title: Targeting the junction of CɛmX and ɛ-migis for the specific depletion of mIgE-expressing B cells.
abstract: Monoclonal antibodies targeting the extracellular region of the human IgE heavy chain membrane-tethering domain have been proposed for treating allergies caused by hyperproliferative monoclonal expansion of IgE-producing B cells. Antibodies against this target are expected to deplete membrane IgE (mIgE) displaying B cells and leave B cells of other immunoglobulin isotypes intact. Because of alternative splicing, the mIgE heavy chain has two isoforms that differ in their membrane-proximal segment. In the long isoform, the CH4 domain is followed by a 67-amino acid-long extracellular portion. Out of these 67 amino acids, the first 52 amino acids following the CH4 domain constitute the CɛmX segment while the rest of the 15 amino acids immediately adjacent to the membrane constitute the ɛ-migis. In the short isoform the CɛmX segment is absent and the CH4 domain is followed only by the 15-amino acid-long ɛ-migis segment. Using antibodies derived from a phage display library, we investigated: (1) ɛ-migis and (2) the junction of CɛmX and ɛ-migis (CɛmX.migis), as potential therapeutic antibody targets. Our results indicate that antibodies obtained from our phage library that target ɛ-migis bind to a variety of human cells irrespective of mIgE expression, possibly due to homology between ɛ-migis and a region of phosphoinositide-binding protein (ARAP3). In contrast, antibodies specific for the CɛmX.migis junctional region, bound specifically to transfected and primary B cells expressing human mIgE and elicited antibody-dependent cellular cytotoxicity and reduction in IgE production. These antibodies did not bind secreted IgE or the mIgE isoform in which CɛmX is absent. These results suggest that CɛmX.migis junctional region is a promising antibody target and the human antibodies we describe warrant further evaluation.
authors: Department of Antibody Discovery and Protein Engineering, MedImmune, LLC, Gaithersburg, MD 20878, USA. chowdhuryp@medimmune.com
authors: Partha S
authors: PS
authors: Chowdhury
authors: Yan
authors: Y
authors: Chen
authors: Chunning
authors: C
authors: Yang
authors: Kimberly E
authors: KE
authors: Cook
authors: Andrew C
authors: AC
authors: Nyborg
authors: Rachel
authors: R
authors: Ettinger
authors: Ronald
authors: R
authors: Herbst
authors: Peter A
authors: PA
authors: Kiener
authors: Herren
authors: H
authors: Wu
mesh_terms: D000888:Antibodies, Anti-Idiotypic / Q000276:immunology*; D000911:Antibodies, Monoclonal; D000920:Antibody-Dependent Cell Cytotoxicity; D001402:B-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology; D049109:Cell Proliferation; D057809:HEK293 Cells; D006801:Humans; D007073:Immunoglobulin E / Q000096:biosynthesis / Q000276:immunology*; D007139:Immunoglobulin epsilon-Chains / Q000276:immunology*; D010716:Phosphatidylinositols / Q000276:immunology; D020033:Protein Isoforms / Q000276:immunology; D011947:Receptors, Antigen, B-Cell / Q000276:immunology* / Q000378:metabolism
chemical_list: D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D007139:Immunoglobulin epsilon-Chains; D010716:Phosphatidylinositols; D020033:Protein Isoforms; D011947:Receptors, Antigen, B-Cell; D007073:Immunoglobulin E