Skip to main content
NCBI home page
Immunology logoLink to Immunology
. 1997 Feb;90(2):176–182. doi: 10.1046/j.1365-2567.1997.00156.x

Relationship between a low toxicity of the mutant A subunit of enterotoxigenic Escherichia coli enterotoxin and its strong adjuvant action.

T Tsuji 1, T Yokochi 1, H Kamiya 1, Y Kawamoto 1, A Miyama 1, Y Asano 1
PMCID: PMC1456747  PMID: 9135544

Abstract

In the work described here it was determined if and how unnicking in the A subunit of Escherichia coli enterotoxin at Arg192 or nearby residues affected biological activities of the toxin. The mutant toxin was constructed to lack the nick site in the A subunit by deleting the tripeptide Arg192-Thr193-Ile194, which is essential for toxicity. The mutant toxin did not exhibit agmatine ADP-ribosyltransferase activity in the presence or absence of the ADP-ribosylation factor and had less diarrhoeal activity and lower induction of cyclic AMP than did LT. The mutant toxin exhibited a much stronger adjuvant action on antibody responses to measles virus, keyhole limpt haemocyanin, bovine immunoglobulin and ovalbumin compared with LT. The altered toxicity of the mutant toxin might be closely related to the potent adjuvant action on antibody responses to antigens. The relationship between two activities is discussed.

