FIGURE 8.

Activation of α7 nAChR inhibits nuclear mobilization and decreases the expression of NF-κB in human NK cells. Human NK cells were stimulated for 48 h with IL-12, IL-18, and IL-15, and further cultured for 18 h with cytokines in the absence (−) or presence of PNU-282987 (P), PNU-282987+α-BTX (α-BTX + P). Then, NK cells were processed for NF-κB p65 detection and analyzed by confocal microscopy (A-D) and FC (E and F). Upper panels, representative fluorescence microscopy images of NK cells in the absence (A) or presence of PNU-282987 (P) (B) or α-BTX + PNU-282987 (C). D, fluorescence was quantified as detailed under “Experimental Procedures” and graphs show mean fluorescence intensity (AU) of NK cells in cytoplasm (gray bars) and nucleus (black bars). The results are the mean ± S.E., and are cumulative of three independent experiments performed with NK cells from different donors (n = 5). At least 100 cells/condition were analyzed. E, representative histograms of the expression of NF-κB p65 determined by FC. Gray histogram, IC mAb; black line, NK cells stimulated with cytokines; dashed line, NK cells stimulated with cytokines and cultured in the presence of PNU-282987; gray line, NK cells stimulated with cytokines and cultured in the presence of α-BTX + PNU-282987. F, expression of NF-κB p65 assessed by FC. Data are shown as mean ± S.E., and are cumulative of three independent experiments performed with NK cells from different donors (n = 3). *, p < 0.05; ***, p < 0.001; one-way ANOVA test with Bonferroni post hoc test.