CD154 is transiently up-regulated on activated CD4+ T cells and plays an important role as a co-stimulatory molecule in T cell/ag-presenting cell interactions through ligation of CD40. Due to its transient expression within hours after activation, CD154 can be used as a marker for activated ag-specific CD4+ T cells. Adding a CD40-blocking antibody during the stimulation of cell suspensions prevents down-regulation of CD154 expression induced by the interaction with CD40 expressed on ag-presenting cells. Combining magnetic cell enrichment and multiparameter flow cytometry analysis of CD154+CD4+ T cells allows the direct ex vivo detection and characterization of rare ag-specific T cells.
Freshly prepare 200 μL of antibody staining mix for each well:
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