EV-TRACK

Search > Results

You searched for: EV230894 (EV-TRACK ID)

Showing 1 - 1 of 1

Results list Study Tree

Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV230894	1/1	Staphylococcus aureus	Newbould 305 (ATCC 29740)	(d)(U)C DG Filtration UF	Tartaglia NR	2018	71%
Study summary Full title Extracellular Vesicles Elicit an Immunostimulatory Response on the Murine Mammary Gland. All authors Tartaglia NR, Breyne K, Meyer E, Cauty C, Jardin J, Chrétien D, Dupont A, Demeyere K, Berkova N, Azevedo V, Guédon E, Le Loir Y Journal Front Cell Infect Microbiol Abstract is a major pathogen responsible for bovine mastitis, the most common and costly disease affecting da (show more...)is a major pathogen responsible for bovine mastitis, the most common and costly disease affecting dairy cattle. naturally releases extracellular vesicles (EVs) during its growth. EVs play an important role in the bacteria-bacteria and bacteria-host interactions and are notably considered as nanocarriers that deliver virulence factors to the host tissues. Whether EVs play a role in a mastitis context is still unknown. In this work, we showed that Newbould 305 (N305), a bovine mastitis isolate, has the ability to generate EVs with a designated protein content. Purified N305-secreted EVs were not cytotoxic when tested on MAC-T and PS, two bovine mammary epithelial cell lines. However, they induced the gene expression of inflammatory cytokines at levels similar to those induced by live N305. The immune response to purified N305-secreted EVs was tested in a mouse model for bovine mastitis and their immunogenic effect was compared to that of live N305, heat-killed N305 and to lipoteichoic acid (LTA). Clinical and histopathological signs were evaluated and pro-inflammatory and chemotactic cytokine levels were measured in the mammary gland 24 h post-inoculation. Live induced a significantly stronger inflammatory response than that of any other condition tested. Nevertheless, N305-secreted EVs induced a dose-dependent neutrophil recruitment and the production of a selected set of pro-inflammatory mediators as well as chemokines. This immune response elicited by intramammary N305-secreted EVs was comparable to that of heat-killed N305 and, partly, by LTA. These results demonstrated that N305-secreted EVs induce a mild inflammatory response distinct from the live pathogen after intramammary injection. Overall, our combined and data suggest that EVs are worth to be investigated to better understand the pathogenesis and are relevant tools to develop strategies against bovine mastitis. (hide) EV-METRIC 71% (96th 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 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 (Differential) (ultra)centrifugation Density gradient Filtration Ultrafiltration Protein markers EV: None non-EV: None Proteomics yes EV density (g/ml) 1.08–1.13 Show all info Study aim Function/Identification of content (omics approaches) Sample Species Staphylococcus aureus Sample Type Cell culture supernatant EV-producing cells Newbould 305 (ATCC 29740) EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: speed (g) 150000 Density gradient Type Discontinuous Lowest density fraction 8% Highest density fraction 68% Speed (g) 100000 Duration (min) 150 Fraction processing Centrifugation Pelleting: speed (g) 150000 Pelleting: adjusted k-factor TDB Filtration steps 0.22µm or 0.2µm Ultra filtration Cut-off size (kDa) 100 Membrane type NS Characterization: Protein analysis Protein Concentration Method Bradford Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) 126 EV concentration Yes Particle yield particles per milliliter of starting sample: 4.00E+09 TRPS Report type Mean Reported size (nm) 67 EV concentration Yes Particle yield particles per milliliter of starting sample: 4.00E+09 EM EM-type Transmission-EM/ Cryo-EM Image type Close-up, Wide-field Report size (nm) 91

				1 - 1 of 1

EV-TRACK ID	EV230894
species	Staphylococcus aureus
sample type	Cell culture
cell type	Newbould 305 (ATCC 29740)
condition	Control condition
separation protocol	dUC/ Density gradient/ Filtration/ Ultrafiltration
Exp. nr.	1
EV-METRIC %	71