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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV130089	1/1	Mus musculus	NAY	(d)(U)C Filtration	Gehrmann U	2013	14%
Study summary Full title Synergistic induction of adaptive antitumor immunity by codelivery of antigen with alpha-galactosylceramide on exosomes All authors Gehrmann U, Hiltbrunner S, Georgoudaki AM, Karlsson MC, Näslund TI, Gabrielsson S Journal Cancer Res Abstract Exosomes and the invariant NKT (iNKT) immune cell ligand ?-galactosylceramide (?GC) may offer novel (show more...)Exosomes and the invariant NKT (iNKT) immune cell ligand ?-galactosylceramide (?GC) may offer novel tools for cancer immunotherapy. In this study, we investigated whether exosomes loaded with ?GC can activate iNKT cells and potentiate a cancer-specific adaptive immune response. ?GC loaded exosomes readily activated iNKT cells both in vitro and in vivo. Exosomes loaded with ?GC plus the model antigen ovalbumin (OVA) induced potent NK and ?? T-cell innate immune responses, and they also synergistically amplified T- and B-cell responses that were OVA specific. In contrast to soluble ?GC, which anergizes iNKT cells, we found that ?GC/OVA-loaded exosomes did not induce iNKT cell anergy but were more potent than soluble ?GC + OVA in inducing adaptive immune responses. In an OVA-expressing mouse model of melanoma, treatment of tumor-bearing mice with ?GC/OVA-loaded exosomes decreased tumor growth, increased antigen-specific CD8(+) T-cell tumor infiltration, and increased median survival, relative to control mice immunized with soluble ?GC + OVA alone. Notably, an additional injection of ?GC/OVA-loaded exosomes further augmented the treatment effects. Our findings show that exosomes loaded with protein antigen and ?GC will activate adaptive immunity in the absence of triggering iNKT-cell anergy, supporting their application in the design of a broad variety of cancer immunotherapy trials. (hide) EV-METRIC 14% (44th 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 Protein markers EV: CD54/ CD81/ CD80/ CD40/ MHC2/ CD9 non-EV: Proteomics no Show all info Study aim Function Sample Species Mus musculus Sample Type Cell culture supernatant EV-harvesting Medium EV Depleted Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 130 Filtration steps 0.22µm or 0.2µm Western Blot Antibody details provided? No Detected EV-associated proteins MHC2/ CD40/ CD54/ CD80 ELISA Antibody details provided? No Detected EV-associated proteins MHC2/ CD40/ CD54/ CD80 Flow cytometry specific beads Antibody details provided? No Antibody dilution provided? No Selected surface protein(s) Yes

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EV-TRACK ID	EV130089
species	Mus musculus
sample type	Cell culture
cell type	NAY
condition	NAY
separation protocol	(d)(U)C Filtration
Exp. nr.	1
EV-METRIC %	14