A new biological agent for treatment of Shiga toxigenic Escherichia coli infections and dysentery in humans


Gastrointestinal disease caused by Shiga toxin-producing bacteria (such as Escherichia coli O157:H7 and Shigella dysenteriae) is often complicated by life-threatening toxin-induced systemic sequelae, including hemolytic–uremic syndrome. Such infections can now be diagnosed very early in the course of the disease, but at present no effective therapeutic intervention is possible. Here, we constructed a recombinant bacterium that displayed a Shiga toxin receptor mimic on its surface, and it adsorbed and neutralized Shiga toxins with very high efficiency. Moreover, oral administration of the recombinant bacterium completely protected mice from challenge with an otherwise 100%-fatal dose of Shiga toxigenic E. coli. Thus, the bacterium shows great promise as a ‘probiotic’ treatment for Shiga toxigenic E. coli infections and dysentery.

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Figure 1: Construction of pJCP-Gb3.
Figure 2: Proposed structure of the LPS of E. coli CWG308:pJCP-Gb3.
Figure 3: Total Stx adsorption/neutralization capacity of E. coli CWG308:pJCP-Gb3.
Figure 4: Immunofluorescent staining.
Figure 5: Survival of mice challenged with virulent STEC.


  1. 1

    O'Brien, A.D. & Holmes, R.K. Shiga and shiga-like toxins. Microbiol. Rev. 51, 206–220 (1987).

    CAS  Google Scholar 

  2. 2

    Karmali, M.A. Infection by verocytotoxin-producing E. coli. Clin. Microbiol. Rev. 2, 15–38 ( 1989).

    CAS  Article  Google Scholar 

  3. 3

    Nataro, J.P. & Kaper, J.B. Diarrheagenic Escherichia coli. Clin. Microbiol. Rev. 11, 142– 201 (1998).

    CAS  Article  Google Scholar 

  4. 4

    Paton, J.C. & Paton, A.W. Pathogenesis and diagnosis of Shiga toxin-producing Escherichia coli infections. Clin. Microbiol. Rev. 11, 450–479 ( 1998).

    CAS  Google Scholar 

  5. 5

    Lingwood, C.A. Role of verotoxin receptors in pathogenesis. Trends Microbiol. 4, 147–153 ( 1996).

    CAS  Article  Google Scholar 

  6. 6

    DeGrandis, S., Law, H., Brunton, J., Gyles, C. & Lingwood, C.A. Globotetraosyl ceramide is recognized by the pig edema disease toxin. J. Biol. Chem. 264, 12520 –12525 (1989).

    CAS  Google Scholar 

  7. 7

    Paton, A.W. et al. Molecular microbiological investigation of an outbreak of hemolytic uremic syndrome caused by dry fermented sausage contaminated with Shiga-like toxin-producing Escherichia coli. J. Clin. Microbiol. 34, 1622–1627 ( 1996).

    CAS  Google Scholar 

  8. 8

    Paton, A.W., Woodrow, M.C., Doyle, R.M., Lanser, J.A. & Paton, J.C. Molecular characterization of a Shiga-toxigenic Escherichia coli O113:H21 strain lacking eae responsible for a cluster of cases of hemolytic-uremic syndrome. J. Clin. Microbiol. 37, 3357–3361 (1999).

    CAS  Google Scholar 

  9. 9

    Wakarchuk, W.W. et al. Structure of an alpha-2,6-sialylated lipooligosaccharide from Neisseria meningitidis immunotype L1. Eur. J. Biochem. 254, 626–633 (1998).

    CAS  Article  Google Scholar 

  10. 10

    Mandrell, R.E. & Apicella, M.A. Lipooligosaccharides (LOS) of mucosal pathogens: molecular mimicry and host modification of LOS . Immunobiology 187, 382– 402 (1993).

    CAS  Article  Google Scholar 

  11. 11

    Risberg, A. et al. Structural analysis of the lipopolysaccharide oligosaccharide epitopes expressed by a capsule deficient strain of Haemophilus influenzae Rd. Eur. J. Biochem. 261, 171– 180 (1999).

