Exopolysaccharides of Lactobacillus reuteri : Their influence on adherence of E. coli to epithelial cells and inflammatory response

Kšonžeková, Petra; Bystrický, Peter; Vlčková, Silvia; Pätoprstý, Vladimı́r; Pulzová, Lucia; Mudroňová, Dagmar; Terezia, Kubaskova; Csank, Tomáš; Tkáčiková, Ľudmila

doi:10.1016/j.carbpol.2015.12.037

Cited by 90 publications

(57 citation statements)

References 36 publications

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“…This has been proved with in vitro (Ksonzeková et al, 2016; Zivkovic et al, 2016) as well as in vivo studies (Fanning et al, 2012; Chen et al, 2014; Nácher-Vázquez et al, 2015) using different intestinal cell lines and animal models, respectively. It was postulated that the mechanism behind this EPS protection could be the formation of a protective “biofilm-like” layer covering the intestinal epithelium and, therefore, preventing the adhesion of the pathogen or its toxins (Ruas-Madiedo et al, 2010; Fanning et al, 2012).…”

Section: Resultsmentioning

confidence: 87%

Gene Replacement and Fluorescent Labeling to Study the Functional Role of Exopolysaccharides in Bifidobacterium animalis subsp. lactis

et al. 2017

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An extracellular layer of exopolysaccharides (EPS) covers the surface of some Bifidobacterium animalis subsp. lactis strains, which could be of relevance for its probiotic performance. In order to understand the functional characteristics of B. animalis subsp. lactis, two isogenic strains that differ in their EPS-producing phenotype, due to a single mutation in the gene Balat_1410, were studied. By means of a double crossover recombination strategy, successfully used for the first time in bifidobacteria, Balat_1410 in the type strain B. animalis subsp. lactis DSM10140 was replaced by a mutated gene containing a non-synonymous mutation previously associated with the appearance of a mucoid-ropy phenotype. Nuclear magnetic resonance and SEC-MALS analyses showed that the novel strain harboring the mutation acquired a ropy phenotype, due to the production of a high molecular weight (HMW)-EPS that is not produced in the wild-type strain. Fluorescence labeling of both strains with two fluorescent proteins, m-Cherry and Green Fluorescent Protein, was achieved by expressing the corresponding genes under the control of a native selected promoter (the elongation factor Tu promoter). Remarkably, qualitative and quantitative fluorescence analyses demonstrated that the ropy strain displays a lower capability to adhere to human intestinal epithelial cells. In addition, the presence of the HMW-EPS reduced the capability of the producing strain to form biofilms upon three different abiotic surfaces. This work also highlights the fact that different EPS confer variable functional characteristics to the bifidobacterial surface, which may be relevant for the performance of B. animalis subsp. lactis as a probiotic. The construction of molecular tools allowing the functional characterization of surface structures in next generation probiotics is still a challenging issue that deserves further attention, given the relevant role that such molecules must play in the interaction with the host.

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Section: Resultsmentioning

confidence: 87%

Gene Replacement and Fluorescent Labeling to Study the Functional Role of Exopolysaccharides in Bifidobacterium animalis subsp. lactis

et al. 2017

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“…Among the beneficial bacteria, Lactobacillus represents one of the best producers of EPS. Exopolysaccharides produced by lactobacilli have not only positive effects on their producers [2,3] but also immunomodulatory effects on the gut mucosal immune system [4][5][6]. Exopolysaccharides stimulate the immune response in intestinal epithelial cells (IECs) through the activation of C-type lectin receptors (CLRs).…”

Section: Introductionmentioning

confidence: 99%

“…We have previously observed that the EPS isolated from Lactobacillus reuteri strain L26 Biocenol possesses cytokine-modulating activity. This EPS is a homopolymer consisting of α-d-glucose (1→3) and (1→6) glycosidic bonds at a ratio of 1.3:1 with a molecular weight (Mw) of 8.2 × 10 5 Da [6]. To date, scarce literature is available that presents a complete picture of the cell response (at the transcriptomic level) to the EPS derived from Lactobacillus.…”

Section: Introductionmentioning

confidence: 99%

Comprehensive mapping of the cell response to E. coli infection in porcine intestinal epithelial cells pretreated with exopolysaccharide derived from Lactobacillus reuteri

