Document Detail

Measuring pH, ROS production, maturation, and degradation in dendritic cell phagosomes using cytofluorometry-based assays.
MedLine Citation:
PMID:  19941126     Owner:  NLM     Status:  MEDLINE    
Phagosomes are complex organelles that form after ingestion by phagocytic cells of pathogens, dying cells, or cell debris. Highly dynamic interactions of phagosomes first with endosomes and then with lysosomes lead to the maturation of phagosomes into phagolysosomes. Contrary to other phagocytes, which degrade ingested particles to amino acids, dendritic cells only partially degrade ingested proteins, preserving short peptides for the onset of adaptive immune responses. We have modified a series of latex bead-based techniques, previously reported, in order to analyze phagosome maturation using flow cytometry. The analysis of the phagosomal pH, degradation, or oxidation relies on techniques based on the fate of specific probes bound to particles to be phagocytosed. These techniques are very sensitive and quantitative.
Ariel Savina; Pablo Vargas; Pierre Guermonprez; Ana-Maria Lennon; Sebastian Amigorena
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  595     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2010  
Date Detail:
Created Date:  2009-11-26     Completed Date:  2010-02-02     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  383-402     Citation Subset:  IM    
Institut Curie, INSERM U653, Immunit?? et Cancer, Paris, France.
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MeSH Terms
Dendritic Cells / cytology*,  metabolism*
Flow Cytometry / methods*
Hydrogen-Ion Concentration
Models, Biological
Phagosomes / metabolism*
Reactive Oxygen Species / metabolism*
Reg. No./Substance:
0/Reactive Oxygen Species

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