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Separation of Specific Antibody-Forming Mouse Cells by their Adherence to Insolubilized Endogenous Hormones

Kenneth L. Melmon, Yacob Weinstein, G. M. Shearer, Henry R. Bourne and Sara Bauminger

Division of Clinical Pharmacology, Department of Pharmacology, University of California Medical Center, San Francisco, California 94143Division of Clinical Pharmacology, Department of Medicine, University of California Medical Center, San Francisco, California 94143Department of Chemical Immunology, The Weizmann Institute of Science, Rehovot, IsraelDepartment of Biodynamics, The Weizmann Institute of Science, Rehovot, Israel

Published January 1974

Spleen cells from mice immunized with sheep red cells were separated by differential adherence to insolubilized histamine, catecholamines, and prostaglandins. The hormones were insolubilized by linking them to Sepharose beads through a protein carrier. We measured hemolytic plaque formation (per million splenic leukocytes) of cells which passed through columns of hormone-carrier-Sepharose beads (i.e., those cells that failed to bind). As compared with control (no column) cells, the number of plaque-forming cells was substantially reduced by passage through histamine, epinephrine, isoproterenol, and prostaglandin-E2 columns. Plaque-forming cells were not significantly reduced by passage through carrier Sepharose (another control) or norepinephrine- and prostaglandin-F-carrier Sepharose columns. Thus, the ability of an insolubilized hormone preparation to subtract plaque-forming cells roughly correlated with the presence of pharmacologic receptors for the corresponding free hormones, as judged by stimulation of cyclic AMP accumulation in the same cells, reported previously. Both 19S and 7S plaque-forming cells were subtracted by columns prepared from pharmacologically active hormones, but none of the insolubilized hormones stimulated accumulation of intracellular cyclic AMP. The cell membrane phenomenon that allows adherence to a given hormone-carrier-bead column may be identical with the cell receptor.

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