Impermeant stilbene disulfonic acids block chemotactic peptide receptor function on human granulocytes

Anion transport is important in a variety of cell functions. 4,4'-Diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) and 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS) are two impermeant agents that have been reported to specifically block the anion channel in erythrocytes. These agents block several responses of human neutrophils to stimulation by immune complexes, the synthetic chemotaxin N-formyl-met-leu-phe (FMLP), and a calcium ionophore. They also alter the function of C3b receptors on the neutrophil surface. We studied the effects of DIDS and SITS on the aggregation of human neutrophils, a process that has been implicated in a number of diverse clinical syndromes. Both DIDS and SITS inhibited granulocyte aggregation induced by FMLP, zymosan-activated plasma, 12-O-tetra-decanoylphorbolmyristate acetate (TPA), and the calcium ionophore A23187. To further study the mechanism of inhibition the effects of DIDS and SITS on FMLP-receptor function were tested. Similar concentrations of the anion channel blockers also inhibited binding of radiolabeled FMLP to its specific receptor on the neutrophil surface. Inhibition of binding was due to a decrease in both the number and affinity of the surface receptors available for FMLP; DIDS did not inactivate FMLP. The effects of stilbene disulfonic acid on cell function may be due to effects of these agents on other cell-surface structures in addition to the anion channel.