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Mediators of Inflammation
Volume 2017 (2017), Article ID 4137563, 11 pages
https://doi.org/10.1155/2017/4137563
Research Article

Expression and Function of Granzymes A and B in Escherichia coli Peritonitis and Sepsis

1Center for Experimental and Molecular Medicine (CEMM), Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105 AZ Amsterdam, Netherlands
2Department of Immunopathology, Sanquin Research, Plesmanlaan 125, 1066 CX Amsterdam, Netherlands
3Department of Pathology, Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105 AZ Amsterdam, Netherlands
4Laboratory for Experimental Oncology and Radiobiology (LEXOR), Academic Medical Center, Meibergdreef 9, 1105 AZ Amsterdam, Netherlands
5Division of Infectious Diseases, Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105 AZ Amsterdam, Netherlands

Correspondence should be addressed to M. Isabel García-Laorden; se.oohay@lgmelahi

Received 12 February 2017; Revised 5 April 2017; Accepted 20 April 2017; Published 12 June 2017

Academic Editor: Mirella Giovarelli

Copyright © 2017 M. Isabel García-Laorden et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Supplementary Data. Supplementary Table 1: Percentage and median fluorescence intensity (MFI) of gzmA and gzmB in diverse lymphocyte populations from WT mice during E. coli peritonitis. Supplementary Table 2: Cell counts of leukocytes, NK cells and granzyme-positive NK cells in peritoneal lavage fluid from wild-type mice during E. coli peritonitis. Supplementary Table 3: Cytokines and chemokine plasma levels of wild-type, gzmA-/-, gzmB-/- and gzmAxB-/- mice during E. coli peritonitis. Supplementary Figure 1: Gating strategy for the analysis of the expression of granzymes A and B by lymphocyte populations in wild-type mice. Intracellular expression of gzmA and B by lymphocyte populations was analysed in peritoneal lavage fluid (PLF) and blood from wild-type mice by flow cytometry. Leukocytes region was gated on the basis of forward (FSC) and side scattering (SSC) characteristics. A. CD8+ T (CD3+CD8+), CD4+ T (CD3+CD4+), γδ T (CD3+γδ TCR+), NK1.1+ T (CD3+NK1.1+) and NK (CD3-NK1.1+) cells were identified by dot-plots, and the percentage of gzm+ cells in each lymphocyte population as well as the median fluorescence intensity (MFI) of the positive expression were determined in histogram plots. B. GzmA+ and gzmB+ cells were identified by histogram plots, and the percentage of cells corresponding to each lymphocyte population within the gzm+ cells were determined in dot-plots. Data shown are of blood from a representative individual (gating of PLF samples was done similarly as for blood). Supplementary Figure 2: Histopathology of liver and lung from wild-type, gzmA-/-, gzmB-/- and gzmAxB-/- mice during E. coli peritonitis. Mice were infected intraperitoneally with 1.3*104 CFU E. coli and sacrificed at 6, 14 and 20h after infection. Data are box-and-whisker diagrams depicting the smallest observation, lower quartile, median, upper quartile and largest observation. N = 7-8 per group at each time point. * P<0.05, ** P<0.01 determined by Mann-Whitney U test.

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