4,4′-Diisothiocyanatostilbene-2,2′-disulfonic acid inhibits CD3-T cell antigen receptor-stimulated Ca2+ influx in human T lymphocytes

• Published in: The Journal of biological chemistry 1988-12, Vol.263 (36), p.19535-19540
• Main Authors: Rosoff, P M, Hall, C, Gramates, L S, Terlecky, S R
• Format: Article
• Language: English
• Subjects: 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid; 4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid - analogs & derivatives; 4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid - pharmacology; Antigens, Differentiation, T-Lymphocyte - metabolism; Antigens, Surface - immunology; Biological and medical sciences; Biological Transport - drug effects; Calcium - metabolism; CD3 Complex; Cell Line; Cell physiology; Egtazic Acid - pharmacology; Fundamental and applied biological sciences. Psychology; Humans; Kinetics; Membrane and intracellular transports; Molecular and cellular biology; Receptors, Antigen, T-Cell - drug effects; Receptors, Antigen, T-Cell - metabolism; Receptors, Antigen, T-Cell - physiology; Stilbenes - pharmacology; T-Lymphocytes - drug effects; T-Lymphocytes - immunology; T-Lymphocytes - metabolism
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(19)77668-6

Stimulation of the CD3-T cell antigen receptor complex on T lymphocytes results in a rapid rise in intracellular calcium from both intra- and extracellular sources. The former is thought to be released from the endoplasmic reticulum in response to inositol trisphosphate, while the latter enters the cells through a membrane potential-sensitive transporter (Oettgen, H. C., Terhorst, C., Cantley, L. C., and Rosoff, P. M. (1985) Cell 40, 583-590). In this report we show that the stilbene disulfonate, DIDS (4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid), inhibited the ability of monoclonal anti-CD3 complex antibodies to stimulate an influx of calcium in the human T lymphocyte cell line, Jurkat. DIDS had no effect on either antibody binding to the receptor or receptor-stimulated phosphatidylinositol turnover. The Ki was approximately 25 microM in the presence of extracellular Cl- and 10 microM when labeling was performed in the absence of Cl-, suggesting that DIDS was competing with Cl- for binding to the cell membrane. The reduced form of DIDS, dihydroDIDS, was only 50% as effective as DIDS itself, and the monoisothiocyanate stilbene, 4-acetamido-4′-isothiocyantostilbene-2,2′-disulfonic acid, was totally ineffective, even to concentrations of 0.750 mM. Removal of extracellular Cl- also inhibited the antibody-stimulated influx of calcium. These data suggest that the function of the CD3-T cell receptor-activated calcium channel/transporter may be dependent on or regulated by extracellular Cl-.