Separation of specific antibody forming mouse cells by their adherence to insolubilized endogenous hormones

Spleen cells from mice immunized with sheep red cells were separated by differential adherence into insolubilized histamine, catecholamines, and prostaglandins. The hormones were insolubilized by linking them to Sepharose beads through a protein carrier. The authors 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 F2α 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 adenosine monophosphate (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.