Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1997 Jul;65(7):2978–2982. doi: 10.1128/iai.65.7.2978-2982.1997

Murine antibody responses to the verotoxin 1 B subunit: demonstration of major histocompatibility complex dependence and an immunodominant epitope involving phenylalanine 30.

D J Bast 1, J Sandhu 1, N Hozumi 1, B Barber 1, J Brunton 1
PMCID: PMC175418  PMID: 9199476

Abstract

Structurally conserved verotoxin 1 (VT1) mutant derivatives, showing reduced receptor binding and cytotoxicity, may serve as natural toxoids to protect against VT-mediated disease. In this study, the antibody responses to the wild-type VT1 B subunit, a B-subunit mutant (Phe30Ala B), and the corresponding holotoxin (Phe30Ala HT) were examined in three inbred mouse strains. BALB/c (H-2d) and CBA (H-2k) mice produced strong antibody responses to both wild-type and mutant B subunits. VT1 B-raised sera reacted more strongly with VT1 B than with Phe30Ala B in enzyme-linked immunosorbent assays, while Phe30Ala B-raised sera reacted equally with VT1 B and Phe30Ala B. C57BL/6 (H-2b) and congenic BALB/c (BALB x B [H-2b]) mice produced no detectable antibody response to either VT1 B or Phe30Ala B. However, an anti-VT1 B antibody response was detected in H-2b mice immunized with biologically active Phe30Ala HT. Based on these observations, we conclude that the VT1 B subunit possesses a B-cell immunodominant epitope formed partly by phenylalanine 30 and that the B-subunit antibody response is dependent on the H-2 haplotype of the mouse strain. Our results also support a potential role for the A subunit in providing the T-cell help necessary to overcome a deficient B-subunit antibody response in H-2b mice.

Full Text

The Full Text of this article is available as a PDF (153.0 KB).

