Complexes of platelet factor 4 and heparin activate Toll-like receptor 4

Abstract

Background: In some patients, the anticoagulant heparin elicits formation of antibodies that can cause the life and/or limb-threatening syndrome known as heparin-induced thrombocytopenia (HIT). HIT antibodies target complexes formed at specific molar ratios of heparin and platelet factor 4 (PF4). The unpredictable occurrence and the mechanism of this atypical immune response to PF4:heparin complexes are poorly understood. Objective: We investigated whether complexes formed at specific PF4:heparin ratios (PHRs) might resemble molecular patterns associated with host defense responses. Methods: We used an in vitro cytokine release assay to determine whether defined PHRs caused cytokine release from human whole blood. Lipopolysaccharide (LPS) was used as a positive assay control, and some experiments included antibodies to block Toll-like receptor 4 (TLR4). Results: PF4:heparin complexes caused release of the biomarker interleukin 8 in whole blood, and the level of response varied with the stoichiometric ratio of PF4 to heparin. The profile of response to LPS and to PF4:heparin complexes varied among blood donors, and the interleukin 8 response to both LPS and PF4:heparin was inhibited by TLR4-blocking antibodies. Conclusions: Specific PF4-heparin complexes can elicit a TLR4-mediated response, suggesting that these complexes can mimic a pathogen-associated molecular pattern, and supporting the suggestion that the HIT immune response represents a misdirected host defense mechanism.