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Cloning and functional characterization of the rabbit C-C chemokine receptor 2

BACKGROUND: CC-family chemokine receptor 2 (CCR2) is implicated in the trafficking of blood-borne monocytes to sites of inflammation and is implicated in the pathogenesis of several inflammatory diseases such as rheumatoid arthritis, multiple sclerosis and atherosclerosis. The major challenge in the...

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Autores principales: Lu, Deshun, Yuan, Xiu-juan, Evans, Robert J, Pappas, Amy T, Wang, He, Su, Eric W, Hamdouchi, Chafiq, Venkataraman, Chandrasekar
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2005
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1182369/
https://www.ncbi.nlm.nih.gov/pubmed/16001983
http://dx.doi.org/10.1186/1471-2172-6-15
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author Lu, Deshun
Yuan, Xiu-juan
Evans, Robert J
Pappas, Amy T
Wang, He
Su, Eric W
Hamdouchi, Chafiq
Venkataraman, Chandrasekar
author_facet Lu, Deshun
Yuan, Xiu-juan
Evans, Robert J
Pappas, Amy T
Wang, He
Su, Eric W
Hamdouchi, Chafiq
Venkataraman, Chandrasekar
author_sort Lu, Deshun
collection PubMed
description BACKGROUND: CC-family chemokine receptor 2 (CCR2) is implicated in the trafficking of blood-borne monocytes to sites of inflammation and is implicated in the pathogenesis of several inflammatory diseases such as rheumatoid arthritis, multiple sclerosis and atherosclerosis. The major challenge in the development of small molecule chemokine receptor antagonists is the lack of cross-species activity to the receptor in the preclinical species. Rabbit models have been widely used to study the role of various inflammatory molecules in the development of inflammatory processes. Therefore, in this study, we report the cloning and characterization of rabbit CCR2. Data regarding the activity of the CCR2 antagonist will provide valuable tools to perform toxicology and efficacy studies in the rabbit model. RESULTS: Sequence alignment indicated that rabbit CCR2 shares 80 % identity to human CCR2b. Tissue distribution indicated that rabbit CCR2 is abundantly expressed in spleen and lung. Recombinant rabbit CCR2 expressed as stable transfectants in U-937 cells binds radiolabeled (125)I-mouse JE (murine MCP-1) with a calculated K(d )of 0.1 nM. In competition binding assays, binding of radiolabeled mouse JE to rabbit CCR2 is differentially competed by human MCP-1, -2, -3 and -4, but not by RANTES, MIP-1α or MIP-1β. U-937/rabbit CCR2 stable transfectants undergo chemotaxis in response to both human MCP-1 and mouse JE with potencies comparable to those reported for human CCR2b. Finally, TAK-779, a dual CCR2/CCR5 antagonist effectively inhibits the binding of (125)I-mouse JE (IC(50 )= 2.3 nM) to rabbit CCR2 and effectively blocks CCR2-mediated chemotaxis. CONCLUSION: In this study, we report the cloning of rabbit CCR2 and demonstrate that this receptor is a functional chemotactic receptor for MCP-1.
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spelling pubmed-11823692005-08-04 Cloning and functional characterization of the rabbit C-C chemokine receptor 2 Lu, Deshun Yuan, Xiu-juan Evans, Robert J Pappas, Amy T Wang, He Su, Eric W Hamdouchi, Chafiq Venkataraman, Chandrasekar BMC Immunol Research Article BACKGROUND: CC-family chemokine receptor 2 (CCR2) is implicated in the trafficking of blood-borne monocytes to sites of inflammation and is implicated in the pathogenesis of several inflammatory diseases such as rheumatoid arthritis, multiple sclerosis and atherosclerosis. The major challenge in the development of small molecule chemokine receptor antagonists is the lack of cross-species activity to the receptor in the preclinical species. Rabbit models have been widely used to study the role of various inflammatory molecules in the development of inflammatory processes. Therefore, in this study, we report the cloning and characterization of rabbit CCR2. Data regarding the activity of the CCR2 antagonist will provide valuable tools to perform toxicology and efficacy studies in the rabbit model. RESULTS: Sequence alignment indicated that rabbit CCR2 shares 80 % identity to human CCR2b. Tissue distribution indicated that rabbit CCR2 is abundantly expressed in spleen and lung. Recombinant rabbit CCR2 expressed as stable transfectants in U-937 cells binds radiolabeled (125)I-mouse JE (murine MCP-1) with a calculated K(d )of 0.1 nM. In competition binding assays, binding of radiolabeled mouse JE to rabbit CCR2 is differentially competed by human MCP-1, -2, -3 and -4, but not by RANTES, MIP-1α or MIP-1β. U-937/rabbit CCR2 stable transfectants undergo chemotaxis in response to both human MCP-1 and mouse JE with potencies comparable to those reported for human CCR2b. Finally, TAK-779, a dual CCR2/CCR5 antagonist effectively inhibits the binding of (125)I-mouse JE (IC(50 )= 2.3 nM) to rabbit CCR2 and effectively blocks CCR2-mediated chemotaxis. CONCLUSION: In this study, we report the cloning of rabbit CCR2 and demonstrate that this receptor is a functional chemotactic receptor for MCP-1. BioMed Central 2005-07-07 /pmc/articles/PMC1182369/ /pubmed/16001983 http://dx.doi.org/10.1186/1471-2172-6-15 Text en Copyright © 2005 Lu et al; licensee BioMed Central Ltd.
spellingShingle Research Article
Lu, Deshun
Yuan, Xiu-juan
Evans, Robert J
Pappas, Amy T
Wang, He
Su, Eric W
Hamdouchi, Chafiq
Venkataraman, Chandrasekar
Cloning and functional characterization of the rabbit C-C chemokine receptor 2
title Cloning and functional characterization of the rabbit C-C chemokine receptor 2
title_full Cloning and functional characterization of the rabbit C-C chemokine receptor 2
title_fullStr Cloning and functional characterization of the rabbit C-C chemokine receptor 2
title_full_unstemmed Cloning and functional characterization of the rabbit C-C chemokine receptor 2
title_short Cloning and functional characterization of the rabbit C-C chemokine receptor 2
title_sort cloning and functional characterization of the rabbit c-c chemokine receptor 2
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1182369/
https://www.ncbi.nlm.nih.gov/pubmed/16001983
http://dx.doi.org/10.1186/1471-2172-6-15
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