Limits...
Early down-regulation of the pro-inflammatory potential of monocytes is correlated to organ dysfunction in patients after severe multiple injury: a cohort study.

Kirchhoff C, Biberthaler P, Mutschler WE, Faist E, Jochum M, Zedler S - Crit Care (2009)

Bottom Line: Whole blood was challenged with lipopolysaccharide (LPS) and subsequently analyzed for intracellular monocyte-related TNF-alpha, IL-1beta, IL-6, and IL-8.The degree of organ dysfunction was assessed using the multiple organ dysfunction syndrome (MODS)-score of Marshall on admission, 24 hours and 72 hours after injury.In contrast to the initial injury severity, there was a significant correlation detectable between the clinical signs of multiple organ dysfunction and the ex vivo cytokine response.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Orthopaedic Surgery and Traumatology, Klinikum Rechts der Isar, Technische Universitaet, 81675 Munich, Germany. chlodwig.kirchhoff@mac.com

ABSTRACT

Introduction: Severe tissue trauma results in a general inflammatory immune response (SIRS) representing an overall inflammatory reaction of the immune system. However, there is little known about the functional alterations of monocytes in the early posttraumatic phase, characterized by the battle of the individual with the initial trauma.

Methods: Thirteen patients with severe multiple injury; injury severity score (ISS) >16 points (17 to 57) were included. The cytokine synthesis profiles of monocytes were characterized on admission, and followed up 6, 12, 24, 48, and 72 hours after severe multiple injury using flow cytometry. Whole blood was challenged with lipopolysaccharide (LPS) and subsequently analyzed for intracellular monocyte-related TNF-alpha, IL-1beta, IL-6, and IL-8. The degree of organ dysfunction was assessed using the multiple organ dysfunction syndrome (MODS)-score of Marshall on admission, 24 hours and 72 hours after injury.

Results: Our data clearly show that the capacity of circulating monocytes to produce these mediators de novo was significantly diminished very early reaching a nadir 24 hours after severe injury followed by a rapid and nearly complete recovery another 48 hours later compared with admission and controls, respectively. In contrast to the initial injury severity, there was a significant correlation detectable between the clinical signs of multiple organ dysfunction and the ex vivo cytokine response.

Conclusions: As our data derived from very narrow intervals of measurements, they might contribute to a more detailed understanding of the early immune alterations recognized after severe trauma. It can be concluded that indeed as previously postulated an immediate hyperactivation of circulating monocytes is rapidly followed by a substantial paralysis of cell function. Moreover, our findings clearly demonstrate that the restricted capacity of monocytes to produce proinflammatory cytokines after severe injury is not only an in vitro phenomenon but also undistinguishable associated with the onset of organ dysfunction in the clinical scenario.

Show MeSH

Related in: MedlinePlus

Severe multiple injury results in a rapid decline of intracellular cytokine synthesis by monocytes within the first 24 hours after trauma. Cytokine de novo synthesis capacity was determined using an ex vivo whole blood approach in response to lipopolysaccharide (LPS). Results are calculated as percentage of cytokine positive CD14+ monocytes. Blood samples were drawn on admission, 6, 12, 24, 48, and 72 hours post trauma. Data are given as boxplots (median, 5th, 95th percentile). n = 13 patients (grey), n = 8 controls (white). # P < 0.05 vs. control group; * P < 0.05 vs. admission values.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2717459&req=5

Figure 1: Severe multiple injury results in a rapid decline of intracellular cytokine synthesis by monocytes within the first 24 hours after trauma. Cytokine de novo synthesis capacity was determined using an ex vivo whole blood approach in response to lipopolysaccharide (LPS). Results are calculated as percentage of cytokine positive CD14+ monocytes. Blood samples were drawn on admission, 6, 12, 24, 48, and 72 hours post trauma. Data are given as boxplots (median, 5th, 95th percentile). n = 13 patients (grey), n = 8 controls (white). # P < 0.05 vs. control group; * P < 0.05 vs. admission values.

Mentions: For flow cytometric analysis an Epics™ XL MCL flow cytometer (Beckman-Coulter, Krefeld, Germany) was used, fitted with an air-cooled 15 mW 488 nm argon ion laser, and filter settings for FITC (525 nm), PE (575 nm), and PE-Cy5 emitting in the deep red (675 nm). Data acquisition on the flow cytometer was obtained with System II™ Software (Beckman-Coulter, Krefeld, Germany). After appropriate instrument settings and spectral compensations, instrument alignment and fluidics were regularly verified using FlowCheck™ beads (Beckman-Coulter, Krefeld, Germany). A minimum of 5000 events was computed in list mode using log-amplified fluorescence signals and linearly amplified side- and forward-scatter signals. The data were analyzed using free WinMDI™ Software (Version 2.8, Bio-Soft Net [17]). A gate was set around the monocyte population, which was most strongly positive for CD14 on side scatter versus PE-Cy5 (CD14) dot plots, in order to exclude lymphocytes and debris from data analysis. Histograms representing the mean fluorescence intensity (MFI) of unstimulated cells were used as a guide for setting cutoff markers to delineate positive and negative populations. Even MFI of CD14 surface receptor expression was determined by a logarithmic scale, whereas results of cytokine de novo synthesis are shown as percentage of cytokine containing CD14+ monocytes after ex vivo stimulation with LPS (Figures 1 and 2).


