Macrophage-derived Chemokine Is a Functional Ligand for the CC Chemokine Receptor 4

Toshio Imai,David Chantry,Carol J Raport,Christi L Wood,Miyuki Nishimura,Ronald Godiska,Osamu Yoshie,Patrick W Gray

doi:10.1074/jbc.273.3.1764

Abstract

Macrophage-derived chemokine (MDC) is a recently identified member of the CC chemokine family. MDC is not closely related to other chemokines, sharing most similarity with thymus- and activation-regulated chemokine (TARC), which contains 37% identical amino acids. Both chemokines are highly expressed in the thymus, with little expression seen in other tissues. In addition, the genes for MDC and TARC are encoded by human chromosome 16. To explore this relationship in greater detail, we have more precisely localized the MDC gene to chromosome 16q13, the same position reported for the TARC gene. We have also examined the interaction of MDC with CC chemokine receptor 4 (CCR4), recently shown to be a receptor for TARC. Using a fusion protein of MDC with secreted alkaline phosphatase, we observed high affinity binding of MDC-secreted alkaline phosphatase to CCR4-transfected L1.2 cells (Kd = 0.18 nM). MDC and TARC competed for binding to CCR4, while no binding competition was observed for six other chemokines (MCP-1, MCP-3, MCP-4, RANTES (regulated on activation normal T cell expressed and secreted), macrophage inflammatory protein-1 alpha, macrophage inflammatory protein-1 beta). MDC was tested for calcium mobilization in L1.2 cells tranfected with seven different CC chemokine receptors. MDC induced a calcium flux in CCR4-transfected cells, but other receptors did not respond to MDC. TARC, which also induced calcium mobilization in CCR4 transfectants, was unable to desensitize the response to MDC. In contrast, MDC fully desensitized a subsequent response to TARC. Both MDC and TARC functioned as chemoattractants for CCR4 transfectants, confirming that MDC is also a functional ligand for CCR4. Since MDC and TARC are both expressed in the thymus, one role for these chemokines may be to attract CCR4-bearing thymocytes in the process of T cell education and differentiation.

Highlights

Chemokines are small secreted proteins that mediate recruitment of leukocytes to sites of inflammation [1, 2]
We show here that Macrophage-derived chemokine (MDC) is a ligand for chemokine receptor 4 (CCR4) and binds with a higher affinity than thymus- and activation-regulated chemokine (TARC)
To determine the ability of MDC to cause signaling through chemokine receptors, we examined calcium mobilization in L1.2 cells expressing CCR1, CCR2B, CCR3, CCR4, CCR5, CCR6, or CCR7 (Fig. 3)

Summary

Introduction

Chemokines are small secreted proteins that mediate recruitment of leukocytes to sites of inflammation [1, 2]. Since MDC and TARC are both expressed in the thymus, one role for these chemokines may be to attract CCR4-bearing thymocytes in the process of T cell education and differentiation. TARC was recently found to be a highly specific ligand for CCR4 [13], and both are co-expressed in the thymus [19, 22].

Results

Conclusion