World Journal of Surgery

, Volume 31, Issue 1, pp 105–115

Interleukin-10 Gene Transfer: Prevention of Multiple Organ Injury in a Murine Cecal Ligation and Puncture Model of Sepsis

  • Burhan Kabay
  • Cetin Kocaefe
  • Atac Baykal
  • Hilmi Özden
  • Cengiz Baycu
  • Zafer Oner
  • Meral Özgüç
  • Iskender Sayek
Article

Abstract

Introduction

The aim of this study was to determine the effect of immunoregulatory cytokine interleukin-10 (IL-10) gene therapy on multiple organ injury (MOI) induced by a cecal ligation and puncture (CLP) model of sepsis in mice.

Methods

Male Balb/c mice subjected to CLP were treated with either an hIL-10-carrying vector or an empty control vector. We assessed the degree of lung, liver, and kidney tissue destruction biochemically by measuring myeloperoxidase (MPO) and malondialdehyde (MDA) activity. Histologic assessments were based on neutrophil infiltration in lung and liver tissue. IL-10 protein expression was examined immunohistochemically, and ultrastructural changes in the liver were studied by transmission electron microscopy. We analyzed the expression of tumor necrosis factor-α (TNFα) mRNA by reverse transcription polymerase chain reaction 3, 8, and 24 hours after CLP in all organs.

Results

Organ damage was significantly reduced by hIL-10 gene transfer, which was associated at the tissue level with reduced MPO activity in the liver, lung, and kidney and decreased leukocyte sequestration and MDA formation in the lung. The liver MDA was not significantly higher in the hIL-10 gene therapy group than in the controls and seemed not to be affected by hIL-10 gene transfer. The reduced portal tract neutrophilic infiltration and preserved ultrastructure of the hepatocytes also showed that tissue function was not impaired. The lung and kidney TNFα mRNA expression was suppressed markedly in the hIL-10 gene therapy group, but liver TNFα mRNA expression varied over time.

Conclusions

These findings showed that IL-10 gene therapy significantly attenuated sepsis-induced MOI.

