Figure 5

Confocal microscopic demonstration of apoptotic cell ingestion and class II localization in lysosomal compartments in differentiating DC. Fig. 5A: Cell populations prepared as described in Figure 1 were evaluated by confocal microscopy after fixation and permeabilization and staining. A representative activated monocyte/DC is shown after CD3 column passage and recombination with the apoptotic CTCL cells as detected by a: membrane class II-FITC (green); b: cytopolasmic APO2-PE (red, white arrows) and c: merged image demonstrating internalization of apoptotic material in a class II positive cell. A representative activated monocyte/DC is shown after CD4 column passage and recombination with viable CTCL cells as detected by d: membrane class II-FITC (green); e: cytopolasmic APO2-PE (red, white arrow) and f: merged image demonstrating absence of internalization of apoptotic material in a class II positive cell. Fig. 5B: Cells prepared as described in Fig. 5A were passed through the CD3 column and stained for a: membrane class II-PE (red); b: lysosomal membrane marker, LAMP (green); and c: merged image showing co-localization of class II molecules in lysosomal compartments. Cells obtained after passage through the CD4 column were stained for d: membrane class II-PE (red); e: lysosomal membrane marker, LAMP (green); and f: merged image showing an absence of co-localization of class II molecules in lysosomal compartments.