Exchange of extracellular domains of CCR1 and CCR5 reveals confined functions in CCL5-mediated cell recruitment

Summary

The chemokine CCL5 recruits monocytes into inflamed tissues by triggering primarily CCR1-mediated arrest on endothelial cells, whereas subsequent spreading is dominated by CCR5. The CCL5-induced arrest can be enhanced by heteromer formation with CXCL4. To identify mechanisms for receptor-specific functions, we employed CCL5 mutants and transfectants expressing receptor chimeras carrying transposed extracellular regions. Mutation of the basic 50s cluster of CCL5, a coordinative site for CCL5 surface presentation, reduced CCR5- but not CCR1-mediated arrest and transmigration. Impaired arrest was restored by exchanging the CCR5-N-terminus for that of CCR1, which supported arrest even without the 50s cluster, whereas mutation of the basic 40s cluster essential for proteoglycan binding of CCL5 could not be rescued. The enhancement of CCL5-induced arrest by CXCL4 was mediated by CCR1 requiring its third extracellular loop. The domain exchanges did not affect formation and co-localisation of receptor dimers, indicating a sensing role of the third extracellular loop for hetero-oligomers in an arrest microenvironment. Our data identify confined targetable regions of CCR1 specialised to facilitate CCL5-induced arrest and enhanced responsiveness to the CXCL4-CCL5 heteromer.

Note: The review process for this manuscript was fully handled by G. Y. H. Lip, Editor in Chief.

