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-1alpha (MIP-1alpha). 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 vivo treatment 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.
Copyright 1998 Academic Press.