Tkáčiková

Mochnáčová

Tyagi

et al. 2020

Vet Res

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Bacterial exopolysaccharides (EPSs) are known to modulate immunity. To date, a plethora of studies have reported the effect of EPSs on intestinal cells; however few works have revealed a complete picture of the signalling events in intestinal epithelial cells induced by bacterial EPSs. Here, using transcriptomics, we comprehensively mapped the biological processes in porcine intestinal epithelial cells challenged with EPS derived from Lactobacillus reuteri alone, enterotoxigenic Escherichia coli (ETEC) or ETEC after pretreatment with EPS. The Gene Ontology analysis of differentially expressed genes (DEGs) showed that ETEC is able to evoke biological processes specifically involved in cell junction reorganization, extracellular matrix degradation, and activation of the innate immune response through the activation of pattern recognition receptors, such as TLRs and CTRs. A total of 495 DEGs were induced in ETEC-challenged cells. On the other hand, EPS pretreatment was able to attenuate overexpression of the genes induced by ETEC infection. The most relevant finding of this study is that EPS has a suppressive effect on the inflammatory response evoked by ETEC infection. On the basis of high-throughput RNA-seq, this report is the first to describe the effects of EPSs derived from L. reuteri used as a pretreatment of global gene expression in porcine epithelial cells.

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“…Furthermore, human moDCs generated in the presence of the soluble factors of L. reuteri DSM 17938 down-modulated LPS-induced IL-6, IL-10, and IL-23 secretion while the secretion of regulatory cytokine TGF-β remained unaffected (Haileselassie et al, 2016). EPS isolated from L. reuteri strain DSM 17938 and L26 Biocenol TM was recently shown to exert up-regulation of the mRNA level of IL-1β, NF-κB, TNF-α, and IL-6 (Kšonžeková et al, 2016). One of the putative L. reuteri surface proteins appeared to be important for the stimulation of THP-1 cells and the activation of NF-κB in U937-3xkB-LUC cells by L. reuteri strains (Jensen et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

Lactobacillus reuteri Surface Mucus Adhesins Upregulate Inflammatory Responses Through Interactions With Innate C-Type Lectin Receptors

et al. 2017

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The vertebrate gut symbiont Lactobacillus reuteri exhibits strain-specific adhesion and health-promoting properties. Here, we investigated the role of the mucus adhesins, CmbA and MUB, upon interaction of L. reuteri ATCC PTA 6475 and ATCC 53608 strains with human monocyte-derived dendritic cells (moDCs). We showed that mucus adhesins increased the capacity of L. reuteri strains to interact with moDCs and promoted phagocytosis. Our data also indicated that mucus adhesins mediate anti- and pro-inflammatory effects by the induction of interleukin-10 (IL-10), tumor necrosis factor alpha (TNF-α), IL-1β, IL-6, and IL-12 cytokines. L. reuteri ATCC PTA 6475 and ATCC 53608 were exclusively able to induce moDC-mediated Th1 and Th17 immune responses. We further showed that purified MUB activates moDCs and induces Th1 polarized immune responses associated with increased IFNγ production. MUB appeared to mediate these effects via binding to C-type lectin receptors (CLRs), as shown using cell reporter assays. Blocking moDCs with antibodies against DC-specific intercellular adhesion molecule 3-grabbing non-integrin (DC-SIGN) or Dectin-2 did not affect the uptake of the MUB-expressing strain, but reduced the production of TNF-α and IL-6 by moDCs significantly, in line with the Th1 polarizing capacity of moDCs. The direct interaction between MUB and CLRs was further confirmed by atomic force spectroscopy. Taken together these data suggest that mucus adhesins expressed at the cell surface of L. reuteri strains may exert immunoregulatory effects in the gut through modulating the Th1-promoting capacity of DCs upon interaction with C-type lectins.

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Exopolysaccharides of Lactobacillus reuteri : Their influence on adherence of E. coli to epithelial cells and inflammatory response

Cited by 90 publications

References 36 publications

Gene Replacement and Fluorescent Labeling to Study the Functional Role of Exopolysaccharides in Bifidobacterium animalis subsp. lactis

Gene Replacement and Fluorescent Labeling to Study the Functional Role of Exopolysaccharides in Bifidobacterium animalis subsp. lactis

Comprehensive mapping of the cell response to E. coli infection in porcine intestinal epithelial cells pretreated with exopolysaccharide derived from Lactobacillus reuteri

Lactobacillus reuteri Surface Mucus Adhesins Upregulate Inflammatory Responses Through Interactions With Innate C-Type Lectin Receptors

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