Production of Hybrid-IgG/IgA Plantibodies with Neutralizing Activity against Shiga Toxin 1
| Autor: | Yasuyuki Imai, Katsuhiro Nakanishi, Hirokazu Kobayashi, Shiori Ichikawa, Sanshiro Narimatsu, Yuki Tobisawa, Yasuo Niwa, Kohta Kurohane |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
Immunoglobulin A
Arabidopsis lcsh:Medicine Gene Expression Shiga Toxin 1 medicine.disease_cause Plantibodies Immunoglobulin G Microbiology fluids and secretions Gene Order medicine Antigens Tumor-Associated Carbohydrate lcsh:Science Antibodies Blocking Escherichia coli Multidisciplinary Cell Death biology Toxin lcsh:R Shiga toxin Plants Genetically Modified Molecular biology Recombinant Proteins J chain biology.protein Plantibody lcsh:Q Expression cassette Protein Binding Research Article |
| Zdroj: | PLoS ONE PLoS ONE, Vol 8, Iss 11, p e80712 (2013) |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0080712 |
| Popis: | Shiga toxin 1 (Stx1) is a virulence factor of enterohemorrhagic Escherichia coli, such as the O157:H7 strain. In the intestines, secretory IgA (SIgA) is a major component of the immune defense against pathogens and toxins. To form SIgA, the production of dimeric IgA that retains biological activity is an important step. We previously established hybrid-IgG/IgA having variable regions of the IgG specific for the binding subunit of Stx1 (Stx1B) and the heavy chain constant region of IgA. If hybrid-IgG/IgA cDNAs can be expressed in plants, therapeutic or preventive effects may be expected in people eating those plants containing a "plantibody". Here, we established transgenic Arabidopsis thaliana expressing dimeric hybrid-IgG/IgA. The heavy and light chain genes were placed under the control of a bidirectional promoter and terminator of the chlorophyll a/b-binding protein of Arabidopsis thaliana (expression cassette). This expression cassette and the J chain gene were subcloned into a single binary vector, which was then introduced into A. thaliana by means of the Agrobacterium method. Expression and assembly of the dimeric hybrid-IgG/IgA in plants were revealed by ELISA and immunoblotting. The hybrid-IgG/IgA bound to Stx1B and inhibited Stx1B binding to Gb3, as demonstrated by ELISA. When Stx1 holotoxin was pre-treated with the resulting plantibody, the cytotoxicity of Stx1 was inhibited. The toxin neutralization was also demonstrated by means of several assays including Stx1-induced phosphatidylserine translocation on the plasma membrane, caspase-3 activation and 180 base-pair DNA ladder formation due to inter-nucleosomal cleavage. These results indicate that edible plants containing hybrid-IgG/IgA against Stx1B have the potential to be used for immunotherapy against Stx1-caused food poisoning. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|