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https://www.readbyqxmd.com/read/29121670/quantitative-membrane-proteomics-reveals-a-role-for-tetraspanin-enriched-microdomains-during-entry-of-human-cytomegalovirus
#1
Kasinath Viswanathan, Marieke C Verweij, Nessy John, Daniel Malouli, Klaus Früh
Human cytomegalovirus (HCMV) depends on and modulates multiple host cell membrane proteins during each stage of the viral life cycle. To gain a global view of the impact of HCMV-infection on membrane proteins, we analyzed HCMV-induced changes in the abundance of membrane proteins in fibroblasts using stable isotope labeling with amino acids (SILAC), membrane fractionation and protein identification by two-dimensional liquid chromatography and tandem mass spectrometry. This systematic approach revealed that CD81, CD44, CD98, caveolin-1 and catenin delta-1 were down-regulated during infection whereas GRP-78 was up-regulated...
2017: PloS One
https://www.readbyqxmd.com/read/29117231/human-pancreatic-islet-derived-extracellular-vesicles-modulate-insulin-expression-in-3d-differentiating-ipsc-clusters
#2
Diana Ribeiro, Eva-Marie Andersson, Nikki Heath, Anette Persson-Kry, Richard Collins, Ryan Hicks, Niek Dekker, Anna Forslöw
It has been suggested that extracellular vesicles (EVs) can mediate crosstalk between hormones and metabolites within pancreatic tissue. However, the possible effect of pancreatic EVs on stem cell differentiation into pancreatic lineages remains unknown. Herein, human islet-derived EVs (h-Islet-EVs) were isolated, characterized and subsequently added to human induced pluripotent stem cell (iPSC) clusters during pancreatic differentiation. The h-islet-EVs had a mean size of 117±7 nm and showed positive expression of CD63 and CD81 EV markers as measured by ELISA...
2017: PloS One
https://www.readbyqxmd.com/read/29116068/-hydrogel-microchip-as-a-tool-for-studying-exosomes-in-human-serum
#3
V I Butvilovskaya, A A Tikhonov, E N Savvateeva, A A Ragimov, E L Salimov, S A Voloshin, D V Sidorov, M A Chernichenko, A P Polyakov, M M Filushin, M V Tsybulskaya, A Yu Rubina
Exosomes are cell-derived vesicles that are secreted by both normal and cancer cells. Over the last decade, a few studies have revealed that exosomes cross talk and/or influence major tumor-related pathways such as angiogenesis and metastasis involving many cell types within the tumor microenvironment. The protein composition of the membrane of an exosome reflects that of the membrane of the cell of origin. Because of this, tumor-derived exosomes differ from exosomes that are derived from normal cells. The detection of tumor exosomes and analysis of their molecular composition hold promise for diagnosis and prognosis of cancer...
September 2017: Molekuliarnaia Biologiia
https://www.readbyqxmd.com/read/29099494/prognostic-value-of-antigen-expression-in-multiple-myeloma-a-pethema-gem-study-on-1-265-patients-enrolled-in-four-consecutive-clinical-trials
#4
P Arana, B Paiva, M-T Cedena, N Puig, L Cordon, M-B Vidriales, N C Gutierrez, F Chiodi, L Burgos, L-L Anglada, J Martinez-Lopez, M-T Hernandez, A-I Teruel, M Gironella, M-A Echeveste, L Rosiñol, R Martinez, A Oriol, J De la Rubia, A Orfao, J Blade, J-J Lahuerta, M-V Mateos, J F S Miguel
Persistence of minimal residual disease (MRD) after treatment for myeloma predicts inferior outcomes, but within MRD-positive patients there is great heterogeneity with both early and very late relapses. Amongst different MRD techniques, flow cytometry provides additional information about antigen expression on tumor cells, which could potentially contribute to stratify MRD-positive patients. We investigated the prognostic value of those antigens required to monitor MRD in 1265 newly-diagnosed patients enrolled in the GEM2000, GEM2005MENOS65, GEM2005MAS65 and GEM2010MAS65 protocols...
November 3, 2017: Leukemia: Official Journal of the Leukemia Society of America, Leukemia Research Fund, U.K
https://www.readbyqxmd.com/read/29090272/a-new-panel-of-epitope-mapped-monoclonal-antibodies-recognising-the-prototypical-tetraspanin-cd81
#5
Joe Grove, Ke Hu, Michelle J Farquhar, Margaret Goodall, Lucas Walker, Mohammed Jamshad, Heidi E Drummer, Roslyn M Bill, Peter Balfe, Jane A McKeating
Background: Tetraspanins are small transmembrane proteins, found in all higher eukaryotes, that compartmentalize cellular membranes through interactions with partner proteins. CD81 is a prototypical tetraspanin and contributes to numerous physiological and pathological processes, including acting as a critical entry receptor for hepatitis C virus (HCV). Antibody engagement of tetraspanins can induce a variety of effects, including actin cytoskeletal rearrangements, activation of MAPK-ERK signaling and cell migration...
