Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1991 Mar;59(3):1065–1073. doi: 10.1128/iai.59.3.1065-1073.1991

Two copies of Shiga-like toxin II-related genes common in enterohemorrhagic Escherichia coli strains are responsible for the antigenic heterogeneity of the O157:H- strain E32511.

C K Schmitt 1, M L McKee 1, A D O'Brien 1
PMCID: PMC258368  PMID: 1997410

Abstract

Thirty-two clinical isolates of Shiga-like toxin (SLT)-producing Escherichia coli associated with single cases or outbreaks of bloody diarrhea, hemorrhagic colitis, the hemolytic uremic syndrome, or edema disease of swine were examined for multiple copies of genes belonging to the slt-I or slt-II toxin families. Five of 19 strains that were known to produce SLT-II or to hybridize to slt-II-specific probes by colony blot were found by Southern hybridization to contain two copies of toxin genes related to slt-II. The genes for two toxins closely related to slt-II were cloned from one of the isolates, Escherichia coli O157:H- strain E32511. One copy of the operon was found to be essentially identical to slt-II; it differed from slt-II by only one nucleotide base. This single nucleotide difference did not affect the predicted amino acid sequence. The predicted amino acid sequence of the A subunit of the second operon was identical to that of SLT-II, but the predicted amino acid sequence of the B subunit was identical to that of the B2F1 toxin VT2ha. We designated this second operon slt-IIc. Neutralization assays using several monoclonal antibodies and polyclonal antiserum prepared against SLT-II showed that SLT-IIc was antigenically related to but distinct from SLT-II.

