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Background: Immune cell dysfunction plays a central role in sepsis-associated immune paralysis. The transfusion of healthy donor immune cells, i.e., granulocyte concentrates (GC) potentially induces tissue damage via local effects of neutrophils. Initial clinical trials using standard donor GC in a strictly extracorporeal bioreactor system for treatment of septic shock patients already provided evidence for beneficial effects with fewer side effects, by separating patient and donor immune cells using plasma filters. In this ex vivo study, we demonstrate the functional characteristics of a simplified extracorporeal therapy system using purified granulocyte preparations. Methods: Purified GC were used in an immune cell perfusion model prefilled with human donor plasma simulating a 6-h treatment. The extracorporeal circuit consisted of a blood circuit and a plasma circuit with 3 plasma filters (PF). PF1 is separating the plasma from the patient's blood. Plasma is then perfused through PF2 containing donor immune cells and used in a dead-end mode. The filtrated plasma is finally retransfused to the blood circuit. PF3 is included in the plasma backflow as a redundant safety measure. The donor immune cells are retained in the extracorporeal system and discarded after treatment. Phagocytosis activity, oxidative burst and cell viability as well as cytokine release and metabolic parameters of purified GCs were assessed. Results: Cells were viable throughout the study period and exhibited well-preserved functionality and efficient metabolic activity. Course of lactate dehydrogenase and free hemoglobin concentration yielded no indication of cell impairment. The capability of the cells to secret various cytokines was preserved. Of particular interest is equivalence in performance of the cells on day 1 and day 3, demonstrating the sustained shelf life and performance of the immune cells in the purified GCs. Conclusion: Results demonstrate the suitability of a simplified extracorporeal system. Furthermore, granulocytes remain viable and highly active during a 6-h treatment even after storage for 3 days supporting the treatment of septic patients with this system in advanced clinical trials. [ABSTRACT FROM AUTHOR]

Objective and design: The development of liver failure is a major problem in septic patients. In this prospective clinical experimental study the hepatotoxicity of plasma from septic and non-septic patients was tested. Methods and subjects: The basic test components consist of human liver cells (HepG2/C3A) used in a standardized microtiter plate assay. After incubation with patient's plasma viability of cells (XTT-test), the cytochrome 1A2 activity and synthesis of micro albumin were measured. Subjects (28) enrolled comprise the septic shock group (SSG, n = 10), the non-septic group (NSG, n = 5) and the healthy volunteers group (HVG, n = 13). Results: The 28-day mortality was 30% in the SSG. The APACHE II-, SOFA-, and SAPS-scores and the values of bilirubin and prothrombin time as INR were significantly higher in the SSG than in the NSG. The cytochrome 1A2 activity and the release of albumin were significantly reduced in HepG2/C3A cells incubated with plasma of the SSG ( p < 0.05). The cytochrome 1A2 activities were higher in survivors compared to non-survivors at the time point 0 and were increasing in survivors and decreasing in non-survivors within 54 h in the SSG. In the SSG there was a significant decrease in IL-10 and IL-8 between inclusion and 54 h. Values of IL-6, TNF alpha and IL-10 were significantly lower in the NSG compared with the values of the SSG at inclusion and after 54 h. Conclusion: The plasma of patients with septic shock impaired cellular functions of HepG2/C3A cells. [ABSTRACT FROM AUTHOR]

The predictive value of circulating free DNA/neutrophil extracellular traps (cf-DNA/NETs) has recently been shown in patients with major trauma for sepsis, multiple organ failure, and mortality. Here we report on the predictive potential of cf-DNA/NETs for mortality in patients with severe burn injury. In a prospective study 32 patients with severe burn injury were included. Blood samples were sequentially obtained on day 1, 3, 5, and 7 after admission. cf-DNA/NETs was directly quantified from plasma by means of rapid fluorescence assay. Time kinetics of cf-DNA/NETs were correlated with clinical data, C-reactive protein (CRP), procalcitonin (PCT), and interleukin (IL)-6. Furthermore sensitivity, specificity, and positive and negative predictive value, as well as receiver operation characteristic (ROC) curves were calculated. Seven patients died within the first month after burn injury. cf-DNA/NETs values from these patients were significantly increased already on day 1 and 3 after admission compared with patients who survived ( p < 0.01). In contrast, PCT levels of nonsurvivors were significantly elevated on day 3 and 5 ( p < 0.01), while CRP and IL-6 did not show any significant difference between survivors and nonsurvivors. At a cutoff of 255 ng/ml, cf-DNA/NETs had sensitivity of 0.8 and specificity of 0.74. ROC revealed largest areas under the curve (AUC) for cf-DNA/NETs on day 1 (0.851) and 3 (0.883) after admission. For all values between day 1 and 7, AUC was 0.815. cf-DNA/NETs seems to be a rapid, valuable marker for prediction of mortality in burn patients. A larger confirmation trial ought to be carried out. [ABSTRACT FROM AUTHOR]

Neutrophil extracellular traps (NETs) have recently been discovered as a central part of antimicrobial innate immunity. In the meanwhile, evidence accumulated that NETs are also generated upon non-infectious stimuli in various clinical settings. In acute or chronic inflammatory disorders aberrantly enhanced NET formation and/or decreased NET degradation seems to correlate with disease outcome. This review summarizes current knowledge about the relation of NETs in a broad spectrum of clinical settings. Specifically, we focus on the importance of NETs as a predictive marker in severely ill patients and further, we speculate about the potential pathophysiology of NETs. [ABSTRACT FROM AUTHOR]