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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV130058	1/1	Trichomonas vaginalis	Parasite	(d)(U)C DG Filtration UF	Twu O	2013	56%
Study summary Full title Trichomonas vaginalis exosomes deliver cargo to host cells and mediate hostratioparasite interactions All authors Twu O, de Miguel N, Lustig G, Stevens GC, Vashisht AA, Wohlschlegel JA, Johnson PJ Journal PLoS Pathog Abstract Trichomonas vaginalis is a common sexually transmitted parasite that colonizes the human urogential (show more...)Trichomonas vaginalis is a common sexually transmitted parasite that colonizes the human urogential tract where it remains extracellular and adheres to epithelial cells. Infections range from asymptomatic to highly inflammatory, depending on the host and the parasite strain. Here, we use a combination of methodologies including cell fractionation, immunofluorescence and electron microscopy, RNA, proteomic and cytokine analyses and cell adherence assays to examine pathogenic properties of T. vaginalis. We have found that T.vaginalis produces and secretes microvesicles with physical and biochemical properties similar to mammalian exosomes. The parasite-derived exosomes are characterized by the presence of RNA and core, conserved exosomal proteins as well as parasite-specific proteins. We demonstrate that T. vaginalis exosomes fuse with and deliver their contents to host cells and modulate host cell immune responses. Moreover, exosomes from highly adherent parasite strains increase the adherence of poorly adherent parasites to vaginal and prostate epithelial cells. In contrast, exosomes from poorly adherent strains had no measurable effect on parasite adherence. Exosomes from parasite strains that preferentially bind prostate cells increased binding of parasites to these cells relative to vaginal cells. In addition to establishing that parasite exosomes act to modulate host?parasite interactions, these studies are the first to reveal a potential role for exosomes in promoting parasite?parasite communication and host cell colonization. (hide) EV-METRIC 56% (90th 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 Parasite 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 DG Filtration UF Adj. k-factor 25.32 (pelleting) Protein markers EV: Tspan1 non-EV: Proteomics yes EV density (g/ml) 1.03-1.25 Show all info Study aim Function Sample Species Trichomonas vaginalis Sample Type Parasite Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 75 Pelleting: rotor type TLA100 Pelleting: adjusted k-factor 25.32 Wash: volume per pellet (ml) 2 Density gradient Lowest density fraction 0.25 Highest density fraction 2 Orientation Bottom-up Speed (g) 100000 Pelleting-wash: volume per pellet (mL) 3 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Western Blot Antibody details provided? No Detected EV-associated proteins Tspan1 ELISA Antibody details provided? No Detected EV-associated proteins Tspan1 Characterization: Particle analysis NTA EM EM-type transmission EM Image type Close-up

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EV-TRACK ID	EV130058
species	Trichomonas vaginalis
sample type	Parasite
condition	NAY
separation protocol	(d)(U)C DG Filtration UF
Exp. nr.	1
EV-METRIC %	56