Heparin and enoxaparin enhance endotoxin-induced tumor necrosis factor-α production in human monocytes

Objective To determine whether heparin or the low-molecular-weight heparin enoxaparin alter lipopolysaccharide (LPS)-induced monocyte activation. Summary Background Data Heparin is widely used in clinical practice to inhibit the coagulation cascade. However, heparin also is a naturally occurring glucosaminoglycan and pleiotropic immunomodulator that binds to a variety of proteins. LPS isa component of gramnegative bacteria and is thought to be responsible for many of the deleterious effects seen in sepsis. The binding of LPS to CD14 induces a signaling cascade that results in the release of many inflammatory mediators, including tumor necrosis factor-alpha (TNF-alpha). Methods Monocytes from healthy volunteers were isolated and cultured in the presence of saline, LPS (10 ng/ml), heparin (0.1 to 1000 mu g/ml), or enoxaparin (0.1 to 1000 mu g/ml). In blocking experiments, cells were pretreated for 60 minutes with the mono-clonal anti-CD14 antibody MY4 (10 mu g/ml) or with isotype-matched control IgG(2) (10 mu g/ml). TNF-alpha values were measured with enzyme-linked immunosorbent assay. Significance was assessed with analysis of variance. Results Heparin (10 to 1000 mu g/ml) and enoxaparin (1000 mu g/ml) significantly enhanced LPS-induced TNF-alpha release. Heparin (1000 mu g/ml) or enoxaparin (1000 mu g/ml) did not produce TNF-alpha in the absence of LPS. Blockade of CD14 abrogated both LPS-induced TNF-alpha release and the effect of heparin or enoxaparin to enhance LPS-induced TNF-alpha release. Conclusions The effect of heparin to enhance LPS-induced TNF-alpha release is a biologic phenomenon that reveals a novel and potentially important host defense mechanism during endotoxemia and sepsis. Binding of LPS to CD14 is necessary to induce this phenomenon, suggesting that both heparin and enoxaparin induce signaling mechanisms that are downstream from the initial binding of LPS on CD14.