Production of Escherichia coli heat labile toxin (LT) B subunit in soybean seed and analysis of its immunogenicity as an oral vaccine

Tomas Moravec; Monica A. Schmidt; Eliot M. Herman; Terry Woodford-Thomas

doi:10.1016/j.vaccine.2006.11.010

Production of Escherichia coli heat labile toxin (LT) B subunit in soybean seed and analysis of its immunogenicity as an oral vaccine

Tomas Moravec, Monica A. Schmidt, Eliot M. Herman, Terry Woodford-Thomas

Research output: Contribution to journal › Article › peer-review

123 Scopus citations

Abstract

The B subunit of the heat labile toxin of enterotoxigenic Escherichia coli (LTB) was used as a model immunogen for production in soybean seed. LTB expression was directed to the endoplasmic reticulum (ER) of seed storage parenchyma cells for sequestration in de novo synthesized inert protein accretions derived from the ER. Pentameric LTB accumulated to 2.4% of the total seed protein at maturity and was stable in desiccated seed. LTB-soybean extracts administered orally to mice induced both systemic IgG and IgA, and mucosal IgA antibody responses, and was particularly efficacious when used in a parenteral prime-oral gavage boost immunization strategy. Sera from immunized mice blocked ligand binding in vitro and immunized mice exhibited partial protection against LT challenge. Moreover, soybean-expressed LTB stimulated the antibody response against a co-administered antigen by 500-fold. These results demonstrate the utility of soybean as an efficient production platform for vaccines that can be used for oral delivery.

Original language	English (US)
Pages (from-to)	1647-1657
Number of pages	11
Journal	Vaccine
Volume	25
Issue number	9
DOIs	https://doi.org/10.1016/j.vaccine.2006.11.010
State	Published - Feb 19 2007
Externally published	Yes

Keywords

Microbial toxin
Plant-based vaccines
Transgenic soybean seed

ASJC Scopus subject areas

Molecular Medicine
General Immunology and Microbiology
General Veterinary
Public Health, Environmental and Occupational Health
Infectious Diseases

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10.1016/j.vaccine.2006.11.010

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title = "Production of Escherichia coli heat labile toxin (LT) B subunit in soybean seed and analysis of its immunogenicity as an oral vaccine",

abstract = "The B subunit of the heat labile toxin of enterotoxigenic Escherichia coli (LTB) was used as a model immunogen for production in soybean seed. LTB expression was directed to the endoplasmic reticulum (ER) of seed storage parenchyma cells for sequestration in de novo synthesized inert protein accretions derived from the ER. Pentameric LTB accumulated to 2.4% of the total seed protein at maturity and was stable in desiccated seed. LTB-soybean extracts administered orally to mice induced both systemic IgG and IgA, and mucosal IgA antibody responses, and was particularly efficacious when used in a parenteral prime-oral gavage boost immunization strategy. Sera from immunized mice blocked ligand binding in vitro and immunized mice exhibited partial protection against LT challenge. Moreover, soybean-expressed LTB stimulated the antibody response against a co-administered antigen by 500-fold. These results demonstrate the utility of soybean as an efficient production platform for vaccines that can be used for oral delivery.",

keywords = "Microbial toxin, Plant-based vaccines, Transgenic soybean seed",

author = "Tomas Moravec and Schmidt, {Monica A.} and Herman, {Eliot M.} and Terry Woodford-Thomas",

note = "Funding Information: The assistance of Dr. Howard Berg (DPSC) in the preparation of high-pressure frozen tissue samples and Dr. Joseph Jez (DPSC) for assistance in size exclusion chromatography are gratefully acknowledged. We thank Dr. John Clements (Tulane University) for his gift of purified bacterial LTB and LT, and for useful advice and discussions, Dr. Amanda Walmsley (Arizona Biodesign Institute, Arizona State University) for providing the codon-optimized version of the LTB gene and Dr. Ed Cahoon (USDA/DPSC) for the pGly vector. Dr. Roger Beachy (DPSC) is gratefully acknowledged for providing stimulating discussions and intellectual support. We thank Dr. Scott Hultgren, Department of Molecular Microbiology, and the Department of Comparative Medicine, for provision of recombinant FimHt and for use of the animal facilities at Washington University School of Medicine. TM is supported by a Marie Curie Fellowship from the European Commission (MOIF CT 2005-008692).",

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language = "English (US)",

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T1 - Production of Escherichia coli heat labile toxin (LT) B subunit in soybean seed and analysis of its immunogenicity as an oral vaccine

AU - Moravec, Tomas

AU - Schmidt, Monica A.

AU - Herman, Eliot M.

AU - Woodford-Thomas, Terry

N1 - Funding Information: The assistance of Dr. Howard Berg (DPSC) in the preparation of high-pressure frozen tissue samples and Dr. Joseph Jez (DPSC) for assistance in size exclusion chromatography are gratefully acknowledged. We thank Dr. John Clements (Tulane University) for his gift of purified bacterial LTB and LT, and for useful advice and discussions, Dr. Amanda Walmsley (Arizona Biodesign Institute, Arizona State University) for providing the codon-optimized version of the LTB gene and Dr. Ed Cahoon (USDA/DPSC) for the pGly vector. Dr. Roger Beachy (DPSC) is gratefully acknowledged for providing stimulating discussions and intellectual support. We thank Dr. Scott Hultgren, Department of Molecular Microbiology, and the Department of Comparative Medicine, for provision of recombinant FimHt and for use of the animal facilities at Washington University School of Medicine. TM is supported by a Marie Curie Fellowship from the European Commission (MOIF CT 2005-008692).

PY - 2007/2/19

Y1 - 2007/2/19

N2 - The B subunit of the heat labile toxin of enterotoxigenic Escherichia coli (LTB) was used as a model immunogen for production in soybean seed. LTB expression was directed to the endoplasmic reticulum (ER) of seed storage parenchyma cells for sequestration in de novo synthesized inert protein accretions derived from the ER. Pentameric LTB accumulated to 2.4% of the total seed protein at maturity and was stable in desiccated seed. LTB-soybean extracts administered orally to mice induced both systemic IgG and IgA, and mucosal IgA antibody responses, and was particularly efficacious when used in a parenteral prime-oral gavage boost immunization strategy. Sera from immunized mice blocked ligand binding in vitro and immunized mice exhibited partial protection against LT challenge. Moreover, soybean-expressed LTB stimulated the antibody response against a co-administered antigen by 500-fold. These results demonstrate the utility of soybean as an efficient production platform for vaccines that can be used for oral delivery.

AB - The B subunit of the heat labile toxin of enterotoxigenic Escherichia coli (LTB) was used as a model immunogen for production in soybean seed. LTB expression was directed to the endoplasmic reticulum (ER) of seed storage parenchyma cells for sequestration in de novo synthesized inert protein accretions derived from the ER. Pentameric LTB accumulated to 2.4% of the total seed protein at maturity and was stable in desiccated seed. LTB-soybean extracts administered orally to mice induced both systemic IgG and IgA, and mucosal IgA antibody responses, and was particularly efficacious when used in a parenteral prime-oral gavage boost immunization strategy. Sera from immunized mice blocked ligand binding in vitro and immunized mice exhibited partial protection against LT challenge. Moreover, soybean-expressed LTB stimulated the antibody response against a co-administered antigen by 500-fold. These results demonstrate the utility of soybean as an efficient production platform for vaccines that can be used for oral delivery.

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Production of Escherichia coli heat labile toxin (LT) B subunit in soybean seed and analysis of its immunogenicity as an oral vaccine

Abstract

Keywords

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