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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV220428	1/1	Homo sapiens	Ascites	(d)(U)C Filtration	Shenoy GN	2018	44%
Study summary Full title Exosomes Associated with Human Ovarian Tumors Harbor a Reversible Checkpoint of T-cell Responses. All authors Shenoy GN, Loyall J, Maguire O, Iyer V, Kelleher RJ, Minderman H, Wallace PK, Odunsi K, Balu-Iyer SV, Bankert RB Journal Cancer Immunol Res Abstract Nano-sized membrane-encapsulated extracellular vesicles isolated from the ascites fluids of ovarian (show more...)Nano-sized membrane-encapsulated extracellular vesicles isolated from the ascites fluids of ovarian cancer patients are identified as exosomes based on their biophysical and compositional characteristics. We report here that T cells pulsed with these tumor-associated exosomes during TCR-dependent activation inhibit various activation endpoints including translocation of NFκB and NFAT into the nucleus, upregulation of CD69 and CD107a, production of cytokines, and cell proliferation. In addition, the activation of virus-specific CD8 T cells that are stimulated with the cognate viral peptides presented in the context of class I MHC is also suppressed by the exosomes. The inhibition occurs without loss of cell viability and coincidentally with the binding and internalization of these exosomes. This exosome-mediated inhibition of T cells was transient and reversible: T cells exposed to exosomes can be reactivated once exosomes are removed. We conclude that tumor-associated exosomes are immunosuppressive and represent a therapeutic target, blockade of which would enhance the antitumor response of quiescent tumor-associated T cells and prevent the functional arrest of adoptively transferred tumor-specific T cells or chimeric antigen receptor T cells. . (hide) EV-METRIC 44% (75th 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 Ascites Sample origin Control condition Focus vesicles exosome 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 (Differential) (ultra)centrifugation Filtration Protein markers EV: Alix/ CD63/ CD81/ Flotillin1/ TSG101/ EpCAM non-EV: GM130 Proteomics no Show all info Study aim Function Sample Species Homo sapiens Sample Type Ascites Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g Between 800 g and 10,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: speed (g) 200000 Filtration steps 0.2 or 0.22 µm Characterization: Protein analysis Protein Concentration Method Not determined Other 1 Antibody array Detected EV-associated proteins Alix/ CD63/ CD81/ Flotillin1/ TSG101/ EpCAM Not detected contaminants GM130 Characterization: Lipid analysis Yes Characterization: Particle analysis NTA Report type Modus Reported size (nm) 60-80 EM EM-type Transmission-EM Image type Wide-field

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EV-TRACK ID	EV220428
species	Homo sapiens
sample type	Ascites
condition	Control condition
separation protocol	dUC/ Filtration
Exp. nr.	1
EV-METRIC %	44