DIFFERENTIAL USAGE OF THE CXC CHEMOKINE RECEPTOR-1 AND RECEPTOR-2 BY INTERLEUKIN-8, GRANULOCYTE CHEMOTACTIC PROTEIN-2 AND EPITHELIAL-CELL-DERIVED NEUTROPHIL ATTRACTANT-78

The inflammatory response is mediated by a family of chemotactic cytok ines, designated chemokines. The receptor usage of the CXC chemokine g ranulocyte chemotactic protein-2 (GCP-2) was compared with that of int erleukin-8 (IL-8) and epithelial-cell-derived neutrophil attractant-78 (ENA-78). Chemokine activities were evaluated by measurement of intra cellular calcium increase and by chemotaxis and binding assays, using CXC chemokine receptor (CXCR)-transfected cell lines. GCP-2 was equall y potent at inducing a rise in [Ca2+](i) in both CXCR1-transfected and CXCR2-transfected cells (minimal effective concentration 3 nM). IL-8 augmented the [Ca2+](i) more efficiently in CXCR1-transfectants than i n CXCR2- transfectants, whereas for ENA-78, threefold higher concentra tions were necessary to obtain a calcium response in CXCR1-transfected cells than in CXCR2-transfectants. GCP-2 desensitized the calcium inc rease induced by IL-8 in both CXCR1-transfected and CXCR2-transfected cells, but ENA-78 only affected the IL-8-induced calcium response in C XCR2-transfectants The half-maximal effective concentrations for migra tion of CXCR2-transfectants in response to GCP-2 and ENA-78 were simil ar (0.1 nM), whereas GCP-2 was tenfold more potent than ENA-78 an CXCR 1-transfectants. Half-maximal migration of CXCR1-transfected and CXCR2 -transfected cells was obtained with IL-8 at concentrations of no more than 0.01 nM. Radiolabeled IL-8 could efficiently be displaced from C XCR2 by IL-8, GCP-2 and ENA-78. In contrast, only IL-8 and GCP-2 but n ot ENA-78, competed for I-125-IL-8 binding to CXCR1. From these data, it can be concluded that, in addition to IL-8, GCP-2, but not ENA-78, efficiently binds to both CXCR1 and CXCR2. The differential receptor u sage of the structurally related ELR+CXC chemokines GCP-2 and ENA-78 i s indicative of a different role in inflammatory reactions.