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. 1993 Sep;92(3):1412–1417. doi: 10.1172/JCI116717

Role of the eaeA gene in experimental enteropathogenic Escherichia coli infection.

M S Donnenberg 1, C O Tacket 1, S P James 1, G Losonsky 1, J P Nataro 1, S S Wasserman 1, J B Kaper 1, M M Levine 1
PMCID: PMC288285  PMID: 8376594

Abstract

Enteropathogenic Escherichia coli (EPEC) infections are a leading cause of infant diarrhea in developing countries. Recently eaeA, a gene necessary for the characteristic intimate attachment of EPEC to epithelial cells in tissue culture, was described. We conducted a randomized, double-blind study to determine the role of the eaeA gene in human EPEC infection. 11 adult volunteers ingested 2 x 10(10) colony-forming units of O127:H6 EPEC strain E2348/69, and an equal number received the same dose of an isogenic eaeA deletion mutant constructed from E2348/69. Volunteers were monitored for the development of diarrhea, fever, and systemic and gastrointestinal complaints. Diarrhea developed in all 11 volunteers who received E2348/69 and in 4 of 11 who received the mutant (P = 0.002). Fever was more common in recipients of the wild-type strain (P = 0.024). Stool volumes were lower in recipients of the mutant. All volunteers seroconverted to E2348/69 LPS, but the geometric mean peak titers of serum IgG and IgA in recipients of the mutant were lower than those of recipients of the wild-type strain. IgA against LPS was detected in the jejunal fluid of six of six recipients of E2348/69 and 5/6 recipients of the mutant. This study unambiguously assigns a role for eaeA as an EPEC virulence gene, but the residual diarrhea seen in recipients of the mutant indicates that other factors are involved.

