Enumeration, phenotyping, and identification of activation events in conjugates between T cells and antigen-presenting cells by flow cytometry.
Microscopic analysis of T cell-antigen-presenting cell (T cell:APC) interactions at the single cell level has been a powerful, but tedious and subjective, technique. In this paper, we describe a rapid and quantitative method to identify T cell:APC conjugates using succinimidyl ester dyes, which irreversibly label free amine groups on the cell surface. The labeled cell conjugates and subsequent activation events are detected by flow cytometry.
Grebe
Kristie M
KM
Integrated Department of Immunology, National Jewish Medical and Research Center and the University of Colorado Health Sciences Center, 1400 Jackson Street, Denver, CO 80206, USA.
Potter
Terry A
TA
D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology* / Q000378:metabolism*; D016827:CD8 Antigens / Q000032:analysis; D002450:Cell Communication / Q000276:immunology*; D002452:Cell Count; D002460:Cell Line; D002463:Cell Membrane Permeability; D005434:Flow Cytometry / Q000379:methods; D006834:Hydrazines / Q000032:analysis; D016130:Immunophenotyping; D008198:Lymph Nodes / Q000166:cytology; D008213:Lymphocyte Activation; D018655:Lymphocyte Count; D051379:Mice; D008822:Mice, Transgenic; D019000:Phosphotyrosine / Q000032:analysis / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism*
C417561:Alexa 488 hydrazide; D016827:CD8 Antigens; D006834:Hydrazines; D019000:Phosphotyrosine