Production of hybrid-IgG/IgA plantibodies with neutralizing activity against Shiga toxin 1.
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...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2013-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3842918?pdf=render |
| _version_ | 1818951618116190208 |
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| author | Katsuhiro Nakanishi Sanshiro Narimatsu Shiori Ichikawa Yuki Tobisawa Kohta Kurohane Yasuo Niwa Hirokazu Kobayashi Yasuyuki Imai |
| author_facet | Katsuhiro Nakanishi Sanshiro Narimatsu Shiori Ichikawa Yuki Tobisawa Kohta Kurohane Yasuo Niwa Hirokazu Kobayashi Yasuyuki Imai |
| author_sort | Katsuhiro Nakanishi |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-20T09:37:20Z |
| format | Article |
| id | doaj.art-d9f3d39499104671bb3a3923fad51d24 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-20T09:37:20Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-d9f3d39499104671bb3a3923fad51d242022-12-21T19:44:58ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-01811e8071210.1371/journal.pone.0080712Production of hybrid-IgG/IgA plantibodies with neutralizing activity against Shiga toxin 1.Katsuhiro NakanishiSanshiro NarimatsuShiori IchikawaYuki TobisawaKohta KurohaneYasuo NiwaHirokazu KobayashiYasuyuki ImaiShiga 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.http://europepmc.org/articles/PMC3842918?pdf=render |
| spellingShingle | Katsuhiro Nakanishi Sanshiro Narimatsu Shiori Ichikawa Yuki Tobisawa Kohta Kurohane Yasuo Niwa Hirokazu Kobayashi Yasuyuki Imai Production of hybrid-IgG/IgA plantibodies with neutralizing activity against Shiga toxin 1. PLoS ONE |
| title | Production of hybrid-IgG/IgA plantibodies with neutralizing activity against Shiga toxin 1. |
| title_full | Production of hybrid-IgG/IgA plantibodies with neutralizing activity against Shiga toxin 1. |
| title_fullStr | Production of hybrid-IgG/IgA plantibodies with neutralizing activity against Shiga toxin 1. |
| title_full_unstemmed | Production of hybrid-IgG/IgA plantibodies with neutralizing activity against Shiga toxin 1. |
| title_short | Production of hybrid-IgG/IgA plantibodies with neutralizing activity against Shiga toxin 1. |
| title_sort | production of hybrid igg iga plantibodies with neutralizing activity against shiga toxin 1 |
| url | http://europepmc.org/articles/PMC3842918?pdf=render |
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