Intensive Care Medicine

, Volume 30, Issue 11, pp 2028–2037

Immune monitoring of patients with septic shock by measurement of intraleukocyte cytokines

Authors

  • Thierry Fumeaux
    • Laboratory of the Division of Medical Intensive Care, Departments of Internal Medicine and of Microbiology and Molecular MedicineUniversity Hospital and Faculty of Medicine
  • Julien Dufour
    • Laboratory of the Division of Medical Intensive Care, Departments of Internal Medicine and of Microbiology and Molecular MedicineUniversity Hospital and Faculty of Medicine
  • Sabine Stern
    • Laboratory of the Division of Medical Intensive Care, Departments of Internal Medicine and of Microbiology and Molecular MedicineUniversity Hospital and Faculty of Medicine
    • Laboratory of the Division of Medical Intensive Care, Departments of Internal Medicine and of Microbiology and Molecular MedicineUniversity Hospital and Faculty of Medicine
Original

DOI: 10.1007/s00134-004-2429-8

Cite this article as:
Fumeaux, T., Dufour, J., Stern, S. et al. Intensive Care Med (2004) 30: 2028. doi:10.1007/s00134-004-2429-8

Abstract

Objective

To assess the immune competence of patients presenting with septic shock by measuring on-line the production of intracellular cytokines by circulating leukocytes.

Design and setting

Prospective study in a 18-bed medical intensive care unit of a university hospital.

Patients and participants

21 patients with septic shock, and 11 volunteers.

Interventions

Single-step isolation of leukocytes from whole blood obtained within the first 24 h after admission. Leukocytes were fixed immediately or after treatment with lipopolysaccharide (LPS) and/or heterologous plasma.

Measurements and results

Leukocytes were permeabilized, and the intracellular cytokine expression of TNF-α and IL-10 was quantified by immunostaining and flow cytometry. LPS treatment significantly increased monocyte intracellular cytokine TNF-α and IL-10 as well as lymphocyte intracellular cytokine IL-10 in normal leukocytes. Septic monocytes and granulocytes had nonstimulated intracellular cytokine TNF-α concentrations lower than those measured in volunteers and were severely hyporesponsive to LPS. These phenotypic changes were correlated with disease severity and could be reproduced by treatment of normal leukocytes with plasma from patients with septic shock.

Conclusions

Intracellular cytokine staining is a simple and rapid method to assess in situ and on-line the inflammatory balance and responsiveness of leukocyte subpopulations and could therefore represent a useful monitoring tool to assess the immune competence of critically ill patients. This study identifies the cellular source of cytokines in whole blood and confirms prior reports showing that septic phagocytes are characterized by a predominant anti-inflammatory phenotype, with hyporesponsiveness to LPS, depending on a plasma deactivation factor.

Keywords

Septic shockFlow cytometryImmune paralysisTumor necrosis factor αInterleukin 10Leukocyte subpopulation

Copyright information

© Springer-Verlag 2004