Project description:Identification of proteins contained in extracellular vesicles of Lactiplantibacillus plantarum PCM 2675. Dataset is related to publication http://dx.doi.org/10.20517/evcna.2024.49. This work was financially supported by the National Science Centre, Poland (no. 2021/43/D/NZ6/01464).
Project description:This study was conducted to analyze phenotypic and proteomic differences of two Lactiplantibacillus plantarum strains (WCFS1, model strain from human saliva, and CIP104448, stool isolate) when a biofilm was produced under static conditions (well researched), or with the addition of flow (novel).
Project description:Cyclic di-adenosine monophosphate (c-di-AMP) is a crucial second messenger used by bacteria to mediate bacterial-host interactions. However, its roles in Lactiplantibacillus plantarum remain poorly understood. Here, we identify RwpL, which consists solely of an HTH domain, as a c-di-AMP receptor in L. plantarum WCFS1. RwpL mediates epithelial adhesion of L. plantarum WCFS1 and functions as both a transcription inhibitor and activator, with transcriptional output determined by the position of its binding site relative to the transcription start site (TSS) of target genes. Specifically, RwpL inhibits wxlA transcription by binding downstream of its TSS, thus reducing WxlA interactions with fibronectin, a matrix protein on the surface of intestinal epithelia. In contrast, RwpL activates transcription of the matrix-degrading metalloprotease gene mmpL by recruiting the sigma factor RpoA upstream of its TSS. Notably, elevated c-di-AMP levels diminish RwpL binding affinity for both DNA and RpoA, thereby reversing its regulatory functions and enhancing L. plantarum adhesion to host epithelial cells. Together, our findings uncover a dual and reversible regulatory mechanism through which c-di-AMP modulates bacterial-host interactions in L. plantarum.
Project description:To investigate intestinal health and its potential disruptors in vitro, representative models are required. Human induced pluripotent stem cell (hiPSC)-derived intestinal epithelial cells (IECs) more closely resemble the in vivo intestinal tissue than conventional in vitro models like human colonic adenocarcinoma Caco-2 cells. However, the potential of IECs to study immune-related responses upon external stimuli has not been investigated in detail yet. The aim of the current study was to evaluate immune-related effects of IECs by challenging them with a pro-inflammatory cytokine cocktail. Subsequently, the effects of Lactiplantibacillus plantarum WCFS1 were investigated in unchallenged and challenged IECs. All exposures were compared to Caco-2 cells and in vivo data where possible. Upon the inflammatory challenge, IECs and Caco-2 cells induced a pro-inflammatory response which was strongest in IECs. Heat-killed L. plantarum exerted the strongest effect on immune parameters in the IEC model, while L. plantarum in the stationary growth phase had most pronounced effects on immune-related gene expression in Caco-2 cells. Unfortunately, comparison to in vivo transcriptomics data showed limited similarities, which could be explained by essential differences in the study setups. Altogether, hiPSC-derived IECs show a high potential as a model to study immune-related responses in the intestinal epithelium in vitro.
Project description:In order to understand LBG derived galacto-manno-oligosaccharides utilization by a probiotic bacterium, Lactobacillus plantarum WCFS1, we have grown Lactobacillus plantarum WCFS1 (in duplicates) till mid log phase (OD600nm ~0.5, 10 h) in carbon free MRS (de Man, Rogosa Sharpe ) media containing either galacto-manno-oligosaccharides, mannose, glucose or galactose (1% w/v) as the sole carbon source.
Project description:Whole genome transcriptional profiling was used to characterize the response of Lactobacillus plantarum WCFS1 human isolate during challenge with oleuropein. Twelve independent experiments were performed and mixed at random in groups of four for total of three RNA samples. The transcriptional profile shows that Lactobacillus plantarum WCFS1 adapts its metabolic capacity to acquire certain carbohydrates and repress the expression of genes involved in fatty acid biosyntheis. The transcriptomic datasets also revealed the downregulation of genes related to the biosynthesis of capsular polysaccharides and genes coding for ABC-type transporters. In addition, induction of oligopeptide permeases is also part of the response of Lactobacillus plantarum WCFS1 to oleuropein.
Project description:Transcriptome profiles of control Lactobacillus plantarum WCFS1 cells were compared with 8% ethanol adapted cells and with 10 min or 30 min 8% ethanol shocked cells.