Chapter title |
Measurement of Export to the Cytosol in Dendritic Cells Using a Cytofluorimetry-Based Assay.
|
---|---|
Chapter number | 13 |
Book title |
Dendritic Cell Protocols
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3606-9_13 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3604-5, 978-1-4939-3606-9
|
Authors |
Omar I. Vivar, Joao G. Magalhaes, Sebastian Amigorena |
Editors |
Elodie Segura, Nobuyuki Onai |
Abstract |
The presentation of exogenous antigens on MHC class I molecules, known as cross-presentation, is a key function of dendritic cells (DCs). Cross-presentation via the cytosolic pathway involves antigen export from endocytic compartments to the cytosol. We have recently developed a cytofluorimetry-based assay to examine the kinetics and the efficiency of antigen export to the cytosol in DC populations. In this assay, DCs are loaded with a FRET-sensitive cytosolic substrate of β-lactamase, CCF4. Following uptake of β-lactamase by the DCs, the enzyme undergoes export to the cytosol leading to cleavage of the FRET dye. This cleavage and switch of fluorescence are analyzed by flow cytometry, allowing a quantitative measurement of this event. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 10 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 4 | 40% |
Other | 1 | 10% |
Student > Master | 1 | 10% |
Researcher | 1 | 10% |
Professor > Associate Professor | 1 | 10% |
Other | 0 | 0% |
Unknown | 2 | 20% |
Readers by discipline | Count | As % |
---|---|---|
Immunology and Microbiology | 3 | 30% |
Agricultural and Biological Sciences | 2 | 20% |
Biochemistry, Genetics and Molecular Biology | 1 | 10% |
Chemistry | 1 | 10% |
Unknown | 3 | 30% |