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You searched for: EV190053 (EV-TRACK ID)

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Experiment number
  • If needed, multiple experiments were identified in a single publication based on differing sample types, separation protocols and/or vesicle types of interest.
Species
  • Species of origin of the EVs.
Separation protocol
  • Gives a short, non-chronological overview of the different steps of the separation protocol.
    • (d)(U)C = (differential) (ultra)centrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
    • IAF = immuno-affinity capture
Details EV-TRACK ID Experiment nr. Species Sample type separation protocol First author Year EV-METRIC
EV190053 1/2 Homo sapiens Cell culture supernatant SEC
SEC (non-commercial)
UF
Benet, Susana 2021 0%

Study summary

Full title
All authors
Susana Benet, Cristina Gálvez, Francis Drobniewski, Irina Kontsevaya, Lilibeth Arias, Marta Monguió‐Tortajada, Itziar Erkizia, Victor Urrea, Ruo‐Yan Ong, Marina Luquin, Maeva Dupont, Jakub Chojnacki, Judith Dalmau, Paula Cardona, Olivier Neyrolles, Geanncarlo Lugo‐Villarino, Christel Vérollet, Esther Julián, Hansjakob Furrer, Huldrych F. Günthard, Paul R. Crocker, Gustavo Tapia, Francesc E. Borràs, Jacques Fellay, Paul J. McLaren, Amalio Telenti, Pere‐Joan Cardona, Bonaventura Clotet, Cristina Vilaplana, Javier Martinez‐Picado, Nuria Izquierdo‐Useros
Journal
J Extracell Vesicles
Abstract
The identification of individuals with null alleles enables studying how the loss of gene function a (show more...)The identification of individuals with null alleles enables studying how the loss of gene function affects infection. We previously described a non‐functional variant in SIGLEC1, which encodes the myeloid‐cell receptor Siglec‐1/CD169 implicated in HIV‐1 cell‐to‐cell transmission. Here we report a significant association between the SIGLEC1 null variant and extrapulmonary dissemination of Mycobacterium tuberculosis (Mtb) in two clinical cohorts comprising 6,256 individuals. Local spread of bacteria within the lung is apparent in Mtb‐infected Siglec‐1 knockout mice which, despite having similar bacterial load, developed more extensive lesions compared to wild type mice. We find that Siglec‐1 is necessary to induce antigen presentation through extracellular vesicle uptake. We postulate that lack of Siglec‐1 delays the onset of protective immunity against Mtb by limiting antigen exchange via extracellular vesicles, allowing for an early local spread of mycobacteria that increases the risk for extrapulmonary dissemination. (hide)
EV-METRIC
0% (median: 25% 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
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
Cell Name
THP-1-derived macrophages
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.
    • (d)(U)C = (differential) (ultra)centrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
    • IAF = immuno-affinity capture
SEC
Size-exclusion chromatography (non-commercial)
UF
Protein markers
EV: CD63/ CD9
non-EV:
Proteomics
no
Show all info
Study aim
Function/Mechanism of uptake/transfer
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
Control condition
EV-producing cells
THP-1-derived macrophages
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
overnight (16h) at >=100,000g
Separation Method
Ultra filtration
Cut-off size (kDa)
100
Membrane type
Regenerated cellulose
Size-exclusion chromatography
Total column volume (mL)
12
Sample volume/column (mL)
0.25
Resin type
Sepharose CL-2B
Other
Name other separation method
Size-exclusion chromatography (non-commercial)
Protein Concentration Method
Nanodrop Absorbance at 280nm
Flow cytometry aspecific beads
Detected EV-associated proteins
CD9/ CD63
Other 1
Flow cytometry (after non-specific association of vesicles to beads)
EV190053 2/2 Homo sapiens Cell culture supernatant SEC
SEC (non-commercial)
UF
Benet, Susana 2021 0%

Study summary

Full title
All authors
Susana Benet, Cristina Gálvez, Francis Drobniewski, Irina Kontsevaya, Lilibeth Arias, Marta Monguió‐Tortajada, Itziar Erkizia, Victor Urrea, Ruo‐Yan Ong, Marina Luquin, Maeva Dupont, Jakub Chojnacki, Judith Dalmau, Paula Cardona, Olivier Neyrolles, Geanncarlo Lugo‐Villarino, Christel Vérollet, Esther Julián, Hansjakob Furrer, Huldrych F. Günthard, Paul R. Crocker, Gustavo Tapia, Francesc E. Borràs, Jacques Fellay, Paul J. McLaren, Amalio Telenti, Pere‐Joan Cardona, Bonaventura Clotet, Cristina Vilaplana, Javier Martinez‐Picado, Nuria Izquierdo‐Useros
Journal
J Extracell Vesicles
Abstract
The identification of individuals with null alleles enables studying how the loss of gene function a (show more...)The identification of individuals with null alleles enables studying how the loss of gene function affects infection. We previously described a non‐functional variant in SIGLEC1, which encodes the myeloid‐cell receptor Siglec‐1/CD169 implicated in HIV‐1 cell‐to‐cell transmission. Here we report a significant association between the SIGLEC1 null variant and extrapulmonary dissemination of Mycobacterium tuberculosis (Mtb) in two clinical cohorts comprising 6,256 individuals. Local spread of bacteria within the lung is apparent in Mtb‐infected Siglec‐1 knockout mice which, despite having similar bacterial load, developed more extensive lesions compared to wild type mice. We find that Siglec‐1 is necessary to induce antigen presentation through extracellular vesicle uptake. We postulate that lack of Siglec‐1 delays the onset of protective immunity against Mtb by limiting antigen exchange via extracellular vesicles, allowing for an early local spread of mycobacteria that increases the risk for extrapulmonary dissemination. (hide)
EV-METRIC
0% (median: 25% 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
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
Cell Name
THP-1-derived macrophages
Sample origin
Mycobacterium tuberculosis infection
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
  • Gives a short, non-chronological overview of the
    different steps of the separation protocol.
    • (d)(U)C = (differential) (ultra)centrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
    • IAF = immuno-affinity capture
SEC
Size-exclusion chromatography (non-commercial)
UF
Protein markers
EV: CD63/ CD9
non-EV:
Proteomics
no
Show all info
Study aim
Function/Mechanism of uptake/transfer
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
Mycobacterium tuberculosis infection
EV-producing cells
THP-1-derived macrophages
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
overnight (16h) at >=100,000g
Separation Method
Ultra filtration
Cut-off size (kDa)
100
Membrane type
Regenerated cellulose
Size-exclusion chromatography
Total column volume (mL)
12
Sample volume/column (mL)
0.25
Resin type
Sepharose CL-2B
Other
Name other separation method
Size-exclusion chromatography (non-commercial)
Protein Concentration Method
Nanodrop Absorbance 280nm
Flow cytometry aspecific beads
Detected EV-associated proteins
CD9/ CD63
Other 1
Flow cytometry (after non-specific association of vesicles to beads)
1 - 2 of 2