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Production of hepatitis B surface antigen in transgenic plants for oral immunization

Nature Biotechnology volume 18, pages 1167–1171 (2000)Cite this article

Abstract

Here we present data showing oral immunogenicity of recombinant hepatitis B surface antigen (HBsAg) in preclinical animal trials. Mice fed transgenic HBsAg potato tubers showed a primary immune response (increases in HBsAg-specific serum antibody) that could be greatly boosted by intraperitoneal delivery of a single subimmunogenic dose of commercial HBsAg vaccine, indicating that plants expressing HBsAg in edible tissues may be a new means for oral hepatitis B immunization. However, attainment of such a goal will require higher HBsAg expression than was observed for the potatoes used in this study. We conducted a systematic analysis of factors influencing the accumulation of HBsAg in transgenic potato, including 5′ and 3′ flanking elements and protein targeting within plant cells. The most striking improvements resulted from (1) alternative polyadenylation signals, and (2) fusion proteins containing targeting signals designed to enhance integration or retention of HBsAg in the endoplasmic reticulum (ER) of plant cells.

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Figure 1: Oral immunization of mice with transgenic potatoes expressing HBsAg.
Figure 2: Structures of expression cassettes.
Figure 3: Analysis of HBsAg protein and mRNA from leaves of potato lines transgenic for pHB104.
Figure 4: HBsAg expression (% TSP) in leaves of potato lines transgenic for various constructs.
Figure 5: RNA from leaves of potatoes transgenic for pHB110 analyzed by northern blot hybridized with HBsAg coding sequence.

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Acknowledgements

The authors wish to thank Qingxian Kong for performing the oral immunization experiments. This work was supported by a grant from the National Institutes of Health No. AI42836 to Y.T., C.J.A., and H.S.M.

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Authors and Affiliations

  1. Boyce Thompson Institute for Plant Research Inc., Tower Rd., Ithaca, 14853-1801, NY

    Liz J. Richter, Charles J. Arntzen & Hugh S. Mason

  2. Department of Immunology, Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, 14263, NY

    Yasmin Thanavala

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  1. Liz J. Richter
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  2. Yasmin Thanavala
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  4. Hugh S. Mason
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Correspondence to Hugh S. Mason.

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Richter, L., Thanavala, Y., Arntzen, C. et al. Production of hepatitis B surface antigen in transgenic plants for oral immunization. Nat Biotechnol 18, 1167–1171 (2000). https://doi.org/10.1038/81153

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