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Selected References

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  1. Baldini M. M., Kaper J. B., Levine M. M., Candy D. C., Moon H. W. Plasmid-mediated adhesion in enteropathogenic Escherichia coli. J Pediatr Gastroenterol Nutr. 1983;2(3):534–538. doi: 10.1097/00005176-198302030-00023. [DOI] [PubMed] [Google Scholar]
  2. Baldwin T. J., Ward W., Aitken A., Knutton S., Williams P. H. Elevation of intracellular free calcium levels in HEp-2 cells infected with enteropathogenic Escherichia coli. Infect Immun. 1991 May;59(5):1599–1604. doi: 10.1128/iai.59.5.1599-1604.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bolton J. E., Field M. Ca ionophore-stimulated ion secretion in rabbit ileal mucosa: relation to actions of cyclic 3',5'-AMP and carbamylcholine. J Membr Biol. 1977 Jun 30;35(2):159–173. doi: 10.1007/BF01869947. [DOI] [PubMed] [Google Scholar]
  4. Cobeljić M., Mel D., Arsić B., Krstić L., Sokolovski B., Nikolovski B., Sopovski E., Kulauzov M., Kalenić S. The association of enterotoxigenic and enteropathogenic Escherichia coli and other enteric pathogens with childhood diarrhoea in Yugoslavia. Epidemiol Infect. 1989 Aug;103(1):53–62. doi: 10.1017/s0950268800030351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Donnenberg M. S., Calderwood S. B., Donohue-Rolfe A., Keusch G. T., Kaper J. B. Construction and analysis of TnphoA mutants of enteropathogenic Escherichia coli unable to invade HEp-2 cells. Infect Immun. 1990 Jun;58(6):1565–1571. doi: 10.1128/iai.58.6.1565-1571.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Donnenberg M. S., Girón J. A., Nataro J. P., Kaper J. B. A plasmid-encoded type IV fimbrial gene of enteropathogenic Escherichia coli associated with localized adherence. Mol Microbiol. 1992 Nov;6(22):3427–3437. doi: 10.1111/j.1365-2958.1992.tb02210.x. [DOI] [PubMed] [Google Scholar]
  7. Donnenberg M. S., Kaper J. B. Construction of an eae deletion mutant of enteropathogenic Escherichia coli by using a positive-selection suicide vector. Infect Immun. 1991 Dec;59(12):4310–4317. doi: 10.1128/iai.59.12.4310-4317.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Donnenberg M. S., Kaper J. B. Enteropathogenic Escherichia coli. Infect Immun. 1992 Oct;60(10):3953–3961. doi: 10.1128/iai.60.10.3953-3961.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Finlay B. B., Rosenshine I., Donnenberg M. S., Kaper J. B. Cytoskeletal composition of attaching and effacing lesions associated with enteropathogenic Escherichia coli adherence to HeLa cells. Infect Immun. 1992 Jun;60(6):2541–2543. doi: 10.1128/iai.60.6.2541-2543.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Francis C. L., Jerse A. E., Kaper J. B., Falkow S. Characterization of interactions of enteropathogenic Escherichia coli O127:H6 with mammalian cells in vitro. J Infect Dis. 1991 Oct;164(4):693–703. doi: 10.1093/infdis/164.4.693. [DOI] [PubMed] [Google Scholar]
  11. Girón J. A., Ho A. S., Schoolnik G. K. An inducible bundle-forming pilus of enteropathogenic Escherichia coli. Science. 1991 Nov 1;254(5032):710–713. doi: 10.1126/science.1683004. [DOI] [PubMed] [Google Scholar]
  12. Gomes T. A., Vieira M. A., Wachsmuth I. K., Blake P. A., Trabulsi L. R. Serotype-specific prevalence of Escherichia coli strains with EPEC adherence factor genes in infants with and without diarrhea in São Paulo, Brazil. J Infect Dis. 1989 Jul;160(1):131–135. doi: 10.1093/infdis/160.1.131. [DOI] [PubMed] [Google Scholar]
  13. Gurwith M., Hinde D., Gross R., Rowe B. A prospective study of enteropathogenic Escherichia coli in endemic diarrheal disease. J Infect Dis. 1978 Mar;137(3):292–297. doi: 10.1093/infdis/137.3.292. [DOI] [PubMed] [Google Scholar]
  14. Isberg R. R., Leong J. M. Multiple beta 1 chain integrins are receptors for invasin, a protein that promotes bacterial penetration into mammalian cells. Cell. 1990 Mar 9;60(5):861–871. doi: 10.1016/0092-8674(90)90099-z. [DOI] [PubMed] [Google Scholar]
  15. Jerse A. E., Kaper J. B. The eae gene of enteropathogenic Escherichia coli encodes a 94-kilodalton membrane protein, the expression of which is influenced by the EAF plasmid. Infect Immun. 1991 Dec;59(12):4302–4309. doi: 10.1128/iai.59.12.4302-4309.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jerse A. E., Yu J., Tall B. D., Kaper J. B. A genetic locus of enteropathogenic Escherichia coli necessary for the production of attaching and effacing lesions on tissue culture cells. Proc Natl Acad Sci U S A. 1990 Oct;87(20):7839–7843. doi: 10.1073/pnas.87.20.7839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kain K. C., Barteluk R. L., Kelly M. T., He X., de Hua G., Ge Y. A., Proctor E. M., Byrne S., Stiver H. G. Etiology of childhood diarrhea in Beijing, China. J Clin Microbiol. 1991 Jan;29(1):90–95. doi: 10.1128/jcm.29.1.90-95.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Knutton S., Baldwin T., Williams P. H., McNeish A. S. Actin accumulation at sites of bacterial adhesion to tissue culture cells: basis of a new diagnostic test for enteropathogenic and enterohemorrhagic Escherichia coli. Infect Immun. 1989 Apr;57(4):1290–1298. doi: 10.1128/iai.57.4.1290-1298.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Levine M. M., Nataro J. P., Karch H., Baldini M. M., Kaper J. B., Black R. E., Clements M. L., O'Brien A. D. The diarrheal response of humans to some classic serotypes of enteropathogenic Escherichia coli is dependent on a plasmid encoding an enteroadhesiveness factor. J Infect Dis. 1985 Sep;152(3):550–559. doi: 10.1093/infdis/152.3.550. [DOI] [PubMed] [Google Scholar]
  21. Long-Krug S. A., Weikel C. S., Tiemens K. T., Hewlett E. L., Levine M. M., Guerrant R. L. Does enteropathogenic Escherichia coli produce heat-labile enterotoxin, heat-stable enterotoxins a or b, or cholera toxin A subunits? Infect Immun. 1984 Nov;46(2):612–614. doi: 10.1128/iai.46.2.612-614.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Manjarrez-Hernandez H. A., Amess B., Sellers L., Baldwin T. J., Knutton S., Williams P. H., Aitken A. Purification of a 20 kDa phosphoprotein from epithelial cells and identification as a myosin light chain. Phosphorylation induced by enteropathogenic Escherichia coli and phorbol ester. FEBS Lett. 1991 Nov 4;292(1-2):121–127. doi: 10.1016/0014-5793(91)80848-w. [DOI] [PubMed] [Google Scholar]
  23. Moon H. W., Whipp S. C., Argenzio R. A., Levine M. M., Giannella R. A. Attaching and effacing activities of rabbit and human enteropathogenic Escherichia coli in pig and rabbit intestines. Infect Immun. 1983 Sep;41(3):1340–1351. doi: 10.1128/iai.41.3.1340-1351.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nataro J. P., Baldini M. M., Kaper J. B., Black R. E., Bravo N., Levine M. M. Detection of an adherence factor of enteropathogenic Escherichia coli with a DNA probe. J Infect Dis. 1985 Sep;152(3):560–565. doi: 10.1093/infdis/152.3.560. [DOI] [PubMed] [Google Scholar]
  25. Robins-Browne R. M., Still C. S., Miliotis M. D., Richardson N. J., Koornhof H. J., Freiman I., Schoub B. D., Lecatsas G., Hartman E. Summer diarrhoea in African infants and children. Arch Dis Child. 1980 Dec;55(12):923–928. doi: 10.1136/adc.55.12.923. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rosenshine I., Donnenberg M. S., Kaper J. B., Finlay B. B. Signal transduction between enteropathogenic Escherichia coli (EPEC) and epithelial cells: EPEC induces tyrosine phosphorylation of host cell proteins to initiate cytoskeletal rearrangement and bacterial uptake. EMBO J. 1992 Oct;11(10):3551–3560. doi: 10.1002/j.1460-2075.1992.tb05438.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rothbaum R., McAdams A. J., Giannella R., Partin J. C. A clinicopathologic study of enterocyte-adherent Escherichia coli: a cause of protracted diarrhea in infants. Gastroenterology. 1982 Aug;83(2):441–454. [PubMed] [Google Scholar]
  28. Tacket C. O., Moseley S. L., Kay B., Losonsky G., Levine M. M. Challenge studies in volunteers using Escherichia coli strains with diffuse adherence to HEp-2 cells. J Infect Dis. 1990 Aug;162(2):550–552. doi: 10.1093/infdis/162.2.550. [DOI] [PubMed] [Google Scholar]
  29. Ulshen M. H., Rollo J. L. Pathogenesis of escherichia coli gastroenteritis in man--another mechanism. N Engl J Med. 1980 Jan 10;302(2):99–101. doi: 10.1056/NEJM198001103020207. [DOI] [PubMed] [Google Scholar]

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