Identification and manipulation of antigen specific T-cells with artificial antigen presenting cells.
T-cells specific for a particular antigen represent a small percentage of the overall T-cell population. Detecting the presence of antigen specific T-cells in patients, animal models or populations of cultured cells has presented a challenge to researchers. The T-cell capture method described here utilizes a truly artificial method of antigen presentation and requires only 50,000 cells for the detection of the major histomcompatibility complex (MHC) class II and antigen restricted T-cells. With this method, liposomes, prepared with readily available materials, are loaded with neutravidin "rafts" comprised of MHC/peptide complexes, anti-CD28, a costimulatory molecule, and anti-LFA-1, an adhesion molecule. These artificial APCs are easily manipulated to include any MHC, antibodies to cell surface markers and/or costimulatory signals of interest thereby enabling not only T-cell identification but also the manipulation of mechanisms of T-cell activation.
Koffeman
Eva
E
Department of Medicine, University of California San Diego, La Jolla, USA.
Keogh
Elissa
E
Klein
Mark
M
Prakken
Berent
B
Albani
Salvatore
S
D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology*; D000941:Antigens / Q000276:immunology*; D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods; D005802:Genes, MHC Class II; D006801:Humans; D008055:Lipids / Q000737:chemistry; D008081:Liposomes / Q000378:metabolism; D010455:Peptides / Q000276:immunology / Q000302:isolation & purification; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*
D000941:Antigens; D008055:Lipids; D008081:Liposomes; D010455:Peptides