Project description:To determine whether calprotectin can elicit any transcriptional response in the probiotic E. coli Nissle 1917(EcN), EcN was treated with 200 ug/g of calprotectin in log phase.
Project description:We performed comparative transcriptomic profiling of E. coli Nissle 1917 (EcN) to determine the effect of microgravity (MG) on cell growth and metabolism.
Project description:RNA sequencing of Escherichia coli Nissle 1917 before and after HOCl treatment was perfomed to identify pathways that may be important in responding to oxidative stress caused by reachive chlorine species (RCS).
Project description:Extracellular membrane vesicles (MVs) released by gut microbiota are key players in the communication with the host. Next-generation sequencing of small RNAs was used to quantify miRNA expression in monocyte-derived dendritic cells after 24 h-stimulation with MVs isolated from two E. coli intestinal isolates, the probiotic E. coli strain Nissle 1917 (EcN) and the commensal ECOR12. Analysis revealed miRNAs differentially expressed in response to MVs compared to immature control dendritic cells (log2fold-change > 0.7 and padj < 0.001). A common set of miRNAs was modulated by MVs from both strains (46 downregulated, 75 upregulated). In addition, these vesicles elicited differential expression of specific miRNAs depending on the producer strain (26 downregulated and 23 upregulated by EcN; 48 downregulated and 49 upregulated by ECOR12).
Project description:Intestinal T cells responses are strongly influenced by microbes. In this study we aimed to adress how different microbes and what microbial factors influce intestinal T cells differentiation and subsequent clonal expansion. Paticularly we choose Clostridium ramosum (Cr) as a strong induced of a subest of intestinal Tregs (Rorc+), a medium inducer or Rorc+ Tregs an T1h7 Escherichia coli Nissle (EcN) and a strain that did not impact either o those cell (Pm). In addition, to study the impact of Escherichia coli Nissle (EcN) ploysaccharide K5 capsule on intestinal T cells activation, we used a EcN strain lacking the K5 capsule (kfiCD). At day 21 post-monocolonization we isolated CD4 T cells from monocolonized mice in both intestinal Lamina Propria and spleen and performed single cells RNA and TCR seq.
Project description:Escherichia coli Nissle 1917 (EcN) is an intestinal probiotic that is effective for the treatment of intestinal disorders, such as inflammatory bowel disease and ulcerative colitis. EcN is a representative Gram-negative probiotic in biomedical research and is an intensively studied probiotic. However, to date, its genome-wide metabolic network model has not been developed. Here, we developed a comprehensive and highly curated EcN metabolic model, referred to as iDK1463, based on genome comparison and phenome analysis. The model was improved and validated by comparing the simulation results with experimental results from phenotype microarray tests. iDK1463 comprises 1463 genes, 1313 unique metabolites, and 2984 metabolic reactions. Phenome data of EcN were compared with those of Escherichia coli intestinal commensal K-12 MG1655. iDK1463 was simulated to identify the genetic determinants responsible for the observed phenotypic differences between EcN and K-12. Further, the model was simulated for gene essentiality analysis and utilization of nutrient sources under anaerobic growth conditions. These analyses provided insights into the metabolic mechanisms by which EcN colonizes and persists in the gut. iDK1463 will contribute to the system-level understanding of the functional capacity of gut microbes and their interactions with microbiota and human hosts, as well as the development of live microbial therapeutics.
Project description:The mass spectrometry proteomics data of outer membrane vesicles of wide-type Escherichia coli Nissle 1917 and engineered Escherichia coli Nissle 1917
Project description:Escherichia coli Nissle 1917 (EcN) is a probiotic used for treatment of intestinal disorders. EcN improves gastrointestinal homeostasis and microbiota balance; however little is known about how this probiotic delivers effector molecules to the host. Outer membrane vesicles (OMVs) are constitutively produced by gram-negative bacteria and have a relevant role in bacteria-host interactions. Here we performed proteomic analysis of EcN OMVs. Using 1D SDSD-PAGE and highly sensitive LC-MS/MS analysis we identified 192 EcN vesicular proteins with high confidence in three independent experiments. Of these proteins, 18 were encoded by strain-linked genes and 57 were common to pathogen-derived OMVs. These proteins may contribute to the ability of this probiotic to colonize the human gut as they fulfil functions related to adhesion to host tissues, immune modulation or bacterial survival in host niches. This study describes the first global OMV proteome of a probiotic strain and provides evidence that probiotic-derived OMVs contain proteins that can target these vesicles to the host and mediate their beneficial effects on intestinal function.
