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Figure 2 | Critical Care

Figure 2

From: Increased serum soluble Fas after major trauma is associated with delayed neutrophil apoptosis and development of sepsis

Figure 2

Inhibition of neutrophil extrinsic apoptosis by sFas. (a) Reduced percentage of apoptotic neutrophils isolated from healthy controls (n = 15, white box), sepsis patients (n = 7, dark gray boxes) and nonsepsis patients (n = 13, light gray boxes) at day 1, day 5 and day 9 after trauma. Boxplots represent the median (heavy line in boxes) and the 25th and 75th percentiles (lower and upper lines of the box, respectively). Whiskers indicate the minimum and maximum values, respectively. *P < 0.05, **P <0.01, ***P < 0.001 vs. control group. (b) Neutrophils from healthy controls were incubated with 50 ng/mL anti-Fas antibody (CH-11) in the presence of serial dilutions of recombinant human soluble Fas (sFas) (range, 0 to 2 μg/mL) for 18 hours. Thereafter cells were lysed in hypotonic solution containing propidium iodide, and the percentage of apoptotic cells was determined by flow cytometry. Data (means ± SEM) from three independent experiments are presented. (c) Control neutrophils were incubated with 200 ng/mL agonistic anti-Fas antibodies (clone CH-11) and pooled serum from four sepsis patients immunoprecipitated by anti-Fas antibodies (clone ZB4) or not. After 18 hours of culture, apoptotic neutrophils with hypodiploid DNA content were quantified by propidium iodide staining and flow cytometry. Data (means ± SEM) from six independent experiments are depicted. (d) Representative histogram of CH-11-induced apoptosis in the presence of patient serum. Region M1 describes the percentage of hypodiploid DNA.

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