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Identification and manipulation of antigen specific T-cells with artificial antigen presenting cells.

Methods in molecular medicine (2007-11-07)
Eva Koffeman, Elissa Keogh, Mark Klein, Berent Prakken, Salvatore Albani
ABSTRACT

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.

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Sigma-Aldrich
Cocktail di inibitori delle proteasi, for use with mammalian cell and tissue extracts, DMSO solution
Sigma-Aldrich
Cholera Toxin B subunit, biotin conjugate, lyophilized powder