Proinflammatory cytokines potentiate thrombospondin-mediated phagocytosis of neutrophils undergoing apoptosis.

Abstract

Apoptosis leads to swift recognition, ingestion, and degradation of intact senescent neutrophils by macrophages. This protects tissues from leakage of noxious contents from dying cells and may promote resolution of inflammation. However, little has been known of the mechanisms that regulate macrophage capacity for apoptotic cells during an inflammatory response. We examined whether proinflammatory cytokines modulated phagocytosis of senescent neutrophils undergoing apoptosis by human monocyte-derived macrophages at 4 days maturity (4d M phi), an in vitro model of neutrophil "disposal" by apoptosis. Pretreatment of 4d M phi with granulocyte-macrophage-CSF increased the proportion of 4d M phi taking up apoptotic PMN in a concentration-dependent fashion by up to approximately 240%. This was by a rapid effect detectable by 4 h and exerted on the M phi, not the PMN. Granulocyte-macrophage-CSF also increased the number of apoptotic PMN taken up by each M phi. IFN-gamma, IL-1 beta, TNF-alpha, and TGF-beta 1 also enhanced phagocytosis, but IL-4 and IL-6 had no effect. In each case, the cytokine-expanded phagocytic subpopulation employed the thrombospondin (TSP)-dependent recognition mechanism defined for mature M phi, in which M phi vitronectin receptor and CD36 cooperate. However, commensurate increases in M phi expression of VnR, TSP, or CD36 were not detectable, indicating that TSP-mediated recognition can be recruited by other mechanisms. Cytokines did not recruit phosphatidylserine-dependent recognition, the other major mechanism by which some macrophage populations ingest apoptotic cells. Thus, M phi phagocytosis of apoptotic neutrophils can be potentiated by proinflammatory cytokines, suggesting a mechanism for negative feedback control of neutrophil number at inflamed sites.