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

• Published in: Nature biotechnology Vol. 18; no. 11; pp. 1167 - 1171
• Main Authors: Mason, Hugh S, Richter, Liz J, Thanavala, Yasmin, Arntzen, Charles J
• Format: Journal Article
• Language: English
• Published: New York, NY Nature 01.11.2000; Nature Publishing Group
• Subjects: Administration, Oral; Animals; Antibodies; Antigens; Biological and medical sciences; Biotechnology; Blotting, Northern; Cholera Toxin - pharmacology; Endoplasmic reticulum; Endoplasmic Reticulum - metabolism; Enzyme-Linked Immunosorbent Assay; Fundamental and applied biological sciences. Psychology; Health. Pharmaceutical industry; Hepatitis; Hepatitis B; Hepatitis B - immunology; Hepatitis B - prevention & control; hepatitis B surface antigen; Hepatitis B Surface Antigens - genetics; Hepatitis B Surface Antigens - immunology; Hepatitis B Surface Antigens - therapeutic use; Hepatitis B virus; Humans; Immune response; Immunization; Immunogenicity; Industrial applications and implications. Economical aspects; Mice; Models, Genetic; Nicotiana - genetics; Plants, Genetically Modified; Plants, Toxic; Plasmids - metabolism; Plastids - metabolism; Poly A - metabolism; Polymerization; Potatoes; Production of active biomolecules; Promoter Regions, Genetic; Proteins; Recombinant Fusion Proteins - metabolism; Recombinant Proteins - therapeutic use; Retention; RNA, Messenger - metabolism; Solanum tuberosum; Solanum tuberosum - genetics; Time Factors; Tobacco; Transformation, Genetic; Transgenic plants; Vaccines; Vaccines - administration & dosage; Vaccines - therapeutic use; Vaccins; Vegetables; Yeast; Animal model; Transgenic animal; Preclinical trial; Dicotyledones; Angiospermae; Production; Transgenic plant; Solanaceae; Recombinant protein; Fusion protein; Immunization; Hepatitis B surface antigen; Immune response; Antibody; Solanum tuberosum; Rodentia; Oral administration; Vaccine; Gene expression; Vertebrata; Mammalia; Immunogenicity; Mouse; Spermatophyta; Tuber plant
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/81153

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.