Quantifying viable virus-specific T cells without a priori knowledge of fine epitope specificity.
Identification of pathogen-specific T cells has been greatly facilitated by the advent of synthetic peptide-major histocompatibility complex (MHC) tetramers. In many cases, however, specific epitopes have not been defined, necessitating detection methods that function independently of exact peptide-MHC specificity. Lymphocytes acquire surface proteins from antigen-presenting cells (APCs), and we have exploited this phenomenon to develop the T-cell recognition of APCs by protein transfer (TRAP) assay. This method is based on biotinylation and streptavidin-fluorochrome labeling of APCs, followed by subsequent acquisition of this label by antigen-specific T cells. The TRAP procedure detects MHC class I-restricted T cells regardless of their cytokine profiles or peptide-MHC affinities, and provides a versatile tool for monitoring the phenomenon of APC membrane acquisition by antigen-specific T cells.
Beadling
Carol
C
Vaccine and Gene Therapy Institute, Oregon Health & Science University, 505 NW 185th Avenue, Beaverton, Oregon 97006, USA.
Slifka
Mark K
MK
D000818:Animals; D000941:Antigens / Q000737:chemistry; D019921:Biotinylation; D002460:Cell Line; D002469:Cell Separation; D016207:Cytokines / Q000378:metabolism; D000939:Epitopes / Q000737:chemistry*; D005434:Flow Cytometry / Q000379:methods*; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D019809:Streptavidin / Q000737:chemistry; D013601:T-Lymphocytes / Q000378:metabolism / Q000821:virology*
D000941:Antigens; D016207:Cytokines; D000939:Epitopes; D019809:Streptavidin