Abstract
Neutrophils are the first responders of the inflammatory response. They are characterized by their potent cytotoxic content but also by their limited lifetime. This short half-life is thought to be a self-protecting mechanism for the host, as highlighted by the numerous pathologies associated with imbalanced neutrophil survival. Neutrophil spontaneous death is the prototype of programmed cell death, harboring all the phenotypic hallmarks of apoptosis and dependent on the activation of the effector caspase-3. However, the pathways regulating neutrophil spontaneous death remain ill-defined. In a recent publication, we determined that in aging neutrophils, the cleavage and activation of caspase-3 was mediated by the serine protease Proteinase 3 (PR3), and was independent of the canonical extrinsic and intrinsic apoptosis pathways. In mature neutrophils, PR3 was stored in granules and progressively released to the cytosol during neutrophil aging. The release of PR3 was dependent on lysosomal membrane permeabilization (LMP). Once in the cytosol, PR3 cleaved procaspase-3 at a site upstream of the caspase-9 cleavage site, leading to caspase-3 activation. Inhibition, knockdown or knockout of PR3 delayed neutrophil apoptosis in vitro and in vivo. The adoptive transfer of both WT and PR3-deficient neutrophils to WT mice revealed that the delayed death of neutrophils lacking PR3 in vivo was due to an altered intrinsic apoptosis/survival pathway and not to difference in the inflammatory microenvironment. The cytosolic inhibitor of serine proteases serpin b1 counterbalanced the activity of PR3 in the cytosol of neutrophils, and the deletion of serpinb1 in neutrophils accelerated their spontaneous death. In summary, our results reveal that PR3 and serpinB1 are part of a newly characterized apoptosis pathway, regulating caspase-3 activation and neutrophil spontaneous death and the survival of neutrophils during inflammation.