Keywords

Arrest - chemotaxis - inflammation - chemokine receptor chimera - RANTES

• References

• 1 Gerard C, Rollins BJ. Chemokines and disease. Nat Immunol 2001; 2: 108-115.
• 2 Proudfoot AE. Chemokine receptors: multifaceted therapeutic targets. Nat Rev Immunol 2002; 2: 106-115.
• 3 Weber C, Noels H. Atherosclerosis: current pathogenesis and therapeutic options. Nat Med 2011; 17: 1410-1422.
• 4 Blanchet X, Langer M, Weber C. et al. Touch of chemokines. Frontiers Immunol 2012; 3: 175.
• 5 Blanpain C, Migeotte I, Lee B. et al. CCR5 binds multiple CC-chemokines: MCP-3 acts as a natural antagonist. Blood 1999; 94: 1899-1905.
• 6 Schall TJ, Proudfoot AE. Overcoming hurdles in developing successful drugs targeting chemokine receptors. Nat Rev Immunol 2011; 11: 355-363.
• 7 Koenen RR, Weber C. Therapeutic targeting of chemokine interactions in atherosclerosis. Nat Rev Drug Discov 2010; 9: 141-153.
• 8 Baltus T, Weber KS, Johnson Z. et al. Oligomerisation of RANTES is required for CCR1-mediated arrest but not CCR5-mediated transmigration of leukocytes on inflamed endothelium. Blood 2003; 102: 1985-1988.
• 9 Weber C, Weber KS, Klier C. et al. Specialized roles of the chemokine receptors CCR1 and CCR5 in the recruitment of monocytes and T(H)1-like/CD45RO(+) T cells. Blood 2001; 97: 1144-1146.
• 10 Czaplewski LG, McKeating J, Craven CJ. et al. Identification of amino acid residues critical for aggregation of human CC chemokines macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and RANTES. Characterisation of active disaggregated chemokine variants. J Biol Chem 1999; 274: 16077-16084.
• 11 Martin L, Blanpain C, Garnier P. et al. Structural and functional analysis of the RANTES-glycosaminoglycans interactions. Biochemistry 2001; 40: 6303-6318.
• 12 Proudfoot AE, Fritchley S, Borlat F. et al. The BBXB motif of RANTES is the principal site for heparin binding and controls receptor selectivity. J Biol Chem 2001; 276: 10620-10626.
• 13 Segerer S, Djafarzadeh R, Grone HJ. et al. Selective binding and presentation of CCL5 by discrete tissue microenvironments during renal inflammation. J Am Soc Nephrol 2007; 18: 1835-1844.
• 14 Segerer S, Johnson Z, Rek A. et al. The basic residue cluster (55)KKWVR(59) in CCL5 is required for in vivo biologic function. Mol Immunol 2009; 46: 2533-2538.
• 15 Johnson Z, Kosco-Vilbois MH, Herren S. et al. Interference with heparin binding and oligomerisation creates a novel anti-inflammatory strategy targeting the chemokine system. J Immunol 2004; 173: 5776-5785.
• 16 Braunersreuther V, Steffens S, Arnaud C. et al. A novel RANTES antagonist prevents progression of established atherosclerotic lesions in mice. Arterioscler Thromb Vasc Biol 2008; 28: 1090-1096.
• 17 von Hundelshausen P, Koenen RR, Sack M. et al. Heterophilic interactions of platelet factor 4 and RANTES promote monocyte arrest on endothelium. Blood 2005; 105: 924-930.
• 18 Koenen RR, von Hundelshausen P, Nesmelova IV. et al. Disrupting functional interactions between platelet chemokines inhibits atherosclerosis in hyperlipidemic mice. Nat Med 2009; 15: 97-103.
• 19 Magrane M, Consortium U. UniProt Knowledgebase: a hub of integrated protein data. Database 2011; 2011 bar009.
• 20 Thompson JD, Higgins DG, Gibson TJ. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 1994; 22: 4673-4680.
• 21 Sarabi A, Kramp BK, Drechsler M. et al. CXCL4L1 inhibits angiogenesis and induces undirected endothelial cell migration without affecting endothelial cell proliferation and monocyte recruitment. J Thromb Haemost 2011; 9: 209-219.
• 22 Proudfoot AE, Handel TM, Johnson Z. et al. Glycosaminoglycan binding and oligomerisation are essential for the in vivo activity of certain chemokines. Proc Natl Acad Sci USA 2003; 100: 1885-1890.
• 23 Vasina EM, Cauwenberghs S, Feijge MA. et al. Microparticles from apoptotic platelets promote resident macrophage differentiation. Cell Death Dis 2011; 2: e211.
• 24 Thiele S, Steen A, Jensen PC. et al. Allosteric and orthosteric sites in CC chemokine receptor (CCR5), a chimeric receptor approach. J Biol Chem 2011; 286: 37543-37554.
• 25 Wu B, Chien EY, Mol CD. et al. Structures of the CXCR4 chemokine GPCR with small-molecule and cyclic peptide antagonists. Science 2010; 330: 1066-1071.
• 26 Grommes J, Alard JE, Drechsler M. et al. Disruption of platelet-derived chemokine heteromers prevents neutrophil extravasation in acute lung injury. Am J Respir Crit Care Med 2012; 185: 628-636.
• 27 Kramp BK, Sarabi A, Koenen RR. et al. Heterophilic chemokine receptor interactions in chemokine signalling and biology. Exp Cell Res 2011; 317: 655-663.
• 28 Mellado M, Rodriguez-Frade JM, Vila-Coro AJ. et al. Chemokine receptor homo- or heterodimerisation activates distinct signalling pathways. Embo J 2001; 20: 2497-2507.
• 29 Wright WR, Parzych K, Crawford D. et al. Inflammatory transcriptome profiling of human monocytes exposed acutely to cigarette smoke. PLoS One 2012; 7: e30120.
• 30 Ren M, Guo Q, Guo L. et al. Polymerisation of MIP-1 chemokine (CCL3 and CCL4) and clearance of MIP-1 by insulin-degrading enzyme. EMBO J 2010; 29: 3952-3966.
• 31 Kenakin T. Functional selectivity and biased receptor signalling. J Pharmacol Exp Therap 2011; 336: 296-302.
• 32 Braunersreuther V, Zernecke A, Arnaud C. et al. Ccr5 but not Ccr1 deficiency reduces development of diet-induced atherosclerosis in mice. Arterioscler Thromb Vasc Biol 2007; 27: 373-379.
• 33 Furuichi K, Gao JL, Horuk R. et al. Chemokine receptor CCR1 regulates inflammatory cell infiltration after renal ischemia-reperfusion injury. J Immunol 2008; 181: 8670-8676.
• 34 Horuk R, Shurey S, Ng HP. et al. CCR1-specific non-peptide antagonist: efficacy in a rabbit allograft rejection model. Immunol Lett 2001; 76: 193-201.
• 35 Horuk R. Chemokine receptor antagonists: overcoming developmental hurdles. Nat Rev Drug Discov 2009; 8: 23-33.
• 36 Bedke J, Kiss E, Schaefer L. et al. Beneficial effects of CCR1 blockade on the progression of chronic renal allograft damage. Am J Transplant 2007; 7: 527-537.