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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV120095	1/1	Homo sapiens	NAY	(d)(U)C Filtration	Wahlgren J	2012	29%
Study summary Full title Activated human T cells secrete exosomes that participate in IL-2 mediated immune response signaling All authors Wahlgren J, Karlson Tde L, Glader P, Telemo E, Valadi H Journal PLoS One Abstract It has previously been shown that nano-meter sized vesicles (30-100 nm), exosomes, secreted by antig (show more...)It has previously been shown that nano-meter sized vesicles (30-100 nm), exosomes, secreted by antigen presenting cells can induce T cell responses thus showing the potential of exosomes to be used as immunological tools. Additionally, activated CD3? T cells can secrete exosomes that have the ability to modulate different immunological responses. Here, we investigated what effects exosomes originating from activated CD3? T cells have on resting CD3? T cells by studying T cell proliferation, cytokine production and by performing T cell and exosome phenotype characterization. Human exosomes were generated in vitro following CD3? T cell stimulation with anti-CD28, anti-CD3 and IL-2. Our results show that exosomes purified from stimulated CD3? T cells together with IL-2 were able to generate proliferation in autologous resting CD3? T cells. The CD3? T cells stimulated with exosomes together with IL-2 had a higher proportion of CD8? T cells and had a different cytokine profile compared to controls. These results indicate that activated CD3? T cells communicate with resting autologous T cells via exosomes. (hide) EV-METRIC 29% (68th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin NAY Focus vesicles exosomes Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (d)(U)C Filtration Adj. k-factor 130.7 (pelleting) Protein markers EV: CD63/ CD81/ ICAM1/ MHC2/ CD9/ MHC1 non-EV: Proteomics no Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-harvesting Medium EV Depleted Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 70 Pelleting: rotor type 70Ti Pelleting: adjusted k-factor 130.7 Filtration steps 0.22µm or 0.2µm Western Blot Antibody details provided? No Detected EV-associated proteins MHC1/ MHC2/ ICAM1 ELISA Antibody details provided? No Detected EV-associated proteins MHC1/ MHC2/ ICAM1 Flow cytometry specific beads Antibody details provided? No Antibody dilution provided? No Selected surface protein(s) Yes Characterization: Particle analysis DLS

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EV-TRACK ID	EV120095
species	Homo sapiens
sample type	Cell culture
cell type	NAY
condition	NAY
separation protocol	(d)(U)C Filtration
Exp. nr.	1
EV-METRIC %	29