Document Detail


Nanoparticle tracking analysis monitors microvesicle and exosome secretion from immune cells.
MedLine Citation:
PMID:  22348503     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Nanoparticle tracking analysis permits the determination of both the size distribution and relative concentration of microvesicles, including exosomes, in the supernatants of cultured cells and biological fluids. We have studied the release of microvesicles from the human lymphoblastoid T-cell lines Jurkat and CEM. Unstimulated, both cell lines release microvesicles in the size range 70-90 nm, which can be depleted from the supernatant by ultracentrifugation at 100 000 g, and by anti-CD45 magnetic beads, and which by immunoblotting also contain the exosome-associated proteins Alix and Tsg101. Incubation with known potentiators of exosome release, the ionophores monensin and A23187, resulted in a significant increase in microvesicle release that was both time and concentration dependent. Mass spectrometric analysis of proteins isolated from ultracentrifuged supernatants of A23187-treated cells revealed the presence of exosome-associated proteins including heat-shock protein 90, tubulin, elongation factor α1, actin and glyceraldehyde 3-phosphate dehydrogenase. Additionally, treatment of peripheral blood monocyte-derived dendritic cells with bacterial lipopolysaccharide displayed an increase in secreted microvesicles. Consequently, nanoparticle tracking analysis can be effectively applied to monitor microvesicle release from cells of the immune system.
Authors:
Chin Y Soo; Yaqiong Song; Ying Zheng; Elaine C Campbell; Andrew C Riches; Frank Gunn-Moore; Simon J Powis
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Immunology     Volume:  136     ISSN:  1365-2567     ISO Abbreviation:  Immunology     Publication Date:  2012 Jun 
Date Detail:
Created Date:  2012-04-24     Completed Date:  2012-06-27     Revised Date:  2014-02-20    
Medline Journal Info:
Nlm Unique ID:  0374672     Medline TA:  Immunology     Country:  England    
Other Details:
Languages:  eng     Pagination:  192-7     Citation Subset:  IM    
Copyright Information:
© 2012 The Authors. Immunology © 2012 Blackwell Publishing Ltd.
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MeSH Terms
Descriptor/Qualifier:
Antigens, CD45 / chemistry
Calcimycin / pharmacology
Calcium-Binding Proteins / chemistry
Cell Cycle Proteins / chemistry
Cell Line
Cell Tracking / methods*
DNA-Binding Proteins / chemistry
Dendritic Cells / drug effects
Endosomal Sorting Complexes Required for Transport / chemistry
Exosomes / drug effects,  immunology*
Humans
Immunomagnetic Separation
Ionophores / pharmacology
Lipopolysaccharides / pharmacology
Monensin / pharmacology
Nanoparticles*
T-Lymphocytes / drug effects,  immunology*
Transcription Factors / chemistry
Grant Support
ID/Acronym/Agency:
ETM/56//Chief Scientist Office
Chemical
Reg. No./Substance:
0/Calcium-Binding Proteins; 0/Cell Cycle Proteins; 0/DNA-Binding Proteins; 0/Endosomal Sorting Complexes Required for Transport; 0/Ionophores; 0/Lipopolysaccharides; 0/PDCD6IP protein, human; 0/Transcription Factors; 0/Tsg101 protein; 37H9VM9WZL/Calcimycin; 906O0YJ6ZP/Monensin; EC 3.1.3.48/Antigens, CD45; EC 3.1.3.48/PTPRC protein, human
Comments/Corrections

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