Project description:<p>We extracted Chlorella vulgaris exosomes by ultraionization method, and analyzed its active substances by non targeted metabonomics detection.</p>
Project description:Foxp3+ regulatory T (Treg) cells prevent inflammatory disease but the mechanistic basis of suppression is not understood completely . Gene silencing by RNA interference can act in a cell-autonomous and non-cell-autonomous manner, providing mechanisms of inter-cellular regulation. Here, we demonstrate that non-cell-autonomous gene silencing, mediated by miRNA-containing exosomes, is a mechanism employed by Treg cells to suppress T cell-mediated disease. Treg cells transferred microRNAs (miRNA) to various immune cells, including T helper 1 (Th1) cells, suppressing Th1 cell proliferation and cytokine secretion. Use of Dicer-deficient or Rab27a and Rab27b double-deficient Treg cells to disrupt miRNA-biogenesis or the exosomal pathway, respectively, established a requirement for miRNAs and exosomes for Treg cell-mediated suppression. Transcriptional analysis and miRNA inhibitor studies showed that exosome-mediated transfer of Let-7d from Treg cell to Th1 cells contributed to suppression and prevention of systemic disease. These studies reveal a mechanism of Treg cell-mediated suppression mediated by miRNA-containing exosomes. Regulatory T cells (CD4+CD25hiFoxp3rfp+, Treg) were isolated from naive mice. RNA as extracted form some Treg cells, while others were cultured in complete IMDM media for 3 days, stimulated with anti-CD3 anti-CD3 (1ug/ml) and anti-CD28 (10ug/ml). Exosomes were recovered from Treg cell supernatant, as described, and RNA was extracted form the purified exosomes. To identify which miRNAs were transferred to Dicer-deficient (KO) cells from Treg cells, we cultured Dicer KO cells alone, or co-cultured Dicer KO cells with Treg cells. RNA was extracted form Dicer KO cells cultured alone or from Dicer KO cells cultured in the presence of Treg cells. 3 x biological replicates were used. Each biological replicate was derived from a pool of 3-5 samples.
Project description:Colostrum and milk have high nutritional value and provide a complete diet for neonates, along with bioactive substances which modulate various functions such as immune defense. Exosomes are membranous vesicles of endocytic origin, recently been considered as major players in cell-cell communication. The mechanisms by which milk components can prime the infant’s active immunity are not entirely clear, and exosomes are suggested to be essential for the infant’s physiological development. We assessed the exosomal proteome profile from milk samples obtained from 10 healthy sows, at day 0 (colostrum), day 7, and 14 post-partum. Exosomes were isolated by ultracentrifugation coupled with size exclusion chromatography, and were characterized by nanoparticle tracking analysis, transmission electron microscopy and Western blotting for exosome markers. Isolated exosomes were in-gel digested and after TMT-labelling of the peptides, they were subjected to LC-MS/MS. Quantitative proteomics analysis revealed different proteome profiles for colostrum exosomes and milk exosomes, and functional analysis highlighted pathways related to immune response, cellular development, and regulation of cellular processes. This study endorses the importance of exosomes as active biocomponents of milk and provides knowledge for future studies exploring their role in regulation of immunity and growth of the newborn.
Project description:Unprogrammed macrophage polarization, is associated with diabetic wound ulcers. Nevertheless, development of corresponding drugs is still a challenge. Here, exosomes are isolated from naive bone marrow-derived macrophages (BMDMs) (M0-Exos), inflammatory BMDMs (M1-Exos), and anti-inflammatory BMDMs (M2-Exos), with the aim of pinpointing the exosomes functionality and identify global miRNAs expression profiles.
