Skip to main content
SpringerLink
Log in

Effect of verocytotoxins (Shiga-like toxins) on human neutrophils in vitro

  • Original Article
  • Published:
Pediatric Nephrology Aims and scope Submit manuscript

Abstract

Neutrophil activation occurs in diarrhoea-associated HUS and correlates with disease severity, implying a role in pathogenesis. Verocytotoxin (Shiga-like toxin) has been shown to stimulate endothelium to release chemokines and express leukocyte adhesion molecules that would lead to indirect neutrophil-endothelial interaction. A direct action of verocytotoxin (VT) on neutrophils has been proposed, although in vitro studies of this are controversial. In this report we examine the effect of verocytotoxin-1 (Shiga-like toxin-1) (VT1) and verocytotoxin-2 (VT2) on human neutrophils in vitro with regard to priming, the release of superoxide and elastase, and chemotaxis. Neutrophils were incubated with VT1 or VT2 and superoxide and elastase release was measured over 120 and 45 minutes respectively. Priming was investigated by pre-treating the neutrophils with VT1 or VT2, exposing them to formyl-met-leu-phe (fMLP) or phorbol myristic acid (PMA) and measuring superoxide release. Neutrophil chemotaxis towards fMLP was assessed with and without pre-incubation with VT1 and VT2. We found that neither of the toxins induced superoxide or elastase release. Priming with VT1 significantly reduced superoxide release when neutrophils were stimulated with fMLP or PMA. VT2 priming gave a reduced superoxide release with PMA but not fMLP. Heat-inactivation of the toxins gave similar results. Pre-treatment of neutrophils with VT1 or VT2 did not affect chemotaxis towards fMLP after a 2-hour incubation period. In conclusion, VT1 and VT2 do not activate primed neutrophils in vitro. Nor do they affect chemotaxis towards fMLP. They may impair neutrophil priming.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. O’Brien AD, Holmes RK (1987) Shiga and Shiga-like toxins. Microbiol Rev 51:206–220

    Google Scholar 

  2. Stein PE, Boodhoo A, Tyrrell GJ, et al ( 1992) Crystal structure of the cell-binding B oligomer of verotoxin-1 from E. coli. Nature 355:748–750

    Article  CAS  PubMed  Google Scholar 

  3. Lingwood CA. (1994) Verotoxin-binding in human renal sections. Nephron 66:21–28

    CAS  PubMed  Google Scholar 

  4. Ergonul Z, Clayton F, Fogo AB, Kohan DE (2003) Shigatoxin-1 binding and receptor expression in human kidneys do not change with age. Pediatr Nephrol 18:246–253

    PubMed  Google Scholar 

  5. Bitzan M, Richardson S, Huang C, et al (1994) Evidence that verotoxins (Shiga-like toxins) from Escherichia coli bind to P blood group antigens of human erythrocytes in vitro. Infect Immun 62:3337–3347

    CAS  PubMed  Google Scholar 

  6. van Setten PA, Monnens LAH, Verstraten RG, et al.(1996) Effects of verotoxin-1 on non-adherent human monocytes: binding characteristics, protein synthesis, and induction of cytokine release. Blood 88:174–183

    PubMed  Google Scholar 

  7. Cooling LL, Walker KE, Gille T, et al (1998) Shiga toxins binds human platelets via globotriaosylceramide (Pk antigen) and a novel platelet glycosphingolipid. Infect Immun 66:4355–4366

    CAS  PubMed  Google Scholar 

  8. Lingwood CA (1996) Role of verotoxin receptors in pathogenesis. Trends Microbiol 4:147–153

    Google Scholar 

  9. te Loo M, Monnens LA, van der Velden TJ, et al (2000) Binding and transfer of verocytotoxin by polymorphonuclear leukocytes in hemolytic uremic syndrome. Blood 95:3396–2402

    PubMed  Google Scholar 

  10. Tesh VL, Samuel JE, Perera LP, et al (1994) Evalution of the role of Shiga and Shiga-like toxins in mediating direct damage to human vascular endothelial cells. J Infect Dis 164:344–352