2017: Wellcome Open Research
https://www.readbyqxmd.com/read/29063370/pathways-of-production-and-delivery-of-hepatocyte-exosomes
#6
Li Chen, Ruju Chen, Sherri Kemper, David R Brigstock
Hepatocyte exosomes (Exo(Hep)) are proposed to mediate physiological or pathophysiological signaling in a variety of hepatic target cells. Exo(Hep) were purified from the medium of primary mouse hepatocytes or AML12 cells and characterized as ~100 nm nanovesicles that were positive for proteins commonly found in exosomes (CD9, CD81, flotillin) or hepatocytes (asialoglycoprotein receptor). Ethanol treatment of hepatocytes caused increased Exo(Hep) release and increased cellular mRNA expression of components involved in intracellular vesicle trafficking (Rab 5a,b,c, Rab 7a, Rab 27a,b) or exosome biogenesis via the ESCRT (HGS, Alix, STAM1, TSG101, VTA1, YKT6) or ceramide (nSmase2) pathways...
October 23, 2017: Journal of Cell Communication and Signaling
https://www.readbyqxmd.com/read/29030127/a-method-for-the-isolation-and-enrichment-of-purified-bovine-milk-exosomes
#7
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/29024461/reproducible-diagnosis-of-chronic-lymphocytic-leukemia-by-flow-cytometry-an-european-research-initiative-on-cll-eric-european-society-for-clinical-cell-analysis-escca-harmonisation-project
#8
Andy C Rawstron, Karl-Anton Kreuzer, Asha Soosapilla, Martin Spacek, Olga Stehlikova, Peter Gambell, Neil McIver-Brown, Neus Villamor, Katherina Psarra, Maria Arroz, Raffaella Milani, Javier de la Serna, M Teresa Cedena, Ozren Jaksic, Josep Nomdedeu, Carol Moreno, Gian Matteo Rigolin, Antonio Cuneo, Preben Johansen, Hans E Johnsen, Richard Rosenquist, Carsten Utoft Niemann, Wolfgang Kern, David Westerman, Marek Trneny, Stephen Mulligan, Michael Doubek, Sarka Pospisilova, Peter Hillmen, David Oscier, Michael Hallek, Paolo Ghia, Emili Montserrat
The diagnostic criteria for CLL rely on morphology and immunophenotype. Current approaches have limitations affecting reproducibility and there is no consensus on the role of new markers. The aim of this project was to identify reproducible criteria and consensus on markers recommended for the diagnosis of CLL. ERIC/ESCCA members classified 14 of 35 potential markers as "required" or "recommended" for CLL diagnosis, consensus being defined as >75% and >50% agreement, respectively. An approach to validate "required" markers using normal peripheral blood was developed...
October 10, 2017: Cytometry. Part B, Clinical Cytometry
https://www.readbyqxmd.com/read/28959386/cerebrospinal-fluid-extracellular-vesicle-enrichment-for-protein-biomarker-discovery-in-neurological-disease-multiple-sclerosis
#9
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/28949447/exosomal-proteins-as-prognostic-biomarkers-in-non-small-cell-lung-cancer
#10
(no author information available yet)
BACKGROUND: Use of exosomes as biomarkers in non-small cell lung cancer (NSCLC) is an intriguing approach in the liquid-biopsy era. Exosomes are nano-sized vesicles with membrane-bound proteins that reflect their originating cell. Prognostic biomarkers are needed to improve patient selection for optimal treatment. We here evaluate exosomes by protein phenotyping as a prognostic biomarker in NSCLC. METHODS: Exosomes from plasma of 276 NSCLC patients were phenotyped using the Extracellular Vesicle Array; 49 antibodies captured the proteins on the exosomes, and a cocktail of biotin-conjugated antibodies binding the general exosome markers CD9, CD81 and CD63 was used to visualise the captured exosomes...
