Figure 6.

Granzyme B and perforin within NK cells activated using BCG-Tice subcellular fractions. PBMCs from healthy donors were incubated with the indicated fractions of BCG-Tice (see Materials and Methods). At day 7, cells in suspension were recovered from the co-culture, centrifuged and analyzed by flow cytometry. The percentage of activation was determined by identifying the resting and activating lymphocyte regions by FSC ^vs SSC and further analyzed to distinguish T cells from NK cells by staining with antibodies against population markers (CD3, CD56). The percentage of activated lymphocytes was plotted for each donor (different colors) (left). The percentage of the CD56^bright population was obtained within the NK cell region and the content of Granzyme B (middle) and perforin (right) were determined by intracellular staining. Statistical analysis was performed using one-way ANOVA (*p < 0.05; **p < 0.01; ***p < 0.001; ns, non-significant) and compared with the untreated culture.