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Isolation of Murine Peritoneal Macrophages to Carry Out Gene Expression Analysis Upon Toll-like Receptors Stimulation
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Peritoneal macrophage priming in cirrhosis is related to ERK phosphorylation and IL-6 secretion

Antonio J Ruiz-Alcaraz1, María Martínez-Esparza, Rocío Caño

  • 1Department of Biochemistry and Molecular Biology B and Immunology, School of Medicine, Universidad de Murcia, Murcia, Spain.

European Journal of Clinical Investigation
|August 25, 2010
PubMed
Abstract

Insights

Peritoneal macrophages in cirrhotic patients are primed for a rapid immune response, even without active bacterial infection. This sustained immune readiness, potentially IL-6 controlled, prepares them for bacterial translocation events.

Area of Science:

  • Immunology
  • Gastroenterology
  • Cell Biology

Background:

  • Bacterial infections and DNA translocation are common in cirrhosis with ascites.
  • These events increase inflammation and worsen prognosis.
  • The study investigates if peritoneal macrophages are primed for a rapid response.

Purpose of the Study:

  • To determine if peritoneal macrophages in cirrhotic patients with ascites are in a chronically primed state.
  • To assess their readiness for responding to bacterial translocation.

Main Methods:

  • Peritoneal macrophages from 25 cirrhotic patients were compared to blood monocytes.
  • Flow cytometry assessed cell-surface markers.
  • Western blot analyzed signaling pathway phosphorylation (ERK1/2, p38, PKB, c-Jun, p65 NFκB).
  • ELISA measured cytokine synthesis (TNF-α, IL-6, IL-10).

Main Results:

  • Peritoneal macrophages showed high CD54, CD86, and HLA-DR expression.
  • Elevated baseline phosphorylation of ERK1/2, PKB, and c-Jun was observed.
  • Increased baseline IL-6 production correlated with ERK phosphorylation (r=0.9, P=0.005).
  • Bacterial stimuli induced higher signaling intermediate phosphorylation than DNA stimuli, but similar p65 NFκB activation.

Conclusions:

  • Ascitic fluid in cirrhotic patients harbors a sustained immune response, even without bacterial antigens.
  • This sustained response likely facilitates rapid defense against bacterial DNA translocation.
  • Interleukin-6 (IL-6) may play a key role in controlling this response during chronic liver inflammation.

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