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(Received for publication, September 12, 1996, and in revised form, March 5, 1997)
From the Cancer Research Campaign Laboratories, The Beatson
Institute for Cancer Research, Garscube Estate, Switchback Road,
Bearsden, Glasgow G61 1BD, United Kingdom
The
Volume 272, Number 19,
Issue of May 9, 1997
pp. 12495-12504
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
Chemokine
Receptor, D6
COMPARISON TO THREE OTHER RELATED MACROPHAGE INFLAMMATORY
PROTEIN-1
RECEPTORS, CCR-1, CCR-3, AND CCR-5
-chemokine macrophage inflammatory
protein-1
(MIP-1) is chemotactic for many hemopoietic cell types
and can inhibit hemopoietic stem cell (HSC) proliferation, effects
mediated through G-protein coupled heptahelical receptors. We have
isolated cDNAs for seven chemokine receptors, CCR-1 to -5, MIP-1RL1, and a novel cDNA, D6. Chinese hamster ovary cells
expressing CCR-1, -3, -5, and D6 bound 125I-murine
MIP-1: the order of affinity was D6 > CCR-5 > CCR-1 > CCR-3. Each bound a distinct subset of other -chemokines: the order of competition for 125I-murine MIP-1 on D6 was
murine MIP-1 > human and murine MIP-1 > human RANTES~JE > human MCP-3 > human MCP-1. Human MIP-1 and the
-chemokines did not compete. Like other chemokine receptors, D6
induced transient increases in [Ca2+] in HEK 293 cells
upon ligand binding. D6 mRNA was abundant in lung and detectable in
many other tissues. Bone marrow cell fractionation demonstrated T-cell
and macrophage/monocyte expression of D6, and CCR-1, -3, and -5. Moreover, we could detect expression of CCR-3, CCR-5, and to a greater
extent D6 in a cell population enriched for HSCs. Thus, we have
characterized four murine chemokine receptors that are likely
involved in mediating the pro-inflammatory functions of MIP-1 and
other chemokines, and we present D6, CCR-3, and CCR-5 as candidate
receptors in MIP-1-induced HSC inhibition.
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