Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1981 Aug;33(2):473–476. doi: 10.1128/iai.33.2.473-476.1981

Vascular permeability activity in Escherichia coli heat-stable enterotoxin.

J P Craig, K Yamamoto, T Takeda, Y Takeda, T Miwatani
PMCID: PMC350721  PMID: 7196884

Abstract

Purified heat-stable enterotoxin (ST) prepared from supernatants of Escherichia coli strain 53402 A-1, isolated from a patient with diarrhea, caused an increase in the permeability of the small blood vessels of the skin of adult rabbits after intracutaneous injection. Increased permeability was manifested by localized accumulation of intravenously injected blue dye at the injection sites. Permeability factor (PF) activity reached a peak 1 h after injection, and recovery of normal permeability was nearly complete in 3 h. Residual PF activity had disappeared by 24 h. Dose-response curves demonstrated a straight-line relationship between the logarithm of the dose and mean blueing diameter over a range of 3 to 12 mm. The PF assay was less sensitive that the suckling mouse assay, and adult rabbits varied in their sensitivity to the PF effect. Five to 100 mouse units of ST were required to consistently evoke strong and unequivocal blue lesions of 7 mm or more in diameter. PF activity was reduced 50 to 70% by heating at 70 degrees C for 30 min, 60 to 80% by boiling for 30 min, and 95 to 98% by autoclaving for 15 min, but it was unaffected by treatment with cholera antitoxin. These findings are consistent with the notion that PF activity is a property of the ST molecule itself. This PF assay cannot be used for the detection of ST in crude culture supernatant fluid because of low sensitivity and the presence of nonspecific PF effects of culture media and other E. coli products.

Full text

PDF
473

Images in this article

Selected References

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

  1. Alderete J. F., Robertson D. C. Purification and chemical characterization of the heat-stable enterotoxin produced by porcine strains of enterotoxigenic Escherichia coli. Infect Immun. 1978 Mar;19(3):1021–1030. doi: 10.1128/iai.19.3.1021-1030.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Craig J. B. Toward the development of a standard reference cholera antitoxin. Dev Biol Stand. 1978;41:415–422. [PubMed] [Google Scholar]
  3. Craig J. P. A permeability factor (toxin) found in cholera stools and culture filtrates and its neutralization by convalescent cholera sera. Nature. 1965 Aug 7;207(997):614–616. doi: 10.1038/207614a0. [DOI] [PubMed] [Google Scholar]
  4. Dean A. G., Ching Y. C., Williams R. G., Harden L. B. Test for Escherichia coli enterotoxin using infant mice: application in a study of diarrhea in children in Honolulu. J Infect Dis. 1972 Apr;125(4):407–411. doi: 10.1093/infdis/125.4.407. [DOI] [PubMed] [Google Scholar]
  5. Donta S. T., Moon H. W., Whipp S. C. Detection of heat-labile Escherichia coli enterotoxin with the use of adrenal cells in tissue culture. Science. 1974 Jan 25;183(4122):334–336. doi: 10.1126/science.183.4122.334. [DOI] [PubMed] [Google Scholar]
  6. Evans D. J., Jr, Chen L. C., Curlin G. T., Evans D. G. Stimulation of adenyl cyclase by Escherichia coli enterotoxin. Nat New Biol. 1972 Apr 5;236(66):137–138. doi: 10.1038/newbio236137a0. [DOI] [PubMed] [Google Scholar]
  7. Evans D. J., Jr, Evans D. G., Gorbach S. L. Production of vascular permeability factor by enterotoxigenic Escherichia coli isolated from man. Infect Immun. 1973 Nov;8(5):725–730. doi: 10.1128/iai.8.5.725-730.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Field M., Graf L. H., Jr, Laird W. J., Smith P. L. Heat-stable enterotoxin of Escherichia coli: in vitro effects on guanylate cyclase activity, cyclic GMP concentration, and ion transport in small intestine. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2800–2804. doi: 10.1073/pnas.75.6.2800. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Guerrant R. L., Brunton L. L., Schnaitman T. C., Rebhun L. I., Gilman A. G. Cyclic adenosine monophosphate and alteration of Chinese hamster ovary cell morphology: a rapid, sensitive in vitro assay for the enterotoxins of Vibrio cholerae and Escherichia coli. Infect Immun. 1974 Aug;10(2):320–327. doi: 10.1128/iai.10.2.320-327.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Moss J., Richardson S. H. Activation of adenylate cyclase by heat-labile Escherichia coli enterotoxin. Evidence for ADP-ribosyltransferase activity similar to that of choleragen. J Clin Invest. 1978 Aug;62(2):281–285. doi: 10.1172/JCI109127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mundell D. H., Anselmo C. R., Wishnow R. M. Factors influencing heat-labile Escherichia coli enterotoxin activity. Infect Immun. 1976 Aug;14(2):383–388. doi: 10.1128/iai.14.2.383-388.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Sack D. A., Sack R. B. Test for enterotoxigenic Escherichia coli using Y-1 adrenal cells in miniculture. Infect Immun. 1975 Feb;11(2):334–336. doi: 10.1128/iai.11.2.334-336.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Staples S. J., Asher S. E., Giannella R. A. Purification and characterization of heat-stable enterotoxin produced by a strain of E. coli pathogenic for man. J Biol Chem. 1980 May 25;255(10):4716–4721. [PubMed] [Google Scholar]
  14. Takeda Y., Takeda T., Yano T., Yamamoto K., Miwatani T. Purification and partial characterization of heat-stable enterotoxin of enterotoxigenic Escherichia coli. Infect Immun. 1979 Sep;25(3):978–985. doi: 10.1128/iai.25.3.978-985.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES