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Efficacy of a food plant-based oral cholera toxin B subunit vaccine

Abstract

Transgenic potatoes were engineered to synthesize a cholera toxin B subunit (CTB) pentamer with affinity for GM1-ganglioside. Both serum and intestinal CTB-specific antibodies were induced in orally immunized mice. Mucosal antibody titers declined gradually after the last immunization but were restored following an oral booster of transgenic potato. The cytopathic effect of cholera holotoxin (CT) on Vero cells was neutralized by serum from mice immunized with transgenic potato tissues. Following intraileal injection with CT, the plant-immunized mice showed up to a 60% reduction in diarrheal fluid accumulation in the small intestine. Protection against CT was based on inhibition of enterotoxin binding to the cell-surface receptor GM1-ganglioside. These results demonstrate the ability of transgenic food plants to generate protective immunity in mice against a bacterial enterotoxin.

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References

  1. Holmgren, J. and Svennerholm, A.-M. 1996. Oral vaccines against cholera and enterotoxigenic Escherichia coli diarrhea, pp. 241–253 in Mucosal vaccines. Kiyono, H., Ogra, P.L., McGhee, J.R. (eds.), Academic Press, San Diego.

    Chapter  Google Scholar 

  2. Finkelstein, R.A. 1995. Why do we not yet have a suitable vaccine against cholera? Adv. Exp. Med. Biol. 371B: 1633–1640.

    CAS  PubMed  Google Scholar 

  3. Mekalanos, J.J. and Sadoff, J.C. 1994. Cholera vaccines: fighting an ancient scourge. Science 265: 1387–1389.

    Article  CAS  PubMed  Google Scholar 

  4. Waldor, M.K. and Mekalanos, J.J. 1996. Progress toward live-attenuated cholera vaccines. 229–240 in Mucosal vaccines. Kiyono, H., Ogra, P.L., McGhee, J.R. (eds.), Academic Press, San Diego.

    Chapter  Google Scholar 

  5. Mason, H.S., Ball, J.M., Shi, J.-J., Jiang, X., Estes, M.K. and Arntzen, C.J. 1996. Expression of Norwalk virus capsid protein in transgenic tobacco and potato and its oral immunogenicity in mice. Proc. Natl. Acad. Sci. USA 93: 5335–5340.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Haq, T.A., Mason, H.S., Clements, J.D. and Arntzen, C.J. 1995. Oral immunization with a recombinant bacterial antigen produced in transgenic plants. Science 268: 714–716.

    Article  CAS  PubMed  Google Scholar 

  7. Thanavala, Y., Yang, Y.-F., Lyons, P., Mason, H.S. and Arntzen, C. 1995. Immunogenicity of transgenic plant-derived hepatitis B surface antigen. Proc. Natl. Acad. Sci. USA 92: 3358–3361.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Mason, H.S. and Arntzen, C.J. 1995. Transgenic plants as vaccine production systems. Trends in Biotechnology 13: 388–392.

    Article  CAS  PubMed  Google Scholar 

  9. Fujita, K. and Finkelstein, R.A. 1972. Antitoxic immunity in experimental cholera: comparison of immunity induced perorally and parenterally in mice. J. Infect. Dis. 125: 647–655.

    Article  CAS  PubMed  Google Scholar 

  10. Elson, C.O. and Ealding, W. 1984. Generalized systemic and mucosal immunity in mice after mucosal stimulation with cholera toxin. J. Immunol. 132: 2736–2741.

    CAS  PubMed  Google Scholar 

  11. Lycke, N., Lindholm, L. and Holmgren, J. 1983. IgA isotype restriction in the mucosal but not in the extramucosal immune response after oral immunizations with cholera toxin or cholera B subunit. Int. Arch. Allergy Appl. Immunol. 72: 119–127.

