Rhodamine efflux and MRK-16 staining in T-cell subsets. (A) Rhodamine efflux in isolated mononuclear cells from a normal patient. Blank histogram (solid line), cells incubated 30 minutes in media alone. Control histogram (dotted line), cells incubated 30 minutes in media with 0.5 μg/mL rhodamine 123. PSC/Efflux histogram (dashed line), cells incubated 30 minutes in media with 0.5 μg/mL rhodamine 123 and 3 μg/mL PSC 833, washed, and incubated in rhodamine-free media with 3 μg/mL PSC 833 for 60 minutes. Efflux histogram (heavy solid line), cells incubated 30 minutes in media with 0.5 μg/mL rhodamine 123, washed, and incubated in rhodamine-free medium for 60 minutes. Staining with the appropriate T-cell marker was performed with each sample. (B) MRK-16 staining of normal mononuclear cells. Blank histogram (solid line), unstained mononuclear cells. IgG Control histogram (dotted line), cells incubated with mouse IgG1, stained with FITC-labeled horse anti-mouse antibody, then stained with the appropriate PE-labeled T-cell antibody. MRK-16 histogram (dashed line), cells were incubated in unlabeled MRK-16 antibody for 30 minutes, stained with FITC-labeled horse anti-mouse, and subsequently labeled with the desired PE-labeled T-cell marker.
