J. Exp. Med.
© The Rockefeller University Press
0022-1007/97/09/825/11 $2.00
Volume 186, Number 6, September 15, 1997 825-835

Cloning and Characterization of a Specific Receptor for the Novel CC Chemokine MIP-3 from Lung Dendritic Cells

By Christine A. Power,^* Dennis J. Church,^* Alexandra Meyer,^* Sami Alouani,^* Amanda E.I. Proudfoot,^* Ian Clark-Lewis, Silvano Sozzani,^§ Alberto Mantovani,^§ and Timothy N.C. Wells^*

From the ^* Geneva Biomedical Research Institute, GlaxoWellcome Research and Development, Geneva, Switzerland;  University of British Columbia, Vancouver, Canada; and ^§ Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy

Dendritic cells are potent antigen-presenting cells involved in the initiation of immune responses. The trafficking of these cells to tissues and lymph nodes is mediated by members of the chemokine family. Recently, a novel CC chemokine known as MIP-3 or liver and activation-regulated chemokine has been identified from the EMBL/GenBank/DDBJ expressed sequence tag database. In the present study, we have shown that the messenger RNA for MIP-3 is expressed predominantly in inflamed and mucosal tissues. MIP-3 produced either synthetically or by human embryonic kidney 293 cells is chemotactic for CD34⁺-derived dendritic cells and T cells, but is inactive on monocytes and neutrophils. MIP-3 was unable to displace the binding of specific CC or CXC chemokines to stable cell lines expressing their respective high affinity receptors, namely CCR1-5 and CXCR1 and CXCR2, suggesting that MIP-3 acts through a novel CC chemokine receptor. Therefore, we used degenerate oligonucleotide-based reverse transcriptase PCR to identify candidate MIP-3 receptors in lung dendritic cells. Our results show that the orphan receptor known as GCY-4, CKRL-3, or STRL-22 is a specific receptor for MIP-3, and that its activation leads to pertussis toxin-sensitive and phospholipase C-dependent intracellular Ca²⁺ mobilization when it is expressed in HEK 293 cells.

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