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. 1995 Jun;95(6):2853–2861. doi: 10.1172/JCI117991

Shigella enterotoxin 1: an enterotoxin of Shigella flexneri 2a active in rabbit small intestine in vivo and in vitro.

A Fasano 1, F R Noriega 1, D R Maneval Jr 1, S Chanasongcram 1, R Russell 1, S Guandalini 1, M M Levine 1
PMCID: PMC295972  PMID: 7769126

Abstract

Culture filtrates of Shigella flexneri 2a strain M4243 grown in iron-depleted medium, caused significant fluid accumulation in rabbit ileal loops. Also, when tested in Ussing chambers, a greater rise in potential difference and short circuit current was seen with such filtrates compared with the medium control. Analogous filtrates from two M4243 derivatives lacking the 140-MD invasiveness plasmid (either M4243avir or BS103) retained 60-65% of the wild-type enterotoxic activity. Ultrafiltration and gel exclusion size fractionation of M4243 filtrate revealed that the activity was approximately 60 kD. SDS-PAGE performed on this fraction showed 18 bands, 5 of which reacted with human convalescent sera. Genes encoding this enterotoxin, named ShET1 for Shigella enterotoxin 1, were cloned from the S. flexneri 2a chromosome, and two separate open reading frames of 534 and 186 bp were sequenced. These observations suggest that S. flexneri 2a elaborates two distinct enterotoxins: ShET1, encoded by genes located on the chromosome, and ShET2, encoded by a gene on the 140-MD invasiveness plasmid. ShET1, which is composed of two distinct subunits and is elaborated in vivo, where it elicits an immune response, may be important in the pathogenesis of diarrheal illness due to S. flexneri 2a.

