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Commensal yeast derived b-1,6-glucan induces regulatory T cells

ABSTRACT: To characterize the effect of MGCP in colonic dendritic cell phenotypic changes, we performed gene expression RNAseq analysis for colonic CD11c+ dendritic cells from mock or MGCP administered germ-free mice

ORGANISM(S): Mus musculus

PROVIDER: GSE126937 | GEO | 2021/04/21

REPOSITORIES: GEO

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Commensal yeast derived b-1,6-glucan induces regulatory T cells

Project description:Commensal yeast derived b-1,6-glucan induces regulatory T cells

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Commensal yeast cell surface polysaccharide activities unleashed upon removal of b-1,3-glucans restrain inflammatory disorders

Project description:To characterize the effect of MGCP in dendritic cell phenotypic changes, we performed gene expression RNAseq analysis from CD8a-CD11b+ dendritic cells that stimulated with mock or MGCP in vitro.

2021-04-21 | GSE150685 | GEO

Yeast β-1,6-glucan is a primary target for the Saccharomyces cerevisiae K2 toxin.

Project description:Certain Saccharomyces cerevisiae strains secrete different killer proteins of double-stranded-RNA origin. These proteins confer a growth advantage to their host by increasing its survival. K2 toxin affects the target cell by binding to the cell surface, disrupting the plasma membrane integrity, and inducing ion leakage. In this study, we determined that K2 toxin saturates the yeast cell surface receptors in 10 min. The apparent amount of K2 toxin, bound to a single cell of wild type yeast under saturating conditions, was estimated to be 435 to 460 molecules. It was found that an increased level of β-1,6-glucan directly correlates with the number of toxin molecules bound, thereby impacting the morphology and determining the fate of the yeast cell. We observed that the binding of K2 toxin to the yeast surface receptors proceeds in a similar manner as in case of the related K1 killer protein. It was demonstrated that the externally supplied pustulan, a poly-β-1,6-glucan, but not the glucans bearing other linkage types (such as laminarin, chitin, and pullulan) efficiently inhibits the K2 toxin killing activity. In addition, the analysis of toxin binding to the intact cells and spheroplasts confirmed that majority of K2 protein molecules attach to the β-1,6-glucan, rather than the plasma membrane-localized receptors. Taken together, our results reveal that β-1,6-glucan is a primary target of K2 toxin and is important for the execution of its killing property.

| S-EPMC4385797 | biostudies-literature

Structural specificities of cell surface β-glucan polysaccharides determine commensal yeast mediated immuno-modulatory activities.

Project description:Yeast is an integral part of mammalian microbiome, and like commensal bacteria, has the potential of being harnessed to influence immunity in clinical settings. However, functional specificities of yeast-derived immunoregulatory molecules remain elusive. Here we find that while under steady state, β-1,3-glucan-containing polysaccharides potentiate pro-inflammatory properties, a relatively less abundant class of cell surface polysaccharides, dubbed mannan/β-1,6-glucan-containing polysaccharides (MGCP), is capable of exerting potent anti-inflammatory effects to the immune system. MGCP, in contrast to previously identified microbial cell surface polysaccharides, through a Dectin1-Cox2 signaling axis in dendritic cells, facilitates regulatory T (Treg) cell induction from naïve T cells. Furthermore, through a TLR2-dependent mechanism, it restrains Th1 differentiation of effector T cells by suppressing IFN-γ expression. As a result, administration of MGCP display robust suppressive capacity towards experimental inflammatory disease models of colitis and experimental autoimmune encephalomyelitis (EAE) in mice, thereby highlighting its potential therapeutic utility against clinically relevant autoimmune diseases.

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Kre6 protein essential for yeast cell wall beta-1,6-glucan synthesis accumulates at sites of polarized growth.

Project description:Saccharomyces cerevisiae Kre6 is a type II membrane protein with amino acid sequence homology with glycoside hydrolase and is essential for β-1,6-glucan synthesis as revealed by the mutant phenotype, but its biochemical function is still unknown. The localization of Kre6, determined by epitope tagging, is a matter of debate. We raised anti-Kre6 rabbit antiserum and examined the localization of Kre6 and its tagged protein by immunofluorescence microscopy, subcellular fractionation in sucrose density gradients, and immunoelectron microscopy. Integration of the results indicates that the majority of Kre6 is in the endoplasmic reticulum; however, a small but significant portion is also present in the secretory vesicle-like compartments and plasma membrane. Kre6 in the latter compartments is observed as strong signals that accumulate at the sites of polarized growth by immunofluorescence. The truncated Kre6 without the N-terminal 230-amino acid cytoplasmic region did not show this polarized accumulation and had a severe defect in β-1,6-glucan synthesis. This is the first evidence of a β-1,6-glucan-related protein showing the polarized membrane localization that correlates with its biological function.