Full text

PDF
1065

Images in this article

1067Image
on p.1067

Selected References

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

  1. Dickie N., Speirs J. I., Akhtar M., Johnson W. M., Szabo R. A. Purification of an Escherichia coli serogroup O157:H7 verotoxin and its detection in North American hemorrhagic colitis isolates. J Clin Microbiol. 1989 Sep;27(9):1973–1978. doi: 10.1128/jcm.27.9.1973-1978.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Downes F. P., Barrett T. J., Green J. H., Aloisio C. H., Spika J. S., Strockbine N. A., Wachsmuth I. K. Affinity purification and characterization of Shiga-like toxin II and production of toxin-specific monoclonal antibodies. Infect Immun. 1988 Aug;56(8):1926–1933. doi: 10.1128/iai.56.8.1926-1933.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Gannon V. P., Teerling C., Masri S. A., Gyles C. L. Molecular cloning and nucleotide sequence of another variant of the Escherichia coli Shiga-like toxin II family. J Gen Microbiol. 1990 Jun;136(6):1125–1135. doi: 10.1099/00221287-136-6-1125. [DOI] [PubMed] [Google Scholar]
  4. Gentry M. K., Dalrymple J. M. Quantitative microtiter cytotoxicity assay for Shigella toxin. J Clin Microbiol. 1980 Sep;12(3):361–366. doi: 10.1128/jcm.12.3.361-366.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Head S. C., Karmali M. A., Roscoe M. E., Petric M., Strockbine N. A., Wachsmuth I. K. Serological differences between verocytotoxin 2 and shiga-like toxin II. Lancet. 1988 Sep 24;2(8613):751–751. doi: 10.1016/s0140-6736(88)90228-0. [DOI] [PubMed] [Google Scholar]
  6. Hohn B., Collins J. A small cosmid for efficient cloning of large DNA fragments. Gene. 1980 Nov;11(3-4):291–298. doi: 10.1016/0378-1119(80)90069-4. [DOI] [PubMed] [Google Scholar]
  7. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  8. Ito H., Terai A., Kurazono H., Takeda Y., Nishibuchi M. Cloning and nucleotide sequencing of Vero toxin 2 variant genes from Escherichia coli O91:H21 isolated from a patient with the hemolytic uremic syndrome. Microb Pathog. 1990 Jan;8(1):47–60. doi: 10.1016/0882-4010(90)90007-d. [DOI] [PubMed] [Google Scholar]
  9. Johnson W. M., Lior H., Bezanson G. S. Cytotoxic Escherichia coli O157:H7 associated with haemorrhagic colitis in Canada. Lancet. 1983 Jan 1;1(8314-5):76–76. doi: 10.1016/s0140-6736(83)91616-1. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Karmali M. A., Steele B. T., Petric M., Lim C. Sporadic cases of haemolytic-uraemic syndrome associated with faecal cytotoxin and cytotoxin-producing Escherichia coli in stools. Lancet. 1983 Mar 19;1(8325):619–620. doi: 10.1016/s0140-6736(83)91795-6. [DOI] [PubMed] [Google Scholar]
  12. Keen N. T., Tamaki S., Kobayashi D., Trollinger D. Improved broad-host-range plasmids for DNA cloning in gram-negative bacteria. Gene. 1988 Oct 15;70(1):191–197. doi: 10.1016/0378-1119(88)90117-5. [DOI] [PubMed] [Google Scholar]
  13. Kleanthous H., Smith H. R., Scotland S. M., Gross R. J., Rowe B., Taylor C. M., Milford D. V. Haemolytic uraemic syndromes in the British Isles, 1985-8: association with verocytotoxin producing Escherichia coli. Part 2: Microbiological aspects. Arch Dis Child. 1990 Jul;65(7):722–727. doi: 10.1136/adc.65.7.722. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Konowalchuk J., Speirs J. I., Stavric S. Vero response to a cytotoxin of Escherichia coli. Infect Immun. 1977 Dec;18(3):775–779. doi: 10.1128/iai.18.3.775-779.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. MacLeod D. L., Gyles C. L. Purification and characterization of an Escherichia coli Shiga-like toxin II variant. Infect Immun. 1990 May;58(5):1232–1239. doi: 10.1128/iai.58.5.1232-1239.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Magnani J. L., Smith D. F., Ginsburg V. Detection of gangliosides that bind cholera toxin: direct binding of 125I-labeled toxin to thin-layer chromatograms. Anal Biochem. 1980 Dec;109(2):399–402. doi: 10.1016/0003-2697(80)90667-3. [DOI] [PubMed] [Google Scholar]
  17. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Marques L. R., Moore M. A., Wells J. G., Wachsmuth I. K., O'Brien A. D. Production of Shiga-like toxin by Escherichia coli. J Infect Dis. 1986 Aug;154(2):338–341. doi: 10.1093/infdis/154.2.338. [DOI] [PubMed] [Google Scholar]
  19. Milford D. V., Taylor C. M., Guttridge B., Hall S. M., Rowe B., Kleanthous H. Haemolytic uraemic syndromes in the British Isles 1985-8: association with verocytotoxin producing Escherichia coli. Part 1: Clinical and epidemiological aspects. Arch Dis Child. 1990 Jul;65(7):716–721. doi: 10.1136/adc.65.7.716. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Newland J. W., Strockbine N. A., Neill R. J. Cloning of genes for production of Escherichia coli Shiga-like toxin type II. Infect Immun. 1987 Nov;55(11):2675–2680. doi: 10.1128/iai.55.11.2675-2680.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  22. O'Brien A. D., Holmes R. K. Shiga and Shiga-like toxins. Microbiol Rev. 1987 Jun;51(2):206–220. doi: 10.1128/mr.51.2.206-220.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. O'Brien A. D., Laveck G. D. Immunochemical and cytotoxic activities of Shigella dysenteriae 1 (shiga) and shiga-like toxins. Infect Immun. 1982 Mar;35(3):1151–1154. doi: 10.1128/iai.35.3.1151-1154.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. O'Brien A. D., Lively T. A., Chang T. W., Gorbach S. L. Purification of Shigella dysenteriae 1 (Shiga)-like toxin from Escherichia coli O157:H7 strain associated with haemorrhagic colitis. Lancet. 1983 Sep 3;2(8349):573–573. doi: 10.1016/s0140-6736(83)90601-3. [DOI] [PubMed] [Google Scholar]
