E. coli pp 275-289 | Cite as

Protocols to Study Effects of Shiga Toxin on Mononuclear Leukocytes

  • Christian Menge
Part of the Methods in Molecular Medicine™ book series (MIMM, volume 73)


Endothelial cells are regarded as the main targets of the Shiga toxins (Stxs) during infections caused by Stx-producing Escherichia coli (STEC). However, several investigations also confirmed an effect of these toxins on immune cell functions in species naturally infected with STEC. Human B-cell lines (1) and tonsillar B-cells (2) are highly susceptible to the cytotoxic activity of Stx1, which also hampers activation and proliferation of bovine B- and T- cell subpopulations in vitro (3). Although Stxs appear to be immunosuppressive, they do not prevent the development of a specific antibody response in STECinfected individuals (4, 5, 6). Thus, the question of an immunosuppressive effect of Stx in the pathogenesis of STEC-mediated diseases needs to be addressed. STEC infections lead to an immunocompromised condition in gnotobiotic pigs and calves (7,8), which is assumed to contribute to the observed persistency of infection (e.g., in calves and humans) (9,10).


  1. 1.
    Mangeney M., Lingwood C. A., Taga S., Caillou B., Tursz T., and Wiels J. (1993) Apoptosis induced in Burkitt’s lymphoma cells via Gb3/CD77, a glycolipid antigen. Cancer Res. 53, 5314–5319.PubMedGoogle Scholar
  2. 2.
    Cohen A., Madrid-Marina V., Estrov Z., Freedman M. H., Lingwood C. A., and Dosch H. M. (1990) Expression of glycolipid receptors to Shiga-like toxin on human B lymphocytes: a mechanism for the failure of long-lived antibody response to dysenteric disease. Int. Immunol. 2, 1–8.CrossRefPubMedGoogle Scholar
  3. 3.
    Menge C., Wieler L. H., Schlapp T., and Baljer G. (1999) Shiga toxin 1 from Escherichia coli blocks activation and proliferation of bovine lymphocyte subpopulations in vitro. Infect. Immun. 67, 2209–2217.PubMedGoogle Scholar
  4. 4.
    Pirro F., Wieler L. H., Failing K., Bauerfeind R., and Baljer G. (1995) Neutralizing antibodies against Shiga-like toxins from Escherichia coli in colostra and sera of cattle. Vet. Microbiol. 43, 131–141.CrossRefPubMedGoogle Scholar
  5. 5.
    Wieler L. H., Franke S., Menge C., Rose M., Bauerfeind R., Karch H., et al. (1995) Investigations on the immunoresponse during edema disease of piglets after weaning by using a recombinant B subunit of Shiga-like-toxin IIe. Dtsch. Tierarztl. Wochenschr. 102, 40–43.PubMedGoogle Scholar
  6. 6.
    Reymond D., Johnson R. P., Karmali M. A., Petric M., Winkler M., Johnson S., et al. (1996) Neutralizing antibodies to Escherichia coli Vero cytotoxin 1 and antibodies to O157 lipopolysacccharide in healthy farm family members and urban residents. J. Clin. Microbiol. 34, 2053–2057.PubMedGoogle Scholar
  7. 7.
    Christopher-Hennings J., Willgohs J. A., Francis D. H., Raman U. A. K., Moxley R. A., and Hurley D. J. (1993) Immunocompromise in gnotobiotic pigs induced by Verotoxin-producing Escherichia coli (O111:NM). Infect. Immun. 61, 2304–2308.PubMedGoogle Scholar
  8. 8.
    Hoffman M., Casey T., and Bosworth B. (1997) Bovine immune response to Escherichia coli O157, Abstracts of the 3rd International Symposium and Workshop on Shiga Toxin (Verocytotoxin)-producing Escherichia coli infections, p. 117.Google Scholar
