1887

Journal of Medical Microbiology logo

Volume 23, Issue 1

Preparation and immunogenicity of a bivalent cell-surface protein-polysaccharide conjugate of Free

Abstract

Summary

Alkali-treated lipopolysaccharides (LPS) from Ogawa and Inaba serotypes of were chemically coupied to cell-surface proteins of . The reaction product was eluted in the void volume when fractionated on a column of Sephacryl S-300. The material did not enter the gel when subjected to polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate (SDS-PAGE). The bivalent protein-polysaccharide conjugate was nonpyrogenic, as determined by the Limulus lysate assay. It was immunogenic and elicited, in rabbits, antibodies against both intact LPS and cell surface proteins, as determined by enzyme linked immunosorbent assay. LPS from Ogawa serotype was resolved into two major bands by SDS-PAGE and that from the Inaba serotype into one major band. Immunoblotting studies indicated that antisera to the protein-polysaccharide conjugate contained antibodies to the major LPS fractions from both serotypes. Antisera to the protein-polysaccharide conjugate tested by crossed-immunoelectrophoresis produced immunoprecipitation with whole-cell sonicates of both biotypes and serotypes of . Such antisera also possessed agglutinating and complement-mediated bactericidal activities towards strains of both biotypes and serotypes. These results suggest that a bivalent cell-surface protein-polysaccharide conjugate of could be developed as a nonpyrogenic vaccine against cholera.

  • Received:
  • Accepted:
  • Published Online:
© J. MED. MICROBIOL. 1987
Loading

Article metrics loading...

/content/journal/jmm/10.1099/00222615-23-1-9
1987-02-01
2024-04-16
Loading full text...

Full text loading...

/deliver/fulltext/jmm/23/1/medmicro-23-1-9.html?itemId=/content/journal/jmm/10.1099/00222615-23-1-9&mimeType=html&fmt=ahah

