Cloning of the murine interferon-inducible protein 10 (IP-10) receptor and its specific expression in lymphoid organs.
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.
Tamaru
M
M
JT Central Pharmaceutical Research Institute, Yokohama, Kanagawa, 236-0004, Japan.
Tominaga
Y
Y
Yatsunami
K
K
Narumi
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D054357:Chemokine CXCL10; D019743:Chemokines, CXC / Q000378:metabolism; D003001:Cloning, Molecular; D003208:Concanavalin A / Q000494:pharmacology; D005260:Female; D008213:Lymphocyte Activation; D008221:Lymphoid Tissue / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009928:Organ Specificity / Q000235:genetics / Q000276:immunology; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D019707:Receptors, Chemokine / Q000096:biosynthesis* / Q000235:genetics* / Q000502:physiology; D011994:Recombinant Proteins / Q000378:metabolism; D013154:Spleen / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology / Q000378:metabolism; D014162:Transfection
D054357:Chemokine CXCL10; D019743:Chemokines, CXC; D012333:RNA, Messenger; D019707:Receptors, Chemokine; D011994:Recombinant Proteins; D003208:Concanavalin A; D002118:Calcium