Project description:L1CAM-captured extracellular vesicles (LCEVs) were isolated and characterized meticulously. Whole-transcriptome of LCEVs was analyzed by lncRNA microarray and RNA-Sequencing. RNAs expressed differently in LCEVs from ASD sera vs. TD sera were screened, analyzed, and further validated.
Project description:Extracellularly released particles, including membrane vesicles, have increasingly been recognized as important for bacterial community functions and host-interaction processes, but their compositions and functional roles differ between species and also between strains of the same species. In this study, we have determined the composition of membrane vesicles and protein particles identified in the cell-free pellets of two strains of Apilactobacillus kunkeei, a defensive symbiont of honeybees. The membrane vesicles were separated from the extracellular particles using density gradient ultracentrifugation. The peaks of the RNA and protein distributions were separated from each other and the highest concentration of RNA was observed in the fractions that contained the membrane vesicles while the highest protein concentration coincided with the fractions that contained extracellular particles. A comparative proteomics analysis by LC-MS/MS showed that 37 proteins with type-I signal peptides were consistently identified across the fractionated samples obtained from the cell-free pellets, of which 29 were orthologs detected in both strains. Functional predictions of the extracellular proteins revealed the presence of glycoside hydrolases, glycosyltransferases, giant proteins and peptidases. The extracellular transcriptomes mapped to a broad set of genes with a similar functional profile as the whole cell transcriptome. This study provides insights into the composition of membrane vesicles and extracellular proteins of a bee-associated symbiont.
Project description:Membrane vesicles released by neoplastic cells into extracellular medium contain potential of carrying arrays of oncogenic molecules including proteins and microRNAs (miRNA). Extracellular (exosome-like) vesicles play a major role in cell-to-cell communication. Thus, the characterization of miRNAs of exosome-like vesicles is imperative in clarifying intercellular signaling as well as identifying disease markers. microarray analysis identified several oncogenic miRNA between the two types vesicles. Exosome-like vesicles were isolated using gradient centrifugation from MCF-7 and MDA-MB 231 cultures. Sample-derived miRNA expression was analyzed on Miltenyi's miRXploreM-bM-^DM-" platform. The RNA was isolated from two replicate samples of MDA-MB231 and MCF7 cell lines. The miRNAs extracted from all four samples were labeled with Hy5 and hybridized against Hy3-labeled Universal Reference miRNA (UR). Candidate miRNAs were identified. In this report, the re-ratios of MDA1 or MDA2 relative to a virtual pool of both MCF7 samples 1 and 2 were used for further analyses. 1- Reratio No. 7 MDA 1 vs MCF7 1+2 reratios of (MDA 1 vs UR) and (MCF7 1 vs UR + MCF7 2 vs UR) 7970041, (7970039 + 7970040) 2- Reratio No. 8 MDA 2 vs MCF7 1+2 reratios of (MDA 2 vs UR) and (MCF7 1 vs UR + MCF7 2 vs UR) 7970042, (7970039 + 7970040) Exosome-like vesicles were isolated using gradient centrifugation from MCF-7 and MDA-MB 231 cultures. Sample-derived miRNA expression was analyzed on Miltenyi's miRXploreM-bM-^DM-" platform. The RNA was isolated from two replicate samples of MDA-MB231 and MCF7 cell lines. The miRNAs extracted from all four samples were labeled with Hy5 and hybridized against Hy3-labeled Universal Reference miRNA (UR). Candidate miRNAs were identified. In this report, the re-ratios of MDA1 or MDA2 relative to a virtual pool of both MCF7 samples 1 and 2 were used for further analyses. 1- Reratio No. 7 MDA 1 vs MCF7 1+2 reratios of (MDA 1 vs UR) and (MCF7 1 vs UR + MCF7 2 vs UR) 7970041, (7970039 + 7970040) 2- Reratio No. 8 MDA 2 vs MCF7 1+2 reratios of (MDA 2 vs UR) and (MCF7 1 vs UR + MCF7 2 vs UR) 7970042, (7970039 + 7970040)
Project description:Intercellular communication is critical for integrating complex signals in multicellular eukaryotes. Vascular endothelial cells and T lymphocytes closely interact during the recirculation and trans-endothelial migration of T cells. In addition to direct cell-cell contact, we show that T cell derived extracellular vesicles can interact with endothelial cells and modulate their cellular functions. Thrombospondin-1 and its receptor CD47 are expressed on exosomes/ectosomes derived from T cells, and these extracellular vesicles are internalized and modulate signaling in both T cells and endothelial cells. Extracellular vesicles released from cells expressing or lacking CD47 differentially regulate activation of T cells induced by engaging the T cell receptor. Similarly, T cell-derived extracellular vesicles modulate endothelial cell responses to vascular endothelial growth factor and tube formation in a CD47-dependent manner. Uptake of T cell derived extracellular vesicles by recipient endothelial cells globally alters gene expression in a CD47-dependent manner. CD47 also regulates the mRNA content of extracellular vesicles in a manner consistent with some of the resulting alterations in target endothelial cell gene expression. Therefore, the thrombospondin-1 receptor CD47 directly or indirectly regulates intercellular communication mediated by the transfer of extracellular vesicles between vascular cells. HuVEC cells were cocultured with exosomes derived either from Jurkat or JinB8 cells culture media. Each condition was done in triplicate. Also, Exosome RNA from Jurkat or JINB8 cells were compared to each other in triplicate.
Project description:Plasmodium falciparum secretes extracellular vesicles that contain RNA. The biological benefit of this secretion to the secreting parasite is not known. Here, we sequenced the RNA content of extracellular vesicles and compared with that of the secreting whole parasites. The data suggests that extracellular vesicles might be part of a post-transcriptional regulatory mechanism that shapes intracellular RNA levels in the parasite.