  26. O'Brien A. O., Lively T. A., Chen M. E., Rothman S. W., Formal S. B. Escherichia coli O157:H7 strains associated with haemorrhagic colitis in the United States produce a Shigella dysenteriae 1 (SHIGA) like cytotoxin. Lancet. 1983 Mar 26;1(8326 Pt 1):702–702. doi: 10.1016/s0140-6736(83)91987-6. [DOI] [PubMed] [Google Scholar]
  27. Oku Y., Yutsudo T., Hirayama T., O'Brien A. D., Takeda Y. Purification and some properties of a Vero toxin from a human strain of Escherichia coli that is immunologically related to Shiga-like toxin II (VT2). Microb Pathog. 1989 Feb;6(2):113–122. doi: 10.1016/0882-4010(89)90014-4. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Perera L. P., Marques L. R., O'Brien A. D. Isolation and characterization of monoclonal antibodies to Shiga-like toxin II of enterohemorrhagic Escherichia coli and use of the monoclonal antibodies in a colony enzyme-linked immunosorbent assay. J Clin Microbiol. 1988 Oct;26(10):2127–2131. doi: 10.1128/jcm.26.10.2127-2131.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rietra P. J., Willshaw G. A., Smith H. R., Field A. M., Scotland S. M., Rowe B. Comparison of Vero-cytotoxin-encoding phages from Escherichia coli of human and bovine origin. J Gen Microbiol. 1989 Aug;135(8):2307–2318. doi: 10.1099/00221287-135-8-2307. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Samuel J. E., Perera L. P., Ward S., O'Brien A. D., Ginsburg V., Krivan H. C. Comparison of the glycolipid receptor specificities of Shiga-like toxin type II and Shiga-like toxin type II variants. Infect Immun. 1990 Mar;58(3):611–618. doi: 10.1128/iai.58.3.611-618.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Schreier P. H., Cortese R. A fast and simple method for sequencing DNA cloned in the single-stranded bacteriophage M13. J Mol Biol. 1979 Mar 25;129(1):169–172. doi: 10.1016/0022-2836(79)90068-8. [DOI] [PubMed] [Google Scholar]
  35. Scotland S. M., Smith H. R., Rowe B. Two distinct toxins active on Vero cells from Escherichia coli O157. Lancet. 1985 Oct 19;2(8460):885–886. doi: 10.1016/s0140-6736(85)90146-1. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. Smith H. R., Day N. P., Scotland S. M., Gross R. J., Rowe B. Phage-determined production of vero cytotoxin in strains of Escherichia coli serogroup O157. Lancet. 1984 Jun 2;1(8388):1242–1243. doi: 10.1016/s0140-6736(84)91729-x. [DOI] [PubMed] [Google Scholar]
  38. Smith H. R., Scotland S. M. Vero cytotoxin-producing strains of Escherichia coli. J Med Microbiol. 1988 Jun;26(2):77–85. doi: 10.1099/00222615-26-2-77. [DOI] [PubMed] [Google Scholar]
  39. Strockbine N. A., Jackson M. P., Sung L. M., Holmes R. K., O'Brien A. D. Cloning and sequencing of the genes for Shiga toxin from Shigella dysenteriae type 1. J Bacteriol. 1988 Mar;170(3):1116–1122. doi: 10.1128/jb.170.3.1116-1122.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Strockbine N. A., Marques L. R., Holmes R. K., O'Brien A. D. Characterization of monoclonal antibodies against Shiga-like toxin from Escherichia coli. Infect Immun. 1985 Dec;50(3):695–700. doi: 10.1128/iai.50.3.695-700.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Strockbine N. A., Marques L. R., Newland J. W., Smith H. W., Holmes R. K., O'Brien A. D. Two toxin-converting phages from Escherichia coli O157:H7 strain 933 encode antigenically distinct toxins with similar biologic activities. Infect Immun. 1986 Jul;53(1):135–140. doi: 10.1128/iai.53.1.135-140.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Tarr P. I., Neill M. A., Clausen C. R., Newland J. W., Neill R. J., Moseley S. L. Genotypic variation in pathogenic Escherichia coli O157:H7 isolated from patients in Washington, 1984-1987. J Infect Dis. 1989 Feb;159(2):344–347. doi: 10.1093/infdis/159.2.344. [DOI] [PubMed] [Google Scholar]
  43. Taylor C. M., White R. H., Winterborn M. H., Rowe B. Haemolytic-uraemic syndrome: clinical experience of an outbreak in the West Midlands. Br Med J (Clin Res Ed) 1986 Jun 7;292(6534):1513–1516. doi: 10.1136/bmj.292.6534.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Wadolkowski E. A., Burris J. A., O'Brien A. D. Mouse model for colonization and disease caused by enterohemorrhagic Escherichia coli O157:H7. Infect Immun. 1990 Aug;58(8):2438–2445. doi: 10.1128/iai.58.8.2438-2445.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Wadolkowski E. A., Sung L. M., Burris J. A., Samuel J. E., O'Brien A. D. Acute renal tubular necrosis and death of mice orally infected with Escherichia coli strains that produce Shiga-like toxin type II. Infect Immun. 1990 Dec;58(12):3959–3965. doi: 10.1128/iai.58.12.3959-3965.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Weinstein D. L., Jackson M. P., Samuel J. E., Holmes R. K., O'Brien A. D. Cloning and sequencing of a Shiga-like toxin type II variant from Escherichia coli strain responsible for edema disease of swine. J Bacteriol. 1988 Sep;170(9):4223–4230. doi: 10.1128/jb.170.9.4223-4230.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Willshaw G. A., Smith H. R., Scotland S. M., Field A. M., Rowe B. Heterogeneity of Escherichia coli phages encoding Vero cytotoxins: comparison of cloned sequences determining VT1 and VT2 and development of specific gene probes. J Gen Microbiol. 1987 May;133(5):1309–1317. doi: 10.1099/00221287-133-5-1309. [DOI] [PubMed] [Google Scholar]
  48. Willshaw G. A., Smith H. R., Scotland S. M., Rowe B. Cloning of genes determining the production of vero cytotoxin by Escherichia coli. J Gen Microbiol. 1985 Nov;131(11):3047–3053. doi: 10.1099/00221287-131-11-3047. [DOI] [PubMed] [Google Scholar]
  49. Yutsudo T., Nakabayashi N., Hirayama T., Takeda Y. Purification and some properties of a Vero toxin from Escherichia coli O157:H7 that is immunologically unrelated to Shiga toxin. Microb Pathog. 1987 Jul;3(1):21–30. doi: 10.1016/0882-4010(87)90034-9. [DOI] [PubMed] [Google Scholar]

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

RESOURCES