Concanavalin A-Induced Histamine Release from Normal Rat Mast Cells

Abstract

Concanavalin A- (con A) induced release of histamine from normal rat mast cells was studied. In the presence of phosphatidylserine (PS) con A induced a concentration and temperature-dependent, noncytotoxic histamine release at con A concentrations ranging from 0.1 to 100 µg/ml. The optimal con A concentration, 100 µg/ml, caused a 27.3% (±2.7 S.E.M.) net histamine release. Release began approximately 30 sec after addition of con A and was complete within 45 min. In the absence of PS, no net con A-induced release occurred. The effect of PS was concentration dependent from 1 to 100 µg/ml. PS alone, however, did not cause histamine release. Binding studies indicated that mast cells bound up to 16 × 10⁶ con A molecules per cell without histamine release. Upon removal of unbound con A and the addition of PS, normal histamine release occurred. α-Methyl-D-mannose (50 mM) prevented both con A binding and histamine release and if added after Con A, caused a rapid cessation of histamine release and a reversal of con A binding. This study indicates several important advantages of the con A-induced histamine release system. Binding of con A to mast cells can be dissociated from histamine release by omitting PS from the medium. Release can then be induced by the addition of PS. α-Methyl-D-mannose can be used to terminate rapidly the ongoing release reaction at any phase of the interaction. This system is potentially very useful for investigation of metabolic events during histamine release.