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A plant-based multicomponent vaccine protects mice from enteric diseases

Nature Biotechnology volume 19, pages 548–552 (2001)Cite this article

Abstract

Cholera toxin (CT) B and A2 subunit complementary DNAs (cDNAs) were fused to a rotavirus enterotoxin and enterotoxigenic Escherichia coli fimbrial antigen genes and transferred into potato. Immunoblot and enzyme-linked immunosorbent assay (ELISA) results indicated that the fusion antigens were synthesized in transformed tuber tissues and assembled into cholera holotoxin-like structures that retained enterocyte-binding affinity. Orally immunized mice generated detectable levels of serum and intestinal antibodies against the pathogen antigens. Elevated levels of interleukin 2 (IL2) and interferon γ (INFγ) detected in immunogen-challenged spleen cells from the immunized mice indicated the presence of a strong Th1 immune response to the three plant-synthesized antigens. This result was supported by flow cytometry analysis of immunized mouse spleen cells that showed a significant increase in CD4+ lymphocyte numbers. Diarrhea symptoms were reduced in severity and duration in passively immunized mouse neonates following rotavirus challenge. The results suggest that food plants can function as vaccines for simultaneous protection against infectious virus and bacterial diseases.

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Figure 1: Expression of cholera toxin fusion proteins in transgenic potato.
Figure 2: Measurement of anti-CTB, NSP4, and CFA/I antibody titers in mice after oral immunization with transgenic potato tuber tissues.
Figure 3: The adjuvant and carrier functions of CTB in CTB-NSP4 fusion protein.
Figure 4: Measurement of cytokine levels in spleen cells from mice immunized with potato tissues containing CTB, NSP4, and CFA/I proteins.
Figure 5: Detection of CD4+ memory cells in spleen cells from mice immunized with transgenic potato tissues.
Figure 6: Protection against rotavirus infection in passively immunized mouse pups.

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Acknowledgements

We would like to thank Dr. John Mekalanos (Harvard University School of Medicine) for the gift of the CTX gene, Dr. Mary Estes and Dr. Carl Zeng (Baylor School of Medicine) for providing rotavirus antiserum specific for the NSP4 22–amino acid epitope and rotavirus SA-11 for rotavirus challenge experiments, and for many helpful discussions. Further, we would like to thank Dr. Myron Levine (Center for Vaccine Development, University of Maryland School of Medicine) for providing us with the E. coli CFA/I gene and the CFA/I fimbrial protein. This work was supported in part by a subcontract to W.H.R.L. by the US Army and the National Medical Technology Testbed Inc. and by an intramural grant awarded to W.H.R.L. by Loma Linda University. The views, opinions, and/or findings contained in this report are strictly those of the authors and should not be construed as a position policy, decision, or endorsement of the federal government or of the National Medical Technology Testbed, Inc.

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  1. Department of Biochemistry and Center for Molecular Biology and Gene Therapy, Loma Linda University, Loma Linda, 92350, CA

    Jie Yu & William H.R. Langridge

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  1. Jie Yu
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Correspondence to William H.R. Langridge.

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Yu, J., Langridge, W. A plant-based multicomponent vaccine protects mice from enteric diseases. Nat Biotechnol 19, 548–552 (2001). https://doi.org/10.1038/89297

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