    Article  CAS  PubMed  Google Scholar 

  12. Arakawa, T., Chong, D.K.X., Merritt, J.L. and Langridge, W.H.R. 1997. Expression of cholera toxin B subunit oligomers in transgenic potato plants. Transgenic Res. 6: 403–413.

    Article  CAS  PubMed  Google Scholar 

  13. Jackson, R.J., Fujihashi, K., Kiyono, H. and McGhee, J.R. 1996. Luminometry: a novel bioluminescent immunoassay enhances the quantitation of mucosal and systemic antibody responses. J. Immunol. Methods 190: 189–197.

    Article  CAS  PubMed  Google Scholar 

  14. Apter, F.M., Lender, W.I., Finkelstein, R.A., Mekalanos, J.J. and Neutra, M.R. 1993. Monoclonal immunoglobulin A antibodies directed against cholera toxin prevent the toxin-induced chloride secretory response and block toxin binding to intestinal epithelial cells in vitro. Infect. Immun. 61: 5271–5278.

    CAS  PubMed  PubMed Central  Google Scholar 

  15. de Vos, T. and Dick, T.A. 1991. A rapid method to determine the isotype and specificity of coproantibodies in mice infected with Trichinella or fed cholera toxin. J. Immunol. Methods 141: 285–288.

    Article  CAS  Google Scholar 

  16. Dertzbaugh, M.T. and Elson, C.O. 1993. Reduction in oral immunogenicity of cholera toxin B subunit by N-terminal peptide addition. Infect. Immun. 61: 384–390.

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Svennerholm, A.M., Jertborn, M., Gothefors, L., Karim, A.M.M.M., Sack, D.A. and Holmgren, J. 1984. Mucosal antitoxic and antibacterial immunity after cholera disease and after immunization with a combined B subunit-whole cell vaccine. J. Infect. Dis. 149: 884–893.

    Article  CAS  PubMed  Google Scholar 

  18. Jertborn, M., Svennerholm, A.M. and Holmgren, J. 1986. Saliva, breast milk, and serum antibody responses as indirect measures of intestinal immunity after oral cholera vaccination or natural disease. J. Clin. Microbiol. 24: 203–209.

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Lycke, N., Bromander, A. and Holmgren, J. 1989. Role of local IgA antitoxin-producing cells for intestinal protection against cholera toxin challenge. Int. Arch. Allergy Appl. Immunol. 88: 273–279.

    Article  CAS  PubMed  Google Scholar 

  20. Clemens, J.D., van Loon, F., Sack, D.A., Chakraborty, J., Rao, M.R., Ahmed, R. et al. 1991. Field trial of oral cholera vaccines in Bangladesh: serum vibriocidal and antitoxic antibodies as markers of the risk of cholera. J. Infect. Dis. 163: 1235–1242.

    Article  CAS  PubMed  Google Scholar 

  21. Clemens, J.D., Sack, D.A., Rao, M.R., Chakraborty, J., Khan, M.R., Kay, B. et al. 1992. Evidence that inactivated oral cholera vaccines both prevent and mitigate Vibrio cholerae O1 infections in a cholera-endemic area. J. Infect. Dis. 166: 1029–1034.

    Article  CAS  PubMed  Google Scholar 

  22. Levine, M.M., Kaper, J.B., Black, R.E. and Clements, M.L. 1983. New knowledge on pathogenesis of bacterial enteric infections as applied to vaccine development. Microbiol. Rev. 47: 510–550.

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Levine, M.M., Nalin, D.R., Craig, J.P., Hoover, D., Bergquist, E.J., Waterman, D. et al. 1979. Immunity of cholera in man: relative role of antibacterial versus antitoxic immunity. Trans. R. Soc. Trap. Med. Hyg. 73: 3–9.

    Article  CAS  Google Scholar 

  24. Langermann, S., Palaszynski, S., Barnhart, M., Auguste, G., Pinkner, J.S., Burlein, J. et al. 1997. Prevention of mucosal Escherichia coli infection by FimH-adhesin-based systemic vaccination. Science 276: 607–611.

