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Human G protein-coupled receptor GPR-9-6/CC chemokine receptor 9 is selectively expressed on intestinal homing T lymphocytes, mucosal lymphocytes, and thymocytes and is required for thymus-expressed chemokine-mediated chemotaxis.

Zabel BA, Agace WW, Campbell JJ, Heath HM, Parent D, Roberts AI, Ebert EC, Kassam N, Qin S, Zovko M, LaRosa GJ, Yang LL, Soler D, Butcher EC, Ponath PD, Parker CM, Andrew DP - J. Exp. Med. (1999)

Bottom Line: TECK (thymus-expressed chemokine), a recently described CC chemokine expressed in thymus and small intestine, was found to mediate chemotaxis of human G protein-coupled receptor GPR-9-6/L1.2 transfectants.In agreement with the nomenclature rules for chemokine receptors, we propose the designation CCR-9 for GPR-9-6.The selective expression of TECK and GPR-9-6 in thymus and small intestine implies a dual role for GPR-9-6/CCR-9, both in T cell development and the mucosal immune response.

View Article: PubMed Central - PubMed

Affiliation: LeukoSite, Inc., Cambridge, Massachusetts 02142, USA.

ABSTRACT
TECK (thymus-expressed chemokine), a recently described CC chemokine expressed in thymus and small intestine, was found to mediate chemotaxis of human G protein-coupled receptor GPR-9-6/L1.2 transfectants. This activity was blocked by anti-GPR-9-6 monoclonal antibody (mAb) 3C3. GPR-9-6 is expressed on a subset of memory alpha4beta7(high) intestinal trafficking CD4 and CD8 lymphocytes. In addition, all intestinal lamina propria and intraepithelial lymphocytes express GPR-9-6. In contrast, GPR-9-6 is not displayed on cutaneous lymphocyte antigen-positive (CLA(+)) memory CD4 and CD8 lymphocytes, which traffic to skin inflammatory sites, or on other systemic alpha4beta7(-)CLA(-) memory CD4/CD8 lymphocytes. The majority of thymocytes also express GPR-9-6, but natural killer cells, monocytes, eosinophils, basophils, and neutrophils are GPR-9-6 negative. Transcripts of GPR-9-6 and TECK are present in both small intestine and thymus. Importantly, the expression profile of GPR-9-6 correlates with migration to TECK of blood T lymphocytes and thymocytes. As migration of these cells is blocked by anti-GPR-9-6 mAb 3C3, we conclude that GPR-9-6 is the principal chemokine receptor for TECK. In agreement with the nomenclature rules for chemokine receptors, we propose the designation CCR-9 for GPR-9-6. The selective expression of TECK and GPR-9-6 in thymus and small intestine implies a dual role for GPR-9-6/CCR-9, both in T cell development and the mucosal immune response.

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Tissue distribution of TECK and GPR-9-6. (A) TECK hybridizations: 32P-labeled TECK DNA was used to probe multitissue Northern blot filters (2 μg poly-A RNA per lane, 24-h exposure; Clontech). (B) GPR-9-6 hybridizations: 32P-labeled GPR-9-6 DNA was used to probe the same filters as in A as well as a multi-cell line Northern blot filter (72-h exposure). (C) Equivalent amounts (0.5 ng) of primary cDNA (Clontech) from colon, small intestine, brain, and thymus, as well as 500 ng genomic DNA were amplified in PCR (30 cycles) using GPR-9-6 and TECK primers. G3PDH primers and G3PDH intron B primers were used as controls.
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Figure 13: Tissue distribution of TECK and GPR-9-6. (A) TECK hybridizations: 32P-labeled TECK DNA was used to probe multitissue Northern blot filters (2 μg poly-A RNA per lane, 24-h exposure; Clontech). (B) GPR-9-6 hybridizations: 32P-labeled GPR-9-6 DNA was used to probe the same filters as in A as well as a multi-cell line Northern blot filter (72-h exposure). (C) Equivalent amounts (0.5 ng) of primary cDNA (Clontech) from colon, small intestine, brain, and thymus, as well as 500 ng genomic DNA were amplified in PCR (30 cycles) using GPR-9-6 and TECK primers. G3PDH primers and G3PDH intron B primers were used as controls.

Mentions: Due to the expression of GPR-9-6 on mucosal homing lymphocytes, we examined the distribution of TECK and GPR-9-6 transcripts in lymphoid and mucosal tissues (Fig. 13). TECK was selectively expressed in thymus and small intestine (Fig. 13 A). GPR-9-6 was expressed at high levels in thymus and weakly in spleen and PBLs (Fig. 13 B). Although we could not detect GPR-9-6 transcripts by Northern blot analysis in small intestine, we were able to detect GPR-9-6 message in small intestine and thymus using the more sensitive technique of RT-PCR (Fig. 13 C). Message for either gene was not detected in brain or colon. Also as predicted from our expression data on cell lines with mAb 3C3, GPR-9-6 was expressed by Molt-13 cells (and Molt-4 cells, data not shown) but not in GPR-9-6− JY, KU812, and EOL cells. In additional Northern blot analysis, TECK and GPR-9-6 were not detected in Th1, Th2, or T regulatory type 1 lymphocytes, LAK (lymphokine-activated NK) cells, monocytes, CD34-derived dendritic cells, monocyte-derived dendritic cells, astrocytes, high umbilical vein endothelial cells, or pulmonary vein endothelial cells (data not shown).


Human G protein-coupled receptor GPR-9-6/CC chemokine receptor 9 is selectively expressed on intestinal homing T lymphocytes, mucosal lymphocytes, and thymocytes and is required for thymus-expressed chemokine-mediated chemotaxis.

Zabel BA, Agace WW, Campbell JJ, Heath HM, Parent D, Roberts AI, Ebert EC, Kassam N, Qin S, Zovko M, LaRosa GJ, Yang LL, Soler D, Butcher EC, Ponath PD, Parker CM, Andrew DP - J. Exp. Med. (1999)

Tissue distribution of TECK and GPR-9-6. (A) TECK hybridizations: 32P-labeled TECK DNA was used to probe multitissue Northern blot filters (2 μg poly-A RNA per lane, 24-h exposure; Clontech). (B) GPR-9-6 hybridizations: 32P-labeled GPR-9-6 DNA was used to probe the same filters as in A as well as a multi-cell line Northern blot filter (72-h exposure). (C) Equivalent amounts (0.5 ng) of primary cDNA (Clontech) from colon, small intestine, brain, and thymus, as well as 500 ng genomic DNA were amplified in PCR (30 cycles) using GPR-9-6 and TECK primers. G3PDH primers and G3PDH intron B primers were used as controls.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2195678&req=5

Figure 13: Tissue distribution of TECK and GPR-9-6. (A) TECK hybridizations: 32P-labeled TECK DNA was used to probe multitissue Northern blot filters (2 μg poly-A RNA per lane, 24-h exposure; Clontech). (B) GPR-9-6 hybridizations: 32P-labeled GPR-9-6 DNA was used to probe the same filters as in A as well as a multi-cell line Northern blot filter (72-h exposure). (C) Equivalent amounts (0.5 ng) of primary cDNA (Clontech) from colon, small intestine, brain, and thymus, as well as 500 ng genomic DNA were amplified in PCR (30 cycles) using GPR-9-6 and TECK primers. G3PDH primers and G3PDH intron B primers were used as controls.
Mentions: Due to the expression of GPR-9-6 on mucosal homing lymphocytes, we examined the distribution of TECK and GPR-9-6 transcripts in lymphoid and mucosal tissues (Fig. 13). TECK was selectively expressed in thymus and small intestine (Fig. 13 A). GPR-9-6 was expressed at high levels in thymus and weakly in spleen and PBLs (Fig. 13 B). Although we could not detect GPR-9-6 transcripts by Northern blot analysis in small intestine, we were able to detect GPR-9-6 message in small intestine and thymus using the more sensitive technique of RT-PCR (Fig. 13 C). Message for either gene was not detected in brain or colon. Also as predicted from our expression data on cell lines with mAb 3C3, GPR-9-6 was expressed by Molt-13 cells (and Molt-4 cells, data not shown) but not in GPR-9-6− JY, KU812, and EOL cells. In additional Northern blot analysis, TECK and GPR-9-6 were not detected in Th1, Th2, or T regulatory type 1 lymphocytes, LAK (lymphokine-activated NK) cells, monocytes, CD34-derived dendritic cells, monocyte-derived dendritic cells, astrocytes, high umbilical vein endothelial cells, or pulmonary vein endothelial cells (data not shown).

Bottom Line: TECK (thymus-expressed chemokine), a recently described CC chemokine expressed in thymus and small intestine, was found to mediate chemotaxis of human G protein-coupled receptor GPR-9-6/L1.2 transfectants.In agreement with the nomenclature rules for chemokine receptors, we propose the designation CCR-9 for GPR-9-6.The selective expression of TECK and GPR-9-6 in thymus and small intestine implies a dual role for GPR-9-6/CCR-9, both in T cell development and the mucosal immune response.

View Article: PubMed Central - PubMed

Affiliation: LeukoSite, Inc., Cambridge, Massachusetts 02142, USA.

ABSTRACT
TECK (thymus-expressed chemokine), a recently described CC chemokine expressed in thymus and small intestine, was found to mediate chemotaxis of human G protein-coupled receptor GPR-9-6/L1.2 transfectants. This activity was blocked by anti-GPR-9-6 monoclonal antibody (mAb) 3C3. GPR-9-6 is expressed on a subset of memory alpha4beta7(high) intestinal trafficking CD4 and CD8 lymphocytes. In addition, all intestinal lamina propria and intraepithelial lymphocytes express GPR-9-6. In contrast, GPR-9-6 is not displayed on cutaneous lymphocyte antigen-positive (CLA(+)) memory CD4 and CD8 lymphocytes, which traffic to skin inflammatory sites, or on other systemic alpha4beta7(-)CLA(-) memory CD4/CD8 lymphocytes. The majority of thymocytes also express GPR-9-6, but natural killer cells, monocytes, eosinophils, basophils, and neutrophils are GPR-9-6 negative. Transcripts of GPR-9-6 and TECK are present in both small intestine and thymus. Importantly, the expression profile of GPR-9-6 correlates with migration to TECK of blood T lymphocytes and thymocytes. As migration of these cells is blocked by anti-GPR-9-6 mAb 3C3, we conclude that GPR-9-6 is the principal chemokine receptor for TECK. In agreement with the nomenclature rules for chemokine receptors, we propose the designation CCR-9 for GPR-9-6. The selective expression of TECK and GPR-9-6 in thymus and small intestine implies a dual role for GPR-9-6/CCR-9, both in T cell development and the mucosal immune response.

Show MeSH
Related in: MedlinePlus