Murine CCR9, a Chemokine Receptor for Thymus-Expressed Chemokine That Is Up-Regulated Following Pre-TCR Signaling1

Chemokines are likely to play an important role in regulating the trafficking of developing T cells within the thymus. By using anti-CD3ε treatment of recombinase-activating gene 2 (Rag2−/−) mice to mimic pre-TCR signaling and drive thymocyte development to the double positive stage, we have identified murine GPR-9-6 as a chemokine receptor whose expression is strongly induced following pre-TCR signaling. GPR-9-6 mRNA is present at high levels in the thymus, and by RT-PCR analysis its expression is induced as normal thymocytes undergo the double negative to double positive transition. Furthermore we show that TECK (thymus-expressed chemokine), a chemokine produced by thymic medullary dendritic cells, is a functional ligand for GPR-9-6. TECK specifically induces a calcium flux and chemotaxis of GPR-9-6-transfected cells. In addition, TECK stimulates the migration of normal double positive thymocytes, as well as Rag2−/− thymocytes following anti-CD3ε treatment. Hence, GPR-9-6 has been designated as CC chemokine receptor 9 (CCR9). Our results suggest that TECK delivers signals through CCR9 important for the navigation of developing thymocytes.

[1]  L. Williams,et al.  Mice Lacking Expression of Secondary Lymphoid Organ Chemokine Have Defects in Lymphocyte Homing and Dendritic Cell Localization , 1999, The Journal of experimental medicine.

[2]  A. Zlotnik,et al.  A developmental pathway involving four phenotypically and functionally distinct subsets of CD3-CD4-CD8- triple-negative adult mouse thymocytes defined by CD44 and CD25 expression. , 1993, Journal of immunology.

[3]  B. Premack,et al.  Activation of dual T cell signaling pathways by the chemokine RANTES. , 1995, Science.

[4]  E. Butcher,et al.  Genetic defect in T lymphocyte‐specific homing into peripheral lymph nodes , 1997, European journal of immunology.

[5]  O. Yoshie,et al.  The T Cell-directed CC Chemokine TARC Is a Highly Specific Biological Ligand for CC Chemokine Receptor 4* , 1997, The Journal of Biological Chemistry.

[6]  C. Power,et al.  Molecular Cloning and Functional Expression of a Novel CC Chemokine Receptor cDNA from a Human Basophilic Cell Line (*) , 1995, The Journal of Biological Chemistry.

[7]  M. Bevan,et al.  Correlating notch signaling with thymocyte maturation. , 1998, Immunity.

[8]  D. Williams,et al.  A thymic stromal cell line supports in vitro development of surface IgM+ B cells and produces a novel growth factor affecting B and T lineage cells. , 1994, Experimental hematology.

[9]  J. White,et al.  Differential chemotactic behavior of developing T cells in response to thymic chemokines. , 1998, Blood.

[10]  V. Stewart,et al.  RAG-2-deficient mice lack mature lymphocytes owing to inability to initiate V(D)J rearrangement , 1992, Cell.

[11]  Bernhard Moser,et al.  The CXC chemokine SDF-1 is the ligand for LESTR/fusin and prevents infection by T-cell-line-adapted HIV-1 , 1996, Nature.

[12]  Richard A Koup,et al.  Homozygous Defect in HIV-1 Coreceptor Accounts for Resistance of Some Multiply-Exposed Individuals to HIV-1 Infection , 1996, Cell.

[13]  E. Coligan.  Current protocols in immunology , 1991 .

[14]  T. Wilson,et al.  The thymic microenvironment. , 1993, Immunology today.

[15]  J. Cyster,et al.  Epstein-Barr Virus–induced Molecule 1 Ligand Chemokine Is Expressed by Dendritic Cells in Lymphoid Tissues and Strongly Attracts Naive T Cells and Activated B Cells , 1998, The Journal of experimental medicine.

[16]  Masahiko Kuroda,et al.  Function of the chemokine receptor CXCR4 in haematopoiesis and in cerebellar development , 1998, Nature.

[17]  K. Shortman,et al.  Different Subpopulations of Developing Thymocytes are Associated with Adherent (Macrophage) or Nonadherent (Dendritic) Thymic Rosettes , 1991, Developmental immunology.

[18]  M. Bevan,et al.  CD8 lineage commitment in the absence of CD8. , 1997, Immunity.

[19]  S. Tonegawa,et al.  Regulation of thymocyte development through CD3: Functional dissociation between p56lck and CD3ζ in early thymic selection , 1995 .

[20]  D. Steiner,et al.  Cloning, expression, and chromosomal mapping of a novel human CC-chemokine receptor (CCR10) that displays high-affinity binding for MCP-1 and MCP-3. , 1997, DNA and cell biology.

