Measuring pH, ROS Production, Maturation, and Degradation in Dendritic Cell Phagosomes Using Cytofluorometry-Based Assays
- 6 November 2009
- book chapter
- Published by Springer Science and Business Media LLC
- Vol. 595, 383-402
- https://doi.org/10.1007/978-1-60761-421-0_25
Abstract
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.Keywords
This publication has 7 references indexed in Scilit:
- The Small GTPase Rac2 Controls Phagosomal Alkalinization and Antigen Crosspresentation Selectively in CD8+ Dendritic CellsImmunity, 2009
- NOX2 Controls Phagosomal pH to Regulate Antigen Processing during Crosspresentation by Dendritic CellsCell, 2006
- The Protease Inhibitor Cystatin C Is Differentially Expressed among Dendritic Cell Populations, but Does Not Control Antigen PresentationThe Journal of Immunology, 2003
- Activation of Lysosomal Function During Dendritic Cell MaturationScience, 2003
- Analysis of Protease Activity in Live Antigen-presenting Cells Shows Regulation of the Phagosomal Proteolytic Contents During Dendritic Cell ActivationThe Journal of Experimental Medicine, 2002
- Structural Basis for Different Inhibitory Specificities of Human Cystatins C and DBiochemistry, 1998
- Maturation Stages of Mouse Dendritic Cells in Growth Factor–dependent Long-Term CulturesThe Journal of Experimental Medicine, 1997