Genetically-engineered plants yield an orally immunogenic PirA-like toxin from Vibrio parahaemolyticus
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Vibrio parahaemolyticus is the main etiological agent of human gastroenteritis by seafood consumption and some strains from this species causing the Acute Hepatopancreatic Necrosis Disease in shrimp have been recently reported. The PirA-like toxin from V. parahaemolyticus (ToxA) has been recently reported as an attractive antigen implicated in subunit vaccine development. Since plants are attractive hosts for the production and delivery of vaccines in the present study plants expressing ToxA were developed to account with a low cost platform for the production and oral delivery of ToxA. Tobacco plants were genetically engineered by Agrobacterium-mediated transformation to stably integrate the ToxA-coding gene into the nuclear genome. Transgenic lines were rescued in kanamycin-containing medium and analyzed by ELISA to determine ToxA yields observing levels up to 9 μg g−1 FW leaf tissues. Western blot analysis confirmed the presence of the ToxA protein in plant extracts. Immunogenicity assessment of the plant-made ToxA was performed in mice, comprising a 4-dose oral immunization scheme; revealing the induction of anti-ToxA humoral responses (IgG in serum and IgA in feces). This study opens the path for the development of low cost plant-based vaccines against Vibrio parahaemolyticus. © 2019 Elsevier B.V.
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Acute Hepatopancreatic Necrosis Disease; Adjuvant; Oral immunization; Plant-based vaccine; ToxA toxin bacterial toxin; immunoglobulin A; immunoglobulin G; immunoglobulin G antibody; kanamycin; PirA like toxin; unclassified drug; bacterial toxin; Agrobacterium; animal experiment; antibody titer; Article; biotechnological production; blood level; controlled study; cost control; enzyme linked immunosorbent assay; feces level; female; genetic engineering; genetic transformation; genome; humoral immunity; immunogenicity; leaf tissue; mouse; Nicotiana tabacum; nonhuman; nuclear genome; plant biotechnology; polymerase chain reaction; protein expression; toxin synthesis; transgenic plant; vaccine production; Vibrio parahaemolyticus; Western blotting; amino acid sequence; animal; biosynthesis; biotechnology; chemistry; genetics; immunology; metabolism; oral drug administration; plant leaf; tobacco; transgenic plant; Vibrio parahaemolyticus; Administration, Oral; Amino Acid Sequence; Animals; Bacterial Toxins; Biotechnology; Immunity, Humoral; Mice; Plant Leaves; Plants, Genetically Modified; Tobacco; Vibrio parahaemolyticus
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