Nature Biotechnology
16, 934 - 938 (1998)
doi:10.1038/nbt1098-934
A plant-based cholera toxin B subunit−insulin fusion protein protects against the development of autoimmune diabetesTakeshi Arakawa1, 2, Jie Yu1, 3, Daniel K. X. Chong1, John Hough4, Paul C. Engen4
& William H. R. Langridge1, 2, 3, *
1Center for Molecular Biology and Gene Therapy, School of Medicine, Loma Linda University, Loma Linda, CA 92350.
2Department of Microbiology and Molecular Genetics, School of Medicine, Loma Linda University, Loma Linda, CA 92350.
3Department of Biochemistry, School of Medicine, Loma Linda University, Loma Linda, CA 92350.
4Department of Anatomy, School of Medicine, Loma Linda University, Loma Linda, CA 92350.
*Corresponding author (e-mail: blangridge@ccmail.llu.edu). Oral administration of disease-specific autoantigens can prevent or delay the onset of autoimmune disease symptoms. We have generated transgenic potato plants that synthesize human insulin, a major insulin-dependent diabetes mellitus autoantigen, at levels up to 0.05% of total soluble protein. To direct delivery of plant-synthesized insulin to the gut-associated lymphoid tissues, insulin was linked to the C-terminus of the cholera toxin B subunit (CTB). Transgenic potato tubers produced 0.1% of total soluble protein as the pentameric CTB−insulin fusion, which retained GM1-ganglioside binding affinity and native antigenicity of both CTB and insulin. Nonobese diabetic mice fed transformed potato tuber tissues containing microgram amounts of the CTB−insulin fusion protein showed a substantial reduction in pancreatic islet inflammation (insulitis), and a delay in the progression of clinical diabetes. Feeding transgenic potato tissues producing insulin or CTB protein alone did not provide a significant reduction in insulitis or diabetic symptoms. The experimental results indicate that food plants are feasible production and delivery systems for immunotolerization against this T cell−mediated autoimmune disease.
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