Endothelin-1 Regulation of Intercellular Adhesion Molecule-1 Expression in Normal and Sclerodermal Fibroblasts

We investigated the potential modulation of cell surface intercellular adhesion molecule-1 (ICAM-1) expression and function by ET-1 in fibroblasts grown from skin biopsies of scleroderma (SSc) patients compared with healthy controls. Surface ICAM-1 expression was quantified by cell-bound ELISA and by FACS analysis. ICAM-1 function was investigated by measuring cell adhesion, and we studied ICAM-1 gene expression using RT-PCR. Fibroblast ET-1 binding sites were measured using 125I-labeled ET-1, and the modulation of ICAM-1 function by ET-1 was determined by measuring the binding of human U937 cells to fibroblasts in the presence of a mixed ETA/B receptor antagonist (bosentan) or a neutralizing anti-ICAM-1 antibody. ICAM-1 expression was significantly higher in SSc fibroblasts compared with normal controls. ET-1 increased ICAM-1 on both normal and SSc fibroblasts to comparable levels. RT-PCR demonstrated that ICAM-1 mRNA was upregulated by ET-1, and results from binding studies showed fibroblasts exposed to ET-1 supported more U937 cells than controls, a process that could be inhibited by bosentan and ICAM-1 neutralizing antibody. Autoradiography showed ET-1 receptors on both normal and SSc fibroblasts. Our findings indicate that SSc fibroblasts express intrinsically elevated levels of surface ICAM-1 and message. ET-1 can induce normal fibroblasts to express some SSc phenotypes and may function as a potent proinflammatory mediator, similar to cytokines, and therefore may also have immunoregulatory functions for immune cells infiltrating and binding to connective tissues.