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Transcriptomic microRNA profiling of dendritic cells in response to gut microbiota secreted vesicles

ABSTRACT: 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).

ORGANISM(S): Homo sapiens

PROVIDER: GSE144355 | GEO | 2020/06/25

REPOSITORIES: GEO

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E. coli Nissle 1917 transcriptional response after treatment with Calprotectin

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2013-12-19 | PXD000367 | Pride

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Project description:Bacteria release nano-sized membrane vesicles (MVs) into the extracellular milieu. Bacterial MVs contain molecular cargo originating from the parent bacterium and have important roles in bacterial survival and pathogenesis. Using 8-plex iTRAQ approaches, we profiled the MV proteome of two Escherichia coli strains - uropathogenic E. coli (UPEC) 536 and probiotic Nissle 1917. For these strains, we compared the difference in the MV proteome between a crude input MV prepared by ultracentrifugation alone with that from MVs that were further purified by either density gradient centrifugation (DGC) or size exclusion chromatography (SEC); and also compared MVs from bacterial cultures that were grown in iron-restricted (R) and ironsupplemented (RF) conditions. We found that overall, outer membrane components were highly enriched, and bacterial inner membrane components were significantly depleted in both UPEC and Nissle MVs, in keeping with an outer membrane origin. In addition, we found enrichment of ribosome-related Gene Ontology terms in UPEC MV, and proteins involved in glycolytic process and ligase activity in Nissle MV. We have identified that three proteins (RbsB of UPEC in R; YoeA of UPEC in RF; BamA of Nissle in R) were consistently enriched in the DGC- and SEC-purified MV samples in comparison to their crude input MV, whereas conversely the 60 kDa chaperonin GroEL was enriched in the crude input MVs for both UPEC and Nissle in R condition. Such proteins may have a potential utility as technical markers for assessing the purity from contaminating non-vesicular protein after MV purification. Several proteins were changed in their abundance depending on the iron availability in the media.

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