lines that were transfected to express LPA5. In the current study, we engineered a mouse that lacks LPA5 in intestinal epithelial cells, Lpar5ΔIEC, and investigated the role of LPA5 in NHE3 regulation and fluid absorption in vivo. The intestine of Lpar5ΔIEC mice appeared morphologically normal, and the stool frequency and fecal water content were unchanged compared with wild-type mice. Basal rates of NHE3 activity and fluid absorption and total NHE3 expression were not changed in Lpar5ΔIEC mice. However, LPA did not activate NHE3 activity or fluid absorption in Lpar5ΔIEC mice, providing direct evidence for the regulatory role of LPA5. NHE3 activation involves trafficking of NHE3 from the terminal web to microvilli, and this mobilization of NHE3 by LPA was abolished in Lpar5ΔIEC mice. Dysregulation of NHE3 was specific to LPA, and insulin and cholera toxin were able to stimulate and inhibit NHE3, respectively, in both wild-type and Lpar5ΔIEC mice. The current study for the first time demonstrates the necessity of LPA5 in LPA-mediated stimulation of NHE3 in vivo. NEW & NOTEWORTHY This study is the first to assess the role of LPA5 in NHE3 regulation and fluid absorption in vivo using a mouse that lacks LPA5 in intestinal epithelial cells, Lpar5ΔIEC. Basal rates of NHE3 activity and fluid absorption, and total NHE3 expression were not changed in Lpar5ΔIEC mice. However, LPA did not activate NHE3 activity or fluid absorption in Lpar5ΔIEC mice, providing direct evidence for the regulatory role of LPA5.

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