    CAS  Article  Google Scholar 

  12. 12

    Masoud, H., Moxon, E.R., Martin, A., Krajcarski, D. & Richards, J.C. Structure of the variable and conserved lipopolysaccharide oligosaccharide epitopes expressed by Haemophilus influenzae serotype b strain Eagan. Biochemistry 36, 2091– 2103 (1997).

    CAS  Article  Google Scholar 

  13. 13

    Gotschlich, E.C. Genetic locus for the biosynthesis of the variable portion of Neisseria gonorrhoeae lipopolysaccharide. J. Exp. Med. 180 , 2181–2190 (1994).

    CAS  Article  Google Scholar 

  14. 14

    Yang, Q.-L. & Gotschlich, E.C. Variation of gonococcal lipooligosaccharide structure is due to alterations in poly-G tracts in lgt genes encoding glycosyl transferases. J. Exp. Med. 183, 323–327 (1996).

    CAS  Article  Google Scholar 

  15. 15

    Jennings, M.P., Hood, D.W., Peak, I.R., Virji, M. & Moxon, E.R. Molecular analysis of a locus for the biosynthesis and phase-variable expression of the lacto-N-neotetraose terminal lipopolysaccharide structure in Neisseria meningitidis. Mol. Microbiol. 18, 729–740 (1995).

    CAS  Article  Google Scholar 

  16. 16

    Heinrichs, D.E., Yethon, J.A., Amor, P.A. & Whitfield, C. The assembly system for the outer core portion of R1- and R4-type lipopolysaccharides of Escherichia coli. J. Biol. Chem. 273, 29497–29505 (1998).

    CAS  Article  Google Scholar 

  17. 17

    Lindgren, S.W., Melton, A.R. & O'Brien, A.D. Virulence of enterohemorrhagic Escherichia coli O91:H21 clinical isolates in an orally infected mouse model. Infect. Immun. 61, 3832–3842 (1993).

    CAS  Google Scholar 

  18. 18

    Armstrong, G.D., Fodor, E. & and Vanmaele, R. Investigation of Shiga-like toxin binding to chemically synthesized oligosaccharide sequences. J. Infect. Dis. 164, 1160–1167 (1991).

    CAS  Article  Google Scholar 

  19. 19

    Takeda, T., Yoshino, K., Adachi, E., Sato, Y. & Yamagata, K. In vitro assessment of a chemically synthesized Shiga toxin receptor analog attached to chromosorb P (Synsorb Pk) as a specific absorbing agent of Shiga toxin 1 and 2. Microbiol. Immunol. 43, 331–337 (1999).

    CAS  Article  Google Scholar 

  20. 20

    Rogers, J.E., Armstrong, G. & O'Brien, A.D. Abstract V149/VII, 114 (3rd International Symposium and Workshop on Shiga Toxin (Verotoxin)-producing Escherichia coli Infections, Baltimore, Maryland, June, 1997). (Lois Joy Galler Foundation for Hemolytic Uremic Syndrome, Melville, New York, 1997).

  21. 21

    Armstrong, G.D. et al. A phase I study of chemically synthesized verotoxin (Shiga-like toxin) Pk-trisaccharide receptors attached to chromosorb for preventing hemolytic-uremic syndrome. J. Infect. Dis. 171, 1042– 1045 (1995).

    CAS  Article  Google Scholar 

  22. 22

    Liljeqvist, S. & Stahl, S. Production of recombinant subunit vaccines: protein immunogens, live delivery systems and nucleic acid vaccines. J. Biotech. 73, 1– 33 (1999).

    CAS  Article  Google Scholar 

  23. 23

    Clements, J.D., Hartzog, N.M. & Lyon, F.L. Adjuvant activity of Escherichia coli heat-labile enterotoxin and effect on the induction of oral tolerance in mice to unrelated protein antigens. Vaccine 6, 269– 277 (1988).

