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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV190032	1/2	Schistosoma mansoni	NA	(d)(U)C	Marije E Kuipers	2020	57%
Study summary Full title DC-SIGN mediated internalisation of glycosylated extracellular vesicles from Schistosoma mansoni increases activation of monocyte-derived dendritic cells All authors Marije E Kuipers, Esther N M Nolte-'t Hoen, Alwin J van der Ham, Arifa Ozir-Fazalalikhan, D Linh Nguyen, Clarize M de Korne, Roman I Koning, John J Tomes, Karl F Hoffmann, Hermelijn H Smits, Cornelis H Hokke Journal J Extracell Vesicles Abstract Helminths like Schistosoma mansoni release excretory/secretory (E/S) products that modulate host imm (show more...)Helminths like Schistosoma mansoni release excretory/secretory (E/S) products that modulate host immunity to enable infection. Extracellular vesicles (EVs) are among these E/S products, yet molecular mechanisms and functionality of S. mansoni EV interaction with host immune cells is unknown. Here we demonstrate that EVs released by S. mansoni schistosomula are internalised by human monocyte-derived dendritic cells (moDCs). Importantly, we show that this uptake was mainly mediated via DC-SIGN (CD209). Blocking DC-SIGN almost completely abrogated EV uptake, while blocking mannose receptor (MR, CD206) or dendritic cell immunoreceptor (DCIR, CLEC4A) had no effect on EV uptake. Mass spectrometric analysis of EV glycans revealed the presence of surface N-glycans with terminal Galβ1-4(Fucα1-3)GlcNAc (LewisX) motifs, and a wide array of fucosylated lipid-linked glycans, including LewisX, a known ligand for DC-SIGN. Stimulation of moDCs with schistosomula EVs led to increased expression of costimulatory molecules CD86 and CD80 and regulatory surface marker PD-L1. Furthermore, schistosomula EVs increased expression of IL-12 and IL-10 by moDCs, which was partly dependent on the interaction with DC-SIGN. These results provide the first evidence that glycosylation of S. mansoni EVs facilitates the interaction with host immune cells and reveals a role for DC-SIGN and EV-associated glycoconjugates in parasite-induced immune modulation. (hide) EV-METRIC 57% (50th 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 Schisostomula (larval stage) culture supernatant Sample origin Control condition Focus vesicles extracellular vesicle 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 Adj. k-factor 213.2 (pelleting) / 213.2 (washing) Protein markers EV: None non-EV: None Proteomics yes Show all info Study aim Mechanism of uptake/transfer, Identification of content (omics approaches) Sample Species Schistosoma mansoni Sample Type Schisostomula (larval stage) culture supernatant 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) 80 Pelleting: rotor type SW 41 Ti Pelleting: speed (g) 120000 Pelleting: adjusted k-factor 213.2 Wash: time (min) 60 Wash: Rotor Type SW 41 Ti Wash: speed (g) 120000 Wash: adjusted k-factor 213.2 Characterization: Protein analysis PMID previous EV protein analysis 26443722 Protein Concentration Method microBCA Protein Yield (µg) 6 Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Size range/distribution Reported size (nm) 30-650 EV concentration Yes Particle yield 23300000000 EM EM-type Transmission-EM/ Cryo-EM Image type Close-up, Wide-field Report size (nm) 35-190;30-715

	EV190032	2/2	Schistosoma mansoni	NA	(d)(U)C	Marije E Kuipers	2020	57%
Study summary Full title DC-SIGN mediated internalisation of glycosylated extracellular vesicles from Schistosoma mansoni increases activation of monocyte-derived dendritic cells All authors Marije E Kuipers, Esther N M Nolte-'t Hoen, Alwin J van der Ham, Arifa Ozir-Fazalalikhan, D Linh Nguyen, Clarize M de Korne, Roman I Koning, John J Tomes, Karl F Hoffmann, Hermelijn H Smits, Cornelis H Hokke Journal J Extracell Vesicles Abstract Helminths like Schistosoma mansoni release excretory/secretory (E/S) products that modulate host imm (show more...)Helminths like Schistosoma mansoni release excretory/secretory (E/S) products that modulate host immunity to enable infection. Extracellular vesicles (EVs) are among these E/S products, yet molecular mechanisms and functionality of S. mansoni EV interaction with host immune cells is unknown. Here we demonstrate that EVs released by S. mansoni schistosomula are internalised by human monocyte-derived dendritic cells (moDCs). Importantly, we show that this uptake was mainly mediated via DC-SIGN (CD209). Blocking DC-SIGN almost completely abrogated EV uptake, while blocking mannose receptor (MR, CD206) or dendritic cell immunoreceptor (DCIR, CLEC4A) had no effect on EV uptake. Mass spectrometric analysis of EV glycans revealed the presence of surface N-glycans with terminal Galβ1-4(Fucα1-3)GlcNAc (LewisX) motifs, and a wide array of fucosylated lipid-linked glycans, including LewisX, a known ligand for DC-SIGN. Stimulation of moDCs with schistosomula EVs led to increased expression of costimulatory molecules CD86 and CD80 and regulatory surface marker PD-L1. Furthermore, schistosomula EVs increased expression of IL-12 and IL-10 by moDCs, which was partly dependent on the interaction with DC-SIGN. These results provide the first evidence that glycosylation of S. mansoni EVs facilitates the interaction with host immune cells and reveals a role for DC-SIGN and EV-associated glycoconjugates in parasite-induced immune modulation. (hide) EV-METRIC 57% (50th 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 Schisostomula (larval stage) culture supernatant Sample origin Control condition Focus vesicles extracellular vesicle 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 Adj. k-factor 213.2 (pelleting) / 83.21 (washing) Protein markers EV: None non-EV: None Proteomics yes Show all info Study aim Mechanism of uptake/transfer, Identification of content (omics approaches) Sample Species Schistosoma mansoni Sample Type Schisostomula (larval stage) culture supernatant 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) 65 Pelleting: rotor type SW 41 Ti Pelleting: speed (g) 120000 Pelleting: adjusted k-factor 213.2 Wash: time (min) 65 Wash: Rotor Type TLS-55 Wash: speed (g) 120000 Wash: adjusted k-factor 83.21 Characterization: Protein analysis PMID previous EV protein analysis 26443722 Protein Concentration Method microBCA Protein Yield (µg) 6 Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Size range/distribution Reported size (nm) 30-650 EV concentration Yes Particle yield 23300000000 EM EM-type Transmission-EM Image type Close-up, Wide-field Report size (nm) 35-190

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EV-TRACK ID	EV190032
species	Schistosoma mansoni
sample type	Schisostomula (larval stage) culture supernatant
condition	Control condition
separation protocol	(d)(U)C	(d)(U)C
Exp. nr.	1	2
EV-METRIC %	57	57