title: Inhibition of B cell receptor-mediated activation of primary human B cells by coengagement of CD19 and FcgammaRIIb with Fc-engineered antibodies.
abstract: The humoral immune response requires antigen-specific B cell activation and subsequent terminal differentiation into plasma cells. Engagement of B cell antigen receptor (BCR) on mature B cells activates an intracellular signaling cascade, including calcium mobilization, which leads to cell proliferation and differentiation. Coengagement by immune complex of BCR with the inhibitory Fc receptor FcgammaRIIb, the only IgG receptor expressed on B cells, inhibits B cell activation signals through a negative feedback loop. We now describe antibodies that mimic the inhibitory effects of immune complex by high-affinity coengagement of FcgammaRIIb and the BCR coreceptor complex on human B cells. We engineered the Fc domain of an anti-CD19 antibody to generate variants with up to approximately 430-fold greater affinity to FcgammaRIIb. Relative to native IgG1, the FcgammaRIIb binding-enhanced (IIbE) variants strongly inhibited BCR-induced calcium mobilization and viability in primary human B cells. Inhibitory effects involved phosphorylation of SH2-containing inositol polyphosphate 5-phosphatase (SHIP), which is known to be involved in FcgammaRIIb-induced negative feedback of B cell activation by immune complex. Coengagement of BCR and FcgammaRIIb by IIbE variants also overcame the anti-apoptotic effects of BCR activation. The use of a single antibody to suppress B cell functions by coengagement of BCR and FcgammaRIIb may represent a novel approach in the treatment of B cell-mediated autoimmune diseases.
authors.affiliation: Xencor, Inc., 111 W. Lemon Avenue, Monrovia, CA 91016, USA.
authors.forename: Seung Y
authors.initials: SY
authors.lastname: Chu
authors.forename: Igor
authors.initials: I
authors.lastname: Vostiar
authors.forename: Sher
authors.initials: S
authors.lastname: Karki
authors.forename: Gregory L
authors.initials: GL
authors.lastname: Moore
authors.forename: Greg A
authors.initials: GA
authors.lastname: Lazar
authors.forename: Erik
authors.initials: E
authors.lastname: Pong
authors.forename: Patrick F
authors.initials: PF
authors.lastname: Joyce
authors.forename: David E
authors.initials: DE
authors.lastname: Szymkowski
authors.forename: John R
authors.initials: JR
authors.lastname: Desjarlais
mesh_terms: D000906:Antibodies / Q000235:genetics / Q000276:immunology*; D018941:Antigens, CD19 / Q000276:immunology*; D017209:Apoptosis; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics*; D000072181:Inositol Polyphosphate 5-Phosphatases; D008213:Lymphocyte Activation; D010744:Phosphoric Monoester Hydrolases / Q000276:immunology; D011947:Receptors, Antigen, B-Cell / Q000276:immunology* / Q000378:metabolism; D017452:Receptors, IgG / Q000235:genetics / Q000276:immunology*; D015398:Signal Transduction; D020349:Surface Plasmon Resonance
chemical_list: D000906:Antibodies; D018941:Antigens, CD19; C080571:Fc gamma receptor IIB; D007141:Immunoglobulin Fc Fragments; D011947:Receptors, Antigen, B-Cell; D017452:Receptors, IgG; D010744:Phosphoric Monoester Hydrolases; D000072181:Inositol Polyphosphate 5-Phosphatases; D002118:Calcium