IL-21 and CpG ODN induce B-CLL cell expression of lysosome-associated molecular protein 1 (LAMP-1, CD107a) and granzyme B. PBMCs from 7 patients with B-CLL were cultured in the presence of CpG ODN, IL-21, or both. Expression of CD107a (LAMP-1) on CD19⁺ B-CLL cells was determined using FACS analysis. (A-B) Dot plots from 1 representative experiment of 7 show percentages of CD19⁺ B-CLL cells with increased side scatter and CD107a expression. The bar graph illustrates relative median fluorescence intensities (MFI) for CD107a expression compared with unstimulated cells. Error bars indicate SEM. (C) PBMCs from 5 patients with B-CLL were isolated and cultured in the presence of IL-21, CpG ODN, or both. For the last 4 hours, 1 μg/mL brefeldin A was added to the cells. Cells were then fixed, permeabilized, and stained with antibodies to anti-CD19 and granzyme B. The boldfaced number in Figure 4C represents the percentage of granzyme B-positive cells when treated with IL-21 and CpG ODN. One representative experiment of 5 is shown. (D) PBMCs from 5 patients with B-CLL were stained with a granzyme B colorimetric substrate, then cultured as outlined for panel C. The bar graph shows the percentage of CD19⁺ cells with substrate that was activated by granzyme B. Error bars indicate SEM.