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https://www.readbyqxmd.com/read/29127410/ultrafiltration-combined-with-size-exclusion-chromatography-efficiently-isolates-extracellular-vesicles-from-cell-culture-media-for-compositional-and-functional-studies
#1
Birke J Benedikter, Freek G Bouwman, Tanja Vajen, Alexandra C A Heinzmann, Gert Grauls, Edwin C Mariman, Emiel F M Wouters, Paul H Savelkoul, Carmen Lopez-Iglesias, Rory R Koenen, Gernot G U Rohde, Frank R M Stassen
Appropriate isolation methods are essential for unravelling the relative contribution of extracellular vesicles (EVs) and the EV-free secretome to homeostasis and disease. We hypothesized that ultrafiltration followed by size exclusion chromatography (UF-SEC) provides well-matched concentrates of EVs and free secreted molecules for proteomic and functional studies. Conditioned media of BEAS-2B bronchial epithelial cells were concentrated on 10 kDa centrifuge filters, followed by separation of EVs and free protein using sepharose CL-4B SEC...
November 10, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29115712/proteomic-profiling-of-exosomes-secreted-by-breast-cancer-cells-with-varying-metastatic-potential
#2
Lahiru Gangoda, Michael Liem, Ching-Seng Ang, Shivakumar Keerthikumar, Christopher G Adda, Belinda S Parker, Suresh Mathivanan
Cancer cells actively release extracellular vesicles, including exosomes, into the surrounding microenvironment. Exosomes play pleiotropic roles in cancer progression and metastasis, including invasion, angiogenesis, and immune modulation. However, the proteome profile of exosomes isolated from cells with different metastatic potential and the role of these exosomes in driving metastasis remains unclear. Here, we conducted a comparative proteomic analysis of exosomes isolated from several genetically related mouse breast tumour lines with different metastatic propensity...
November 8, 2017: Proteomics
https://www.readbyqxmd.com/read/29114223/the-human-amnion-epithelial-cell-secretome-decreases-hepatic-fibrosis-in-mice-with-chronic-liver-fibrosis
#3
Majid Alhomrani, Jeanne Correia, Marcus Zavou, Bryan Leaw, Nathan Kuk, Rong Xu, Mohamed I Saad, Alexander Hodge, David W Greening, Rebecca Lim, William Sievert
Background: Hepatic stellate cells (HSCs) are the primary collagen-secreting cells in the liver. While HSCs are the major cell type involved in the pathogenesis of liver fibrosis, hepatic macrophages also play an important role in mediating fibrogenesis and fibrosis resolution. Previously, we observed a reduction in HSC activation, proliferation, and collagen synthesis following exposure to human amnion epithelial cells (hAEC) and hAEC-conditioned media (hAEC-CM). This suggested that specific factors secreted by hAEC might be effective in ameliorating liver fibrosis...
2017: Frontiers in Pharmacology
https://www.readbyqxmd.com/read/29108957/matrix-vesicles-from-chondrocytes-and-osteoblasts-their-biogenesis-properties-functions-and-biomimetic-models
#4
REVIEW
Massimo Bottini, Saida Mebarek, Karen L Anderson, Agnieszka Strzelecka-Kiliszek, Lukasz Bozycki, Ana Maria Sper Simão, Maytê Bolean, Pietro Ciancaglini, Joanna Bandorowicz Pikula, Slawomir Pikula, David Magne, Niels Volkmann, Dorit Hanein, José Luis Millán, Rene Buchet
BACKGROUND: Matrix vesicles (MVs) are released from hypertrophic chondrocytes and from mature osteoblasts, the cells responsible for endochondral and membranous ossification. Under pathological conditions, they can also be released from cells of non-skeletal tissues such as vascular smooth muscle cells. MVs are extracellular vesicles of approximately 100-300nm diameter harboring the biochemical machinery needed to induce mineralization. SCOPE OF THE REVIEW: The review comprehensively delineates our current knowledge of MV biology and highlights open questions aiming to stimulate further research...