[21]  E. Butcher,et al.  Chemokines and the arrest of lymphocytes rolling under flow conditions. , 1998, Science.

[22]  Michael J. Bevan,et al.  Itm2a Is Induced during Thymocyte Selection and T Cell Activation and Causes Downregulation of Cd8 When Overexpressed in Cd4+Cd8+ Double Positive Thymocytes , 1999, The Journal of experimental medicine.

[23]  B. Kyewski.  Seeding of thymic microenvironments defined by distinct thymocyte- stromal cell interactions is developmentally controlled , 1987, The Journal of experimental medicine.

[24]  E. Jenkinson,et al.  Factors regulating stem cell recruitment to the fetal thymus. , 1999, Journal of immunology.

[25]  T. Bonner,et al.  Brief Definitive Report Identification of CCR8: A Human Monocyte and Thymus Receptor for the CC Chemokine I-309 , 2022 .

[26]  N. Taylor,et al.  Macrophage inflammatory protein-1β induces migration and activation of human thymocytes , 1998 .

[27]  Á. Zaballos,et al.  Molecular cloning, functional characterization and mRNA expression analysis of the murine chemokine receptor CCR6 and its specific ligand MIP‐3α 1 , 1998 .

[28]  G. Anderson,et al.  Cellular interactions in thymocyte development. , 1996, Annual review of immunology.

[29]  J. Farber,et al.  Cloning of STRL22, a new human gene encoding a G-protein-coupled receptor related to chemokine receptors and located on chromosome 6q27. , 1997, Genomics.

[30]  S. Nishikawa,et al.  Defects of B-cell lymphopoiesis and bone-marrow myelopoiesis in mice lacking the CXC chemokine PBSF/SDF-1 , 1996, Nature.

[31]  H. Yonekawa,et al.  A novel mutant gene involved in T-lymphocyte-specific homing into peripheral lymphoid organs on mouse chromosome 4. , 1998, Blood.

[32]  N. Copeland,et al.  TECK: a novel CC chemokine specifically expressed by thymic dendritic cells and potentially involved in T cell development. , 1997, Immunity.

[33]  G. Anderson,et al.  Fibroblast dependency during early thymocyte development maps to the CD25+ CD44+ stage and involves interactions with fibroblast matrix molecules , 1997, European journal of immunology.

[34]  A. Mantovani,et al.  Identification of the CC chemokines TARC and macrophage inflammatory protein‐1β as novel functional ligands for the CCR8 receptor , 1998, European journal of immunology.

[35]  D. Steiner,et al.  Molecular Cloning and Expression of a Novel Rat CC-Chemokine Receptor (rCCR10rR) That Binds MCP-1 and MIP-1β with High Affinity , 1997 .

[36]  R. Nibbs,et al.  Cloning and characterization of a novel murine beta chemokine receptor, D6. Comparison to three other related macrophage inflammatory protein-1alpha receptors, CCR-1, CCR-3, and CCR-5. , 1997, The Journal of biological chemistry.

[37]  J. Renauld,et al.  I-309/T cell activation gene-3 chemokine protects murine T cell lymphomas against dexamethasone-induced apoptosis. , 1996, Journal of immunology.

[38]  S. Tonegawa,et al.  Restoration of early thymocyte differentiation in T-cell receptor beta-chain-deficient mutant mice by transmembrane signaling through CD3 epsilon. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[39]  P. Rabinovitch,et al.  Measurement of Intracellular Calcium Ions by Flow Cytometry , 1997, Current protocols in cytometry.

[40]  F. Alt,et al.  CD3ε-mediated signals rescue the development of CD4+CD8+ thymocytes in RAG-2−/− mice in the absence of TCR β chain expression , 1994 .

[41]  E. Kieff,et al.  Epstein-Barr virus-induced genes: first lymphocyte-specific G protein-coupled peptide receptors , 1993, Journal of virology.

[42]  R. Burgstahler,et al.  Expression of the chemokine receptor BLR2/EBI1 is specifically transactivated by Epstein-Barr virus nuclear antigen 2. , 1995, Biochemical and biophysical research communications.

[43]  J. Unkeless.  Characterization of a monoclonal antibody directed against mouse macrophage and lymphocyte Fc receptors , 1979, The Journal of experimental medicine.

[44]  R Bonecchi,et al.  Differential regulation of chemokine receptors during dendritic cell maturation: a model for their trafficking properties. , 1998, Journal of immunology.

[45]  H. Nomiyama,et al.  Novel lymphocyte‐specific CC chemokines and their receptors , 1997, Journal of leukocyte biology.

[46]  J. Sodroski,et al.  The lymphocyte chemoattractant SDF-1 is a ligand for LESTR/fusin and blocks HIV-1 entry , 1996, Nature.

