title: Protection of Human Colon Cells from Shiga Toxin by Plant-based Recombinant Secretory IgA.
abstract: Shiga toxin is a major virulence factor of food-poisoning caused by Escherichia coli such as O157:H7. Secretory immunoglobulin (Ig) A (SIgA) is supposed to prevent infection of the mucosal surface and is a candidate agent for oral immunotherapy. We previously established a recombinant monoclonal antibody (mAb) consisting of variable regions from a mouse IgG mAb specific for the binding subunit of Shiga toxin 1 (Stx1) and the Fc region of mouse IgA. Here we produced a secretory form of the recombinant IgA (S-hyIgA) with transgenic Arabidopsis thaliana plant. All the S-hyIgA cDNAs (heavy, light, J chain and secretory component) were expressed under the control of a bidirectional promoter of a chlorophyll a/b-binding protein of A. thaliana without using a viral promoter. The plant-based S-hyIgA exhibited antigen binding, and was modified with plant-specific N-linked sugar chains. The Ig heavy chain and secretory components were observed in an intracellular protein body-like structure of the transgenic leaves on immuno-electron microscopy. An extract of the transgenic leaves neutralized the cytotoxicity of Stx1 toward butyrate-treated Caco-2 cells, a human colon carcinoma cell line. These results will contribute to the development of edible therapeutic antibodies such as those for the treatment of mucosal infection.
authors.affiliation: Laboratory of Microbiology and Immunology, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka City, Shizuoka 422-8526, Japan.
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Nakanishi
authors.affiliation: Laboratory of Microbiology and Immunology, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka City, Shizuoka 422-8526, Japan.
authors.forename: Shota
authors.initials: S
authors.lastname: Morikane
authors.affiliation: Laboratory of Microbiology and Immunology, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka City, Shizuoka 422-8526, Japan.
authors.forename: Shiori
authors.initials: S
authors.lastname: Ichikawa
authors.affiliation: Laboratory of Microbiology and Immunology, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka City, Shizuoka 422-8526, Japan.
authors.forename: Kohta
authors.initials: K
authors.lastname: Kurohane
authors.affiliation: Laboratory of Plant Molecular Improvement, Graduate Division of Nutritional and Environmental Sciences, University of Shizuoka, Shizuoka City, Shizuoka 422-8526, Japan.
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Niwa
authors.affiliation: Department of Anatomy, Kyorin University School of Medicine, Mitaka, Tokyo 181-8612, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Akimoto
authors.affiliation: Laboratory for Electron Microscopy, Kyorin University School of Medicine, Mitaka, Tokyo 181-8612, Japan.
authors.forename: Sachie
authors.initials: S
authors.lastname: Matsubara
authors.affiliation: Department of Anatomy, Kyorin University School of Medicine, Mitaka, Tokyo 181-8612, Japan.
authors.forename: Hayato
authors.initials: H
authors.lastname: Kawakami
authors.affiliation: Laboratory of Plant Molecular Improvement, Graduate Division of Nutritional and Environmental Sciences, University of Shizuoka, Shizuoka City, Shizuoka 422-8526, Japan.
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Kobayashi
authors.affiliation: Laboratory of Microbiology and Immunology, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka City, Shizuoka 422-8526, Japan.
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Imai
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology* / Q000494:pharmacology; D017360:Arabidopsis / Q000235:genetics* / Q000276:immunology; D018938:Caco-2 Cells; D019453:Escherichia coli O157 / Q000187:drug effects / Q000276:immunology* / Q000472:pathogenicity; D006801:Humans; D007070:Immunoglobulin A / Q000096:biosynthesis / Q000276:immunology / Q000494:pharmacology*; D007167:Immunotherapy; D007239:Infections / Q000188:drug therapy* / Q000276:immunology / Q000382:microbiology; D051379:Mice; D030821:Plants, Genetically Modified / Q000235:genetics / Q000276:immunology; D022621:Shiga Toxin / Q000037:antagonists & inhibitors / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D007070:Immunoglobulin A; D022621:Shiga Toxin