References

  1. Axelsen N. H. 1973; Intermediate gel in crossed and in fused rocket immunoelectrophoresis. Scandinavian Journal of Immunology 2: Suppl no. 1 71–77
     [Google Scholar]
  2. De S. N., Chatterje D. N. 1953; An experimental study of the mechanism of action of Vibrio cholerae on the intestinal mucous membrane. Journal of Pathology and Bacteriology 66:559–562
     [Google Scholar]
  3. Feeley J. C. 1970; Cholera vaccines. In Principles and practice of cholera control (WHO Public Health Papers no. 40) World Health Organization; Geneva: pp 87–93
     [Google Scholar]
  4. Goldman R. C., Leive L. 1980; Heterogeneity of antigenic-side- chain length in lipopolysaccharide from Escherichia coli 0111 and Salmonella typhimurium LT2. European Journal of Biochemistry 107:145–153
     [Google Scholar]
  5. Joo I. 1974; Cholera vaccines. In Barua D. Burrows W. (eds) Cholera. W B. Saunders Co; Philadelphia: pp 333–355
     [Google Scholar]
  6. Kabir S. 1976; Electrophoretic studies of smooth and rough Salmonella lipopolysaccharides on polyacrylamide gels. Microbios Letters 1:79–85
     [Google Scholar]
  7. Kabir S. 1980; Composition and immunochemical properties of outer membrane proteins of Vibrio cholerae. Journal of Bacteriology 144:382–389
     [Google Scholar]
  8. Kabir S. 1982; Characterization of the lipopolysaccharide from Vibrio cholerae 395 (Ogawa). Infection and Immunity 38:1263–1272
     [Google Scholar]
  9. Kabir S. 1983a; Immunochemical properties of the major outer membrane protein of Vibrio cholerae. Infection and Immunity 39:452–455
     [Google Scholar]
  10. Kabir S. 1983b; The serological properties of the cell surface proteins of Vibrio cholerae. Journal of General Microbiology 129:2199–2206
     [Google Scholar]
  11. Kabir S., Mann P. 1980; Immunological properties of the cell envelope components of Vibrio cholerae. Journal of General Microbiology 119:517–525
     [Google Scholar]
  12. King J., Laemmli U. K. 1971; Polypeptides of the tail fibres of bacteriophage T4. Journal of Molecular Biology 62:465–477
     [Google Scholar]
  13. Kreeftenberg J. G., Loggen H. G., van Ramshoorst J. D., Beuvery E. C. 1977; The Limulus amoebocyte lysate test: micromethod and application in the control of sera and vaccines. Developments in Biological Standardization 34:15–20
     [Google Scholar]
  14. Levine M. M. 1979; Immunity to cholera in man: relative role of antibacterial versus antitoxic immunity. Transactions of the Royal Society of Tropical Medicine and Hygiene 73:3–9
     [Google Scholar]
  15. Majumdar A. S., Dutta P., Dutta D., Ghose A. C. 1981; Antibacterial and antitoxin responses in the serum and milk of cholera patients. Infection and Immunity 32:1–8
     [Google Scholar]
  16. Mall T., Gyr K. 1984; Episode resembling immune complex disease after cholera vaccination. Transactions of the Royal Society of Tropical Medicine and Hygiene 78:106–107
     [Google Scholar]
  17. Marcus S., Nelson J. R. 1977; Tests alternative to the rabbit bioassay for pyrogens. Developments in Biological Standardization 34:45–55
     [Google Scholar]
  18. Mosley W. H. 1970; The 1968-1969. cholera-vaccine field trials in rural East Pakistan. Effectiveness of monovalent Ogawa and Inaba vaccines and a purified Inaba antigen, with comparative results of serological and animal protection tests. Journal of Infectious Diseases 121: (suppl) 1–9
     [Google Scholar]
  19. Raziuddin S. 1978; Toxic and immunological properties of the lipopolysaccharides (O-antigens) from Vibrio el tor. Im- munochemistry 15:611–614
     [Google Scholar]
  20. Redmond J. W. 1978; The 4-amino sugars present in the lipopolysaccharides of Vibrio cholerae and related vibrios. Biochi- mica et Biophysica Acta 542:378–384
     [Google Scholar]
  21. Sadolf J. C., Futrovsky S. L., Sideberry H. F., Iglewski B. H., Seid R. C. 1982; Detoxified lipopolysaccharide-protein conjugates. In Robbins J. B. (eds) Bacterial vaccines (Seminars in infectious Diseases 4). Thieme-Stratton; New York: pp 346–354
     [Google Scholar]
  22. Schrader H. 1975; Ein Fall von Guillain Barre-Syndrome nach cholera-schutzimpfung. Journal of Neurology 209:69–74
     [Google Scholar]
  23. Sen A. K., Mukherjee A. K., Guhathakurta B., Dutta A., Sasmal D. 1979; Structural investigations on the lipopolysaccharide isolated from Vibrio cholerae Inaba 569B. Carbohydrate Research 72:191–199
     [Google Scholar]
  24. Solomons T. W. G. 1976 Organic Chemistry John Wiley; London: p 512
     [Google Scholar]
  25. Weeke B. 1973; Crossed immunoelectrophoresis. Scandinavian Journal of Immunology 2: Suppl 1 47–56
     [Google Scholar]
  26. Westphal O., Liideritz O., Bister F. 1952; Uber die Extraktion von Bakterien mit Phenol-wasser. Zeitschrift fur Naturforschung Teil B 7:148–155
     [Google Scholar]
  27. Westphal O., Jann K. 1965; Bacterial lipopolysaccharides. Extraction with phenol-water and further applications of the procedure. Methods in Carbohydrate Chemistry 5:83–91
     [Google Scholar]
  28. Whang H. Y., Mayer H., Neter E. 1971; Differential effects on immunogenicity and antigenicity of heat, freezing and alkali treatment of bacterial antigens. Journal of Immunology 106:1552–1558
     [Google Scholar]
  29. World Health Organization 1983; Cholera 1963–1982. In World health statistics annual World Health Organization; Geneva: pp 787–790
     [Google Scholar]
  30. Zacharius R. M., Zell T. E., Morrison J. H., Woodlock J. J. 1969; Glycoprotein staining following electrophoresis on acrylamide gels. Analytical Biochemistry 30:148–152
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/00222615-23-1-9
Loading
Preparation and immunogenicity of a bivalent cell-surface protein-polysaccharide conjugate of Vibrio cholerae
J. Med. Microbiol. 23, 9 (1987); https://doi.org/10.1099/00222615-23-1-9
/content/journal/jmm/10.1099/00222615-23-1-9
/content/journal/jmm/10.1099/00222615-23-1-9
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error