Cross-Linking of IgE-receptor complexes at the cell surface: Synthesis and characterization of a long bivalent hapten that is capable of triggering mast cells and rat basophilic leukemia cells

Abstract

The ability of a series of bivalent haptens to bind and cross-link immunoglobulin E (IgE) in solution and on the surface of cells was examined. Several short (< 30 Å) dinitrophenyl (DNP) haptens were found to bind tightly to and cross-link a monoclonal anti-DNP IgE in solution, but these failed to trigger substantial release of 3H-serotonin from sensitized rat basophilic leukemia (RBL) cells or rat peritoneal mast cells. A longer bivalent hapten, ~ 50 Å in length, consisting of two DNP-aminocaproyl- l-tyrosine (DCT) groups coupled to the α-amino groups of l-cystine was synthesized and characterized. This bivalent hapten [(DCT) 2-cystine], binds very tightly to the same monoclonal anti-DNP IgE in solution and cross-links these antibodies to form higher mol. wt aggregates as judged by gel filtration and binding studies. It also stimulates degranulation of both RBL and mast cells sensitized with two different monoclonal anti-DNP IgE antibodies, with the mast cells exhibiting generally greater responsiveness to this ligand. The (DCT) 2-cystine bivalent hapten appears to have the structural features necessary for carrying out detailed binding studies with receptor-bound IgE on the cell surface.