Analysis of the interaction between rat immunoglobulin E and rat mast cells using anti-peptide antibodies.
Polyclonal antisera with pre-determined specificities for a range of rat IgE epitopes were produced by immunizing rabbits with KLH-conjugates of five different synthetic peptides representing sequences 378-396, 414-428, 491-503, 522-535 and 560-571 in the CH3 and CH4 domains of rat IgE. Each rabbit elicited peptide-specific antibodies which were capable of binding affinity-purified rat IgE (IR162) (titres 1/1000-1/10,000) and IgE in rat immunocytoma serum (IR162) either immobilized on microtitre-plates or in free-solution as assessed by ELISA. Heating a solution of rat IgE at 56 degrees C for 1 hr, a treatment known to abolish the cytophilic activity of rat IgE and also induce irreversible conformational changes in the CH3 and CH4 domains, resulted in enhanced binding of the immunoglobulin to antibodies directed against IgE sequences represented by two of the synthetic peptides 414-428 and 491-503, but not to the three other peptides. The five anti-peptide sera together with two previously studied antisera specific for rat IgE sequences 459-472 and 542-557 were tested in functional assays designed to investigate the mode of interaction between rat IgE and its receptor on rat mast cells. Each anti-peptide serum was capable of inhibiting the binding of IgE to mast cells and furthermore, able to initiate the secretion of histamine from cells sensitized with rat IgE in an "anti-IgE"-induced manner. In view of the evidence implicating the CH3 and/or CH4 domains as the location of the mast cell receptor-site on rat IgE, we propose a model to describe the mode of interaction between IgE and its mast cell receptor.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001667:Binding, Competitive; D006636:Histamine Release; D007106:Immune Sera / Q000276:immunology; D007073:Immunoglobulin E / Q000378:metabolism*; D007143:Immunoglobulin Heavy Chains / Q000276:immunology; D008407:Mast Cells / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology; D051381:Rats; D011919:Rats, Inbred Strains; D011961:Receptors, Fc / Q000378:metabolism*; D017455:Receptors, IgE
D000596:Amino Acids; D007106:Immune Sera; D007143:Immunoglobulin Heavy Chains; D010446:Peptide Fragments; D011961:Receptors, Fc; D017455:Receptors, IgE; D007073:Immunoglobulin E