Identification of a common receptor for three types of rat cytokine-induced neutrophil chemoattractants (CINCs).
Rat cytokine-induced neutrophil chemoattractant-1 (CINC-1), CINC-2 and CINC-3/macrophage inflammatory protein-2 (MIP-2), members of the CXC chemokine family, are potent chemotactic factors for neutrophils. In order to identify the receptor for CINCs, rat CXC chemokine receptor 2 (CXCR2) was cloned and expressed in HEK293 cells. CINC-1, CINC-2 and CINC-3 induced calcium mobilizations dose-dependently in CXCR2-transfected cells, whereas formyl-methionyl-leucyl-phenylalanine (FMLP) did not. CINC-3 induced enhancement of cytoplasmic calcium concentration more potently than CINC-1 and CINC-2, and desensitized calcium transients induced by CINC-1 and CINC-2, which were essentially identical to those observed in rat neutrophils. In addition, anti-CXCR2 serum inhibited neutrophil chemotactic activities of three types of CINCs almost completely. The mutant CINC-3, whose amino-terminal amino acid sequence (SELR) was replaced to AAR, lost chemotactic activity of its own but inhibited that of CINC-1 and CINC-2 potently, and that of CINC-3 weakly. The results indicate that rat CXCR2 on neutrophils is the unique receptor for all three types of CINCs, and CINC-1/-2 and CINC-3 exert different biological activities through the common receptor.
Shibata
F
F
Department of Physiological Chemistry, Toyama Medical and Pharmaceutical University, Sugitani, Toyama 930-0194, Japan. fshibata@ms.toyama-mpu.ac.jp
Konishi
K
K
Nakagawa
H
H
D000818:Animals; D015152:Blotting, Northern; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D054360:Chemokine CXCL1; D054426:Chemokine CXCL2; D019743:Chemokines, CXC; D002630:Chemotactic Factors / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology / Q000502:physiology; D002633:Chemotaxis / Q000187:drug effects; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D006133:Growth Substances / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology / Q000502:physiology; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D008099:Liver / Q000378:metabolism; D016296:Mutagenesis; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000378:metabolism; D016133:Polymerase Chain Reaction; D051381:Rats; D023063:Receptors, Interleukin-8B / Q000235:genetics / Q000378:metabolism; D013053:Spectrophotometry; D013997:Time Factors; D014162:Transfection
C475829:CXCL1 protein, human; D054360:Chemokine CXCL1; D054426:Chemokine CXCL2; D019743:Chemokines, CXC; D002630:Chemotactic Factors; C513297:Cxcl1 protein, rat; C513296:Cxcl2 protein, rat; C513294:Cxcl3 protein, rat; D018076:DNA, Complementary; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D023063:Receptors, Interleukin-8B; D009240:N-Formylmethionine Leucyl-Phenylalanine; D002118:Calcium