Project description:<p>Bacteria secrete exosomes containing stress-related molecules such as proteins and nucleic acids, which facilitate adaptation to challenging environments. In order to elucidate the role of A. acidoterrestris exosomes in its acid stress response, this study combined proteomics and metabolomics to analyze the function and biological pathway of exosomes in A. acidoterrestris against acid stress The results of exosome proteomics showed that spore coat protein D, SecA, YidC, chemotactic protein CheA, methyl-accepting chemotactic protein (MCP), serine-type D-alanine-D-alanine carboxypeptidase protein, and lipid II glutamine synthetase subunit GatD protein were significantly up-regulated. These proteins are mainly enriched in ABC transporters, two-component systems, quorum sensing, peptidoglycan biosynthesis and other pathways. The results of exosomal metabolomics showed that 83 and 68 differential metabolites were detected in positive and negative ion modes, respectively. These metabolites were mainly enriched in ABC transporters, carbohydrate metabolism and amino acid metabolism. This study showed that the exosome proteins and metabolites secreted by A. acidoterrestris under acid stress changed significantly, revealing the overall response of A. acidoterrestris to acid stress, and pointing out the possible future direction for the effective control and application of A. acidoterrestris</p>
Project description:<p>Bacteria secrete exosomes containing stress-related molecules such as proteins and nucleic acids, which facilitate adaptation to challenging environments. In order to elucidate the role of A. acidoterrestris exosomes in its acid stress response, this study combined proteomics and metabolomics to analyze the function and biological pathway of exosomes in A. acidoterrestris against acid stress The results of exosome proteomics showed that spore coat protein D, SecA, YidC, chemotactic protein CheA, methyl-accepting chemotactic protein (MCP), serine-type D-alanine-D-alanine carboxypeptidase protein, and lipid II glutamine synthetase subunit GatD protein were significantly up-regulated. These proteins are mainly enriched in ABC transporters, two-component systems, quorum sensing, peptidoglycan biosynthesis and other pathways. The results of exosomal metabolomics showed that 83 and 68 differential metabolites were detected in positive and positive ion modes, respectively. These metabolites were mainly enriched in ABC transporters, carbohydrate metabolism and amino acid metabolism. This study showed that the exosome proteins and metabolites secreted by A. acidoterrestris under acid stress changed significantly, revealing the overall response of A. acidoterrestris to acid stress, and pointing out the possible future direction for the effective control and application of A. acidoterrestris</p>
Project description:microRNA profiles of Exosomes from Pooled NPC Patients serum comparing Control Exosomes from Healthy donors serum Two-condition experiment, Exosomes from Pooled Healthy donors serum vs. Exosomes from Pooled NPC Patients serum. Biological replicates: 1 Exosomes from Pooled Healthy donors serum, 1 Exosomes from Pooled NPC Patients serum,
Project description:Unhealable diabetic wounds need to be addressed with the help of newer, more efficacious strategies. Exosomes combined with biomaterials for sustained delivery of therapeutic agents are expected to bring new hope for chronic wound treatment. Here, the engineered exosomes modified for efficiently loading miR-146a and attaching to silk fibroin patch were demonstrated to promote diabetic wound healing. The transcriptomics analysis suggested anti-inflammatory and regenerative effects with SGM-miR146a-Exo@SFP treatment.
Project description:Exosomes are nanosized extracellular vesicles with lipid bilayer membranes and contain various contents, including lipids, miRNAs and proteins, all of which are widely involved in signaling pathways and genetic information processes. Exosomes derived from adipose tissues (AT-Exos) have been identified as a crucialmedium in the transmission of information from adipose tissue to itself and to other organs, including exosomes derived from inguinal white adipose tissue (iWAT-Exos), visceral white adipose tissue (vWAT-Exos) and brown adipose tissue (BAT-Exos). Here we reported that dietary conditions and tissue origins of exosome can affect the composition and function of miRNAs in AT-Exos, mainly including lipid metabolism and inflammatory signaling pathways associated with metabolic imbalance.
Project description:Exosomes research has been strongly promoted by the discovery of different classes of RNAs, mainly miRNAs, which are transfered to target cells and modulate various simgnaling pathways in target cells. Inflamed endothelial cells derived exosomes, acting as carriers for intercellular information exchange in atherosclerosis, are worthy of further research to investigate the disease pathophysiological mechanisms. This study aimed to investigate the differentially microRNA expression between normal and inflamed endothelial cells derived exosomes.