November 3, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/29093555/exosomes-derived-from-hiv-1-infected-dcs-mediate-viral-trans-infection-via-fibronectin-and-galectin-3
#5
Rutuja Kulkarni, Anil Prasad
Exosomes are membrane enclosed nano-sized vesicles actively released into the extracellular milieu that can harbor genomic, proteomic and lipid cargos. Functionally, they are shown to regulate cell-cell communication and transmission of pathogens. Though studies have implicated a role for exosomes in HIV-1 pathogenesis, their mechanisms are not well defined. Here, we characterized exosomes derived from uninfected or HIV-1 infected T-cells and DCs. We demonstrate substantial differences in morphological, molecular and biogenesis machinery between exosomes derived from these two immune cell types...
November 1, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29078295/intercellular-mrna-trafficking-via-membrane-nanotube-like-extensions-in-mammalian-cells
#6
Gal Haimovich, Christopher M Ecker, Margaret C Dunagin, Elliott Eggan, Arjun Raj, Jeffrey E Gerst, Robert H Singer
RNAs have been shown to undergo transfer between mammalian cells, although the mechanism behind this phenomenon and its overall importance to cell physiology is not well understood. Numerous publications have suggested that RNAs (microRNAs and incomplete mRNAs) undergo transfer via extracellular vesicles (e.g., exosomes). However, in contrast to a diffusion-based transfer mechanism, we find that full-length mRNAs undergo direct cell-cell transfer via cytoplasmic extensions characteristic of membrane nanotubes (mNTs), which connect donor and acceptor cells...
October 24, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29070527/extracellular-vesicles-released-by-cardiomyocytes-in-a-doxorubicin-induced-cardiac-injury-mouse-model-contain-protein-biomarkers-of-early-cardiac-injury
#7
Chontida Yarana, Dustin Carroll, Jing Chen, Luksana Chaiswing, Yanming Zhao, Teresa Noel, Michael Alstott, Younsoo Bae, Emily V Dressler, Jeffrey A Moscow, D Allan Butterfield, Haining Zhu, Daret K St Clair
PURPOSE: Cardiac injury is a major cause of death in cancer survivors, and biomarkers for it are detectable only after tissue injury has occurred. Extracellular vesicles (EVs) remove toxic biomolecules from tissues and can be detected in the blood. Here, we evaluate the potential of using circulating EVs as early diagnostic markers for long-term cardiac injury. EXPERIMENTAL DESIGN: Using a mouse model of doxorubicin (DOX)-induced cardiac injury, we quantified serum EVs, analyzed proteomes, measured oxidized protein levels in serum EVs released after DOX treatment, and investigated the alteration of EV content...
October 25, 2017: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
https://www.readbyqxmd.com/read/29067621/erratum-to-an-adaptable-polyethylene-glycol-based-workflow-for-proteomic-analysis-of-extracellular-vesicles
#8
Stephanie N Hurwitz, David G Meckes
No abstract text is available yet for this article.
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29061044/ubiquitin-conjugation-probed-by-inflammation-in-myeloid-derived-suppressor-cell-extracellular-vesicles
#9
Katherine R Adams, Sitara Chauhan, Divya B Patel, Virginia K Clements, Yan Wang, Steven M Jay, Nathan J Edwards, Suzanne Ostrand-Rosenberg, Catherine Fenselau
Ubiquitinated proteins carried by the extracellular vesicles (EV) released by myeloid-derived suppressor cells (MDSC) have been investigated using proteomic strategies to examine the effect of tumor-associated inflammation. EV were collected from MDSC directly following isolation from tumor-bearing mice with low and high inflammation. Among the 1092 proteins (high inflammation) and 925 proteins (low inflammation) identified, more than 50% were observed as ubiquitinated proteoforms. More than three ubiquitin-attachment sites were characterized per ubiquitinated protein, on average...
November 10, 2017: Journal of Proteome Research
https://www.readbyqxmd.com/read/29042682/il-12-stimulates-ctls-to-secrete-exosomes-capable-of-activating-bystander-cd8-t-cells
#10
Lei Li, Steven M Jay, Yan Wang, Shu-Wei Wu, Zhengguo Xiao
An effective cytotoxic T lymphocyte (CTL) response against intracellular pathogens is generally accomplished by immense CTL expansion and activation, which can destroy infected cells. Vigorous immune responses can lead to activation of bystander CD8(+) T cells, but the contribution from antigen-specific CTLs is not well understood. We found that CTLs secrete extracellular vesicles following antigen stimulation. These CTL-derived vesicles contain CTL proteins and exhibit markers and size profiles consistent with exosomes...