References

  1. 1.
    Natanson C. Selected treatment strategies for septic shock based on proposed mechanisms of pathogenesis. Ann Intern Med 1994;120:771–783PubMedGoogle Scholar
  2. 2.
    Bhatia M, Moochhala S. Role of inflammatory mediatory mediators in the pathophysiology of acute respiratory distress syndrome. J Pathol 2004;202:145–156PubMedCrossRefGoogle Scholar
  3. 3.
    Poll T, Deventer SJH. Cytokines and anticytokines in the pathogenesis of sepsis. Surg Clin North Am 1999;13:413–426Google Scholar
  4. 4.
    Gerard C, Bruyns C, Marchant A, et al. IL-10 reduces the release of TNF and prevents lethality in experimental endotoxemia. J Exp Med 1993;177:547–550PubMedCrossRefGoogle Scholar
  5. 5.
    Cassatella MA, Meda L, Bonaro S, et al. IL-10 inhibits the release of proinflammatory cytokines from human polymorphonuclear leucocytes: evidence for an autocrine role of TNF and IL-1 beta in mediating the production of IL-8 triggered by lipopolysaccharide. J Exp Med 1993;178:2207–2211PubMedCrossRefGoogle Scholar
  6. 6.
    Marchant A, Bruyns C, Vandenabeele P, et al. IL-10 controls IFN-gamma and TNF production during experimental endotoxemia. Eur J Immunol 1994;24:1167–1171PubMedGoogle Scholar
  7. 7.
    De Waal Malefyt R, Abrams J, Bennet B, et al. IL-10 inhibits cytokine synthesis by human monocytes: an autoregulatory role of IL-10 produced by monocytes. J Exp Med 1991;174:1209–1220PubMedCrossRefGoogle Scholar
  8. 8.
    Hart PH, Hunt EK, Bonder CS, et al. Regulation of surface and soluble TNF receptor expression on human monocytes and synovial fluid macrophages by IL-4 and IL-10. J Immunol 1996;157:3672–3680PubMedGoogle Scholar
  9. 9.
    Cassatella MA. The neutrophil: one of the cellular targets of IL-10. Int J Clin Lab Res 1998;28:148–161PubMedCrossRefGoogle Scholar
  10. 10.
    Abraham E. Neutrophils and acute lung injury. Crit Care Med 2003;31:195–199CrossRefGoogle Scholar
  11. 11.
    Koksoy C, Kuzu A, Ergun H, et al. Role of tumour necrosis factor in lung injury caused by intestinal ischaemia-reperfusion. Br J Surg 2001;88:464–468PubMedCrossRefGoogle Scholar
  12. 12.
    Cutrin JC, Boveris A, Zingaro B, et al. In situ determination by surface chemiluminescence of temporal relationships between evolving warm ischemia-reperfusion injury in rat liver and phagocyte activation and recruitment. Hepatology 2000;31:622–632PubMedCrossRefGoogle Scholar
  13. 13.
    Mihara M, Uchiyama M. Determination of malonaldehyde precursor in tissues by thiobarbituric acid test. Anal Biochem 1978;86:271–278PubMedCrossRefGoogle Scholar
  14. 14.
    Lowry OH, Rosebrough NJ, Farr AL, et al. Protein measurement with the folin phenol reagent. J Biol Chem 1951;193:265–275PubMedGoogle Scholar
  15. 15.
    Demling R, Nayak U, Ikegami K, et al. Comparison between lung and liver peroxidation and mortality after zymosan peritonitis in rats. Shock 1994;2:222–227PubMedCrossRefGoogle Scholar
  16. 16.
    van der Poll T, Marchant A, Buurman WA, et al. Endogenous IL-10 protects mice from death during septic peritonitis. J Immunol 1995;155:5367–5401Google Scholar
  17. 17.
    Latifi SQ, O’Riordan MA, Levine AD. IL-10 controls the onset of irreversible septic shock. Infect Immun 2002;70:4441–4446PubMedCrossRefGoogle Scholar
  18. 18.
    Remick DG, Garg SJ, Newcomb DE, et al. Exogeneous IL-10 fails to decrease the mortality or morbidity of sepsis. Crit Care Med 1998;26:895–904PubMedCrossRefGoogle Scholar
  19. 19.
    Huhn RD, Radwanski E, O’Connell SM, et al. Pharmacokinetics and immunomodulatory properties of intravenously administered recombinant human interleukin-10 in healthy volunteers. Blood 1996;87:699–705PubMedGoogle Scholar
  20. 20.
    Chernoff AE, Granowitz EV, Shapiro L, et al. A randomized controlled trial of IL-10 in humans; inhibition of inflammatory cytokine production and immune responses. J Immunol 1995;154:5492–5499PubMedGoogle Scholar
  21. 21.
    Nathan C, Sporn M. Cytokine in context. J Cell Biol 1991;113:981–986PubMedCrossRefGoogle Scholar
  22. 22.
    Chun S, Dahessia M, Lee S, et al. Immune modulation by IL-10 gene transfer via viral vector and plasmid DNA: implication for gene therapy. Cell Immunol 1999;194:194–204PubMedCrossRefGoogle Scholar
  23. 23.
