Recombinant nontoxinogenic Vibrio cholerae strains as attenuated cholera vaccine candidates


An ideal vacine does not yet exist to prevent cholera, a significant health problem in many less developed countries. Vibrio cholerae, the agent of epidemic and endemic cholera, colonizes the small bowel and secretes a potent enterotoxin that consists of a single A subunit, which stimulates adenylate cyclase activity, and five identical B summits which bind to the ganglioside GM1 receptor of intestinal mucosal cells1. Previous studies in man indicate that toxoid-derived antitoxic immunity by itself is insufficient to provide effective, long-lasting protection against cholera2–4. Using recombinant DNA techniques we have now constructed a live, attenuated V. cholerae strain by deleting genes encoding the enterotoxin. Restriction enzyme fragments encoding cholera toxin were deleted in vitro from cloned vibrio chromosomal DNA and the resulting mutations introduced into the chromosome of a vibrio strain of proven immunogenicity. Recently, Mekalanos and coworkers5 have reported attenuated V. cholerae strains constructed by similar methods. It appears that recombinant DNA techniques offer a promising approach to the development of effective cholera vaccines.

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  1. 1

    Holmgren, J. Nature 292, 413–417 (1981).

  2. 2

    Curlin, G., Levine, R., Azia, K. M. A., Mizanur Rahman, A. C. M. & Verway, W. F. Proc. 11 th Joint US–Japan Cholera Conf., New Orleans, 314–329 (Dept of Health, Education and Welfare, Bethesda, 1976).

  3. 3

    Noriki, H. Proc. 12th Joint US–Japan Cholera Conf., Sapporo, 302–310 (Fuji, Tokyo, 1977).

  4. 4

    Levine, M. M. et al. Trans. R. Soc. trop. Med. Hyg. 73, 3–9 (1979).

  5. 5

    Mekalanos, J. J. et al. Nature 306, 551–557 (1983).

  6. 6

    Levine, M. M. in Choiera and Related Diarrhoeas: 43rd Nobel Symp., Stockholm, 195–203 (Karger, Basel, 1980).

  7. 7

    Levine, M. M. et al. J. Infect. Dis. 143, 818–820 (1981).

  8. 8

    Levine, M. M. et al. in Acute Enteric Infections in Children New Prospects for Treatment and Prevention (eds Holme, T., Holmgren, J., Merson, M. H. & Mollby, R.) 449–459 (Elsevier, Amsterdam, 1981).

  9. 9

    Levine, M. M. et al. Infect. Immun. 43, 512–522 (1984).

  10. 10

    Glass, R. I. et al. Am. J. Epidemiol. 116, 959–970 (1982).

  11. 11

    Kaper, J. B. & Levine, M. M. Lancet ii, 1162–1163 (1981).

  12. 12

    Lockman, H. & Kaper, J. B. J. biol. Chem. 258, 13722–13726 (1983).

  13. 13

    Ruvkun, G. B. & Ausubel, F. M. Nature 289, 85–88 (1981).

  14. 14

    Ditta, G., Stanfield, S., Corbin, D. & Helinski, D. R. Proc. natn. Acad. Sci. U.S.A. 77, 7347–7351 (1980).

  15. 15

    Jacob, A. E. et al. in DNA Insertion Elements, Plasmids, and Episomes (eds Bukhari, A. I., Shapiro, J. A. & Adhya, S. L.) 607–638 (Cold Spring Harbor Laboratory, New York, 1977).

  16. 16

    Honda, T. & Finkelstein, R. A. Proc. natn. Acad. Sci. U.S.A. 76, 2052–2056 (1979).

  17. 17

    Birnboim, H. C. & Doly, J. Nucleic Acids Res. 7, 1513–1523 (1979).

  18. 18

    Parker, R. C., Watson, R. M. & Vinograd, J. Proc. natn. Acad. Sci. U.S.A. 74, 851–855 (1977).

  19. 19

    Sack, D. A. & Sack, R. B. Infect. Immun. 11, 334–336 (1975).

  20. 20

    Sack, D. A., Huda, S., Neogi, P. K. B., Daniel, R. R. & Spira, W. M. J. clin. Microbiol. 11, 35–40 (1980).

  21. 21

    Moseley, S. L. et al. J. infect. Dis. 142, 892–898 (1980).

  22. 22

    Maniatis, T., Jeffry, A. & Kleid, A. G. Proc. natn. Acad. Sci. U.S.A. 72, 1184–1188 (1975).

  23. 23

    Bagdasarian, M., Bagdasarian, M. M., Coleman, S. & Timmis, K. N. Plasmids of Medical, Environmental and Commercial Importance (eds Timmis, K. N. & Puhler, A.) 411–422 (Elsevier, Amsterdam, 1979).

  24. 24

    Brenner, D. J., Fanning, G. R., Johnson, K. E., Citarella, R. V. & Falkow, S. J. Bact. 98, 637–650 (1969).

  25. 25

    Southern, E. M. J. molec. Biol. 98, 503–517 (1975).

  26. 26

    Miozzari, G. F. & Yanofsky, C. Nature 276, 684–689 (1978).

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