Full text

PDF
176

Images in this article

178Figure 1
on p.178
180Figure 4
on p.180

Selected References

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

  1. Bromander A. K., Kjerrulf M., Holmgren J., Lycke N. Cholera toxin enhances alloantigen presentation by cultured intestinal epithelial cells. Scand J Immunol. 1993 Apr;37(4):452–458. doi: 10.1111/j.1365-3083.1993.tb03318.x. [DOI] [PubMed] [Google Scholar]
  2. Bromander A., Holmgren J., Lycke N. Cholera toxin stimulates IL-1 production and enhances antigen presentation by macrophages in vitro. J Immunol. 1991 May 1;146(9):2908–2914. [PubMed] [Google Scholar]
  3. Chen K. S., Strober W. Cholera holotoxin and its B subunit enhance Peyer's patch B cell responses induced by orally administered influenza virus: disproportionate cholera toxin enhancement of the IgA B cell response. Eur J Immunol. 1990 Feb;20(2):433–436. doi: 10.1002/eji.1830200230. [DOI] [PubMed] [Google Scholar]
  4. Dallas W. S., Gill D. M., Falkow S. Cistrons encoding Escherichia coli heat-labile toxin. J Bacteriol. 1979 Sep;139(3):850–858. doi: 10.1128/jb.139.3.850-858.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dickinson B. L., Clements J. D. Dissociation of Escherichia coli heat-labile enterotoxin adjuvanticity from ADP-ribosyltransferase activity. Infect Immun. 1995 May;63(5):1617–1623. doi: 10.1128/iai.63.5.1617-1623.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Douce G., Turcotte C., Cropley I., Roberts M., Pizza M., Domenghini M., Rappuoli R., Dougan G. Mutants of Escherichia coli heat-labile toxin lacking ADP-ribosyltransferase activity act as nontoxic, mucosal adjuvants. Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1644–1648. doi: 10.1073/pnas.92.5.1644. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Giraudon P., Wild T. F. Correlation between epitopes on hemagglutinin of measles virus and biological activities: passive protection by monoclonal antibodies is related to their hemagglutination inhibiting activity. Virology. 1985 Jul 15;144(1):46–58. doi: 10.1016/0042-6822(85)90303-4. [DOI] [PubMed] [Google Scholar]
  8. Holmgren J. Actions of cholera toxin and the prevention and treatment of cholera. Nature. 1981 Jul 30;292(5822):413–417. doi: 10.1038/292413a0. [DOI] [PubMed] [Google Scholar]
  9. Holmgren J., Lycke N., Czerkinsky C. Cholera toxin and cholera B subunit as oral-mucosal adjuvant and antigen vector systems. Vaccine. 1993 Sep;11(12):1179–1184. doi: 10.1016/0264-410x(93)90039-z. [DOI] [PubMed] [Google Scholar]
  10. Ito W., Ishiguro H., Kurosawa Y. A general method for introducing a series of mutations into cloned DNA using the polymerase chain reaction. Gene. 1991 Jun 15;102(1):67–70. doi: 10.1016/0378-1119(91)90539-n. [DOI] [PubMed] [Google Scholar]
  11. Jackson R. J., Fujihashi K., Xu-Amano J., Kiyono H., Elson C. O., McGhee J. R. Optimizing oral vaccines: induction of systemic and mucosal B-cell and antibody responses to tetanus toxoid by use of cholera toxin as an adjuvant. Infect Immun. 1993 Oct;61(10):4272–4279. doi: 10.1128/iai.61.10.4272-4279.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kramer S. M., Cremer N. E. Biological activity of a monoclonal antibody to a measles virus haemagglutinin epitope detected late in infection. J Gen Virol. 1984 Mar;65(Pt 3):577–583. doi: 10.1099/0022-1317-65-3-577. [DOI] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Lycke N., Bromander A. K., Ekman L., Karlsson U., Holmgren J. Cellular basis of immunomodulation by cholera toxin in vitro with possible association to the adjuvant function in vivo. J Immunol. 1989 Jan 1;142(1):20–27. [PubMed] [Google Scholar]
  15. Lycke N., Holmgren J. Strong adjuvant properties of cholera toxin on gut mucosal immune responses to orally presented antigens. Immunology. 1986 Oct;59(2):301–308. [PMC free article] [PubMed] [Google Scholar]
  16. Lycke N., Tsuji T., Holmgren J. The adjuvant effect of Vibrio cholerae and Escherichia coli heat-labile enterotoxins is linked to their ADP-ribosyltransferase activity. Eur J Immunol. 1992 Sep;22(9):2277–2281. doi: 10.1002/eji.1830220915. [DOI] [PubMed] [Google Scholar]
  17. McGee D. W., Elson C. O., McGhee J. R. Enhancing effect of cholera toxin on interleukin-6 secretion by IEC-6 intestinal epithelial cells: mode of action and augmenting effect of inflammatory cytokines. Infect Immun. 1993 Nov;61(11):4637–4644. doi: 10.1128/iai.61.11.4637-4644.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. McKenzie S. J., Halsey J. F. Cholera toxin B subunit as a carrier protein to stimulate a mucosal immune response. J Immunol. 1984 Oct;133(4):1818–1824. [PubMed] [Google Scholar]
  19. Moss J., Stanley S. J., Vaughan M., Tsuji T. Interaction of ADP-ribosylation factor with Escherichia coli enterotoxin that contains an inactivating lysine 112 substitution. J Biol Chem. 1993 Mar 25;268(9):6383–6387. [PubMed] [Google Scholar]
  20. Muñoz E., Zubiaga A. M., Merrow M., Sauter N. P., Huber B. T. Cholera toxin discriminates between T helper 1 and 2 cells in T cell receptor-mediated activation: role of cAMP in T cell proliferation. J Exp Med. 1990 Jul 1;172(1):95–103. doi: 10.1084/jem.172.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Okuno Y., Ueda S., Kurimura T., Suzuki N., Yamanishi K. Studies on further attenuated liver measles vaccine. VII. Development and evaluation of CAM-70 measles virus vaccine. Biken J. 1971 Sep;14(3):253–258. [PubMed] [Google Scholar]
  22. Tamura S. I., Asanuma H., Ito Y., Hirabayashi Y., Suzuki Y., Nagamine T., Aizawa C., Kurata T., Oya A. Superior cross-protective effect of nasal vaccination to subcutaneous inoculation with influenza hemagglutinin vaccine. Eur J Immunol. 1992 Feb;22(2):477–481. doi: 10.1002/eji.1830220228. [DOI] [PubMed] [Google Scholar]
  23. Tamura S., Samegai Y., Kurata H., Nagamine T., Aizawa C., Kurata T. Protection against influenza virus infection by vaccine inoculated intranasally with cholera toxin B subunit. Vaccine. 1988 Oct;6(5):409–413. doi: 10.1016/0264-410x(88)90140-5. [DOI] [PubMed] [Google Scholar]
  24. Tsuji T., Inoue T., Miyama A., Okamoto K., Honda T., Miwatani T. A single amino acid substitution in the A subunit of Escherichia coli enterotoxin results in a loss of its toxic activity. J Biol Chem. 1990 Dec 25;265(36):22520–22525. [PubMed] [Google Scholar]
  25. Uesaka Y., Otsuka Y., Lin Z., Yamasaki S., Yamaoka J., Kurazono H., Takeda Y. Simple method of purification of Escherichia coli heat-labile enterotoxin and cholera toxin using immobilized galactose. Microb Pathog. 1994 Jan;16(1):71–76. doi: 10.1006/mpat.1994.1007. [DOI] [PubMed] [Google Scholar]
  26. Van der Heijden P. J., Bianchi A. T., Dol M., Pals J. W., Stok W., Bokhout B. A. Manipulation of intestinal immune responses against ovalbumin by cholera toxin and its B subunit in mice. Immunology. 1991 Jan;72(1):89–93. [PMC free article] [PubMed] [Google Scholar]
  27. Xu-Amano J., Kiyono H., Jackson R. J., Staats H. F., Fujihashi K., Burrows P. D., Elson C. O., Pillai S., McGhee J. R. Helper T cell subsets for immunoglobulin A responses: oral immunization with tetanus toxoid and cholera toxin as adjuvant selectively induces Th2 cells in mucosa associated tissues. J Exp Med. 1993 Oct 1;178(4):1309–1320. doi: 10.1084/jem.178.4.1309. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Immunology are provided here courtesy of British Society for Immunology

RESOURCES