1. Cloning of the murine interferon-inducible protein 10 (IP-10) receptor and its specific expression in lymphoid organs
- Author
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Kimio Yatsunami, Shosaku Narumi, Yuko Tominaga, and Masahiro Tamaru
- Subjects
Chemokine receptor CCR5 ,Lymphoid Tissue ,Molecular Sequence Data ,Biophysics ,C-C chemokine receptor type 7 ,C-C chemokine receptor type 6 ,CCR8 ,Lymphocyte Activation ,Transfection ,Biochemistry ,Mice ,Interleukin-4 receptor ,Concanavalin A ,Animals ,5-HT5A receptor ,Amino Acid Sequence ,RNA, Messenger ,Cloning, Molecular ,Molecular Biology ,Cells, Cultured ,Common gamma chain ,biology ,Base Sequence ,Cell Biology ,Molecular biology ,Recombinant Proteins ,Rats ,Chemokine CXCL10 ,Mice, Inbred C57BL ,Organ Specificity ,Interleukin-21 receptor ,biology.protein ,Calcium ,Female ,Receptors, Chemokine ,Chemokines, CXC ,Spleen ,T-Lymphocytes, Cytotoxic - Abstract
To isolate the interferon-inducible protein 10 (IP-10) receptor gene, we searched for cells that respond to IP-10. Among several human and murine T cell lines, only CTLL2 cells ( a murine cytotoxic T cell line) responded to IP-10 with transient elevation of intracellular Ca2+. The murine IP-10 receptor gene has been cloned from cDNA derived from CTLL2 cells using the reverse transcriptase-polymerase chain reaction protocol with two degenerate primers corresponding to conserved regions of chemokine receptors. The cDNA encoding the murine IP-10 receptor has an open reading frame of 1101 bp corresponding to a protein of 367 amino acids that exhibits 86 % identity with the human IP-10 receptor. It mediates Ca2+mobilization in response to IP-10, but does not recognize other rodent chemokines, including GRO, RANTES, monocyte chemoattractant protein-1 (MCP-1), and macrophage inflammatory protein-1α (MIP-1α). Northern blot analysis revealed that murine IP-10 and its receptor mRNA were constitutively expressed in the spleen and thymus from normal mouse, while IP-10 and its receptor mRNA were derived from stromal cells and lymphocytes in both tissues, respectively.In vivotreatment with concanavalin A (Con A) for 12 hrs revealed that splenocytes significantly induce IP-10 receptor mRNA expression and show a good chemotactic response to IP-10. Therefore, it is supposed that IP-10 and its receptor are important for lymphocyte trafficking to lymphoid organs and that the IP-10 receptor on lymphocytes is rapidly inducible on inflammation or in immunological events.
- Published
- 1998