Vanessa L. Ott, PhD, Dana C. Fong, PhD, John C. Cambier, PhD

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Presentation transcript:

FcγRIIB as a potential molecular target for intravenous gamma globulin therapy Vanessa L. Ott, PhD, Dana C. Fong, PhD, John C. Cambier, PhD Journal of Allergy and Clinical Immunology Volume 108, Issue 4, Pages S95-S98 (October 2001) DOI: 10.1067/mai.2001.117822 Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 1 Coaggregation of FcγRIIB and the BCR results in FcγRIIB recruitment of SHIP and Grb2. The FcγRIIB-deficient B-cell line IIA1.6, either untransfected (IIA1.6), transfected with wild-type FcγRIIB (WT), or transfected with FcγRIIB that lacks the C-terminal 16 amino acids (CT314) were used in this experiment. Cells were either left untreated (u) or stimulated with equimolar amounts of either rabbit anti-mouse F(ab')2 (to aggregate the BCR) or rabbit anti-mouse IgG (to coaggregate the BCR and FcγRIIB) for 5 minutes at 37°C. Cells were lysed in 1% NP-40 lysis buffer for 10 minutes on ice, followed by centrifugation to remove the insoluble material. FcγRIIB was isolated from cleared lysates by immunoprecipitation with an anti-FcγRIIB antibody and protein A-conjugated Sepharose beads. Immune complexes were resolved by sodium dodecyl sulfate (SDS)–polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride (PVDF) membrane. FcγRIIB, SHIP, and Grb2 were detected by immunoblotting with specific antibodies, followed by horseradish peroxidase (HRP)–conjugated secondary antibodies and detection with enhanced chemiluminescence (ECL) and autoradiography. Journal of Allergy and Clinical Immunology 2001 108, S95-S98DOI: (10.1067/mai.2001.117822) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 2 Schematic of BCR signal transduction and inhibition by FcγRIIB. PtdIns , phosphatidylinositol; Ins , inositol; BCR , B-cell receptor; PLC γ, phospholipase Cγ; PI3-K , phosphatidylinositol 3-kinase; SHP , SH2 domain-containing tyrosine phosphatase; SHIP , SH2 domain-containing inositol 5-phosphatase. Journal of Allergy and Clinical Immunology 2001 108, S95-S98DOI: (10.1067/mai.2001.117822) Copyright © 2001 Mosby, Inc. Terms and Conditions