title: Immune monitoring of patients with septic shock by measurement of intraleukocyte cytokines.
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.
  
  
  METHODS
  Prospective study in a 18-bed medical intensive care unit of a university hospital.
  
  
  METHODS
  21 patients with septic shock, and 11 volunteers.
  
  
  METHODS
  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.
  
  
  RESULTS
  Leukocytes were permeabilized, and the intracellular cytokine expression of TNF-alpha and IL-10 was quantified by immunostaining and flow cytometry. LPS treatment significantly increased monocyte intracellular cytokine TNF-alpha and IL-10 as well as lymphocyte intracellular cytokine IL-10 in normal leukocytes. Septic monocytes and granulocytes had nonstimulated intracellular cytokine TNF-alpha 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.
authors.affiliation: Department of Internal Medicine, University Hospital and Faculty of Medicine, 24 rue Micheli-du-Crest, 1211 Geneva 14, Switzerland.
authors.forename: Thierry
authors.initials: T
authors.lastname: Fumeaux
authors.forename: Julien
authors.initials: J
authors.lastname: Dufour
authors.forename: Sabine
authors.initials: S
authors.lastname: Stern
authors.forename: Jérôme
authors.initials: J
authors.lastname: Pugin
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000704:Analysis of Variance; D015415:Biomarkers; D016022:Case-Control Studies; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D016753:Interleukin-10 / Q000097:blood*; D007962:Leukocytes / Q000276:immunology*; D008070:Lipopolysaccharides; D008297:Male; D008875:Middle Aged; D011237:Predictive Value of Tests; D011446:Prospective Studies; D012772:Shock, Septic / Q000175:diagnosis* / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10