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Immunodesign of experimental sepsis by cecal ligation and puncture

Nature Protocols volume 4, pages 31–36 (2009)Cite this article

Abstract

Sepsis remains a prevalent clinical challenge and the underlying pathophysiology is still poorly understood. To investigate the complex molecular mechanisms of sepsis, various animal models have been developed, the most frequently used being the cecal ligation and puncture (CLP) model in rodents. In this model, sepsis originates from a polymicrobial infectious focus within the abdominal cavity, followed by bacterial translocation into the blood compartment, which then triggers a systemic inflammatory response. A requirement of this model is that it is performed with high consistency to obtain reproducible results. Evidence is now emerging that the accompanying inflammatory response varies with the severity grade of sepsis, which is highly dependent on the extent of cecal ligation. In this protocol, we define standardized procedures for inducing sepsis in mice and rats by applying defined severity grades of sepsis through modulation of the position of cecal ligation. The CLP procedure can be performed in as little as 10 min for each animal by an experienced user, with additional time required for subsequent postoperative care and data collection.

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Figure 1: Critical steps in the CLP procedure in mice.
Figure 2: Anatomical locations for cecal ligation of rodents and map of arteries and ileocecal valve.
Figure 3: Survival rates after CLP in rodents as a function of rite of cecal ligation.

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References

  1. Dombrovskiy, V.Y., Martin, A.A., Sunderram, J. & Paz, H.L. Rapid increase in hospitalization and mortality rates for severe sepsis in the United States: a trend analysis from 1993 to 2003. Crit. Care. Med. 35, 1244–1250 (2007).

    Article  Google Scholar 

  2. Kochanek, K.D. & Smith, B.L. Deaths: preliminary data for 2002. Natl. Vital. Stat. Rep. 52, 1–47 (2004).

    PubMed  Google Scholar 

  3. Rittirsch, D., Hoesel, L.M. & Ward, P.A. The disconnect between animal models of sepsis and human sepsis. J. Leukoc. Biol. 81, 137–143 (2007).

    Article  CAS  Google Scholar 

  4. Remick, D.G., Newcomb, D.E., Bolgos, G.L. & Call, D.R. Comparison of the mortality and inflammatory response of two models of sepsis: lipopolysaccharide vs. cecal ligation and puncture. Shock 13, 110–116 (2000).

    Article  CAS  Google Scholar 

  5. Deitch, E.A. Rodent models of intra-abdominal infection. Shock 24 (Suppl. 1): 19–23 (2005).

    Article  Google Scholar 

  6. Buras, J.A., Holzmann, B. & Sitkovsky, M. Animal models of sepsis: setting the stage. Nat. Rev. Drug Discov. 4, 854–865 (2005).

    Article  CAS  Google Scholar 

  7. Wichterman, K.A., Baue, A.E. & Chaudry, I.H. Sepsis and septic shock—a review of laboratory models and a proposal. J. Surg. Res. 29, 189–201 (1980).

    Article  CAS  Google Scholar 

  8. Hubbard, W.J. et al. Cecal ligation and puncture. Shock 24 (Suppl 1): 52–57 (2005).

    Article  Google Scholar 

  9. Benjamim, C.F., Hogaboam, C.M. & Kunkel, S.L. The chronic consequences of severe sepsis. J. Leukoc. Biol. 75, 408–412 (2004).

    Article  CAS  Google Scholar 

  10. Xiao, H., Siddiqui, J. & Remick, D.G. Mechanisms of mortality in early and late sepsis. Infect. Immun. 74, 5227–5235 (2006).

    Article  CAS  Google Scholar 

  11. Singleton, K.D. & Wischmeyer, P.E. Distance of cecum ligated influences mortality, tumor necrosis factor-alpha and interleukin-6 expression following cecal ligation and puncture in the rat. Eur. Surg. Res. 35, 486–491 (2003).