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Selected References

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

  1. Andrews G. P., Hromockyj A. E., Coker C., Maurelli A. T. Two novel virulence loci, mxiA and mxiB, in Shigella flexneri 2a facilitate excretion of invasion plasmid antigens. Infect Immun. 1991 Jun;59(6):1997–2005. doi: 10.1128/iai.59.6.1997-2005.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  3. Clements J. D., Flint D. C., Klipstein F. A. Immunological and physicochemical characterization of heat-labile enterotoxins isolated from two strains of Escherichia coli. Infect Immun. 1982 Nov;38(2):806–809. doi: 10.1128/iai.38.2.806-809.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DuPont H. L., Hornick R. B., Dawkins A. T., Snyder M. J., Formal S. B. The response of man to virulent Shigella flexneri 2a. J Infect Dis. 1969 Mar;119(3):296–299. doi: 10.1093/infdis/119.3.296. [DOI] [PubMed] [Google Scholar]
  5. Fasano A., Kay B. A., Russell R. G., Maneval D. R., Jr, Levin M. M. Enterotoxin and cytotoxin production by enteroinvasive Escherichia coli. Infect Immun. 1990 Nov;58(11):3717–3723. doi: 10.1128/iai.58.11.3717-3723.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fontaine A., Arondel J., Sansonetti P. J. Role of Shiga toxin in the pathogenesis of bacillary dysentery, studied by using a Tox- mutant of Shigella dysenteriae 1. Infect Immun. 1988 Dec;56(12):3099–3109. doi: 10.1128/iai.56.12.3099-3109.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Guandalini S., Fasano A., Migliavacca M., Marchesano G., Ferola A., Rubino A. Effects of berberine on basal and secretagogue-modified ion transport in the rabbit ileum in vitro. J Pediatr Gastroenterol Nutr. 1987 Nov-Dec;6(6):953–960. doi: 10.1097/00005176-198711000-00023. [DOI] [PubMed] [Google Scholar]
  9. Hale T. L., Sansonetti P. J., Schad P. A., Austin S., Formal S. B. Characterization of virulence plasmids and plasmid-associated outer membrane proteins in Shigella flexneri, Shigella sonnei, and Escherichia coli. Infect Immun. 1983 Apr;40(1):340–350. doi: 10.1128/iai.40.1.340-350.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Keusch G. T., Donohue-Rolfe A., Jacewicz M. Shigella toxin(s): description and role in diarrhea and dysentery. Pharmacol Ther. 1981;15(3):403–438. doi: 10.1016/0163-7258(81)90052-8. [DOI] [PubMed] [Google Scholar]
  11. Kinsey M. D., Formal S. B., Dammin G. J., Giannella R. A. Fluid and electrolyte transport in rhesus monkeys challenged intracecally with Shigella flexneri 2a. Infect Immun. 1976 Aug;14(2):368–371. doi: 10.1128/iai.14.2.368-371.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kotloff K. L., Herrington D. A., Hale T. L., Newland J. W., Van De Verg L., Cogan J. P., Snoy P. J., Sadoff J. C., Formal S. B., Levine M. M. Safety, immunogenicity, and efficacy in monkeys and humans of invasive Escherichia coli K-12 hybrid vaccine candidates expressing Shigella flexneri 2a somatic antigen. Infect Immun. 1992 Jun;60(6):2218–2224. doi: 10.1128/iai.60.6.2218-2224.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kétyi I., Málovics I., Vertényi A., Kontrohr T., Pácsa S., Kuch B. Heat-stable enterotoxin produced by Shigella flexneri. Acta Microbiol Acad Sci Hung. 1978;25(3):165–171. [PubMed] [Google Scholar]
  14. Kétyi I., Vertényi A., Málovics I., Kontrohr T., Pácsa S. Unique features of heat-stable enterotoxin of Shigella flexneri. Acta Microbiol Acad Sci Hung. 1978;25(3):219–227. [PubMed] [Google Scholar]
  15. Kétyi I., Vertényi A., Pácsa S., Kocsis B. Enterotoxin production by Shigella flexneri type 2A, strain no. M42-43. Acta Microbiol Acad Sci Hung. 1978;25(4):319–325. [PubMed] [Google Scholar]
  16. 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]
  17. Levine M. M., Bergquist E. J., Nalin D. R., Waterman D. H., Hornick R. B., Young C. R., Sotman S. Escherichia coli strains that cause diarrhoea but do not produce heat-labile or heat-stable enterotoxins and are non-invasive. Lancet. 1978 May 27;1(8074):1119–1122. doi: 10.1016/s0140-6736(78)90299-4. [DOI] [PubMed] [Google Scholar]
  18. Levine M. M. Escherichia coli that cause diarrhea: enterotoxigenic, enteropathogenic, enteroinvasive, enterohemorrhagic, and enteroadherent. J Infect Dis. 1987 Mar;155(3):377–389. doi: 10.1093/infdis/155.3.377. [DOI] [PubMed] [Google Scholar]
  19. Levine M. M., Kaper J. B., Lockman H., Black R. E., Clements M. L., Falkow S. Recombinant DNA risk assessment studies in humans: efficacy of poorly mobilizable plasmids in biologic containment. J Infect Dis. 1983 Oct;148(4):699–709. doi: 10.1093/infdis/148.4.699. [DOI] [PubMed] [Google Scholar]
  20. Lospalluto J. J., Finkelstein R. A. Chemical and physical properties of cholera exo-enterotoxin (choleragen) and its spontaneously formed toxoid (choleragenoid). Biochim Biophys Acta. 1972 Jan 26;257(1):158–166. doi: 10.1016/0005-2795(72)90265-6. [DOI] [PubMed] [Google Scholar]
  21. Makino S., Sasakawa C., Yoshikawa M. Genetic relatedness of the basic replicon of the virulence plasmid in shigellae and enteroinvasive Escherichia coli. Microb Pathog. 1988 Oct;5(4):267–274. doi: 10.1016/0882-4010(88)90099-x. [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., Thompson M. R., Formal S. B. Production of Shigella dysenteriae type 1-like cytotoxin by Escherichia coli. J Infect Dis. 1982 Dec;146(6):763–769. doi: 10.1093/infdis/146.6.763. [DOI] [PubMed] [Google Scholar]
  24. O'Brien A. D., Thompson M. R., Gemski P., Doctor B. P., Formal S. B. Biological properties of Shigella flexneri 2A toxin and its serological relationship to Shigella dysenteriae 1 toxin. Infect Immun. 1977 Mar;15(3):796–798. doi: 10.1128/iai.15.3.796-798.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Olsnes S., Reisbig R., Eiklid K. Subunit structure of Shigella cytotoxin. J Biol Chem. 1981 Aug 25;256(16):8732–8738. [PubMed] [Google Scholar]
  26. Pál T., Formal S. B., Hale T. L. Characterization of virulence marker antigen of Shigella spp. and enteroinvasive Escherichia coli. J Clin Microbiol. 1989 Mar;27(3):561–563. doi: 10.1128/jcm.27.3.561-563.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rogers H. J. Iron-Binding Catechols and Virulence in Escherichia coli. Infect Immun. 1973 Mar;7(3):445–456. doi: 10.1128/iai.7.3.445-456.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rout W. R., Formal S. B., Giannella R. A., Dammin G. J. Pathophysiology of Shigella diarrhea in the rhesus monkey: intestinal transport, morphological, and bacteriological studies. Gastroenterology. 1975 Feb;68(2):270–278. [PubMed] [Google Scholar]
  29. Sansonetti P. J., Kopecko D. J., Formal S. B. Involvement of a plasmid in the invasive ability of Shigella flexneri. Infect Immun. 1982 Mar;35(3):852–860. doi: 10.1128/iai.35.3.852-860.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Seidah N. G., Donohue-Rolfe A., Lazure C., Auclair F., Keusch G. T., Chrétien M. Complete amino acid sequence of Shigella toxin B-chain. A novel polypeptide containing 69 amino acids and one disulfide bridge. J Biol Chem. 1986 Oct 25;261(30):13928–13931. [PubMed] [Google Scholar]
  31. Stoll B. J., Glass R. I., Huq M. I., Khan M. U., Banu H., Holt J. Epidemiologic and clinical features of patients infected with Shigella who attended a diarrheal disease hospital in Bangladesh. J Infect Dis. 1982 Aug;146(2):177–183. doi: 10.1093/infdis/146.2.177. [DOI] [PubMed] [Google Scholar]
  32. Taylor D. N., Echeverria P., Pál T., Sethabutr O., Saiborisuth S., Sricharmorn S., Rowe B., Cross J. The role of Shigella spp., enteroinvasive Escherichia coli, and other enteropathogens as causes of childhood dysentery in Thailand. J Infect Dis. 1986 Jun;153(6):1132–1138. doi: 10.1093/infdis/153.6.1132. [DOI] [PubMed] [Google Scholar]
  33. Taylor D. N., Echeverria P., Sethabutr O., Pitarangsi C., Leksomboon U., Blacklow N. R., Rowe B., Gross R., Cross J. Clinical and microbiologic features of Shigella and enteroinvasive Escherichia coli infections detected by DNA hybridization. J Clin Microbiol. 1988 Jul;26(7):1362–1366. doi: 10.1128/jcm.26.7.1362-1366.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Venkatesan M. M., Buysse J. M., Kopecko D. J. Characterization of invasion plasmid antigen genes (ipaBCD) from Shigella flexneri. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9317–9321. doi: 10.1073/pnas.85.23.9317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Vial P. A., Robins-Browne R., Lior H., Prado V., Kaper J. B., Nataro J. P., Maneval D., Elsayed A., Levine M. M. Characterization of enteroadherent-aggregative Escherichia coli, a putative agent of diarrheal disease. J Infect Dis. 1988 Jul;158(1):70–79. doi: 10.1093/infdis/158.1.70. [DOI] [PubMed] [Google Scholar]

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