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Identification of CCR8: a human monocyte and thymus receptor for the CC chemokine I-309.

Tiffany HL, Lautens LL, Gao JL, Pease J, Locati M, Combadiere C, Modi W, Bonner TI, Murphy PM - J. Exp. Med. (1997)

Bottom Line: Signaling was sensitive to pertussis toxin, suggesting coupling to a Gi-type G protein.These properties parallel those of endogenous I-309 receptors expressed in an HL-60 clone 15 cell line model.The apparent monogamous relationship between I-309 and CCR8 is unusual among known CC chemokines and known CC chemokine receptors.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA.

ABSTRACT
The human CC chemokine I-309 is a potent monocyte chemoattractant and inhibits apoptosis in thymic cell lines. Here, we identify a specific human I-309 receptor, and name it CCR8 according to an accepted nomenclature system. The receptor has seven predicted transmembrane domains, is expressed constitutively in monocytes and thymus, and is encoded by a previously reported gene of previously unknown function named, alternatively, CY6, TER1, and CKR-L1. After transfection with the CY6 open reading frame, a mouse pre-B cell line exhibited calcium flux and chemotaxis in response to I-309 (EC50 = 2 nM for each), whereas 20 other chemokines were inactive. Signaling was sensitive to pertussis toxin, suggesting coupling to a Gi-type G protein. These properties parallel those of endogenous I-309 receptors expressed in an HL-60 clone 15 cell line model. The apparent monogamous relationship between I-309 and CCR8 is unusual among known CC chemokines and known CC chemokine receptors. CCR8 may regulate monocyte chemotaxis and thymic cell line apoptosis.

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RNA distribution  of CCR8. Northern blots containing total RNA from the  sources indicated above each  lane were hybridized with a  CCR8 ORF probe under high  stringency conditions. M, monocyte/macrophages (PBMCs that  remained adherent to plastic after  18-h overnight culture); L, lymphocytes (nonadherent PBMCs); N, neutrophils. The blots  were exposed for 3 d to x-ray  film using an intensifying screen.
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Figure 1: RNA distribution of CCR8. Northern blots containing total RNA from the sources indicated above each lane were hybridized with a CCR8 ORF probe under high stringency conditions. M, monocyte/macrophages (PBMCs that remained adherent to plastic after 18-h overnight culture); L, lymphocytes (nonadherent PBMCs); N, neutrophils. The blots were exposed for 3 d to x-ray film using an intensifying screen.

Mentions: Using a multitissue Northern blot, mRNA was detected at high levels in thymus, and at lower levels in spleen, but not in 14 other tissues tested (data not shown). Using Northern blots from the same supplier, the same RNA distribution pattern has been reported by Napolitano et al. and Zaballos et al. (15, 16). Napolitano et al. (15) also detected transcripts in the MOLT-4 T cell line and the NK3.3 NK cell line, but not in primary NK cells, monocytes, neutrophils, or PHA/PMA-activated PBMCs. We were able to detect a 4.6-kb mRNA band in total RNA from adherent monocytes, consistent with the size in thymus, but not in neutrophil or lymphocyte samples (Fig. 1). Zaballos et al. (16) also detected mRNA in monocyte/macrophages, as well as in lymph node, and CD4+, CD8+, and CD19+ lymphocytes.


Identification of CCR8: a human monocyte and thymus receptor for the CC chemokine I-309.

Tiffany HL, Lautens LL, Gao JL, Pease J, Locati M, Combadiere C, Modi W, Bonner TI, Murphy PM - J. Exp. Med. (1997)

RNA distribution  of CCR8. Northern blots containing total RNA from the  sources indicated above each  lane were hybridized with a  CCR8 ORF probe under high  stringency conditions. M, monocyte/macrophages (PBMCs that  remained adherent to plastic after  18-h overnight culture); L, lymphocytes (nonadherent PBMCs); N, neutrophils. The blots  were exposed for 3 d to x-ray  film using an intensifying screen.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2198957&req=5

Figure 1: RNA distribution of CCR8. Northern blots containing total RNA from the sources indicated above each lane were hybridized with a CCR8 ORF probe under high stringency conditions. M, monocyte/macrophages (PBMCs that remained adherent to plastic after 18-h overnight culture); L, lymphocytes (nonadherent PBMCs); N, neutrophils. The blots were exposed for 3 d to x-ray film using an intensifying screen.
Mentions: Using a multitissue Northern blot, mRNA was detected at high levels in thymus, and at lower levels in spleen, but not in 14 other tissues tested (data not shown). Using Northern blots from the same supplier, the same RNA distribution pattern has been reported by Napolitano et al. and Zaballos et al. (15, 16). Napolitano et al. (15) also detected transcripts in the MOLT-4 T cell line and the NK3.3 NK cell line, but not in primary NK cells, monocytes, neutrophils, or PHA/PMA-activated PBMCs. We were able to detect a 4.6-kb mRNA band in total RNA from adherent monocytes, consistent with the size in thymus, but not in neutrophil or lymphocyte samples (Fig. 1). Zaballos et al. (16) also detected mRNA in monocyte/macrophages, as well as in lymph node, and CD4+, CD8+, and CD19+ lymphocytes.

Bottom Line: Signaling was sensitive to pertussis toxin, suggesting coupling to a Gi-type G protein.These properties parallel those of endogenous I-309 receptors expressed in an HL-60 clone 15 cell line model.The apparent monogamous relationship between I-309 and CCR8 is unusual among known CC chemokines and known CC chemokine receptors.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA.

ABSTRACT
The human CC chemokine I-309 is a potent monocyte chemoattractant and inhibits apoptosis in thymic cell lines. Here, we identify a specific human I-309 receptor, and name it CCR8 according to an accepted nomenclature system. The receptor has seven predicted transmembrane domains, is expressed constitutively in monocytes and thymus, and is encoded by a previously reported gene of previously unknown function named, alternatively, CY6, TER1, and CKR-L1. After transfection with the CY6 open reading frame, a mouse pre-B cell line exhibited calcium flux and chemotaxis in response to I-309 (EC50 = 2 nM for each), whereas 20 other chemokines were inactive. Signaling was sensitive to pertussis toxin, suggesting coupling to a Gi-type G protein. These properties parallel those of endogenous I-309 receptors expressed in an HL-60 clone 15 cell line model. The apparent monogamous relationship between I-309 and CCR8 is unusual among known CC chemokines and known CC chemokine receptors. CCR8 may regulate monocyte chemotaxis and thymic cell line apoptosis.

Show MeSH
Related in: MedlinePlus