Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1989 Dec;57(12):3823–3827. doi: 10.1128/iai.57.12.3823-3827.1989

Comparative immunogenicities of Vi polysaccharide-protein conjugates composed of cholera toxin or its B subunit as a carrier bound to high- or lower-molecular-weight Vi.

S C Szu 1, X R Li 1, R Schneerson 1, J H Vickers 1, D Bryla 1, J B Robbins 1
PMCID: PMC259911  PMID: 2807549

Abstract

The effect of molecular weight or size of the components on the immunogenicity of polysaccharide-protein conjugates prepared with the native Vi capsular polysaccharide (Vi) (approximately 3 x 10(3) kilodaltons) or lower-molecular-weight Vi (Vis; approximately 46 kilodaltons) abound to cholera toxin (CT) or to its B subunit (CTB) was studied. In mice, Vi-CT, Vi-CTB, and Vis-CTB elicited higher Vi antibody levels than the Vi alone (P less than 0.0001). Vi-CT and Vi-CTB were more immunogenic than Vis-CTB (P less than 0.01). CT or Vi-CT elicited higher levels of CT antibodies, as measured by enzyme-linked immunosorbent assay, than did CTB or Vi-CTB. In rhesus monkeys, the Vi conjugates elicited higher Vi antibody levels than the Vi alone (P less than 0.01). Vi-CTB elicited higher levels of Vi antibody after each injection than did Vis-CTB. Similar levels of CT antibodies, as measured by enzyme-linked immunosorbent assay, were elicited by all three conjugates. In contrast, Vi-CT elicited higher levels of neutralizing antibodies than Vi-CTB or Vis-CTB when either CT or the related heat-labile toxin of Escherichia coli was used as the antigen. These results indicate that the holotoxin and the native Vi provide the most immunogenic components for conjugates designed to induce both Vi and CT antibodies.

Full text

PDF
3823

Selected References

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

  1. Acharya I. L., Lowe C. U., Thapa R., Gurubacharya V. L., Shrestha M. B., Cadoz M., Schulz D., Armand J., Bryla D. A., Trollfors B. Prevention of typhoid fever in Nepal with the Vi capsular polysaccharide of Salmonella typhi. A preliminary report. N Engl J Med. 1987 Oct 29;317(18):1101–1104. doi: 10.1056/NEJM198710293171801. [DOI] [PubMed] [Google Scholar]
  2. Anderson P. W., Pichichero M. E., Insel R. A., Betts R., Eby R., Smith D. H. Vaccines consisting of periodate-cleaved oligosaccharides from the capsule of Haemophilus influenzae type b coupled to a protein carrier: structural and temporal requirements for priming in the human infant. J Immunol. 1986 Aug 15;137(4):1181–1186. [PubMed] [Google Scholar]
  3. Carlsson J., Drevin H., Axén R. Protein thiolation and reversible protein-protein conjugation. N-Succinimidyl 3-(2-pyridyldithio)propionate, a new heterobifunctional reagent. Biochem J. 1978 Sep 1;173(3):723–737. doi: 10.1042/bj1730723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clements J. D., Finkelstein R. A. Isolation and characterization of homogeneous heat-labile enterotoxins with high specific activity from Escherichia coli cultures. Infect Immun. 1979 Jun;24(3):760–769. doi: 10.1128/iai.24.3.760-769.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Donta S. T., Moon H. W., Whipp S. C. Detection of heat-labile Escherichia coli enterotoxin with the use of adrenal cells in tissue culture. Science. 1974 Jan 25;183(4122):334–336. doi: 10.1126/science.183.4122.334. [DOI] [PubMed] [Google Scholar]
  6. ELLMAN G. L. Tissue sulfhydryl groups. Arch Biochem Biophys. 1959 May;82(1):70–77. doi: 10.1016/0003-9861(59)90090-6. [DOI] [PubMed] [Google Scholar]
  7. Elson C. O., Ealding W. Generalized systemic and mucosal immunity in mice after mucosal stimulation with cholera toxin. J Immunol. 1984 Jun;132(6):2736–2741. [PubMed] [Google Scholar]
  8. Esposito V. M., Feeley J. C., Leeder W. D. Immunological response of the rabbit to Vi antigen. J Bacteriol. 1969 Jul;99(1):13–17. doi: 10.1128/jb.99.1.13-17.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Esposito V. M., Feeley J. C., Leeder W. D., Pittman M. Immunological response of three mouse strains to typhoid vaccine and Vi antigen. J Bacteriol. 1969 Jul;99(1):8–12. doi: 10.1128/jb.99.1.8-12.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fattom A., Vann W. F., Szu S. C., Sutton A., Li X., Bryla D., Schiffman G., Robbins J. B., Schneerson R. Synthesis and physicochemical and immunological characterization of pneumococcus type 12F polysaccharide-diphtheria toxoid conjugates. Infect Immun. 1988 Sep;56(9):2292–2298. doi: 10.1128/iai.56.9.2292-2298.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Finkelstein R. A., Burks M. F., Zupan A., Dallas W. S., Jacob C. O., Ludwig D. S. Epitopes of the cholera family of enterotoxins. Rev Infect Dis. 1987 May-Jun;9(3):544–561. doi: 10.1093/clinids/9.3.544. [DOI] [PubMed] [Google Scholar]
  12. Gill D. M., Clements J. D., Robertson D. C., Finkelstein R. A. Subunit number and arrangement in Escherichia coli heat-labile enterotoxin. Infect Immun. 1981 Sep;33(3):677–682. doi: 10.1128/iai.33.3.677-682.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gotschlich E. C., Rey M., Etienne J., Sanborn W. R., Triau R., Cvjetanović B. The immunological responses observed in field studies in Africa with group A meningococcal vaccines. Prog Immunobiol Stand. 1971;5:485–491. [PubMed] [Google Scholar]
  14. Holmgren J., Söderlind O., Wadström T. Cross-reactivity between heat labile enterotoxins of Vibrio cholerae and Escherichia coli in neutralization tests in rabbit ileum and skin. Acta Pathol Microbiol Scand B Microbiol Immunol. 1973 Dec;81(6):757–762. doi: 10.1111/j.1699-0463.1973.tb02272.x. [DOI] [PubMed] [Google Scholar]
  15. KABAT E. A., BEZER A. E. The effect of variation in molecular weight on the antigenicity of dextran in man. Arch Biochem Biophys. 1958 Dec;78(2):306–318. doi: 10.1016/0003-9861(58)90354-0. [DOI] [PubMed] [Google Scholar]
  16. Klugman K. P., Gilbertson I. T., Koornhof H. J., Robbins J. B., Schneerson R., Schulz D., Cadoz M., Armand J. Protective activity of Vi capsular polysaccharide vaccine against typhoid fever. Lancet. 1987 Nov 21;2(8569):1165–1169. doi: 10.1016/s0140-6736(87)91316-x. [DOI] [PubMed] [Google Scholar]
  17. LANDY M. Studies on Vi antigen. VI. Immunization of human beings with purified Vi antigen. Am J Hyg. 1954 Jul;60(1):52–62. doi: 10.1093/oxfordjournals.aje.a119703. [DOI] [PubMed] [Google Scholar]
  18. LANDY M. Studies on Vi antigen. VII. Characteristics of the immune response in the mouse. Am J Hyg. 1957 Jan;65(1):81–93. doi: 10.1093/oxfordjournals.aje.a119858. [DOI] [PubMed] [Google Scholar]
  19. Lai C. Y. Bacterial protein toxins with latent ADP-ribosyl transferases activities. Adv Enzymol Relat Areas Mol Biol. 1986;58:99–140. doi: 10.1002/9780470123041.ch3. [DOI] [PubMed] [Google Scholar]
  20. Levin D. M., Wong K. H., Reynolds H. Y., Sutton A., Northrup R. S. Vi antigen from Salmonella typhosa and immunity against typhoid fever. 11. Safety and antigenicity in humans. Infect Immun. 1975 Dec;12(6):1290–1294. doi: 10.1128/iai.12.6.1290-1294.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Marchlewicz B. A., Finkelstein R. A. Immunological differences among the cholera/coli family of enterotoxins. Diagn Microbiol Infect Dis. 1983 Jun;1(2):129–138. doi: 10.1016/0732-8893(83)90042-1. [DOI] [PubMed] [Google Scholar]
  22. Martin D. G., Jarvis F. G., Milner K. C. Physicochemical and biological properties of sonically treated Vi antigen. J Bacteriol. 1967 Nov;94(5):1411–1416. doi: 10.1128/jb.94.5.1411-1416.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mäkelä O., Péterfy F., Outschoorn I. G., Richter A. W., Seppälä I. Immunogenic properties of alpha (1----6) dextran, its protein conjugates, and conjugates of its breakdown products in mice. Scand J Immunol. 1984 Jun;19(6):541–550. doi: 10.1111/j.1365-3083.1984.tb00965.x. [DOI] [PubMed] [Google Scholar]
  24. Nozawa R. T., Yokota T., Kuwahara S. Assay method for Vibrio cholerae and Escherichia coli enterotoxins by automated counting of floating chinese hamster ovary cells in culture medium. J Clin Microbiol. 1978 May;7(5):479–485. doi: 10.1128/jcm.7.5.479-485.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pierce N. F. Protection against challenge with Escherichia coli heat-labile enterotoxin by immunization of rats with cholera toxin/toxoid. Infect Immun. 1977 Nov;18(2):338–341. doi: 10.1128/iai.18.2.338-341.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Robb M., Nichols J. C., Whoriskey S. K., Murphy J. R. Isolation of hybridoma cell lines and characterization of monoclonal antibodies against cholera enterotoxin and its subunits. Infect Immun. 1982 Oct;38(1):267–272. doi: 10.1128/iai.38.1.267-272.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Robbins J. D., Robbins J. B. Reexamination of the protective role of the capsular polysaccharide (Vi antigen) of Salmonella typhi. J Infect Dis. 1984 Sep;150(3):436–449. doi: 10.1093/infdis/150.3.436. [DOI] [PubMed] [Google Scholar]
  28. Sandefur P. D., Peterson J. W. Neutralization of Salmonella toxin-induced elongation of Chinese hamster ovary cells by cholera antitoxin. Infect Immun. 1977 Mar;15(3):988–992. doi: 10.1128/iai.15.3.988-992.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schneerson R., Robbins J. B., Chu C., Sutton A., Vann W., Vickers J. C., London W. T., Curfman B., Hardegree M. C., Shiloach J. Serum antibody responses of juvenile and infant rhesus monkeys injected with Haemophilus influenzae type b and pneumococcus type 6A capsular polysaccharide-protein conjugates. Infect Immun. 1984 Sep;45(3):582–591. doi: 10.1128/iai.45.3.582-591.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Stone A. L., Szu S. C. Application of optical properties of the Vi capsular polysaccharide for quantitation of the Vi antigen in vaccines for typhoid fever. J Clin Microbiol. 1988 Apr;26(4):719–725. doi: 10.1128/jcm.26.4.719-725.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Szu S. C., Schneerson R., Robbins J. B. Rabbit antibodies to the cell wall polysaccharide of Streptococcus pneumoniae fail to protect mice from lethal challenge with encapsulated pneumococci. Infect Immun. 1986 Nov;54(2):448–455. doi: 10.1128/iai.54.2.448-455.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Szu S. C., Stone A. L., Robbins J. D., Schneerson R., Robbins J. B. Vi capsular polysaccharide-protein conjugates for prevention of typhoid fever. Preparation, characterization, and immunogenicity in laboratory animals. J Exp Med. 1987 Nov 1;166(5):1510–1524. doi: 10.1084/jem.166.5.1510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Szu S. C., Zon G., Schneerson R., Robbins J. B. Ultrasonic irradiation of bacterial polysaccharides. Characterization of the depolymerized products and some applications of the process. Carbohydr Res. 1986 Sep 1;152:7–20. doi: 10.1016/s0008-6215(00)90283-0. [DOI] [PubMed] [Google Scholar]
  34. Tacket C. O., Ferreccio C., Robbins J. B., Tsai C. M., Schulz D., Cadoz M., Goudeau A., Levine M. M. Safety and immunogenicity of two Salmonella typhi Vi capsular polysaccharide vaccines. J Infect Dis. 1986 Aug;154(2):342–345. doi: 10.1093/infdis/154.2.342. [DOI] [PubMed] [Google Scholar]
  35. Uchida T., Pappenheimer A. M., Jr, Greany R. Diphtheria toxin and related proteins. I. Isolation and properties of mutant proteins serologically related to diphtheria toxin. J Biol Chem. 1973 Jun 10;248(11):3838–3844. [PubMed] [Google Scholar]
  36. Wong K. H., Feeley J. C., Northrup R. S., Forlines M. E. Vi antigen from Salmonella typhosa and immunity against typhoid fever. I. Isolation and immunologic properties in animals. Infect Immun. 1974 Feb;9(2):348–353. doi: 10.1128/iai.9.2.348-353.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES