Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1995 Sep;177(18):5310–5315. doi: 10.1128/jb.177.18.5310-5315.1995

Expression of Shigella dysenteriae serotype 1 O-antigenic polysaccharide by Shigella flexneri aroD vaccine candidates and different S. flexneri serotypes.

I C Fält 1, E K Schweda 1, S Klee 1, M Singh 1, E Floderus 1, K N Timmis 1, A A Lindberg 1
PMCID: PMC177324  PMID: 7545156

Abstract

The potential utility of Shigella flexneri aroD vaccine candidates for the development of bi- or multivalent vaccines has been explored by the introduction of the genetic determinants rfp and rfb for heterologous O antigen polysaccharide from Shigella dysenteriae serotype 1. The serotype Y vaccine strain SFL124 expressed the heterologous antigen qualitatively and quantitatively well, qualitatively in the sense of the O antigen polysaccharide being correctly linked to the S. flexneri lipopolysaccharide R3 core oligosaccharide and quantitatively in the sense that typical yields were obtained, with ratios of homologous to heterologous O antigen being 4:1 for one construct and 1:1 for another. Moreover, both polysaccharide chains were shown to be linked to position O-4 of the subterminal D-glucose residue of the R3 core. In contrast to the hybrid serotype Y SFL124 derivatives, analogous derivatives of serotype 2a vaccine strain SFL1070 did not elaborate a complete heterologous O antigen. Such derivatives, and analogous derivatives of rough, O antigen-negative mutants of SFL1070, formed instead a hybrid lipopolysaccharide molecule consisting of the S. flexneri lipid A R3 core with a single repeat unit of the S. dysenteriae type 1 O antigen. Introduction of the determinants for the S. dysenteriae type 1 O antigen into a second serotype 2a strain and into strains representing other serotypes of S. flexneri, revealed the following for the expression of the heterologous O antigen: serotypes 1a, 1b, 2a, and 5a did not produce the heterologous O antigen, whereas serotypes 2b, 3a, 3b, 4a, 4b, 5b, and X did.

