Eotaxin Selectively Binds Heparin: AN INTERACTION THAT PROTECTS EOTAXIN FROM PROTEOLYSIS AND POTENTIATES CHEMOTACTIC ACTIVITY IN VIVO

• Published in: The Journal of biological chemistry 2007-05, Vol.282 (20), p.15238-15247
• Main Authors: Ellyard, Julia I, Simson, Ljubov, Bezos, Anna, Johnston, Kellie, Freeman, Craig, Parish, Christopher R
• Format: Article
• Language: English
• Subjects: Animals; Anticoagulants - chemistry; Anticoagulants - metabolism; Anticoagulants - pharmacology; Cathepsin G; Cathepsins - metabolism; Chemokine CCL11; Chemokines, CC - chemistry; Chemokines, CC - metabolism; Chemokines, CC - pharmacology; Chemotaxis, Leukocyte - drug effects; Endothelium, Vascular - metabolism; Endothelium, Vascular - pathology; Eosinophils - metabolism; Eosinophils - pathology; Fibrinolysin - metabolism; Heparin - chemistry; Heparin - metabolism; Heparin - pharmacology; Heparitin Sulfate - chemistry; Heparitin Sulfate - metabolism; Heparitin Sulfate - pharmacology; Inflammation - metabolism; Inflammation - pathology; Male; Mast Cells - metabolism; Mast Cells - pathology; Mice; Mice, Inbred BALB C; Mice, Transgenic; Models, Biological; Protein Binding - drug effects; Protein Processing, Post-Translational - drug effects; Serine Endopeptidases - metabolism; Th2 Cells - metabolism; Th2 Cells - pathology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M608046200

An important feature of chemokines is their ability to bind to the glycosaminoglycan (GAG) side chains of proteoglycans, predominately heparin and heparan sulfate. To date, all chemokines tested bind to immobilized heparin in vitro, as well as cell surface heparan sulfate in vitro and in vivo. These interactions play an important role in modulating the action of chemokines by facilitating the formation of stable chemokine gradients within the vascular endothelium and directing leukocyte migration, by protecting chemokines from proteolysis, by inducing chemokine oligomerization, and by facilitating transcytosis. Despite the importance of eotaxin in eosinophil differentiation and recruitment being well established, little is known about the interaction between eotaxin and GAGs and the functional consequences of such an interaction. Here we report that eotaxin binds selectively to immobilized heparin with high affinity (Kd = 1.23 x 10⁻⁸ M), but not to heparan sulfate or a range of other GAGs. The interaction of eotaxin with heparin does not promote eotaxin oligomerization but protects eotaxin from proteolysis directly by plasmin and indirectly by cathepsin G and elastase. In vivo, co-administration of eotaxin and heparin is able to significantly enhance eotaxin-mediated eosinophil recruitment in a mouse air-pouch model. Furthermore, when heparin is co-administered with eotaxin at a concentration that does not normally result in eosinophil infiltration, eosinophil recruitment occurs. In contrast, heparin does not enhance eotaxin-mediated eosinophil chemotaxis in vitro, suggesting protease protection or haptotactic gradient formation as the mechanism by which heparin enhances eotaxin action in vivo. These results suggest a role for mast cell-derived heparin in the recruitment of eosinophils, reinforcing Th2 polarization of inflammatory responses.