Epitope mapping of the site(s) of binding of Fc epsilon RII/CD23 within human IgE. Determination of the B lymphocyte-binding sites by use of synthetic peptides and anti-peptide antibodies representative of linear Fc sequences.
The work undertaken has investigated the structure-function relationship between IgE and its low affinity receptor on B lymphocytes. To identify sites on the IgE molecule which interact with the low affinity receptor (Fc epsilon RII/CD23), 10 different peptide sequences within the CH2, CH3 and CH4 domains of human IgE were selected according to charge, overall hydrophobicity and possible accessibility on native IgE sequences. Peptides representative of these were synthesized by the solid phase procedure; and their cytophilic activities were examined by determining their capacity to inhibit the binding of radiolabelled or erythrocyte-bound IgE to a Fc epsilon RII/CD23 positive B cell line (RPMI-8866). Moreover, these linear sequences were rendered immunogenic by conjugation to a protein carrier (KLH) and used to produced polyclonal antibodies in rabbits. The reactivity of the anti-peptide antibodies with both free peptides and native IgE bound to a solid phase, as well as their capacity to inhibit binding of IgE to a Fc epsilon RII/CD23 positive cell line, were investigated. Results from such use of peptides and anti-peptide antibodies indicate that two sequences, representative of residues 364-383 and 401-415, could be involved in the binding of IgE to both membrane-bound and soluble form Fc epsilon RII/CD23; indicating that the B lymphocyte-binding site on human IgE may be restricted to the CH3 domain.
Ghaderi
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AA
Department of Microbiology and Immunology, Shiraz University of Medical Science, Iran.
Stanworth
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DR
D000818:Animals; D000937:Antigen-Antibody Reactions; D001402:B-Lymphocytes / Q000276:immunology*; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D000939:Epitopes / Q000276:immunology*; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology*; D007141:Immunoglobulin Fc Fragments / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology; D011817:Rabbits; D017455:Receptors, IgE / Q000276:immunology*; D012397:Rosette Formation
D000939:Epitopes; D007141:Immunoglobulin Fc Fragments; D010446:Peptide Fragments; D017455:Receptors, IgE; D007073:Immunoglobulin E