title: Recombinant human monoclonal igA antibody against CEA to recruit neutrophils to CEA-expressing cells.
abstract: IgA is able to trigger antibody-dependent cellular cytotoxicity (ADCC) by recruiting neutrophils expressing the Fc receptor for the CHalpha chain. We herein describe the preparation of a human recombinant anti-CEA IgA antibody to kill carcinoembryonic antigen (CEA)-expressing tumor cells via ADCC by neutrophils. A single chain Fv (scFv) gene was constructed using a cDNA library of a hybridoma clone that produces a human anti-CEA monoclonal IgG4 (C2-45). The scFv gene, linked with a CHalpha gene, was inserted into the pBK283 vector, which was cotransfected into BmN4 insect cells with the wild-virus BmNPV. After cloning and amplification, the recombinated virus was injected into silkworm larvae. The resulting human recombinant IgA, designated as 45scFvLCHalpha, was purified from hemolymph by Ni-affinity chromatography and characterized by ELISA, Western blotting, and the ADCC assay. 45scFvLCHalpha with an IgA antigenicity was bound to CEA and showed effective killing of the CEA-expressing cells in the presence of IFN-gamma-activated neutrophils. These data suggest the recombinant anti-CEA IgA antibody recruiting neutrophils maybe a useful means for the antibody-based immunotherapy of human CEA-expressing tumors.
authors.affiliation: Molecular Oncology Center, Faculty of Medicine, Fukuoka University, Fukuoka 814-0180, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Zhao
authors.forename: Motomu
authors.initials: M
authors.lastname: Kuroki
authors.forename: Hirotomo
authors.initials: H
authors.lastname: Shibaguchi
authors.forename: Lin
authors.initials: L
authors.lastname: Wang
authors.forename: Qun
authors.initials: Q
authors.lastname: Huo
authors.forename: Noboru
authors.initials: N
authors.lastname: Takami
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Tanaka
authors.forename: Tetsushi
authors.initials: T
authors.lastname: Kinugasa
authors.forename: Masahide
authors.initials: M
authors.lastname: Kuroki
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D000920:Antibody-Dependent Cell Cytotoxicity; D012831:Bombyx; D016466:CHO Cells; D002272:Carcinoembryonic Antigen / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D045744:Cell Line, Tumor; D006224:Cricetinae; D003412:Cricetulus; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007070:Immunoglobulin A / Q000235:genetics / Q000276:immunology*; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology; D007167:Immunotherapy / Q000379:methods; D007371:Interferon-gamma / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D002272:Carcinoembryonic Antigen; D007070:Immunoglobulin A; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; D007371:Interferon-gamma