Abstract
In this work asd gene of Shigella flexneri 2a strain T32 was replaced by Vibrio cholerae toxin B subunit (ctxB) gene with DNA recombination in vivo and in vitro. The resulting derivative of T32, designed as FWL01, could stably express CtxB, but its growth in LB medium depended on the presence of diaminopimelic acid (DAP). Then form I plasmid of Shigella sonnei strain S7 was labeled with strain T32 asd gene and mobilized into FWL01. Thus a trivalent candidate oral vaccine strain, designed as FSW01, was constructed. In this candidate strain, a balanced-lethal system was constituted between the host strain and the form I plasmid expressing S. sonnei O antigen. Therefore the candidate strain can express stably not only its own O antigen but also CtxB and O antigen of S. sonnei in the absence of any antibiotic. Experiments showed that FSW01 did not invade HeLa cells or cause keratoconjunctivitis in guinea pigs. However, rabbits immunized FSW01 can elicit significant immune responses. In mice and rhesus monkey models, vaccinated animals were protected against the challenges of wild S. flexneri 2a strain 2457T and S. sonnei strain S9.
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References
Linderberg, A. A., Pal, T., Strategies for development of potential candidate Shigella vaccine, Vaccine, 1993, 11: 168–179.
WHO, Vaccine research and development: new strategies for accelerating vaccine development, Wkly. Epidemiol. Rec., 1997, 72: 73–79.
Nakayama, K., Kelly, S. M., Curtiss III, R., Construction of an asd+ expression-cloning vector: stable maintenance and high level expression of cloned genes in a Salmonella vaccine strain, Bio/technology, 1988, 6: 693–697.
Rui, X. L., Xu, Y. Q., Wu, X. D. et al., Construction of a trivalent candidate Shigella vaccine strain with host-vector balanced-lethal system, Science in China, Ser. C, 1997, 40(1): 52–59.
Rui, X. L., Xu, Y. Q., Wang, H. et al., Construction of stable and non-resistant bivalent vaccine candidate strain against Shigella flexneri 2a and Shigella sonnei, Chin. J. Biotechnol (in Chinese), 1996, 12: 134–139.
Qian, F., Rui, X. L., Su, G. F. et al., Expression of Shigella sonnei form I O antigen and Vibrio cholera toxin B subunit gene in S. flexneri 2a strain and investigation of their immunoprotective response in mice, Chinese Journal of Genetics (Acta Genetica Sinica) (in Chinese), 1996, 23(4): 322–328.
Herrero, M., Lorenzo, V., Timmis, K., Transposon vectors containing non-antibiotic resistance selection markers for cloning and stable chromosomal insertion of foreign genes in gram-negative bacteria, J. Bacteriol., 1990, 172(11): 6557–6567.
Miller, V. L., Mekalanos, J. J., A novel suicide vector and its use in construction of insertion mutations: osmoregulation of outer membrane proteins and virulence determinants in Vibro cholerae requires toxR, J. Bacteriol., 1988, 170(6): 2575–2583.
Wertman, K. F., Wyman, A. R., Botstein, D., Host/vector interactions which affect the viability of recombinant phage lambda clones, Gene, 1986, 49: 253–262.
Liu, W., Wang, B. R., Studies on the 120 Md plasmid and its derivative plasmid in Shigella sonnei, Chin. Microbiol. Immunol, (in Chinese), 1990, 10: 205–208.
Venkatesan, M., Fernandez-Prada, C., Buysse, J. M. et al., Virulence phenotype and genetic characteristic of the T32-ISTRATI Shigella flexneri 2a vaccine strain, Vaccine, 1991, 9(5): 358–363.
Gennaro, M. L., Greenaway, P. J., Broadbent, D. A., The expression of biologically active cholera toxin in Escherichia coli, Nucl. Acids. Res., 1982, 10(16): 4883–4890.
Harayama, S., Tsuda, M., Iino, T., High frequency mobilization of the chromosome of Escherichia coli by a mutant of plasmid RP4 temperature-sensitive for maintenance, Mol. Gen. Genet., 1980, 180: 47–56.
Sambrook, J., Fritsch, E. F., Maniatis, T., Molecular Cloning: A Laboratory Manual, 2nd ed., New York: New York Cold Spring Habor Press, 1989.
Hone, D., Harris, A. M., Chatfield, S. et al., Construction of genetically defined double aro mutants of Salmonella typhi, Vaccine, 1991, 9(11): 810–816.
Sack, D. A., Huda, S., Neogi, P. K. B. et al., Microtiter ganglioside enzyme-linked immunosorbent assay for Vibro and Escherichia coli heat-labile enterotoxins and antitoxin, J. Clin. Microbiol., 1980, 11(1): 35–40.
Maurelli, A. T., Black, B., Curtiss III, R., Temperature-dependent expression of virulence genes in Shigella species, Infect. Immun., 1984, 43(1): 195–201.
Nassif, X., Mazert, M. C., Mounier, J. et al., Evaluation with an iuc::Tn10 mutant of the role of aerobactin production in the virulence of Shigella flexneri, Infect. Immun., 1987, 55(9): 1963–1969.
Makino, S., Sasakawa, C., Yoshikawa, M., Genetic relatedness of the basic replicon of the virulence plasmid in Shigellae and enteroinvasive Escherichia coli, Microb. Pathog., 1988, 5: 267–274.
Wang, B. R., Song, S. Z., Chen, J. R. et al., A bivalent strain with the characteristics of Shigella flexneri 2a and sonnei, Chin. Microbiol. Immunol. (in Chinese), 1987, 7: 373–377.
Mu, Z. Q., Xing, L., Mao, P. J., Use of transponson Tn501 in construction of bivalent strain of dysentery bacillus, Chin. Microbiol. Immunol. (in Chinese), 1990, 10: 92–94.
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Wang, H., Feng, E., Lin, Y. et al. Construction of a trivalent candidate vaccine against Shigella species with DNA recombination. Sci. China Ser. C.-Life Sci. 45, 10–20 (2002). https://doi.org/10.1360/02yc9002
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DOI: https://doi.org/10.1360/02yc9002