Full Text

The Full Text of this article is available as a PDF (283.6 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Carlin N. I., Lindberg A. A. Monoclonal antibodies specific for Shigella flexneri lipopolysaccharides: clones binding to type IV, V, and VI antigens, group 3,4 antigen, and an epitope common to all Shigella flexneri and Shigella dysenteriae type 1 stains. Infect Immun. 1987 Jun;55(6):1412–1420. doi: 10.1128/iai.55.6.1412-1420.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dmitriev B. A., Knirel Y. A., Kochetkov N. K. Somatic antigens of shigella. Structural investigation on the O-specific polysaccharide chain of Shigella dysenteriae type 1 lipopolysaccharide. Eur J Biochem. 1976 Jul 15;66(3):559–566. doi: 10.1111/j.1432-1033.1976.tb10582.x. [DOI] [PubMed] [Google Scholar]
  3. Fellay R., Frey J., Krisch H. Interposon mutagenesis of soil and water bacteria: a family of DNA fragments designed for in vitro insertional mutagenesis of gram-negative bacteria. Gene. 1987;52(2-3):147–154. doi: 10.1016/0378-1119(87)90041-2. [DOI] [PubMed] [Google Scholar]
  4. Fält I. C., Schweda E. K., Weintraub A., Sturm S., Timmis K. N., Lindberg A. A. Expression of the Shigella dysenteriae type-1 lipopolysaccharide repeating unit in Escherichia coli K12/Shigella dysenteriae type-1 hybrids. Eur J Biochem. 1993 Apr 1;213(1):573–581. doi: 10.1111/j.1432-1033.1993.tb17796.x. [DOI] [PubMed] [Google Scholar]
  5. HAKOMORI S. A RAPID PERMETHYLATION OF GLYCOLIPID, AND POLYSACCHARIDE CATALYZED BY METHYLSULFINYL CARBANION IN DIMETHYL SULFOXIDE. J Biochem. 1964 Feb;55:205–208. [PubMed] [Google Scholar]
  6. Hitchcock P. J., Brown T. M. Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J Bacteriol. 1983 Apr;154(1):269–277. doi: 10.1128/jb.154.1.269-277.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Jansson P. E., Lindberg A. A., Lindberg B., Wollin R. Structural studies on the hexose region of the core in lipopolysaccharides from Enterobacteriaceae. Eur J Biochem. 1981 Apr;115(3):571–577. doi: 10.1111/j.1432-1033.1981.tb06241.x. [DOI] [PubMed] [Google Scholar]
  8. Kenne L., Lindberg B., Petersson K. Basic structure of the oligosaccharide repeating-unit of the Shigella flexneri O-antigens. Carbohydr Res. 1977 Jul;56(2):363–370. doi: 10.1016/s0008-6215(00)83357-1. [DOI] [PubMed] [Google Scholar]
  9. Kenne L., Lindberg B., Petersson K., Katzenellenbogen E., Romanowska E. Structural studies of Shigella flexneri O-antigens. Eur J Biochem. 1978 Nov 2;91(1):279–284. doi: 10.1111/j.1432-1033.1978.tb20963.x. [DOI] [PubMed] [Google Scholar]
  10. Kärnell A., Cam P. D., Verma N., Lindberg A. A. AroD deletion attenuates Shigella flexneri strain 2457T and makes it a safe and efficacious oral vaccine in monkeys. Vaccine. 1993;11(8):830–836. doi: 10.1016/0264-410x(93)90358-5. [DOI] [PubMed] [Google Scholar]
  11. Kärnell A., Li A., Zhao C. R., Karlsson K., Nguyen B. M., Lindberg A. A. Safety and immunogenicity study of the auxotrophic Shigella flexneri 2a vaccine SFL1070 with a deleted aroD gene in adult Swedish volunteers. Vaccine. 1995 Jan;13(1):88–99. doi: 10.1016/0264-410x(95)80017-8. [DOI] [PubMed] [Google Scholar]
  12. Kärnell A., Stocker B. A., Katakura S., Reinholt F. P., Lindberg A. A. Live oral auxotrophic Shigella flexneri SFL124 vaccine with a deleted aroD gene: characterization and monkey protection studies. Vaccine. 1992;10(6):389–394. doi: 10.1016/0264-410x(92)90069-v. [DOI] [PubMed] [Google Scholar]
  13. Kärnell A., Sweiha H., Lindberg A. A. Auxotrophic live oral Shigella flexneri vaccine protects monkeys against challenge with S. flexneri of different serotypes. Vaccine. 1992;10(3):167–174. doi: 10.1016/0264-410x(92)90007-7. [DOI] [PubMed] [Google Scholar]
  14. Lesse A. J., Campagnari A. A., Bittner W. E., Apicella M. A. Increased resolution of lipopolysaccharides and lipooligosaccharides utilizing tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis. J Immunol Methods. 1990 Jan 24;126(1):109–117. doi: 10.1016/0022-1759(90)90018-q. [DOI] [PubMed] [Google Scholar]
  15. Li A., Kärnell A., Huan P. T., Cam P. D., Minh N. B., Trâm L. N., Quy N. P., Trach D. D., Karlsson K., Lindberg G. Safety and immunogenicity of the live oral auxotrophic Shigella flexneri SFL124 in adult Vietnamese volunteers. Vaccine. 1993;11(2):180–189. doi: 10.1016/0264-410x(93)90015-p. [DOI] [PubMed] [Google Scholar]
  16. Li A., Pál T., Forsum U., Lindberg A. A. Safety and immunogenicity of the live oral auxotrophic Shigella flexneri SFL124 in volunteers. Vaccine. 1992;10(6):395–404. doi: 10.1016/0264-410x(92)90070-z. [DOI] [PubMed] [Google Scholar]
  17. Lindberg A. A., Karnell A., Pál T., Sweiha H., Hultenby K., Stocker B. A. Construction of an auxotrophic Shigella flexneri strain for use as a live vaccine. Microb Pathog. 1990 Jun;8(6):433–440. doi: 10.1016/0882-4010(90)90030-t. [DOI] [PubMed] [Google Scholar]
  18. Lindberg A. A., Kärnell A., Stocker B. A., Katakura S., Sweiha H., Reinholt F. P. Development of an auxotrophic oral live Shigella flexneri vaccine. Vaccine. 1988 Apr;6(2):146–150. doi: 10.1016/s0264-410x(88)80018-5. [DOI] [PubMed] [Google Scholar]
  19. O'Callaghan D., Charbit A. High efficiency transformation of Salmonella typhimurium and Salmonella typhi by electroporation. Mol Gen Genet. 1990 Aug;223(1):156–158. doi: 10.1007/BF00315809. [DOI] [PubMed] [Google Scholar]
  20. Payne S. M., Finkelstein R. A. Detection and differentiation of iron-responsive avirulent mutants on Congo red agar. Infect Immun. 1977 Oct;18(1):94–98. doi: 10.1128/iai.18.1.94-98.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sturm S., Jann B., Jann K., Fortnagel P., Timmis K. N. Genetic and biochemical analysis of Shigella dysenteriae 1 O antigen polysaccharide biosynthesis in Escherichia coli K-12: structure and functions of the rfb gene cluster. Microb Pathog. 1986 Jun;1(3):307–324. doi: 10.1016/0882-4010(86)90056-2. [DOI] [PubMed] [Google Scholar]
  22. Sturm S., Timmis K. N. Cloning of the rfb gene region of Shigella dysenteriae 1 and construction of an rfb-rfp gene cassette for the development of lipopolysaccharide-based live anti-dysentery vaccines. Microb Pathog. 1986 Jun;1(3):289–297. doi: 10.1016/0882-4010(86)90054-9. [DOI] [PubMed] [Google Scholar]
  23. Verma N. K., Lindberg A. A. Construction of aromatic dependent Shigella flexneri 2a live vaccine candidate strains: deletion mutations in the aroA and the aroD genes. Vaccine. 1991 Jan;9(1):6–9. doi: 10.1016/0264-410x(91)90308-s. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES