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. 1976 Jun;13(6):1692–1698. doi: 10.1128/iai.13.6.1692-1698.1976

Preparation of a purified antigenic cholera toxoid.

R Germanier, E Fürer, S Varallyay, T M Inderbitzin
PMCID: PMC420821  PMID: 823107

Abstract

Purified cholera enterotoxin was prepared by methods described by Finkelstein and Lo Spalluto (1970). This toxin was detoxified by treatment with heat and formaldehyde. Heating cholera toxin at 60 C for 25 min resulted in the formation of a polymer named procholeragenoid by Finkelstein et al. (1971). The weak toxic activity of this product was removed by treatment with formalin. No residual toxicity could be demonstrated in formalinized procholeragenoid by the rabbit ileal loop assay and the highly sensitive rabbit skin tests. This toxoid was nevertheless at least as antigenic in the rabbit as was the toxin. No reversion to toxicity was observed in vivo and in vitro at 4 C. The toxicity of formalinized procholeragenoid never exceeded 1/5,000 to 1/10,000 that of the toxin.

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Selected References

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  1. Cash R. A., Music S. I., Libonati J. P., Craig J. P., Pierce N. F., Hornick R. B. Response of man to infection with Vibrio cholerae. II. Protection from illness afforded by previous disease and vaccine. J Infect Dis. 1974 Oct;130(4):325–333. doi: 10.1093/infdis/130.4.325. [DOI] [PubMed] [Google Scholar]
  2. Craig J. P., Eichner E. R., Hornick R. B. Cutaneous responses to cholera skin toxin in man. I. Responses in unimmunized American males. J Infect Dis. 1972 Mar;125(3):203–215. doi: 10.1093/infdis/125.3.203. [DOI] [PubMed] [Google Scholar]
  3. Finkelstein R. A. Antitoxic immunity in experimental cholera: observations with purified antigens and the ligated ileal loop model. Infect Immun. 1970 May;1(5):464–467. doi: 10.1128/iai.1.5.464-467.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Finkelstein R. A., Fujita K., LoSpalluto J. J. Procholeragenoid: an aggregated intermediate in the formation of choleragenoid. J Immunol. 1971 Oct;107(4):1043–1051. [PubMed] [Google Scholar]
  5. Finkelstein R. A., LoSpalluto J. J. Pathogenesis of experimental cholera. Preparation and isolation of choleragen and choleragenoid. J Exp Med. 1969 Jul 1;130(1):185–202. doi: 10.1084/jem.130.1.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Finkelstein R. A., LoSpalluto J. J. Production of highly purified choleragen and choleragenoid. J Infect Dis. 1970 May;121(Suppl):63+–63+. doi: 10.1093/infdis/121.supplement.s63. [DOI] [PubMed] [Google Scholar]
  7. Finkelstein R. A. Monospecific equine antiserum against cholera exo-enterotoxin. Infect Immun. 1970 Dec;2(6):691–697. doi: 10.1128/iai.2.6.691-697.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fujita K., Finkelstein R. A. Antitoxic immunity in experimental cholera: comparison of immunity induced perorally and parenterally in mice. J Infect Dis. 1972 Jun;125(6):647–655. doi: 10.1093/infdis/125.6.647. [DOI] [PubMed] [Google Scholar]
  9. Holmgren J., Andersson A., Wallerstrom G., Ouchterlony O. Experimental studies on cholera immunization. II. Evidence for protective antitoxic immunity mediated by serum antibodies as well as local antibodies. Infect Immun. 1972 May;5(5):662–667. doi: 10.1128/iai.5.5.662-667.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kasai G. J., Burrows W. The titration of cholera toxin and antitoxin in the rabbit ileal loop. J Infect Dis. 1966 Dec;116(5):606–614. doi: 10.1093/infdis/116.5.606. [DOI] [PubMed] [Google Scholar]
  11. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  12. Mancini G., Carbonara A. O., Heremans J. F. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry. 1965 Sep;2(3):235–254. doi: 10.1016/0019-2791(65)90004-2. [DOI] [PubMed] [Google Scholar]
  13. Northrup R. S., Chisari F. V. Response of monkeys to immunization with cholera toxoid, toxin, and vaccine: reversion of cholera toxoid. J Infect Dis. 1972 May;125(5):471–479. doi: 10.1093/infdis/125.5.471. [DOI] [PubMed] [Google Scholar]
  14. Pierce N. F., Kaniecki E. A., Northrup R. S. Protection against experimental cholera by antitoxin. J Infect Dis. 1972 Dec;126(6):606–616. doi: 10.1093/infdis/126.6.606. [DOI] [PubMed] [Google Scholar]
  15. Rappaport R. S., Bonde G., McCann T., Rubin B. A., Tint H. Development of a purified cholera toxoid. II. Preparation of a stable, antigenic toxoid by reaction of purified toxin with glutaraldehyde. Infect Immun. 1974 Feb;9(2):304–317. doi: 10.1128/iai.9.2.304-317.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Saletti M., Ricci A. Experiments with cholera toxin detoxified with glutaraldehyde. Bull World Health Organ. 1974;51(6):633–639. [PMC free article] [PubMed] [Google Scholar]

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