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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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GPR-9-6 is a chemokine receptor for TECK. (A) GPR-9-6 transfectants were examined for chemotactic responses to various concentrations of TECK and I-TAC ranging from 10 to 1,000 nM. (B) The effect of anti–GPR-9-6 mAb 3C3 and pertussis toxin (PTX) on the migration was examined. All mAb treatments were for 10 min at 4°C before the migration assay. (C) Thymocytes were examined for their chemotaxis to various concentrations of TECK. (D) The effect of anti–GPR-9-6 mAb 3C3 and an IgG2b control on TECK- and SDF-1α–induced thymocyte migration was also examined. The effect of anti–GPR-9-6 mAb and anti-CCR4 mAb 2B10 on the chemotaxis of CD4 lymphocytes to TECK (E) and TARC (F) was also tested (n = 2).
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Figure 8: GPR-9-6 is a chemokine receptor for TECK. (A) GPR-9-6 transfectants were examined for chemotactic responses to various concentrations of TECK and I-TAC ranging from 10 to 1,000 nM. (B) The effect of anti–GPR-9-6 mAb 3C3 and pertussis toxin (PTX) on the migration was examined. All mAb treatments were for 10 min at 4°C before the migration assay. (C) Thymocytes were examined for their chemotaxis to various concentrations of TECK. (D) The effect of anti–GPR-9-6 mAb 3C3 and an IgG2b control on TECK- and SDF-1α–induced thymocyte migration was also examined. The effect of anti–GPR-9-6 mAb and anti-CCR4 mAb 2B10 on the chemotaxis of CD4 lymphocytes to TECK (E) and TARC (F) was also tested (n = 2).

Mentions: From a panel of chemokines including MCP-1–4, MIP-1αβ, MIP-3αβ, eotaxin-1–3, RANTES, I-309, thymus and activation-regulated chemokine (TARC), MDC, liver-expressed chemokine (LEC), CKα2, secondary lymphoid-tissue chemokine (SLC), human CC chemokine 1, fractalkine, lymphotactin, monokine induced by IFN-γ (MIG), IP-10, I-TAC, B cell–attracting chemokine 1 (BCL-1), IL-8, gro-αβψ, leukotactin, stromal cell–derived factor (SDF)-1α/β, MIP-3, and MIP-4, only TECK induced chemotaxis of GPR-9-6/L1.2 transfectants (Fig. 8 A, and data not shown). TECK-mediated chemotaxis of L1.2/GPR-9-6 transfectants was inhibited by anti–GPR-9-6 mAb 3C3 (Fig. 8 B), while pretreatment of the transfectants with pertussis toxin also inhibited migration to TECK (Fig. 8 B). The observation that pertussis toxin abolished TECK-mediated transfectant chemotaxis indicates that chemokine receptor GPR-9-6 couples to the Gi class of G proteins. TECK did not act on any of the other transfectants tested (CCR1,2,4–7 and CXCR1–4), with the exception of CCR3/L1.2 transfectants, for which high concentrations of TECK were weakly chemotactic (data not shown). Also in examining our panel of cell lines, we found that only GPR-9-6+ Molt-4 and Molt-13 cell lines migrated to TECK (Table ). Migration of Molt-4 and Molt-13 cells to TECK was blocked by anti–GPR-9-6 mAb 3C3 (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)

GPR-9-6 is a chemokine receptor for TECK. (A) GPR-9-6 transfectants were examined for chemotactic responses to various concentrations of TECK and I-TAC ranging from 10 to 1,000 nM. (B) The effect of anti–GPR-9-6 mAb 3C3 and pertussis toxin (PTX) on the migration was examined. All mAb treatments were for 10 min at 4°C before the migration assay. (C) Thymocytes were examined for their chemotaxis to various concentrations of TECK. (D) The effect of anti–GPR-9-6 mAb 3C3 and an IgG2b control on TECK- and SDF-1α–induced thymocyte migration was also examined. The effect of anti–GPR-9-6 mAb and anti-CCR4 mAb 2B10 on the chemotaxis of CD4 lymphocytes to TECK (E) and TARC (F) was also tested (n = 2).
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Figure 8: GPR-9-6 is a chemokine receptor for TECK. (A) GPR-9-6 transfectants were examined for chemotactic responses to various concentrations of TECK and I-TAC ranging from 10 to 1,000 nM. (B) The effect of anti–GPR-9-6 mAb 3C3 and pertussis toxin (PTX) on the migration was examined. All mAb treatments were for 10 min at 4°C before the migration assay. (C) Thymocytes were examined for their chemotaxis to various concentrations of TECK. (D) The effect of anti–GPR-9-6 mAb 3C3 and an IgG2b control on TECK- and SDF-1α–induced thymocyte migration was also examined. The effect of anti–GPR-9-6 mAb and anti-CCR4 mAb 2B10 on the chemotaxis of CD4 lymphocytes to TECK (E) and TARC (F) was also tested (n = 2).
Mentions: From a panel of chemokines including MCP-1–4, MIP-1αβ, MIP-3αβ, eotaxin-1–3, RANTES, I-309, thymus and activation-regulated chemokine (TARC), MDC, liver-expressed chemokine (LEC), CKα2, secondary lymphoid-tissue chemokine (SLC), human CC chemokine 1, fractalkine, lymphotactin, monokine induced by IFN-γ (MIG), IP-10, I-TAC, B cell–attracting chemokine 1 (BCL-1), IL-8, gro-αβψ, leukotactin, stromal cell–derived factor (SDF)-1α/β, MIP-3, and MIP-4, only TECK induced chemotaxis of GPR-9-6/L1.2 transfectants (Fig. 8 A, and data not shown). TECK-mediated chemotaxis of L1.2/GPR-9-6 transfectants was inhibited by anti–GPR-9-6 mAb 3C3 (Fig. 8 B), while pretreatment of the transfectants with pertussis toxin also inhibited migration to TECK (Fig. 8 B). The observation that pertussis toxin abolished TECK-mediated transfectant chemotaxis indicates that chemokine receptor GPR-9-6 couples to the Gi class of G proteins. TECK did not act on any of the other transfectants tested (CCR1,2,4–7 and CXCR1–4), with the exception of CCR3/L1.2 transfectants, for which high concentrations of TECK were weakly chemotactic (data not shown). Also in examining our panel of cell lines, we found that only GPR-9-6+ Molt-4 and Molt-13 cell lines migrated to TECK (Table ). Migration of Molt-4 and Molt-13 cells to TECK was blocked by anti–GPR-9-6 mAb 3C3 (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