    Zhu N, Liggitt D, Liu Y, et al. Systemic gene expression after intravenous DNA delivery into adult mice. Science 1993;261:209–211PubMedCrossRefGoogle Scholar
  24. 24.
    Simberg D, Weisman S, Talmon Y, et al. DOTAP (and other cationic lipids): chemistry, biophysics, and transfection. Crit Rev Ther Drug Carrier Syst 2004;21:257–317PubMedCrossRefGoogle Scholar
  25. 25.
    Pedroso de Lima MC, Neves S, Filipe A, et al. Cationic liposomes for gene delivery: from biophysics to biological applications. Curr Med Chem 2003;10:1221–1231PubMedCrossRefGoogle Scholar
  26. 26.
    Hui KM, Ang PT, Huang L, et al. Phase I study of immunotherapy of cutaneous metastases of human carcinoma using allogeneic and xenogeneic MHC DNA-liposome complexes. Gene Ther 1997;4:783–790PubMedCrossRefGoogle Scholar
  27. 27.
    Rini BI, Selk LM, Vogelzang NJ. Phase I study of direct intralesional gene transfer of HLA-B7 into metastatic renal carcinoma lesions. Clin Cancer Res1999;5:2766–2772PubMedGoogle Scholar
  28. 28.
    Yoshida J, Mizuno M, Wakabayashi T. Interferon beta gene therapy for cancer: basic research to clinical application. Cancer Sci 2004;95:858–865PubMedCrossRefGoogle Scholar
  29. 29.
    Sakhuja K, Reddy PS, Ganesh S, et al. Optimization of the generation and propagation of gutless adenoviral vectors. Hum Gene Ther 2003;14:243–254PubMedCrossRefGoogle Scholar
  30. 30.
    Deitch EA. Animal models of sepsis and shock: a review and lessons learned. Shock 1998;9:1–11PubMedCrossRefGoogle Scholar
  31. 31.
    Sayek I. Animal models for intra-abdominal infection. Hepatogastroenterology 1997;44:923–926PubMedGoogle Scholar
  32. 32.
    Takakuwa T, Endo S, Shirakura Y, et al. Interleukin–10 gene transfer improves the survival rate of mice inoculated with E. coli. Crit Care Med 2000;28:2685–2689PubMedCrossRefGoogle Scholar
  33. 33.
    Oberholzer C, Oberholzer A, Bahjat FR, et al. Targeted adenovirus-induced expression of IL-10 decreases thymic apoptosis and improves survival in murine sepsis. Proc Natl Acad Sci 2001;25:11503–11508CrossRefGoogle Scholar
  34. 34.
    Zhou XM, Yagihashi A, Hirata K, et al. Downregulation of cytokine-induced neutrophil chemoattractant and prolongation of rat liver allograft survival by interleukin-10. Surg Today 1998;28:184–191CrossRefGoogle Scholar
  35. 35.
    Fry DE, Pearlstein L, Fulton RL, et al. Multiple system organ failure: the role of uncontrolled infection. Arch Surg 1980;115:136–140PubMedGoogle Scholar
  36. 36.
    Armstrong L, Milla AB. Relative production of tumour necrosis factor alpha and IL-10 in adult respiratory distress syndrome. Thorax 1997;52:442–446PubMedCrossRefGoogle Scholar
  37. 37.
    Wang P, Chaudry IH. Mechanism of hepatocellular dysfunction during hyperdynamic sepsis. Am J Physiol 1996;270:927–938Google Scholar
  38. 38.
    Ertel W, Morrison MH, Wang P, et al. The complex pattern of cytokines in sepsis: association between prostaglandins, cachectin and interleukins. Ann Surg 1991;214:141–148PubMedCrossRefGoogle Scholar
  39. 39.
    Mayumi T, Chan CK, Clemens MG, et al. Zonal heterogeneity of hepatic injury following shock/resuscitation: relationship of xanthine oxidase activity to localization of neutrophil accumulation and central lobular necrosis. Shock 1996;5:324–332PubMedCrossRefGoogle Scholar
  40. 40.
    Zhang JX, Pegoli W Jr, Clemens MG. Endothelin-1 induces direct constriction of hepatic sinusoids. Am J Physiol 1994;266:624–632Google Scholar

Copyright information

© Société Internationale de Chirurgie 2006

Authors and Affiliations

  • Burhan Kabay
    • 1
    • 6
  • Cetin Kocaefe
    • 2
  • Atac Baykal
    • 3
  • Hilmi Özden
    • 4
  • Cengiz Baycu
    • 5
  • Zafer Oner
    • 3
  • Meral Özgüç
    • 2
  • Iskender Sayek
    • 3
  1. 1.Department of General SurgeryPamukkale University, School of MedicineDenizliTurkey
  2. 2.Department of Medical BiologyHacettepe University, School of MedicineAnkaraTurkey
  3. 3.Department of General SurgeryHacettepe University, School of MedicineAnkaraTurkey
  4. 4.Department of AnatomyOsmangazi University, School of MedicineEskisehirTurkey
  5. 5.Department of Histology and EmbryologyOsmangazi University, School of MedicineEskisehirTurkey
  6. 6.YenisehirTurkey

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