Protection of Human Colon Cells from Shiga Toxin by Plant-based Recombinant Secretory IgA

Sci Rep. 2017 Apr 3:7:45843. doi: 10.1038/srep45843.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis
  • Antibodies, Monoclonal / immunology*
  • Antibodies, Monoclonal / pharmacology
  • Arabidopsis / genetics*
  • Arabidopsis / immunology
  • Caco-2 Cells
  • Escherichia coli O157 / drug effects
  • Escherichia coli O157 / immunology*
  • Escherichia coli O157 / pathogenicity
  • Humans
  • Immunoglobulin A / biosynthesis
  • Immunoglobulin A / immunology
  • Immunoglobulin A / pharmacology*
  • Immunotherapy
  • Infections / drug therapy*
  • Infections / immunology
  • Infections / microbiology
  • Mice
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / immunology
  • Shiga Toxin / antagonists & inhibitors
  • Shiga Toxin / immunology

Substances

  • Antibodies, Monoclonal
  • Immunoglobulin A
  • Shiga Toxin