Proteoglycans are potent modulators of the biological responses of eosinophils to chemokines.
Chemokines play critical roles in governing the recruitment and activation of eosinophils at sites of allergic inflammation, particularly the asthmatic lung. However, we know little of how chemokine function is regulated post-translationally. Proteoglycans, consisting of a core protein and glycosaminoglycan (GAG) side chains, are cell surface molecules and components of the extracellular matrix that are able to bind chemokines, whilst heparin is a GAG with therapeutic value in asthma. We examined whether soluble GAG could alter the actions of chemokines in assays of eosinophil activation. Heparin inhibited intracellular calcium flux, respiratory burst and chemotactic responses of eosinophils to CCL11, but not to the chemoattractant C5a, and inhibited binding of CCL11 to CCR3. Heparin also inhibited eosinophil stimulation by CCL11, CCL24, CCL7, CCL13 and CCL5 to differing degrees, which broadly correlated with their relative affinities for heparin. Heparan sulfate and dermatan sulfate, but not chondroitin sulfate, also inhibited the actions of CCL11 and CCL13 in assays of cellular shape change and chemotaxis. Following treatment with the sulfation inhibitor chlorate or proteoglycanases, no inhibition of CCL11-induced activity was observed using either eosinophils or a CCR3-expressing transfectant cell line. This suggests that cell surface proteoglycans are not necessary for signaling via CCR3. However, the GAG context in which chemokines are expressed is likely to represent an important level of regulation of allergic inflammation.
Leukocyte Biology Section, Biomedical Sciences Division, Faculty of Medicine, Imperial College London, London, GB. f.culley@imperial.ac.uk
Fiona J
FJ
Culley
Emma J
EJ
Fadlon
Antje
A
Kirchem
Timothy J
TJ
Williams
Peter J
PJ
Jose
James E
JE
Pease
D002118:Calcium / Q000378:metabolism; D054413:Chemokine CCL11; D018925:Chemokines / Q000494:pharmacology*; D019742:Chemokines, CC / Q000037:antagonists & inhibitors; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D002704:Chlorates / Q000494:pharmacology; D015936:Complement C5a / Q000037:antagonists & inhibitors; D004804:Eosinophils / Q000187:drug effects* / Q000502:physiology; D006493:Heparin / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D018945:Monocyte Chemoattractant Proteins / Q000037:antagonists & inhibitors; D011509:Proteoglycans / Q000502:physiology*; D054397:Receptors, CCR3; D019707:Receptors, Chemokine / Q000378:metabolism; D016897:Respiratory Burst
C515456:CCL11 protein, human; C099677:CCL13 protein, human; C515350:CCR3 protein, human; D054413:Chemokine CCL11; D018925:Chemokines; D019742:Chemokines, CC; D002704:Chlorates; D018945:Monocyte Chemoattractant Proteins; D011509:Proteoglycans; D054397:Receptors, CCR3; D019707:Receptors, Chemokine; D015936:Complement C5a; D006493:Heparin; D002118:Calcium