October 17, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29030127/a-method-for-the-isolation-and-enrichment-of-purified-bovine-milk-exosomes
#11
Kanchan Vaswani, Yong Qin Koh, Fatema B Almughlliq, Hassendrini N Peiris, Murray D Mitchell
Exosomes are nanovesicles that play important roles in intercellular communication as they carry information to target cells. Isolation of high purity exosomes will aid in studying the exosomal cargo and quantity as well as how cell-specific messages are carried. We describe a new method incorporating size exclusion chromatography (SEC) to enrich milk-derived exosomes from extracellular vesicles (EVs). This involved the initial isolation of EVs from bovine milk via milk processing and ultracentrifugation; followed by a new method to enrich exosomes using SEC...
October 10, 2017: Reproductive Biology
https://www.readbyqxmd.com/read/29025360/best-practice-of-identification-and-proteomic-analysis-of-extracellular-vesicles-in-human-health-and-disease
#12
Barbara W Sódar, Árpád Kovács, Tamás Visnovitz, Éva Pállinger, Károly Vékey, Gabriella Pocsfalvi, Lilla Turiák, Edit I Buzás
Extracellular vesicles are emerging sources of biomarkers for modern preventive and precision medicine. Extracellular vesicles in body fluids offer a unique opportunity for integrative biomarker approaches due to their complex biocargo that includes proteins, lipids, nucleic acids and metabolites. Mass spectrometry-based proteomics data suggest that a significant portion of human proteins are sorted into extracellular vesicles and amenable for biomarker discovery schemes. Areas covered: this review focuses on key aspects of isolation, quality control and subsequent analysis of blood plasma- and conditioned medium-derived extracellular vesicle proteins, and summarizes the current state-of-the-art in the field...
October 26, 2017: Expert Review of Proteomics
https://www.readbyqxmd.com/read/28992005/characterization-of-exosomes-from-body-fluids-of-dairy-cows
#13
Y Q Koh, H N Peiris, K Vaswani, S Meier, C R Burke, K A Macdonald, J R Roche, F Almughlliq, B J Arachchige, S Reed, M D Mitchell
Exosomes are a specific subpopulation of extracellular vesicles that are widely released by cells of different origins with divergent functions that make their way into body fluids that can be conveniently sampled. In the current study, we isolated and evaluated exosomes from concurrently collected samples of milk, plasma, saliva, and urine from a group of 6 pregnant Holstein-Friesian dairy cows (aged 7 mo, 174 to 203 d of gestation). The cows had BCS of 3.5 to 5.25 (on a scale of 1 to 10), and the milk production for the season to the time of sampling ranged between 5,118 and 6,959 kg...
September 2017: Journal of Animal Science
https://www.readbyqxmd.com/read/28991197/differential-proteome-analysis-of-extracellular-vesicles-from-breast-cancer-cell-lines-by-chaperone-affinity-enrichment
#14
Steven G Griffiths, Michelle T Cormier, Aled Clayton, Alan A Doucette
The complexity of human tissue fluid precludes timely identification of cancer biomarkers by immunoassay or mass spectrometry. An increasingly attractive strategy is to primarily enrich extracellular vesicles (EVs) released from cancer cells in an accelerated manner compared to normal cells. The Vn96 peptide was herein employed to recover a subset of EVs released into the media from cellular models of breast cancer. Vn96 has affinity for heat shock proteins (HSPs) decorating the surface of EVs. Reflecting their cells of origin, cancer EVs displayed discrete differences from those of normal phenotype...
