title: Peritoneal wash contents used to predict mortality in a murine sepsis model.
abstract: BACKGROUND
  Cecal ligation and puncture (CLP) is considered the gold standard for inducing abdominal sepsis in mice. However, the model lacks source control, a component of sepsis management in humans. Using a CLP-excision model, we characterized peritoneal cytokines and cells and hypothesized these analyses would allow us to predict survival.
  
  
  METHODS
  Fifty-eight mice were first subjected to CLP. Twenty hours later, the necrotic cecums were debrided, abdominal cavity lavaged, and intraperitoneal antibiotics administered. Peritoneal cytokines and leukocytes collected from the peritoneal lavage were analyzed. These immune parameters were used to generate receiver operator characteristic curves. In separate experiments, the accuracy of the model was verified with a survival cohort. Finally, we collected the peritoneal lavage and analyzed both serum and peritoneal cytokines, bacterial load, and leukocyte functionality.
  
  
  RESULTS
  Peritoneal interleukin (IL)-6 levels and neutrophil CD11b intensity were observed to be significantly different in mice that lived versus those who died. In separate experiments, mice predicted to live (P-LIVE) had decreased bacterial loads, systemic IL-10, and neutrophil oxidative burst and increased peritoneal inflammatory monocyte numbers and phagocytosis.
  
  
  CONCLUSIONS
  This study couples a clinically relevant sepsis model with methodology to limit pathogen spread. Using surgical waste, stratification of the mice into groups P-LIVE and predicted to die was possible with a high degree of accuracy and specificity. In mice P-LIVE, increased inflammatory monocyte recruitment and phagocytosis were associated with decreased systemic IL-10 and bacterial loads.
authors.affiliation: Division of Research, Department of Surgery, College of Medicine, University of Cincinnati, Cincinnati, Ohio.
authors.forename: Joshua W
authors.initials: JW
authors.lastname: Kuethe
authors.affiliation: Division of Research, Department of Surgery, College of Medicine, University of Cincinnati, Cincinnati, Ohio.
authors.forename: Emily F
authors.initials: EF
authors.lastname: Midura
authors.affiliation: Division of Research, Department of Surgery, College of Medicine, University of Cincinnati, Cincinnati, Ohio.
authors.forename: Teresa C
authors.initials: TC
authors.lastname: Rice
authors.affiliation: Division of Research, Department of Surgery, College of Medicine, University of Cincinnati, Cincinnati, Ohio. Electronic address: charles.caldwell@uc.edu.
authors.forename: Charles C
authors.initials: CC
authors.lastname: Caldwell
mesh_terms: D000818:Animals; D001202:Ascitic Fluid / Q000276:immunology / Q000378:metabolism / Q000382:microbiology; D058491:Bacterial Load; D015415:Biomarkers / Q000378:metabolism; D002432:Cecum / Q000601:surgery; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D007962:Leukocytes / Q000378:metabolism; D008026:Ligation; D008297:Male; D051379:Mice; D010533:Peritoneal Lavage; D011379:Prognosis; D011677:Punctures; D012372:ROC Curve; D018805:Sepsis / Q000175:diagnosis / Q000378:metabolism / Q000382:microbiology / Q000401:mortality*
chemical_list: D015415:Biomarkers; D016207:Cytokines
keywords: Bacterial clearance; Cecal ligation and puncture; IL-6; Inflammation; Leukocyte; Neutrophil activation