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A Recombinant Live Oral Cholera Vaccine

Bio/Technology volume 2, pages 345–349 (1984)Cite this article

Abstract

A pathogenic strain of Vibrio cholerae was attenuated by deletion of DNA sequences encoding the A1 subunit of the cholera enterotoxin. A restriction endonuclease fragment encoding the A1, but not the A2 or B sequences was deleted in vitro from cloned cholera toxin genes. The mutation was then recombined into the chromosome of V.cholerae Ogawa 395, a pathogenic strain that confers complete immunity to subsequent infection following an initial clinical infection. The resulting strain, which produces the immunogenic but nontoxic B subunit of cholera toxin and is unaltered in other critical antigens, represents a promising candidate for an attenuated live oral cholera vaccine.

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References

  1. Levine, M.M., Nalin, D.R., Craig, J.P., Hoover, D., Bergquist, E.J., Waterman, D., Holley, H.P., Hornick, R.B., Pierce, N.P., and Libonati, J.P. 1979. Immunity to cholera in man: relative role of antibacterial versus antitoxic immunity. Trans. Roy. Soc. Trop. Med. Hyg. 73: 3–9.

    Article  CAS  Google Scholar 

  2. Levine, M.M. 1980. Immunity to cholera as evaluated volunteers, pp. 195–203. In Cholera and Related Diarrhoeas. O. Ouchterlony and J. Holmgren (eds.), Karger, Basel

    Google Scholar 

  3. Levine, M.M., Black, R.E., Clements, M.L., Cisneros, L., Nalin, D.R., and Young, C.R. 1981. Duration of infection-derived immunity to cholera. J. Infect. Dis. 143: 818–820.

    Article  CAS  Google Scholar 

  4. Levine, M.M., Black, R.E., Clements, M.L., Nalin, D.R., Cisneros, L., and Finkelstein, R.A. 1981. Volunteer studies in development of vaccines against cholera and enterotoxigenic Escherichia coli: a review, pp. 449–459. In Acute Enteric Infections in Children: New Prospects for Treatment and Prevention. T. Holme, J. Holmgren, M. H. Mer-son, and R. Mollby (eds.), Elsevier, Amsterdam

    Google Scholar 

  5. Glass, R.I., Becker, S., Huq, M.I., Stoll, B.J., Khan, M.U., Merson, M.H., Lee, J.V., and Black, R.E. 1982. Endemic cholera in rural Bangladesh, 1966-1980. Am. J. Epidemiol. 116: 959–970.

    Article  CAS  Google Scholar 

  6. Kleid, D.G., Yansura, D., Small, B., Dowbenko, D., Moore, D.M., Grubman, M.J., McKercher, P.D., Morgan, D.O., Robertson, B.H., and Bachrach, H.L. 1981. Cloned viral protein vaccine for foot-and-mouth disease: responses in cattle and swine. Science 214: 1125–1129.

    Article  CAS  Google Scholar 

  7. Bachrach, H.L. 1982. Recombinant DNA technology for the preparation of subunit vaccines. J. Am. Vet. Med. Assoc. 181: 992–999.

    CAS  PubMed  Google Scholar 

  8. 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 

  9. Curlin, G., Levine, R., Azia, K.M.A., Mizanur Rahman, A.C.M., and Verway, W.F. 1975. Field trial of cholera toxoid, pp. 314–329. Proc. 11th Joint Conference on Cholera, Washington, D.C

  10. Noriki, H. 1976. Evaluation of toxoid field trials in the Philippines, pp. 302–310. Proc. 12th Joint Conference on Cholera, Tokyo

  11. Lockman, H. and Kaper, J.B. 1983. Nucleotide sequence analysis of the A2 and B subunits of Vibrio cholerae enterotoxin. J. Biol. Chem. 258: 13722–13726.

    CAS  PubMed  Google Scholar 

  12. Kaper, J.B., Moseley, S.L., and Falkow, S. 1981. Molecular characterization of environmental and nontoxinogenic strains of Vibrio cholerae. Infect. Immun. 32: 661–667.

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Mekalanos, J.J. 1983. Duplication and amplification of toxin genes in Vibrio cholerae. Cell 35: 253–263.

    Article  CAS  Google Scholar 

  14. Gennaro, M.L. and Greenway, P.J. 1983. Nucleotide sequences within the cholera toxin operon. Nucl. Acids. Res. 11: 3855–3861.