October 8, 2017: Proteomes
https://www.readbyqxmd.com/read/28986800/isolation-of-exosomes-for-the-purpose-of-protein-cargo-analysis-with-the-use-of-mass-spectrometry
#15
Monika Pietrowska, Sonja Funk, Marta Gawin, Łukasz Marczak, Agata Abramowicz, Piotr Widłak, Theresa Whiteside
Exosomes are intercellular messengers with a high potential for diagnostic and therapeutic utility. It is believed that exosomes present in body fluids are responsible for providing signals which inhibit immune cells, interfere with antitumor immunity, and thus influence the response to treatment and its effect. One of the most interesting issues in exosome studies is proper addressing of their cargo composed of nucleic acids and proteins. Effective and selective isolation of extracellular vesicles and identification of proteins present in exosomes has turned out to be a challenging aspect of their exploration...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28986585/quantitation-of-putative-colorectal-cancer-biomarker-candidates-in-serum-extracellular-vesicles-by-targeted-proteomics
#16
Takashi Shiromizu, Hideaki Kume, Mimiko Ishida, Jun Adachi, Masayuki Kano, Hisahiro Matsubara, Takeshi Tomonaga
At the moment, there is no sensitive clinical test for detecting early-stage colorectal cancer (CRC). Target proteomics has enabled high-throughput verification of hundreds of biomarker candidate proteins. Using this technology, we verified 725 previously reported CRC biomarker candidate proteins that are functionally correlated with CRC in extracellular vesicles (EVs) from patients. Of these, 356 proteins were quantified, and 34 peptides (22 proteins) showed significant differences in the serum EVs between healthy controls and CRC patients of two independent cohorts (n = 77 and 84)...
October 6, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28959386/cerebrospinal-fluid-extracellular-vesicle-enrichment-for-protein-biomarker-discovery-in-neurological-disease-multiple-sclerosis
#17
Joanne L Welton, Samantha Loveless, Timothy Stone, Chris von Ruhland, Neil P Robertson, Aled Clayton
The discovery of disease biomarkers, along with the use of "liquid biopsies" as a minimally invasive source of biomarkers, continues to be of great interest. In inflammatory diseases of the central nervous system (CNS), cerebrospinal fluid (CSF) is the most obvious biofluid source. Extracellular vesicles (EVs) are also present in CSF and are thought to be potential "biomarker treasure chests". However, isolating these CSF-derived EVs remains challenging. This small-scale pilot study developed and tested a protocol to enrich for CSF-EVs, both in relapsing remitting multiple sclerosis (RRMS) CSF and controls...
2017: Journal of Extracellular Vesicles
https://www.readbyqxmd.com/read/28955033/plasma-membrane-derived-extracellular-microvesicles-mediate-non-canonical-intercellular-notch-signaling
#18
Qiyu Wang, Quan Lu
ARMMs (arrestin domain-containing protein 1 (ARRDC1)-mediated microvesicles) are extracellular vesicles that bud directly at the plasma membrane; however, little is known about the molecular composition and physiological function of these vesicles. Here we report that ARMMs contain active NOTCH receptors and mediate a non-canonical intercellular NOTCH signaling. We identify over 100 proteins that are significantly enriched in ARMMs, including ARRDC1, TSG101 and multiple ESCRT complex proteins. About a third of ARMMs-enriched proteins are plasma membrane proteins, including the NOTCH2 receptor...
September 27, 2017: Nature Communications
https://www.readbyqxmd.com/read/28949504/assessment-of-extracellular-vesicles-purity-using-proteomic-standards
#19
Tingting Wang, Kyle W Anderson, Illarion V Turko
The increasing interest in extracellular vesicles (EVs) research is fueled by reports indicating their unique role in intercellular communication and potential connection to the development of common human diseases. The unique role assumes unique protein and nucleic acid cargo. Unfortunately, accurate analysis of EVs cargo faces a challenge of EVs isolation. Generally used isolation techniques do not separate different subtypes of EVs and even more, poorly separate EVs from non-EVs contaminants. Further development of EVs isolation protocols urgently needs a quantitative method of EVs purity assessment...
October 17, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28944300/proteomic-analysis-of-extracellular-vesicles-from-a-plasmodium-falciparum-kenyan-clinical-isolate-defines-a-core-parasite-secretome
#20
Abdirahman Abdi, Lu Yu, David Goulding, Martin K Rono, Philip Bejon, Jyoti Choudhary, Julian Rayner
BACKGROUND: Many pathogens secrete effector molecules to subvert host immune responses, to acquire nutrients, and/or to prepare host cells for invasion. One of the ways that effector molecules are secreted is through extracellular vesicles (EVs) such as exosomes. Recently, the malaria parasite P. falciparum has been shown to produce EVs that can mediate transfer of genetic material between parasites and induce sexual commitment. Characterizing the content of these vesicles may improve our understanding of P...
2017: Wellcome Open Research
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