The CC chemokine eotaxin (CCL11) is a partial agonist of CC chemokine receptor 2b.
Despite sharing considerable homology with the members of the monocyte chemoattractant protein (MCP) family, the CC chemokine eotaxin (CCL11) has previously been reported to signal exclusively via the receptor CC chemokine receptor 3 (CCR3). Using the monocyte cell line THP-1, we investigated the relative abilities of eotaxin and MCPs 1-4 to induce CCR2 signaling, employing assays of directed cell migration and intracellular calcium flux. Surprisingly, 1 microm concentrations of eotaxin were able to recruit THP-1 cells in chemotaxis assays, and this migration was sensitive to antagonism of CCR2 but not CCR3. Radiolabeled eotaxin binding assays performed on transfectants bearing CCR2b or CCR3 confirmed eotaxin binding to CCR2 with a K(d) of 7.50 +/- 3.30 nm, compared with a K(d) of 1.68 +/- 0.91 nm at CCR3. In addition, whereas 1 microm concentrations of eotaxin were able to recruit CCR2b transfectants, substimulatory concentrations of eotaxin inhibited MCP-1-induced chemotaxis of CCR2b transfectants and also inhibited MCP-1-induced intracellular calcium flux of THP-1 cells. Collectively, these findings suggest that eotaxin is a partial agonist of the CCR2b receptor. A greater understanding of the interaction of CCR2 with all of its ligands, both full and partial agonists, may aid the rational design of specific antagonists that hold great promise as future therapeutic treatments for a variety of inflammatory disorders.
Martinelli
R
R
Leukocyte Biology Section, Biomedical Sciences Division, Faculty of Medicine, Sir Alexander Fleming Building, Imperial College of Science, Technology, and Medicine, South Kensington, London SW7 2AZ, United Kingdom.
Sabroe
I
I
LaRosa
G
G
Williams
T J
TJ
Pease
J E
JE
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002465:Cell Movement; D002469:Cell Separation; D054413:Chemokine CCL11; D019742:Chemokines, CC / Q000737:chemistry* / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D006801:Humans; D007700:Kinetics; D008024:Ligands; D051379:Mice; D008969:Molecular Sequence Data; D011485:Protein Binding; D054390:Receptors, CCR2; D019707:Receptors, Chemokine / Q000819:agonists*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D014162:Transfection
D000911:Antibodies, Monoclonal; C515456:CCL11 protein, human; C515334:CCR2 protein, human; C515457:Ccl11 protein, mouse; C515335:Ccr2 protein, mouse; D054413:Chemokine CCL11; D019742:Chemokines, CC; D008024:Ligands; D054390:Receptors, CCR2; D019707:Receptors, Chemokine; D002118:Calcium