    Article  CAS  Google Scholar 

  15. Mekalanos, J.J., Swartz, D.J., Pearson, G.D.N., Harford, N., Groyne, F., and de Wilde, M. 1983. Cholera toxin genes: nucleotide sequence, deletion analysis, and vaccine development. Nature 306: 551–557.

    Article  CAS  Google Scholar 

  16. Gill, D.M. 1976. The arrangement of subunits in cholera toxin. Biochemistry 15: 1242–1248.

    Article  CAS  Google Scholar 

  17. Gill, D.M., and Meren, R. 1978. ADP-ribosylation of membrane proteins catalyzed by cholera toxin: basis of the activation of adenylate cyclase. Proc. Natl. Acad. Sci. U.S.A. 75: 3050–3054.

    Article  CAS  Google Scholar 

  18. Ruvkun, G.R. and Ausubel, F.M. 1981. A general method for site-directed mutagenesis in prokaryotes. Nature 289: 85–88.

    Article  CAS  Google Scholar 

  19. Levine, M.M., Black, R.E., Clements, M.L., Young, C.R., Lanata, C., Sears, S., Honda, T., and Finkelstein, R. 1983. Texas Star-SR: attenuated Vibrio cholerae oral vaccine candidate. Develop. Biol. Stand. 53: 59–65.

    CAS  Google Scholar 

  20. Ditta, G., Stanfield, S., Corbin, D., and Helinski, D.R. 1980. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc. Natl. Acad. Sci. U.S.A. 77: 7347–7351.

    Article  CAS  Google Scholar 

  21. Miller, J.M. 1972. Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  22. Brenner, D.J., Fanning, G.R., Johnson, K.E., Citarella, R.V., and Falkow, S. 1969. Polynucleotide sequence relationships among members of Enterobacteriaceae. J. Bacteriol. 98: 637–650.

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Birnboim, H.C., and Doly, J. 1979. A rapid alkaline extraction procedure for screening recombinant DNA. Nucl. Acids. Res. 7: 1513–1523.

    Article  CAS  Google Scholar 

  24. Maniatis, T., Fritsch, E.F., and Sambrook, J. 1982. Molecular cloning. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  25. Bolivar, F. 1978. Construction and characterization of new cloning vehicles. III. Derivatives of plasmid pBR322 carrying unique Eco RI sites for selection of Eco RI generated recombinant DNA molecules. Gene 4: 121–136.

    Article  CAS  Google Scholar 

  26. Bolivar, F., Rodriguez, R.L., Green, P.J., Betlach, M.C., Heynecker, H.L., Boyer, H.W., Crosa, J.M., and Falkow, S. 1977. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2: 95–113.

    Article  CAS  Google Scholar 

  27. Southern, E.M. 1975. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98: 503–517.

    Article  CAS  Google Scholar 

  28. Maniatis, T., Jeffrey, A., and Kleid, D.G. 1975. Nucleotide sequence of the rightward operator of phage lambda. Proc. Natl. Acad. Sci. U.S.A. 72: 1184–1188.

    Article  CAS  Google Scholar 

  29. Sack, D.A. and Sack, R.B. 1975. Test for enterotoxigenic Escherichia coli using Y1 adrenal cells in miniculture. Infect. Immun. 11: 334–336.

    CAS  PubMed  PubMed Central  Google Scholar 

  30. Sack, D.A., Huda, S., Neogi, P.K.B., Daniel, R.R., and Spira, W.M. 1980. Microtiter ganglioside enzyme-linked immunosorbent assay for Vibrio and Escherichia coli heat-labile enterotoxins and antitoxin. J. Clin. Microbiol. 11: 35–40.

    CAS  PubMed  PubMed Central  Google Scholar 

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Authors and Affiliations

  1. Center for Vaccine Development, University of Maryland School of Medicine, 10 S. Pine Street, Baltimore, Maryland, 21201

    James B. Kaper, Hank Lockman, Mary M. Baldini & Myron M. Levine

Authors
  1. James B. Kaper
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  2. Hank Lockman
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  3. Mary M. Baldini
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  4. Myron M. Levine
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Kaper, J., Lockman, H., Baldini, M. et al. A Recombinant Live Oral Cholera Vaccine. Nat Biotechnol 2, 345–349 (1984). https://doi.org/10.1038/nbt0484-345

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