    Google Scholar 

  11. Louise CB, Obrig TG (1992) Shiga toxin-associated hemolytic uremic syndrome: combined cytotoxic effects of Shiga-toxin, interleukin-1β and tumor necrosis factor alpha on human vascular endothelial cells in vitro. Infect Immun 59:1536-1543

    Google Scholar 

  12. van de Kar NC, Kooistra T, Vermeer M, et al (1992) Tumor necrosis factor alpha and interleukin-1 induce expression of the verocytotoxin receptor globotriaosylceramide on human endothelial cells: implications for the pathogenesis of the haemolytic uremic syndrome. Blood 80:2755–2764

    PubMed  Google Scholar 

  13. Thorpe CM, Hurley BP, Lincicome LL, et al (1999) Shiga toxins stimulate secretion of interleukin-8 from intestinal epithelial cells. Infect Immun 67:5985–5993

    CAS  PubMed  Google Scholar 

  14. Morigi M, Micheletti G, Figliuzzi M, et al (1995) Verotoxin-1 promotes leukocyte adhesion to cultured endothelial cells under physiologic flow conditions. Blood 86:4553–4558

    CAS  PubMed  Google Scholar 

  15. Zoja C, Angioletti S, Donadelli R, et al (2002) Shiga toxin-2 triggers endothelial leukocyte adhesion and transmigration via NF-kB dependent upregulation of IL-8 and MCP-1. Kidney Int 62:846–856

    Article  CAS  PubMed  Google Scholar 

  16. Ishii H, Takada K, Higuchi T, et al (2000) Verotoxin-1 induces tissue factor expression in human umbilical vein endothelial cells through activation of NF-kB/Rel and AP-1. Thromb Haemost 84:712–721

    CAS  PubMed  Google Scholar 

  17. Chandler WL, Jelacic S, Boster DR, et al (2002) Prothrombotic coagulation abnormalities preceding the haemolytic-uremic syndrome. N Engl J Med 346:23–32

    Article  CAS  PubMed  Google Scholar 

  18. King AJ, Sundaram S, Cendoroglo M, et al (1999) Shiga toxin induces superoxide production in polymorphonuclear cells with subsequent impairment of phagocytosis and responsiveness to phorbol esters. J Infect Dis 179:503–507

    Article  CAS  PubMed  Google Scholar 

  19. Aoki Y, Takeda T (2002) Effect of Shiga toxins on granulocyte function. Microb Pathog 32: 279–285

    Google Scholar 

  20. Bajaj MS, Kew RR, Webster RO, et al (1992) Priming of human neutrophil functions by tumor necrosis factor—enhancement of superoxide anion generation, degranulation, and chemotaxis to chemoattractants C5a and F-Met-Leu-Phe. Inflammation 16: 241–50

    Article  CAS  PubMed  Google Scholar 

  21. Sengelov H, Follin P, Kjeldsen L, et al (1995) Mobilization of granules and secretory granules during in vivo exudation of human neutrophils. J Immunol 154:4157–4165

    CAS  PubMed  Google Scholar 

  22. O’Flaherty JT, Rossi AG, Redman JF, et al (1991) Tumor necrosis factor- regulates expression of receptors for formyl-methionyl.leucyl-phenylalanine, leukotriene B4, and platelet-activating factor. Dissociation from priming in human polymorphonuclear neutrophils. J Immunol 147:3842–3847

    PubMed  Google Scholar 

  23. Zimmerli W, Seligmann B, Gallin JI (1986) Exudation primes human and guinea pig neutrophils for subsequent responsiveness to the chemotactic peptide N-formylmethionylleucylphenylalanine and increases component C3b. J Clin Invest 77:925–933

    CAS  PubMed  Google Scholar 

  24. Condliffe AM, Chilvers RR, Haslett C, et al (1996) Priming differentially regulates neutrophil adhesion molecule expression/function. Immunology 89:105–111

    Article  CAS  PubMed  Google Scholar 

  25. Kuhns DB, Priel DAL, Gallin JL (1995) Loss of l-Selectin (CD62L) on human neutrophils following exudation in vivo. Cell Immunol 164:306–310

    Article  CAS  PubMed  Google Scholar 

  26. Liu J, He The Y, Zhang Z, et al (2002) Prolongation of functional life-span of neutrophils by recombinant verotoxin 2. Chin Med J 115:900–903

    CAS  PubMed  Google Scholar 

  27. Liu J, Akahoshi T, Sasahana T, et al (1999) Inhibition of neutrophil apoptosis by verotoxin 2 derived from Escherichia coli O157:H7. Infect Immun 67:6203–6205

    CAS  PubMed  Google Scholar 

  28. Cutler JE, Munoz JJ (1974) A simple method for studies on chemotaxis. Proc Soc Exp Biol Med 147(2):471–474

    CAS  PubMed  Google Scholar 

  29. Nelson RD, Quie PG, Simmons RL (1975) Chemotaxis under agarose: a new simple method for measuring chemotaxis and spontaneous migration of human polymorphonuclear leucocytes and monocytes. J Immunol 115(6):1650–1656

    CAS  PubMed  Google Scholar 

  30. Pavia AT, Nichols CR, Green DP, et al. (1990) Hemolytic-uremic syndrome during an outbreak of Escherichia coli O157:H7 infections in institutions for mentally retarded persons: clinical and epidemiologic observations. J Pediatr 116:544–551

    CAS  PubMed  Google Scholar 

  31. Walters MD, Matthei IU, and Kay R, et al (1989) The polymorphonuclear leucocyte count in childhood haemolytic uremic syndrome. Pediatr Nephrol 3:130–134

    Article  CAS  PubMed  Google Scholar 

  32. Fitzpatrick MM, Shah V, Filler G, et al (1992) Neutrophil activation in the haemolytic uremic syndrome: free and complexed elastase in plasma. Pediatr Nephrol 6(1):50–53

    Article  CAS  PubMed  Google Scholar 

  33. Ishikawa N, Kamitsuji H, Murakami T, et al (2000) Plasma levels of granulocyte elastase-α1-proteinase inhibitor complex in children with haemolytic uremic syndrome caused by verotoxin-producing Escherichia coli. Pediatr Int 42:637–641

    Article  CAS  PubMed  Google Scholar 

  34. Inward CD, Howie AJ, and Fitzpatrick MM, et al (1997) Renal histopathology in fatal cases of diarrhoea-associated haemolytic uraemic syndrome. British Association for Paediatric Nephrology. Pediatr Nephrol 11(5):556–559

    Article  CAS  PubMed  Google Scholar 

  35. Fitzpatrick MM, Shah V, Trompeter RS, et al (1992) Interleukin-8 and polymorphonuclear leucocyte activation in haemolytic uremic syndrome of childhood. Kidney Int 42(4):952–962

    Google Scholar 

  36. Kettritz R, Falk RJ, Jennette JC, Gaido ML (1997) Neutrophhil superoxide release is required for spontaneous and FMLP-mediated but not TNF alpha-mediated apoptosis. J Am Soc Nephrol 8(7):1091–1100

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

Dr J.U. Holle was supported by the Renal Research Fund of the Birmingham Children’s Hospital Trust Funds.

Author information

Authors and Affiliations

  1. Department of Renal Immunobiology, The Medical School, University of Birmingham, Birmingham, B15 2TT, UK

    Julie U. Holle, Julie M. Williams, Lorraine Harper & Caroline O. S. Savage

  2. Birmingham Children’s Hospital, Birmingham, B4 6NH, UK

    C. Mark Taylor

Authors
  1. Julie U. Holle
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Julie M. Williams
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lorraine Harper
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Caroline O. S. Savage
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. C. Mark Taylor
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to C. Mark Taylor.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Holle, J.U., Williams, J.M., Harper, L. et al. Effect of verocytotoxins (Shiga-like toxins) on human neutrophils in vitro. Pediatr Nephrol 20, 1237–1244 (2005). https://doi.org/10.1007/s00467-005-1945-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00467-005-1945-2

Keywords

Search

Navigation