C5a stimulation induces caspase-1 activation and mature IL-1 production in human peripheral blood mononuclear cells

The complement component C5a contributes to the recruitment of immune cells to inflamed tissues and local inflammation. The proinflammatory cytokine interleukin (IL)-1 beta is also related to inflammatory disorders through inflammasome activation. However, the association between inflammasome activation and C5a is unclear. Human peripheral blood mononuclear cells (PBMCs) were stimulated with C5a and measured for IL-1 beta secretion by enzyme-linked immunosorbent assay (ELISA). The pro-IL-1 beta expression in cell lysates was also examined by Western blot analysis. Similarly, magnetic bead-isolated CD14(+) monocyte-depleted and lymphocyte-depleted PBMCs were stimulated with C5a, and immunoblot analysis was performed using an anti-cleaved-IL-1 beta (p17) antibody. FACS was performed to detect caspase-1-activated cells. C5a-stimulated PBMCs produced IL-1 beta in C5a concentration-dependent manner. The protein levels of pro-IL-1 beta in the cell lysates were significantly increased. Furthermore, the cleaved-IL-1 beta (p17) was faintly detected in the same lysates. Active caspase-1 was demonstrated in C5a-simulated CD14(+) monocytes by FACS. Cleaved-IL-1 beta (p17) was demonstrated in the supernatant of C5a-stimulated PBMCs. Lymphocyte-depleted PBMCs stimulated with C5a but monocyte-depleted PBMCs produced cleaved-IL-1 beta (p17). C5a induced the production of mature IL-1 beta in PBMCs. The IL-1 beta production is mediated mainly by caspase-1 activation in CD14(+) monocytes. These results suggest that C5a alone potentiates mature IL-1 beta production mainly in monocytes.