Early down-regulation of the pro-inflammatory potential of monocytes is correlated to organ dysfunction in patients after severe multiple injury: a cohort study.

Kirchhoff C, Biberthaler P, Mutschler WE, Faist E, Jochum M, Zedler S - Crit Care (2009)

Severe multiple injury results in a rapid decline of intracellular cytokine synthesis by monocytes within the first 24 hours after trauma. Cytokine de novo synthesis capacity was determined using an ex vivo whole blood approach in response to lipopolysaccharide (LPS). Results are calculated as percentage of cytokine positive CD14+ monocytes. Blood samples were drawn on admission, 6, 12, 24, 48, and 72 hours post trauma. Data are given as boxplots (median, 5th, 95th percentile). n = 13 patients (grey), n = 8 controls (white). # P < 0.05 vs. control group; * P < 0.05 vs. admission values.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2717459&req=5

Figure 1: Severe multiple injury results in a rapid decline of intracellular cytokine synthesis by monocytes within the first 24 hours after trauma. Cytokine de novo synthesis capacity was determined using an ex vivo whole blood approach in response to lipopolysaccharide (LPS). Results are calculated as percentage of cytokine positive CD14+ monocytes. Blood samples were drawn on admission, 6, 12, 24, 48, and 72 hours post trauma. Data are given as boxplots (median, 5th, 95th percentile). n = 13 patients (grey), n = 8 controls (white). # P < 0.05 vs. control group; * P < 0.05 vs. admission values.
Mentions: For flow cytometric analysis an Epics™ XL MCL flow cytometer (Beckman-Coulter, Krefeld, Germany) was used, fitted with an air-cooled 15 mW 488 nm argon ion laser, and filter settings for FITC (525 nm), PE (575 nm), and PE-Cy5 emitting in the deep red (675 nm). Data acquisition on the flow cytometer was obtained with System II™ Software (Beckman-Coulter, Krefeld, Germany). After appropriate instrument settings and spectral compensations, instrument alignment and fluidics were regularly verified using FlowCheck™ beads (Beckman-Coulter, Krefeld, Germany). A minimum of 5000 events was computed in list mode using log-amplified fluorescence signals and linearly amplified side- and forward-scatter signals. The data were analyzed using free WinMDI™ Software (Version 2.8, Bio-Soft Net [17]). A gate was set around the monocyte population, which was most strongly positive for CD14 on side scatter versus PE-Cy5 (CD14) dot plots, in order to exclude lymphocytes and debris from data analysis. Histograms representing the mean fluorescence intensity (MFI) of unstimulated cells were used as a guide for setting cutoff markers to delineate positive and negative populations. Even MFI of CD14 surface receptor expression was determined by a logarithmic scale, whereas results of cytokine de novo synthesis are shown as percentage of cytokine containing CD14+ monocytes after ex vivo stimulation with LPS (Figures 1 and 2).

Bottom Line: Whole blood was challenged with lipopolysaccharide (LPS) and subsequently analyzed for intracellular monocyte-related TNF-alpha, IL-1beta, IL-6, and IL-8.The degree of organ dysfunction was assessed using the multiple organ dysfunction syndrome (MODS)-score of Marshall on admission, 24 hours and 72 hours after injury.In contrast to the initial injury severity, there was a significant correlation detectable between the clinical signs of multiple organ dysfunction and the ex vivo cytokine response.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Orthopaedic Surgery and Traumatology, Klinikum Rechts der Isar, Technische Universitaet, 81675 Munich, Germany. chlodwig.kirchhoff@mac.com

ABSTRACT

Introduction: Severe tissue trauma results in a general inflammatory immune response (SIRS) representing an overall inflammatory reaction of the immune system. However, there is little known about the functional alterations of monocytes in the early posttraumatic phase, characterized by the battle of the individual with the initial trauma.

Methods: Thirteen patients with severe multiple injury; injury severity score (ISS) >16 points (17 to 57) were included. The cytokine synthesis profiles of monocytes were characterized on admission, and followed up 6, 12, 24, 48, and 72 hours after severe multiple injury using flow cytometry. Whole blood was challenged with lipopolysaccharide (LPS) and subsequently analyzed for intracellular monocyte-related TNF-alpha, IL-1beta, IL-6, and IL-8. The degree of organ dysfunction was assessed using the multiple organ dysfunction syndrome (MODS)-score of Marshall on admission, 24 hours and 72 hours after injury.

Results: Our data clearly show that the capacity of circulating monocytes to produce these mediators de novo was significantly diminished very early reaching a nadir 24 hours after severe injury followed by a rapid and nearly complete recovery another 48 hours later compared with admission and controls, respectively. In contrast to the initial injury severity, there was a significant correlation detectable between the clinical signs of multiple organ dysfunction and the ex vivo cytokine response.

Conclusions: As our data derived from very narrow intervals of measurements, they might contribute to a more detailed understanding of the early immune alterations recognized after severe trauma. It can be concluded that indeed as previously postulated an immediate hyperactivation of circulating monocytes is rapidly followed by a substantial paralysis of cell function. Moreover, our findings clearly demonstrate that the restricted capacity of monocytes to produce proinflammatory cytokines after severe injury is not only an in vitro phenomenon but also undistinguishable associated with the onset of organ dysfunction in the clinical scenario.

Show MeSH
Related in: MedlinePlus