Selected References

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

  1. Acheson D. W., Levine M. M., Kaper J. B., Keusch G. T. Protective immunity to Shiga-like toxin I following oral immunization with Shiga-like toxin I B-subunit-producing Vibrio cholerae CVD 103-HgR. Infect Immun. 1996 Jan;64(1):355–357. doi: 10.1128/iai.64.1.355-357.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Banic D. M., Delplace P., Mazingue C., Camus D. H-2b restriction of the immune response to the p126 Plasmodium falciparum antigen. Clin Exp Immunol. 1994 Mar;95(3):472–478. doi: 10.1111/j.1365-2249.1994.tb07021.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bast D. J., Brunton J. L., Karmali M. A., Richardson S. E. Toxicity and immunogenicity of a verotoxin 1 mutant with reduced globotriaosylceramide receptor binding in rabbits. Infect Immun. 1997 Jun;65(6):2019–2028. doi: 10.1128/iai.65.6.2019-2028.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boyd B., Richardson S., Gariepy J. Serological responses to the B subunit of Shiga-like toxin 1 and its peptide fragments indicate that the B subunit is a vaccine candidate to counter action of the toxin. Infect Immun. 1991 Mar;59(3):750–757. doi: 10.1128/iai.59.3.750-757.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clark C., Bast D., Sharp A. M., St Hilaire P. M., Agha R., Stein P. E., Toone E. J., Read R. J., Brunton J. L. Phenylalanine 30 plays an important role in receptor binding of verotoxin-1. Mol Microbiol. 1996 Feb;19(4):891–899. doi: 10.1046/j.1365-2958.1996.427962.x. [DOI] [PubMed] [Google Scholar]
  6. Elson C. O., Ealding W. Genetic control of the murine immune response to cholera toxin. J Immunol. 1985 Aug;135(2):930–932. [PubMed] [Google Scholar]
  7. Endo Y., Tsurugi K., Yutsudo T., Takeda Y., Ogasawara T., Igarashi K. Site of action of a Vero toxin (VT2) from Escherichia coli O157:H7 and of Shiga toxin on eukaryotic ribosomes. RNA N-glycosidase activity of the toxins. Eur J Biochem. 1988 Jan 15;171(1-2):45–50. doi: 10.1111/j.1432-1033.1988.tb13756.x. [DOI] [PubMed] [Google Scholar]
  8. Fontana M. R., Manetti R., Giannelli V., Magagnoli C., Marchini A., Olivieri R., Domenighini M., Rappuoli R., Pizza M. Construction of nontoxic derivatives of cholera toxin and characterization of the immunological response against the A subunit. Infect Immun. 1995 Jun;63(6):2356–2360. doi: 10.1128/iai.63.6.2356-2360.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hirabayashi Y., Tamura S. I., Suzuki Y., Nagamine T., Aizawa C., Shimada K., Kurata T. H-2-unrestricted adjuvant effect of cholera toxin B subunit on murine antibody responses to influenza virus haemagglutinin. Immunology. 1991 Mar;72(3):329–335. [PMC free article] [PubMed] [Google Scholar]
  10. Hofmann S. L. Southwestern Internal Medicine Conference: Shiga-like toxins in hemolytic-uremic syndrome and thrombotic thrombocytopenic purpura. Am J Med Sci. 1993 Dec;306(6):398–406. doi: 10.1097/00000441-199312000-00009. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Karmali M. A. Infection by verocytotoxin-producing Escherichia coli. Clin Microbiol Rev. 1989 Jan;2(1):15–38. doi: 10.1128/cmr.2.1.15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Karmali M. A., Petric M., Lim C., Fleming P. C., Arbus G. S., Lior H. The association between idiopathic hemolytic uremic syndrome and infection by verotoxin-producing Escherichia coli. J Infect Dis. 1985 May;151(5):775–782. doi: 10.1093/infdis/151.5.775. [DOI] [PubMed] [Google Scholar]
  14. Mangeney M., Lingwood C. A., Taga S., Caillou B., Tursz T., Wiels J. Apoptosis induced in Burkitt's lymphoma cells via Gb3/CD77, a glycolipid antigen. Cancer Res. 1993 Nov 1;53(21):5314–5319. [PubMed] [Google Scholar]
  15. O'Brien A. D., LaVeck G. D. Purification and characterization of a Shigella dysenteriae 1-like toxin produced by Escherichia coli. Infect Immun. 1983 May;40(2):675–683. doi: 10.1128/iai.40.2.675-683.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. O'Brien A. D., Tesh V. L., Donohue-Rolfe A., Jackson M. P., Olsnes S., Sandvig K., Lindberg A. A., Keusch G. T. Shiga toxin: biochemistry, genetics, mode of action, and role in pathogenesis. Curr Top Microbiol Immunol. 1992;180:65–94. doi: 10.1007/978-3-642-77238-2_4. [DOI] [PubMed] [Google Scholar]
