title: Cell surface expression of a human IgG Fc chimera activates macrophages through Fc receptors.
abstract: Antibody-dependent cell-mediated cytotoxicity plays an important role in the macrophage-mediated destruction of target cells. While the selectivity is based on antibody specificity, the lytic attack is triggered by Fc receptor-mediated respiratory burst. To mimic IgG opsonization, a chimeric antibody-like molecule, containing human IgG1 Fc, was expressed on the surface of mammalian cells. The transmembrane domain of the human transferrin receptor was fused in-frame to the N-terminus of the second and third domains of human IgG1 heavy-chain constant region. This fusion molecule was designed to take advantage of the type II membrane anchor property of the transferrin receptor to express the Fc portion of the molecule in a reverse orientation, such that the Fc portion projected away from the cell surface. This is in contrast to the conventional cell surface IgG, which is anchored by a C-terminal type I transmembrane domain. The cell surface expressed reverse Fc no longer activated complement, but retained Fc receptor-binding capability and activated superoxide production by macrophages. This activity was completely blocked by an FcgammaR I-specific monoclonal antibody.
authors.affiliation: Department of Immunology, CytoTherapeutics, Inc., Lincoln, RI 02865, USA.
authors.forename: P F
authors.initials: PF
authors.lastname: Stabila
authors.forename: S C
authors.initials: SC
authors.lastname: Wong
authors.forename: F A
authors.initials: FA
authors.lastname: Kaplan
authors.forename: W
authors.initials: W
authors.lastname: Tao
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology; D006224:Cricetinae; D017931:DNA Primers; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000378:metabolism*; D007074:Immunoglobulin G / Q000235:genetics / Q000378:metabolism*; D008262:Macrophage Activation / Q000276:immunology*; D011961:Receptors, Fc / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D011961:Receptors, Fc; D011993:Recombinant Fusion Proteins