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. 2014 Jun 2;9(6):e97451. doi: 10.1371/journal.pone.0097451

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© 2014 Beyer et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

PMC Copyright notice

A) Spleens, livers and lungs of septic saline-treated (CASP+saline) and septic TRAIL-treated (CASP+TRAIL) were analyzed 20 h after induction of CASP. Sections were stained for Ly6G. Ly6G-positive cells of respective organs (n = 5/group for each organ) were counted in three HPFs and the mean was calculated. The number of neutrophils per HPF is depicted. Box plots and outliers are shown. The infiltration of neutrophils within the septic organs is significantly decreased by TRAIL-treatment in sepsis. Results are representative of two independent experiments. B) The number of apoptotic cells within the spleen, liver and lungs was determined by immunohistochemistry (n = 5/group for each organ, mean of 3 HPFs). TUNEL-straining was performed 20 hours after CASP. Box plots and outliers are shown. TRAIL-treatment decreased the number of apoptotic cells. Results are representative of two experiments performed independently. C) Apoptotic neutrophils were detected by staining Ly6G and TUNEL. The number of apoptotic neutrophils within the respective septic organs 20 hrs after induction of CASP was counted in three HPFs and the mean was calculated (n = 5/group for each organ). Additionally, the number of total apoptotic cells per HPF was counted. The ratio of apoptotic neutrophils over all apoptotic cells was calculated for each HPF. Box plots and outliers are depicted. TRAIL-treatment increased the fraction of apoptotic neutrophils 20 hrs after induction of CASP within the septic organs. D) Representative immunohistochemical analysis of Ly6G (green) and TUNEL (red) in spleens of septic mice 20 hrs after induction of CASP with (right) and without (left) TRAIL-treatment. Apoptotic neutrophils appear yellow. *p<0.05.