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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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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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