    Article  CAS  PubMed  Google Scholar 

  25. Svennerholm, L. 1976. Interaction of cholera toxin and ganglioside G(M1). Adv. Exp. Med. Biol. 71: 191–204.

    Article  CAS  PubMed  Google Scholar 

  26. Czerkinsky, C., Russell, M.W., Lycke, N., Lindblad, M. and Holmgren, J. 1989. Oral administration of a streptococcal antigen coupled to cholera toxin B subunit evokes strong antibody responses in salivary glands and extramucosal tissues. Infect. Immun. 57: 1072–1077.

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Dertzbaugh, M.T. and Elson, C.O. 1993. Comparative effectiveness of the cholera toxin B subunit and alkaline phosphatase as carriers for oral vaccines. Infect. Immun. 61: 48–55.

    CAS  PubMed  PubMed Central  Google Scholar 

  28. Holmgren, J., Czerkinsky, C., Lycke, N. and Svennerholm, A.-M. 1994. Strategies for the induction of immune responses at mucosal surfaces making use of cholera toxin B subunit as immunogen, carrier, and adjuvant. Am. J. Trap. Med. Hyg. 50: 42–54.

    CAS  Google Scholar 

  29. Bergerot, I., Ploix, C., Petersen, J., Moulin, V., Rask, C., Fabien, N., et al. 1997. A cholera toxoid-insulin conjugate as an oral vaccine against spontaneous autoimmune diabetes. Proc. Natl. Acad. Sci. USA 94: 4610–4614.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Sun, J.-B., Rask, C., Olsson, T., Holmgren, J. and Czerkinsky, C. 1996. Treatment of experimental autoimmune encephalomyelitis by feeding myelin basic protein conjugated to cholera toxin B subunit. Proc. Natl. Acad. Sci. USA 93: 7196–7201.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Sun, J.-B., Holmgren, J. and Czerkinsky, C. 1994. Cholera toxin B subunit: an efficient transmucosal carrier-delivery system for induction of peripheral immunological tolerance. Proc. Natl. Acad. Sci. USA 91: 10795–10799.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Koncz, C., Olsson, O., Langridge, W.H.R., Schell, J. and Szalay, A.A. 1987. Expression and assembly of functional bacterial luciferase in plants. Proc. Natl. Acad. Sci. USA 84: 131–135.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Stavric, S., Speirs, J.I., Konowalchuk, J. and Jeffrey, D. 1978. Stimulation of cyclic AMP secretion in Vero cells by enterotoxins of Escherichia coli and Vibrio cholerae . Infect. Immun. 21: 514–517.

    CAS  PubMed  PubMed Central  Google Scholar 

  34. Punyashthiti, K. and Finkelstein, R.A. 1971. Enteropathogenicity of Escherichia coli. Evaluation of mouse intestinal loops. Infect. Immun. 4: 473–478.

    CAS  PubMed  PubMed Central  Google Scholar 

  35. Escher, A., O'Kane, D.J., Lee, J. and Szalay, A.A. 1989. Bacterial luciferase alpha-beta fusion protein is fully active as a monomer and highly sensitive in vivo to elevated temperature. Proc. Natl. Acad. Sci. USA 86: 6528–6532.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Langridge, W.H.R., Fitzgerald, K.L., Koncz, C., Schell, J. and Szalay, A.A. 1989. Dual promoter of Agrobacterium tumefaciens mannopine synthase gene is regulated by plant growth hormones. Proc. Natl. Acad. Sci. USA 86: 3219–3223.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Correspondence to William H. R. Langridge.

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Arakawa, T., Chong, D. & Langridge, W. Efficacy of a food plant-based oral cholera toxin B subunit vaccine. Nat Biotechnol 16, 292–297 (1998). https://doi.org/10.1038/nbt0398-292

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