Protocol abstract
Nature Protocols 1, - 2536 - 2542 (2006)
Published online: 29 December 2006 | doi:10.1038/nprot.2006.400
Subject Categories: Cell and tissue culture | Isolation, purification and separation
A simple trogocytosis-based method to detect, quantify, characterize and purify antigen-specific live lymphocytes by flow cytometry, via their capture of membrane fragments from antigen-presenting cells
Sandrine Daubeuf1, Anne-Laure Puaux1, Etienne Joly1 & Denis Hudrisier1,2
Abstract
We have developed a method exploiting the phenomenon of trogocytosis to detect lymphocytes reacting specifically with target cells by flow cytometry. Trogocytosis is a process by which lymphocytes capture fragments of the plasma membrane from the antigen-presenting cells (APCs) expressing their cognate antigen. For this method, a label (such as a fluorescent lipid or biotin) is first incorporated in the membrane of APCs. These labeled cells are then co-cultured for a few hours with a population of cells containing the lymphocytes to be detected. After this period of stimulation, lymphocytes that have performed trogocytosis are identified by their acquisition of the label initially present on the APC membrane using flow cytometry. A major advantage of this method is its compatibility with the simultaneous detection of phenotypic and/or functional markers on the lymphocytes. Furthermore, cells can be recovered alive and active after detection of trogocytosis, and are therefore available for further characterization or even conceivably for therapeutic purposes.
- IPBS, CNRS UMR5089, Toulouse, France.
- Université Paul Sabatier, Toulouse, France.
Correspondence to: Denis Hudrisier1,2 e-mail: Denis.Hudrisier@ipbs.fr
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A-Z product listing
- Allophycocyanin(SA-APC)
- anti-human anti-bodies(BD-Pharmingen)
- anti-mouse(BD-Pharmingen)
- Cell sorter(Coulter Epics ALTRA)
- DiI(Aldrich)
- Diluent C for general membrane labeling(Sigma)