  9. 9.
    Cray W. C. and Moon H. W. (1995) Experimental infection of calves and adult cattle with Escherichia coli O157:H7. Appl. Environ. Microbiol. 61, 1586–1590.PubMedGoogle Scholar
  10. 10.
    Karch H., Rüssmann H., Schmidt H., Schwarzkopf A., and Heesemann J. (1995) Long-term shedding and clonal turnover of enterohemorrhagic Escherichia coli O157 in diarrheal diseases. J. Clin. Microbiol. 33, 1602–1605.PubMedGoogle Scholar
  11. 11.
    Klapproth J. M., Scaletsky I. C. A., McNamara B. P., Lai L.-C., Malstrom C., James S. P., et al. (2000) A large toxin from pathogenic Escherichia coli strains that inhibits lymphocyte activation. Infect. Immun. 68, 2148–2155.CrossRefPubMedGoogle Scholar
  12. 12.
    Makino K., Ishii K., Yasunaga T., Hattori M., Yokoyama K., Yutsudo C. H., et al. (1998) Complete nucleotide sequences of 93-kb and 3.3-kb plasmids of an enterohemorrhagic Escherichia coli O157:H7 derived from Sakai outbreak. DNA Res. 5, 1–9.CrossRefPubMedGoogle Scholar
  13. 13.
    Burland V., Shao Y., Perna N. T., Plunkett G., Sofia H. J., and Blattner F. R. (1998) The complete DNA sequence and analysis of the large virulence plasmid of Escherichia coli O157:H7. Nucleic Acids Res. 26, 4196–4204.CrossRefPubMedGoogle Scholar
  14. 14.
    Nicholls L., Grant T. H., and Robins-Browne R. M. (2000) Identification of a novel genetic locus that is required for in vitro adhesion of a clinical isolate of enterohaemorrhagic Escherichia coli to epithelial cells. Mol. Microbiol. 35, 275–288.CrossRefPubMedGoogle Scholar
  15. 15.
    Higgins L. M., Frankel G., Connerton I., Goncalves N. S., Dougan G., and MacDonald T. T. (1999) Role of bacterial intimin in colonic hyperplasia and inflammation. Science 285, 588–591.CrossRefPubMedGoogle Scholar
  16. 16.
    Tada H., Shiho O., Kuroshima K., Koyama M., and Tsukamoto K. (1986) An improved colorimetric assay for Interleukin 2. J. Immunol. Methods 93, 157–165.CrossRefPubMedGoogle Scholar
  17. 17.
    Jacewicz M., Clausen H., Nudelman E., Donohue-Rolfe A., and Keusch G. T. (1986) Pathogenesis of Shigella diarrhea XI: Isolation of a Shigella toxin-binding glycolipid from rabbit jejunum and HeLa-cells and its identification as globotriaosylceramide. J. Exp. Med. 163, 1391–1404.CrossRefPubMedGoogle Scholar
  18. 18.
    DeGrandis S., Law H., Brunton J., Gyles C., and Lingwood C. A. (1989) Globotetraosylceramide is recognized by the pig edema disease toxin. J. Biol. Chem. 264, 12,520–12,525.PubMedGoogle Scholar
  19. 19.
    Wiels J., Fellous M., and Tursz T. (1981) Monoclonal antibody against a Burkitt lymphoma-associated antigen. Proc. Natl. Acad. Sci. USA 78, 6485–6488.CrossRefPubMedGoogle Scholar
  20. 20.
    Nicoletti I., Migliorati G., Pagliacci M.C., Grignani F., and Riccardi C. (1991) A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J. Immunol. Methods 139, 271–279.CrossRefPubMedGoogle Scholar

Copyright information

© Humana Press Inc. 2003

Authors and Affiliations

  • Christian Menge
    • 1
  1. 1.Institut f&#00FC;r Hygiene und Infektionskrankheiten der TiereJustus-Liebig-Universit&#00E4t GiessenGiessenGermany

Personalised recommendations