    CAS  Article  Google Scholar 

  24. 24

    Moran, A.P., Prendergast, M.M. & Appelmelk, B.J. Molecular mimicry of host structures by bacterial lipopolysaccharides and its contribution to disease. FEMS Immunol. Med. Microbiol. 16, 105–115 (1996).

    CAS  Article  Google Scholar 

  25. 25

    Maniatis, T., Fritsch, E.F. & Sambrook, J. in Molecular cloning: A Laboratory Manual p. 68 (Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, 1982).

    Google Scholar 

  26. 26

    Darveau, R.P. & Hancock, R.E. Procedure for isolation of bacterial lipopolysaccharides from both smooth and rough Pseudomonas aeruginosa and Salmonella typhimurium strains. J. Bacteriol. 155, 831–838 (1983).

    CAS  Google Scholar 

  27. 27

    Morona, R., Brown, M.H., Yeadon, J., Heuzenroeder, M.W. & Manning, P.A. Effect of lipopolysaccharide core synthesis mutations on the production of Vibrio cholerae O-antigen in Escherichia coli K-12. FEMS Microbiol. Lett. 663, 279 –285 (1991).

    Article  Google Scholar 

  28. 28

    Klauser, T., Pohlner, J. & Meyer, T.F. Extracellular transport of cholera toxin B subunit using Neisseria IgA protease beta-domain: conformation-dependent outer membrane translocation. EMBO J. 9, 1991– 1999 (1990).

    CAS  Article  Google Scholar 

  29. 29

    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. 58, 2438–2445 ( 1990).

    CAS  Google Scholar 

  30. 30

    Wadolkowski, E.A., Sung, L.M., Burris, J.A., Samuel, J.A. & 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. 58, 3959– 3965 (1990).

    CAS  Google Scholar 

  31. 31

    O'Brien, A.D., et al. Shiga-like toxin-converting phages from Escherichia coli strains that cause hemorrhagic colitis or infantile diarrhea. Science 226, 694–696 ( 1984).

    CAS  Article  Google Scholar 

  32. 32

    Yanisch-Perron, C., Vieira, J. & Messing, J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13 mp18 and pUC19 vectors. Gene 33, 103–119 (1985).

    CAS  Article  Google Scholar 

  33. 33

    Paton, A.W., Paton, J.C. & Manning, P.A. Polymerase chain reaction amplification, cloning and sequencing of variant Escherichia coli Shiga-like toxin type II operons . Microb. Pathog. 15, 77– 82 (1993).

    CAS  Article  Google Scholar 

  34. 34

    Paton, A.W., Beutin, L. & Paton, J.C. Heterogeneity of the amino-acid sequences of Escherichia coli Shiga-like toxin type-I operons. Gene 153, 71–74 (1995).

    CAS  Article  Google Scholar 

  35. 35

    Ito, H., Terai, A., Kurazono, H., Takeda, Y. & Nishibuchi, M. Cloning and nucleotide sequencing of Vero toxin 2 variant genes from E. coli O91:H21 isolated from a patient with HUS. Microb. Pathogen. 8, 47–60 (1990).

    CAS  Article  Google Scholar 

  36. 36

    Jobling, M.G. & Holmes, R.K. Construction of vectors with the p15a replicon, kanamycin resistance, inducible lacZαand pUC18 or pUC19 multiple cloning sites. Nucleic Acids Res. 18, 5315–5316 (1990).

    CAS  Article  Google Scholar 

  37. 37

    Vinogradov, E.V. et al. The structures of the carbohydrate backbones of the lipopolysaccharides from Escherichia coli rough mutants F470 (R1 core type) and F576 (R2 coretype). Eur. J. Biochem. 261, 629– 639 (1999).

    CAS  Article  Google Scholar 

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The assistance of L. van den Bosch and E. Parker is acknowledged. This work was supported by grants from the National Health and Medical Research Council of Australia, and the Channel Seven Children's Research Foundation.

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Correspondence to James C. Paton.

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Paton, A., Morona, R. & Paton, J. A new biological agent for treatment of Shiga toxigenic Escherichia coli infections and dysentery in humans. Nat Med 6, 265–270 (2000). https://doi.org/10.1038/73111

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