Macrophage phagocytosis of apoptotic neutrophils is critically regulated by the opposing actions of pro-inflammatory and anti-inflammatory agents: key role for TNF-alpha

Apoptosis of inflammatory cells and their subsequent clearance (efferocytosis) by macrophages (Mphis) are key mechanisms orchestrating successful resolution of inflammation. Although the powerful proinflammatory agents lipopolysaccharide (LPS) and tumor necrosis factor alpha (TNF-alpha) influence rates of inflammatory cell apoptosis, little is known about their effects on efferocytosis. We have demonstrated that LPS and TNF-alpha potently inhibit efferocytosis of neutrophils by monocyte-derived Mphis. Inhibition was both concentration and time dependent, although the effect of TNF-alpha was more rapid. We have found that soluble TNF receptor-I attenuated LPS inhibition of phagocytosis, indicating that TNF-alpha production is critical. Inhibition of efferocytosis by LPS was found to be positively associated with Mphi production of TNF-alpha, but negatively with interleukin-10 (IL-10) release. A critical role for IL-10 in the regulation of phagocytosis was suggested by 2 important findings: LPS inhibition was observed more rapidly in the presence of an anti-human IL-10 receptor-alpha antibody, and efferocytosis by IL-10-deficient Mphis was markedly reduced compared to wild-type Mphis. Furthermore, exogenous IL-10 and glucocorticoids reversed inhibitory effects of LPS on efferocytosis via suppression of TNF-alpha production. We suggest that efferocytosis is regulated in an autocrine manner by pro- and anti-inflammatory mediators, and the inflammatory milieu determines whether inflammation successfully resolves.