December 2016: Molecular Oncology
https://www.readbyqxmd.com/read/28934733/exosomes-from-human-induced-pluripotent-stem-cell-derived-mesenchymal-stromal-cells-hipsc-mscs-protect-liver-against-hepatic-ischemia-reperfusion-injury-via-activating-sphingosine-kinase-and-sphingosine-1-phosphate-signaling-pathway
#11
Yingdong Du, Dawei Li, Conghui Han, Haoyu Wu, Longmei Xu, Ming Zhang, Jianjun Zhang, Xiaosong Chen
BACKGROUND/AIMS: This study aimed to evaluate the effects of exosomes produced by human-induced pluripotent stem cell-derived mesenchymal stromal cells (hiPSC-MSCs-Exo) on hepatic ischemia-reperfusion (I/R) injury, as well as the underlying mechanisms. METHODS: Exosomes derived from hiPSC-MSCs were isolated and characterized both biochemically and biophysically. hiPSC-MSCs-Exo were injected systemically into a murine ischemia/reperfusion injury model via the inferior vena cava, and then the therapeutic effects were evaluated...
2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/28920599/magnetic-nanoparticle-enhanced-surface-plasmon-resonance-biosensor-for-extracellular-vesicle-analysis
#12
Agnes T Reiner, Nicolas-Guillermo Ferrer, Priyamvada Venugopalan, Ruenn Chai Lai, Sai Kiang Lim, Jakub Dostálek
The sensitive analysis of small lipid extracellular vesicles (EVs) by using a grating-coupled surface plasmon resonance (GC-SPR) biosensor has been reported. In order to enable the analysis of trace amounts of EVs present in complex liquid samples, the target analyte is pre-concentrated on the sensor surface by using magnetic nanoparticles and its affinity binding is probed by wavelength interrogation of SPR. The GC-SPR has been demonstrated to allow for the implementation of efficient pulling of EVs to the sensor surface by using magnetic nanoparticles and an external magnetic field gradient applied through the sensor chip...
September 18, 2017: Analyst
https://www.readbyqxmd.com/read/28903311/development-of-hepatoma-derived-bidirectional-oval-like-cells-as-a-model-to-study-host-interactions-with-hepatitis-c-virus-during-differentiation
#13
Masahiko Ito, Suofeng Sun, Takasuke Fukuhara, Ryosuke Suzuki, Miho Tamai, Toyohiko Yamauchi, Kenji Nakashima, Yoh-Ichi Tagawa, Shigetoshi Okazaki, Yoshiharu Matsuura, Takaji Wakita, Tetsuro Suzuki
Directed differentiation of human stem cells including induced pluripotent stem cells into hepatic cells potentially leads to acquired susceptibility to hepatitis C virus (HCV) infection. However, cellular determinants that change their expression during cell reprogramming or hepatic differentiation and are pivotal for supporting the HCV life cycle remain unclear. In this study, by introducing a set of reprogramming factors, we established HuH-7-derived oval-like cell lines, Hdo-17 and -23, which possess features of bipotential liver precursors...
August 15, 2017: Oncotarget
https://www.readbyqxmd.com/read/28886485/development-of-an-epitope-based-vaccine-inhibiting-immune-cells-rolling-and-migration-against-atherosclerosis-using-in-silico-approaches
#14
Mehdi Tourani, Ahmad Karkhah, Ali Najafi
Atherosclerosis is a chronic inflammatory disease characterized by formation of pro-oxidative lipids in large and medium-sized vessels. Over the years, many treatments and drugs have entered the market to improve atherosclerosis and autoantigen-mediated active immunization is currently considered as a beneficial method. Therefore, this study was conducted to design a novel epitope-based vaccine against atherosclerosis employing CD99, CD81 and CD99L2 antigens. In this way, structural vaccinology approaches were used to design a novel multi-epitope vaccine against atherosclerosis...
September 1, 2017: Computational Biology and Chemistry
https://www.readbyqxmd.com/read/28880937/differential-expression-of-tetraspanin-superfamily-members-in-dendritic-cell-subsets
#15
Malou Zuidscherwoude, Kuntal Worah, Alie van der Schaaf, Sonja I Buschow, Annemiek B van Spriel
Dendritic cells (DCs), which are essential for initiating immune responses, are comprised of different subsets. Tetraspanins organize dendritic cell membranes by facilitating protein-protein interactions within the so called tetraspanin web. In this study we analyzed expression of the complete tetraspanin superfamily in primary murine (CD4+, CD8+, pDC) and human DC subsets (CD1c+, CD141+, pDC) at the transcriptome and proteome level. Different RNA and protein expression profiles for the tetraspanin genes across human and murine DC subsets were identified...