  17. Ostroff S. M., Tarr P. I., Neill M. A., Lewis J. H., Hargrett-Bean N., Kobayashi J. M. Toxin genotypes and plasmid profiles as determinants of systemic sequelae in Escherichia coli O157:H7 infections. J Infect Dis. 1989 Dec;160(6):994–998. doi: 10.1093/infdis/160.6.994. [DOI] [PubMed] [Google Scholar]
  18. Pizza M., Fontana M. R., Giuliani M. M., Domenighini M., Magagnoli C., Giannelli V., Nucci D., Hol W., Manetti R., Rappuoli R. A genetically detoxified derivative of heat-labile Escherichia coli enterotoxin induces neutralizing antibodies against the A subunit. J Exp Med. 1994 Dec 1;180(6):2147–2153. doi: 10.1084/jem.180.6.2147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ramotar K., Boyd B., Tyrrell G., Gariepy J., Lingwood C., Brunton J. Characterization of Shiga-like toxin I B subunit purified from overproducing clones of the SLT-I B cistron. Biochem J. 1990 Dec 15;272(3):805–811. doi: 10.1042/bj2720805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Richardson S. E., Karmali M. A., Becker L. E., Smith C. R. The histopathology of the hemolytic uremic syndrome associated with verocytotoxin-producing Escherichia coli infections. Hum Pathol. 1988 Sep;19(9):1102–1108. doi: 10.1016/s0046-8177(88)80093-5. [DOI] [PubMed] [Google Scholar]
  21. Richardson S. E., Rotman T. A., Jay V., Smith C. R., Becker L. E., Petric M., Olivieri N. F., Karmali M. A. Experimental verocytotoxemia in rabbits. Infect Immun. 1992 Oct;60(10):4154–4167. doi: 10.1128/iai.60.10.4154-4167.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Riley L. W., Remis R. S., Helgerson S. D., McGee H. B., Wells J. G., Davis B. R., Hebert R. J., Olcott E. S., Johnson L. M., Hargrett N. T. Hemorrhagic colitis associated with a rare Escherichia coli serotype. N Engl J Med. 1983 Mar 24;308(12):681–685. doi: 10.1056/NEJM198303243081203. [DOI] [PubMed] [Google Scholar]
  23. Ryd M., Verma N., Lindberg A. A. Induction of a humoral immune response to a Shiga toxin B subunit epitope expressed as a chimeric LamB protein in a Shigella flexneri live vaccine strain. Microb Pathog. 1992 Jun;12(6):399–407. doi: 10.1016/0882-4010(92)90002-6. [DOI] [PubMed] [Google Scholar]
  24. Sandvig K., Olsnes S., Brown J. E., Petersen O. W., van Deurs B. Endocytosis from coated pits of Shiga toxin: a glycolipid-binding protein from Shigella dysenteriae 1. J Cell Biol. 1989 Apr;108(4):1331–1343. doi: 10.1083/jcb.108.4.1331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Scotland S. M., Willshaw G. A., Smith H. R., Rowe B. Properties of strains of Escherichia coli belonging to serogroup O157 with special reference to production of Vero cytotoxins VT1 and VT2. Epidemiol Infect. 1987 Dec;99(3):613–624. doi: 10.1017/s0950268800066462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Stein P. E., Boodhoo A., Tyrrell G. J., Brunton J. L., Read R. J. Crystal structure of the cell-binding B oligomer of verotoxin-1 from E. coli. Nature. 1992 Feb 20;355(6362):748–750. doi: 10.1038/355748a0. [DOI] [PubMed] [Google Scholar]
  27. Su G. F., Brahmbhatt H. N., Wehland J., Rohde M., Timmis K. N. Construction of stable LamB-Shiga toxin B subunit hybrids: analysis of expression in Salmonella typhimurium aroA strains and stimulation of B subunit-specific mucosal and serum antibody responses. Infect Immun. 1992 Aug;60(8):3345–3359. doi: 10.1128/iai.60.8.3345-3359.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Su G. F., Brahmbhatt H. N., de Lorenzo V., Wehland J., Timmis K. N. Extracellular export of Shiga toxin B-subunit/haemolysin A (C-terminus) fusion protein expressed in Salmonella typhimurium aroA-mutant and stimulation of B-subunit specific antibody responses in mice. Microb Pathog. 1992 Dec;13(6):465–476. doi: 10.1016/0882-4010(92)90013-e. [DOI] [PubMed] [Google Scholar]
  29. Tyrrell G. J., Ramotar K., Toye B., Boyd B., Lingwood C. A., Brunton J. L. Alteration of the carbohydrate binding specificity of verotoxins from Gal alpha 1-4Gal to GalNAc beta 1-3Gal alpha 1-4Gal and vice versa by site-directed mutagenesis of the binding subunit. Proc Natl Acad Sci U S A. 1992 Jan 15;89(2):524–528. doi: 10.1073/pnas.89.2.524. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES