Abstract
A naturally cholera toxin gene negative Vibrio cholerae (O1, E1 Tor, Ogawa) strain, named IEM101, was isolated in China. The human volunteer tests showed that this strain was safe, able to colonize the intestinal mucosa, and able to induce a strong immune response. Also other studies indicated that it was an efficient live vector to deliver heterologous antigens. In this article, a thymidylate synthase gene (thyA)-defined mutant was constructed using homologous recombination. Except for the morphological changes in minimal medium and slightly reduced colonization capacity, mutant strain IEM101-T maintained most of the desirable features as the wild-type strain IEM101 in terms of growth rate and immunogenicity. However, the mutant was more biosafe than its parent strain. In conclusion, IEM101-T may be a promising strain to develop live vaccine candidate of cholera or an attractive vaccine vector to deliver heterologous antigens in vivo.
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Tauxe, R., Seminario, L., Taipia, R., and Libel, M. O. (1994) The Latin American epidemic. In: Vibrio cholerae and Cholera: Molecular to Global Perspectives (Wachsmuth, I. K., Blake, P. A., and Olsvik, Ø eds.) ASM Press, Washington, DC, pp. 321–344.
Zhang, D., Xu, Z., Sun, W., and Karaolis, D. K. (2003) The vibrio pathogenicity island-encoded mop protein modulates the pathogenesis and reactogenicity of epidemic Vibrio cholerae. Infect. Immunol. 71, 510–515.
Mekalanos, J., Goldberg, I., Miller, V., et al. (1985) Genetic construction of cholera vaccine prototypes. In: Lerner, R., Chanock, R., and Brown, F., eds. Vaccines 85, Cold Spring Harbor, pp. 101–105.
Okada, T., Yanagisawa, K., and Ryan, F. J. (1960) Elective production of thymine-less mutants. Nature 188, 340–341.
Farmer, J. L. and Rothman, F. (1965) Transformable thymine-requiring mutant of Bacillus subtilis. J. Bacteriol. 89, 262–263.
Ross, P., O’Gara, F., and Condon, S. (1990) Cloning and characterization of the thymidylate synthase gene from Lactococcus lactis subsp. lactis. Appl. Environ. Microbiol. 56, 2156–2163.
Ross, P., O’Gara, F., and Condon, S. (1990) Thymidylate synthetase gene from Lactococcus lactis as a genetic marker: an alternative to antibiotic resistance genes. Appl. Environ. Microbiol. 56, 2164–2169.
Levine, M. M., Kaper, J. B., Herrington, D., et al. (1988) Volunteer studies of deletion mutants of Vibrio cholerae O1 prepared by recombinant techniques. Infect. Immun. 56, 161–167.
Valle, E., Ledon, T., Cedre, B., et al. (2000) Construction and characterization of a nonproliferative El Tor cholera vaccine candidate derived from strain 638. Infect. Immun. 68, 6411–6418.
Liu, Y. Q., Qi, G. M., Wang, S. X., et al. (1995) A natural vaccine candidate strain against cholera. Biomed. Environ. Sci. 8, 350–358.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Michalski, J., Galen, J. E., Fasano, A., and Kaper, J. B. (1993) CVD110, an attenuated Vibrio cholerae O1 El Tor live oral vaccine strain. Infect. Immunol. 61, 4462–4468.
Hone, D. M., Harris, A. M., Chatfield, S., Dougan, G., and Levine, M. M. (1991) Construction of genetically defined double aro mutants of Salmonella typhi. Vaccine 9, 810–816.
Tison, D., Murray, P. R., and Baron, E. J. (eds.) (1999) Manual of Clinical Microbiology. Washington, DC, ASM Press, pp. 497–506.
Cedré B., L. Garcá, L., Garcá, H., Fariñs, M., Talavera, A., and Infante, J. F. (1998) Intestinal colonization of the infant mouse model by attenuated and virulent Vibrio cholerae strains. Arch. Med. Res. 29, 231–234.
Attridge, S. R. (1995) Thymine auxotrophy as an attenuating marker in Vibrio cholerae. Microb. Pathog. 19, 11–18.
Liang, W., Wang, S., Yu, F., et al. (2003) Construction and evaluation of a safe, live, oral Vibrio cholerae vaccine candidate, IEM108. Infect. Immun. 71, 5498–5504.
Miller, V. L. and Mekalanos J. J. (1988) A novel suicide vector and its use in construction of insertion mutations: osmoregulation of outer membrane proteins and virulence determinants in Vibrio cholerae requires toxR. J. Bacteriol. 170, 2575–2583.
Owen, R. L., Pierce, N. F., Apple, R. T., and Cray, W. C. Jr. (1996) M cell transport of Vibrio cholerae from the intestinal lumen into Peyer’s patches: a mechanism for antigen sampling and for microbial transepithelial migration. J. Infect. Dis. 153, 1108–1118.
Winner, L. 3rd, Mack, J., Weltzin, R., Mekalanos, J. J., Kraehenbuhl, J. P., and Neutra, M. R. (1991) New model for analysis of mucosal immunity: intestinal secretion of specific monoclonal immunoglobulin A from hybridoma tumors protects against Vibrio cholerae infection. Infect. Immun. 59, 977–982.
Chen, I., Finn, T. M., Yanqing, L., Guoming, Q., Rappuoli, R., and Pizza, M. (1998) A recombinant live attenuated strain of Vibrio cholerae induces immunity against tetanus toxin and Bordetella pertussis tracheal colonization factor. Infect. Immun. 66, 1648–1653.
Fontana, M. R., Monaci, E., Yanqing, L., et al. (2001) IEM 101, a naturally attenuated Vibrio cholerae strain as carrier for genetically detoxified derivatives of cholera toxin. Vaccine 19, 75–85.
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Yu, F., Qi, G., Liu, Y. et al. Construction and characterization of a thyA mutant derived from cholera vaccine candidate IEM101. Mol Biotechnol 29, 191–196 (2005). https://doi.org/10.1385/MB:29:3:191
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DOI: https://doi.org/10.1385/MB:29:3:191