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2-16 - Inhibitory FcR(FcRIIB Activating receptors...

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Inhibitory Fc γ R (Fc γ RIIB) Fc γ RIIB ITIM Inhibitory receptor Immunoreceptor tyrosine-based inhibitory motif Fc γ RI α α γ 2 γ 2 Fc γ RIII α γ 2 Fc γ RIV Activating receptors ITIM P P ITIM can be phosphorylated by Src family tyr kinases SHIP CH 2 CH 2 CH P P P HO OH O O P Phosphatidylinositol-3,4,5- triphosphate (PIP 3 ) CH 2 CH 2 CH P P HO OH P O O Phosphatidylinositol-3,4- biphosphate OH SHIP phosphatase CH 2 CH 2 CH P P P HO OH O O OH IP 3 DAG PI3K Phosphatidylinositol-4,5- biphosphate PLC
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Loss of Fc γ RII increases inflammation. Mice Intravenous injection of Ovalbumin Introduce anti-ovalbumin IgG into the lung IgG-Ova immune complex induces inflammation (Arthur reaction in the lung) Hemorrhage Neutrophil infiltration Increased inflammatory cytokine production Enhanced phagocytosis of IgG-opsonized RBC Enhanced inflammation in Fc γ RII deficient mice
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Activating Fc γ R Inhibitory Fc γ R C5a Interferon γ Strong activation by immune complex TGF- β IL-10 IL-4 Weak activation by immune complex Regulation of Fc receptor expression Different activities of IgG subtypes IgG2a 2.9x10 7 0.4x10 6 IIB IV Ka IgG2b 1.7x10 7 2.2x10 6 IIB IV IgG1 0.3x10 6 3.3x10 6 IIB III IgG3 IIB A/I=70 A/I=7 A/I=0.1 Fc γ R-mediated activation +++ + -
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Antibody therapy Immunization Polyclonal antibody in serum Isolate individual hybridoma clones Monoclonal antibody Ig production Short-lived in culture B cells in spleen recognize different epitopes Myeloma (mouse tumor derived from plasma cells) No Ig production Ig secretion apparatus Immortal Antigen Fusion of B cells with myeloma hybridoma PEG (polyethylene glycol) Ig production Ig secretion apparatus Immortal
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Hybridoma screening Plate cells into 96-well plates
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