| S-EPMC3044999 | biostudies-literature

RNA-seq data of intestinal epithelial cells and lamina propria dendritic cells

Project description:We report that Klebsiella pneumoniae promote Th1 cell induction in colon. To examine the influence of Klebsiella on colonic epithelial cells (ECs) and lamina propria CD11c+ dendritic cells (DCs), we performed RNA seq on them. Germ free mice were orally inoculated with Kp-2H7 or BAA-2552 and total RNA was isolated from colonic ECs and DCs 1 week after inoculation. Furthermore, we examined the involvement of TLRs in induction of Th1 cells using Myd88 KO, Trif KO, Myd88/Trif DKO mice. These deficient germ free mice were orally inoculated with Kp-2H7 and total RNA was isolated from colonic ECs 3 weeks after inoculation.

2017-10-20 | GSE101678 | GEO

Kre6 (yeast 1,6-β-transglycosylase) homolog, PhTGS, is essential for β-glucan synthesis in the haptophyte <i>Pleurochrysis haptonemofera</i>.

Project description:Haptophytes synthesize unique β-glucans containing more β-1,6-linkages than β-1,3 linkages, as a storage polysaccharide. To understand the mechanism of the synthesis, we investigated the roles of Kre6 (yeast 1,6-β-transglycosylase) homologs, PhTGS, in the haptophyte Pleurochrysis haptonemofera. RNAi of PhTGS repressed β-glucan accumulation and simultaneously induced lipid production, suggesting that PhTGS is involved in β-glucan synthesis and that the knockdown leads to the alteration of the carbon metabolic flow. PhTGS was expressed more in light, where β-glucan was actively produced by photosynthesis, than in the dark. The crude extract of E. coli expressing PhKre6 demonstrated its activity to incorporate 14C-UDP-glucose into β-glucan of P. haptonemofera. These findings suggest that PhTGS functions in storage β-glucan synthesis specifically in light, probably by producing the β-1,6-branch.

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Development of a novel ?-1,6-glucan-specific detection system using functionally-modified recombinant endo-?-1,6-glucanase.

Project description:?-1,3-d-Glucan is a ubiquitous glucose polymer produced by plants, bacteria, and most fungi. It has been used as a diagnostic tool in patients with invasive mycoses via a highly-sensitive reagent consisting of the blood coagulation system of horseshoe crab. However, no method is currently available for measuring ?-1,6-glucan, another primary ?-glucan structure of fungal polysaccharides. Herein, we describe the development of an economical and highly-sensitive and specific assay for ?-1,6-glucan using a modified recombinant endo-?-1,6-glucanase having diminished glucan hydrolase activity. The purified ?-1,6-glucanase derivative bound to the ?-1,6-glucan pustulan with a KD of 16.4 nm We validated the specificity of this ?-1,6-glucan probe by demonstrating its ability to detect cell wall ?-1,6-glucan from both yeast and hyphal forms of the opportunistic fungal pathogen Candida albicans, without any detectable binding to glucan lacking the long ?-1,6-glucan branch. We developed a sandwich ELISA-like assay with a low limit of quantification for pustulan (1.5 pg/ml), and we successfully employed this assay in the quantification of extracellular ?-1,6-glucan released by >250 patient-derived strains of different Candida species (including Candida auris) in culture supernatant in vitro We also used this assay to measure ?-1,6-glucan in vivo in the serum and in several organs in a mouse model of systemic candidiasis. Our work describes a reliable method for ?-1,6-glucan detection, which may prove useful for the diagnosis of invasive fungal infections.

| S-EPMC7170528 | biostudies-literature

Immune response to intra-ocular injection of particulate β-(1,3)(1,6) glucan

Project description:Description of cellular and molecular landscape of vitreal immune cells following optic nerve crush injury plus intra-ocular injection of particulate β-glucan after 3days, in WT and Itgam-/- mice

2024-07-03 | GSE244627 | GEO

Commensal bacteria protect against food allergen sensitization

Project description:Analysis of colonic epithelial cell gene expression in germ-free, Bacteroides uniformis-colonized, and Clostridia-colonized gnotobiotic mice. Bacteria were isolated from our SPF mouse facility and were used to selectively colonize germ-free mice. Germ free mice were left germ free or were colonized with Bacteroides uniformis or a consortium of Clostridia. Total RNA was extraced from colonic epithelial cells.

2014-08-15 | E-GEOD-60039 | biostudies-arrayexpress

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