[47]  R. Nibbs,et al.  Cloning and Characterization of a Novel Promiscuous Human β-Chemokine Receptor D6* , 1997, The Journal of Biological Chemistry.

[48]  M. Bevan,et al.  Positive selection of thymocytes. , 1995, Annual review of immunology.

[49]  H. Nomiyama,et al.  Molecular Cloning of a Novel Human CC Chemokine EBI1-ligand Chemokine That Is a Specific Functional Ligand for EBI1, CCR7* , 1997, The Journal of Biological Chemistry.

[50]  R Murray,et al.  Induction of TCR gene rearrangements in uncommitted stem cells by a subset of IL-7 producing, MHC class-II-expressing thymic stromal cells. , 1997, Immunity.

[51]  R. Nibbs,et al.  Cloning and Characterization of a Novel Murine β Chemokine Receptor, D6 , 1997, The Journal of Biological Chemistry.

[52]  A. Bradley,et al.  Ulcerative colitis and adenocarcinoma of the colon in Gαi2-deficient mice , 1995, Nature Genetics.

[53]  B Dewald,et al.  Human chemokines: an update. , 1997, Annual review of immunology.

[54]  I. Weissman.  THYMUS CELL MATURATION , 1973, The Journal of experimental medicine.

[55]  T. Honjo,et al.  Signal sequence trap: a cloning strategy for secreted proteins and type I membrane proteins. , 1993, Science.

[56]  K. Shortman,et al.  Identification of early stages of T lymphocyte development in the thymus cortex and medulla. , 1985, Journal of immunology.

[57]  M. Bevan,et al.  RORgamma t, a novel isoform of an orphan receptor, negatively regulates Fas ligand expression and IL-2 production in T cells. , 1998, Immunity.

[58]  H. Kikutani,et al.  Molecular cloning and structure of a pre-B-cell growth-stimulating factor. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[59]  I. Weissman,et al.  A cell-surface molecule involved in organ-specific homing of lymphocytes , 1983, Nature.

[60]  R. Eddy,et al.  Cloning of human and mouse EBI1, a lymphoid-specific G-protein-coupled receptor encoded on human chromosome 17q12-q21.2. , 1994, Genomics.

[61]  G. Anderson,et al.  MHC class II-positive epithelium and mesenchyme cells are both required for T-cell development in the thymus , 1993, Nature.

[62]  Jason G. Cyster,et al.  Chemokines and the Homing of Dendritic Cells to the T Cell Areas of Lymphoid Organs , 1999, The Journal of experimental medicine.

[63]  K. Rock,et al.  Presentation of exogenous antigens by macrophages: analysis of major histocompatibility complex class I and II presentation and regulation by cytokines , 1994, European journal of immunology.

[64]  H. Fehling,et al.  Structure and function of the pre-T cell receptor. , 1997, Annual review of immunology.

[65]  Á. Zaballos,et al.  Cutting edge: identification of the orphan chemokine receptor GPR-9-6 as CCR9, the receptor for the chemokine TECK. , 1999, Journal of immunology.

[66]  C. Caux,et al.  Selective Recruitment of Immature and Mature Dendritic Cells by Distinct Chemokines Expressed in Different Anatomic Sites , 1998, The Journal of experimental medicine.

[67]  M. Julius,et al.  Thymus independent T cell development and selection in the intestinal epithelium. , 1994, Annual review of immunology.

[68]  P. Fink,et al.  Positive selection of thymocytes. , 1995, Annual review of immunology.

[69]  F. Alt,et al.  Thymocyte development in the absence of pre-T cell receptor extracellular immunoglobulin domains. , 1998, Science.

[70]  Kouji Matsushima,et al.  The chemokine receptor CXCR4 is essential for vascularization of the gastrointestinal tract , 1998, Nature.

[71]  P. Gray,et al.  Macrophage-derived Chemokine Is a Functional Ligand for the CC Chemokine Receptor 4* , 1998, The Journal of Biological Chemistry.

[72]  M. Bevan,et al.  Virus-mediated delivery of antigenic epitopes into dendritic cells as a means to induce CTL. , 1997, Journal of immunology.

[73]  K. Blank,et al.  T cell receptor genes and T cell development in virus-transformed early T cell lines. , 1990, Journal of Immunology.

[74]  J. Westwick,et al.  Chemokines and T lymphocytes: more than an attraction. , 1998, Immunity.

[75]  P. Murphy.  The molecular biology of leukocyte chemoattractant receptors. , 1994, Annual review of immunology.

[76]  E. Butcher,et al.  Cutting edge: developmental switches in chemokine responses during T cell maturation. , 1999, Journal of immunology.

[77]  R. Perlmutter,et al.  A pertussis toxin‐sensitive process controls thymocyte emigration , 1991, European journal of immunology.