    Article  CAS  Google Scholar 

  12. Baker, C.C., Chaudry, I.H., Gaines, H.O. & Baue, A.E. Evaluation of factors affecting mortality rate after sepsis in a murine cecal ligation and puncture model. Surgery 94, 331–335 (1983).

    CAS  PubMed  Google Scholar 

  13. Rittirsch, D. et al. Functional roles for C5a receptors in sepsis. Nat. Med. 14, 551–557 (2008).

    Article  CAS  Google Scholar 

  14. Wichterman, K.A., Baue, A.E. & Chaudry, I.H. Sepsis and septic shock—a review of laboratory models and a proposal. J. Surg. Res. 29, 189–201 (1980).

    Article  CAS  Google Scholar 

  15. Heuer, J.G. et al. Cecal ligation puncture with total parenteral nutrition: a clinically relevant model of the metabolic, hormonal, and inflammatory dysfunction associated with critical illness. J. Surg. Res. 121, 178–186 (2004).

    Article  CAS  Google Scholar 

  16. Smith, E.F. III, et al. Fluid resuscitation improves survival of endotoxemic or septicemic rats: possible contribution of tumor necrosis factor. Pharmacology 46, 254–267 (1993).

    Article  CAS  Google Scholar 

  17. Law, M. & Remick, D. Fluid resuscitation and antibiotics enhance survival by unknown mechanisms. Shock 25, 6 (2006).

    Google Scholar 

  18. Xiao, H., Siddiqui, J. & Remick, D.G. Mechanisms of mortality in early and late sepsis. Infect. Immun. 74, 5227–5235 (2006).

    Article  CAS  Google Scholar 

  19. Latifi, S.Q., O'Riordan, M.A. & Levine, A.D. Interleukin-10 controls the onset of irreversible septic shock. Infect. Immun. 70, 4441–4446 (2002).

    Article  CAS  Google Scholar 

  20. Coria-Avila, G.A., Gavrila, A.M., Menard, S., Ismail, N. & Pfaus, J.G. Cecum location in rats and the implications for intraperitoneal injections. Lab. Anim. (NY) 36, 25–30 (2007).

    Article  Google Scholar 

  21. Godshall, C.J., Scott, M.J., Peyton, J.C., Gardner, S.A. & Cheadle, W.G. Genetic background determines susceptibility during murine septic peritonitis. J. Surg. Res. 102, 45–49 (2002).

    Article  CAS  Google Scholar 

  22. Hollenberg, S.M., et al. Characterization of a hyperdynamic murine model of resuscitated sepsis using echocardiography. Am. J. Respir. Crit. Care Med. 164, 891–895 (2001).

    Article  CAS  Google Scholar 

  23. Newcomb, D., Bolgos, G., Green, L. & Remick, D.G. Antibiotic treatment influences outcome in murine sepsis: mediators of increased morbidity. Shock 10, 110–117 (1998).

    Article  CAS  Google Scholar 

  24. Flierl, M.A. et al. Adverse functions of IL-17A in experimental sepsis. FASEB J. 22, 2198–2205. Epub 2008 Feb 25.

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported by NIH grants GM-61656, GM-029507 and HL-31963 (to P.A.W.).

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Authors and Affiliations

  1. Department of Pathology, University of Michigan Medical School, 1301 Catherine Road, Ann Arbor, 48109, Michigan, USA

    Daniel Rittirsch, Michael A Flierl & Peter A Ward

  2. Department of Traumatology, Hand, Plastic and Reconstructive Surgery, University Hospital Ulm, Steinhövelstrasse 9, Ulm, D-89075, Germany

    Markus S Huber-Lang

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  1. Daniel Rittirsch
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Correspondence to Peter A Ward.

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Rittirsch, D., Huber-Lang, M., Flierl, M. et al. Immunodesign of experimental sepsis by cecal ligation and puncture. Nat Protoc 4, 31–36 (2009). https://doi.org/10.1038/nprot.2008.214

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