2017: PloS One
https://www.readbyqxmd.com/read/28878075/characterization-of-the-quasi-enveloped-hepatitis-e-virus-particles-released-by-the-cellular-exosomal-pathway
#16
Shigeo Nagashima, Masaharu Takahashi, Tominari Kobayashi, Tsutomu Nishizawa, Takashi Nishiyama, Putu Prathiwi Primadharsini, Hiroaki Okamoto
Our previous studies demonstrated that membrane-associated hepatitis E virus (HEV) particles-now considered "quasi-enveloped particles"-are present in the multivesicular body with intraluminal vesicles (exosomes) in infected cells and that the release of HEV virions is related to the exosomal pathway. In this study, we characterized exosomes purified from the culture supernatants of HEV-infected PLC/PRF/5 cells. Purified CD63-, CD9-, or CD81-positive exosomes derived from the culture supernatants of HEV-infected cells that had been cultivated in serum-free medium were found to contain HEV RNA and the viral capsid (ORF2) and ORF3 proteins, as determined by reverse transcription-PCR (RT-PCR) and Western blotting, respectively...
November 15, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28871089/cd81-association-with-samhd1-enhances-hiv-1-reverse-transcription-by-increasing-dntp-levels
#17
Vera Rocha-Perugini, Henar Suárez, Susana Álvarez, Soraya López-Martín, Gina M Lenzi, Felipe Vences-Catalán, Shoshana Levy, Baek Kim, María A Muñoz-Fernández, Francisco Sánchez-Madrid, Maria Yáñez-Mó
In this study, we report that the tetraspanin CD81 enhances human immunodeficiency virus (HIV)-1 reverse transcription in HIV-1-infected cells. This is enabled by the direct interaction of CD81 with the deoxynucleoside triphosphate phosphohydrolase SAMHD1. This interaction prevents endosomal accumulation and favours the proteasome-dependent degradation of SAMHD1. Consequently, CD81 depletion results in SAMHD1 increased expression, decreasing the availability of deoxynucleoside triphosphates (dNTP) and thus HIV-1 reverse transcription...
November 2017: Nature Microbiology
https://www.readbyqxmd.com/read/28840975/cardioprotection-mediated-by-exosomes-is-impaired-in-the-setting-of-type-ii-diabetes-but-can-be-rescued-by-the-use-of-non-diabetic-exosomes-in-vitro
#18
Sean M Davidson, Jaime A Riquelme, Kaloyan Takov, Jose M Vicencio, Claire Boi-Doku, Vanessa Khoo, Christian Doreth, Dina Radenkovic, Sergio Lavandero, Derek M Yellon
Many patients with ischaemic heart disease also have diabetes. As myocardial infarction is a major cause of mortality and morbidity in these patients, treatments that increase cell survival in response to ischaemia and reperfusion are needed. Exosomes-nano-sized, lipid vesicles released from cells-can protect the hearts of non-diabetic rats. We previously showed that exosomal HSP70 activates a cardioprotective signalling pathway in cardiomyocytes culminating in ERK1/2 and HSP27 phosphorylation. Here, we investigated whether the exosomal cardioprotective pathway remains intact in the setting of type II diabetes...
August 25, 2017: Journal of Cellular and Molecular Medicine
https://www.readbyqxmd.com/read/28828654/extracellular-vesicle-isolation-and-analysis-by-western-blotting
#19
Emma J K Kowal, Dmitry Ter-Ovanesyan, Aviv Regev, George M Church
Extracellular vesicles (EVs) are released by mammalian cells and are thought to be important mediators of intercellular communication. There are many methods for isolating EVs from cell culture media, but the most popular methods involve purification based on ultracentrifugation . Here, we provide a detailed protocol for isolating EVs by differential ultracentrifugation and analyzing EV proteins (such as the tetraspanins CD9 , CD63 and CD81 ) by western blotting.
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28759649/the-tetraspanin-cd9-facilitates-mers-coronavirus-entry-by-scaffolding-host-cell-receptors-and-proteases
#20
James T Earnest, Michael P Hantak, Kun Li, Paul B McCray, Stanley Perlman, Tom Gallagher
Infection by enveloped coronaviruses (CoVs) initiates with viral spike (S) proteins binding to cellular receptors, and is followed by proteolytic cleavage of receptor-bound S proteins, which prompts S protein-mediated virus-cell membrane fusion. Infection therefore requires close proximity of receptors and proteases. We considered whether tetraspanins, scaffolding proteins known to facilitate CoV infections, hold receptors and proteases together on cell membranes. Using knockout cell lines, we found that the tetraspanin CD9, but not the tetraspanin CD81, formed cell-surface complexes of dipeptidyl peptidase 4 (DPP4), the MERS-CoV receptor, and the type II transmembrane serine protease (TTSP) member TMPRSS2, a CoV-activating protease...
July 2017: PLoS Pathogens
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