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Antimicrobial Peptides Are Highly Abundant and Active in Postoperative Pleural Drainage Fluids  Konrad Hoetzenecker, MD, PhD, Moritz Hochdaninger, MD,

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Presentation on theme: "Antimicrobial Peptides Are Highly Abundant and Active in Postoperative Pleural Drainage Fluids  Konrad Hoetzenecker, MD, PhD, Moritz Hochdaninger, MD,"— Presentation transcript:

1 Antimicrobial Peptides Are Highly Abundant and Active in Postoperative Pleural Drainage Fluids 
Konrad Hoetzenecker, MD, PhD, Moritz Hochdaninger, MD, Denise Traxler, Maria Gschwandtner, PhD, Mohammad Mahdi Kasiri, MD, Andreas Mitterbauer, MD, Thomas Schweiger, MD, Balazs Hegedus, MD, Walter Klepetko, MD, Erwin Tschachler, MD, Hendrik J. Ankersmit, MD, Michael Mildner, PhD  The Annals of Thoracic Surgery  Volume 98, Issue 3, Pages (September 2014) DOI: /j.athoracsur Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions

2 Fig 1 Study design and patient recruitment. (AB = antibiotic; ELISA = enzyme-linked immunosorbent assay; NP = normal pleura; PCR = polymerase chain reaction.) The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur ) Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions

3 Fig 2 Postoperative pleural fluids showed evidence of profound antimicrobial activity against different pathogens. (A) High bactericidal activity against gram-positive bacteria (Staphylococcus aureus, Streptococcus pneumoniae, and Streptococcus pyogenes) and gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa) was found in microdilution assays. (B) Levels of different antimicrobial peptides (AMPs) in postoperative pleural fluids (18 hours) were determined by enzyme-linked immunosorbent assay (ELISA). Human beta-defensin (HBD)1, HBD2, RNAse5, RNAse7, S100A7, S100A8/A9, and cathelicidin were highly abundant in the pleural effusion after major lung operations. The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur ) Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions

4 Fig 3 (A) Flow cytometry analyses revealed that the cellular content in postoperative fluids invariably consists of CD45+ cells. (B) The majority of these cells were granulocytes (> 90%), which were partly replaced by infiltrating monocytes within the first 36 hours after the operation. (C) Polymerase chain reaction (PCR) evaluations of pleural fluids (lavage), pleural specimens, and a pleural mesothelial cell culture (normal pleura (NP)-1 and NP-2) showed a predominantly pleural origin of S100A7 in contrast to a mainly leukocytic origin of human beta-defensin (HBD)1, RNAses, and S100A8/A9. (7-AAD = 7-amino-actinomycin-D.) The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur ) Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions

5 Fig 4 Immunohistochemical evaluation of pleural samples underlined the polymerase chain reaction (PCR) analyses. Pleural resident leukocytes showed a positive staining pattern for S100A8/A9, human beta-defensin (HBD)1, HBD2, and RNAses, whereas S100A7 was exclusively found in the epithelial lining of the pleural mesothelium (×200 magnification). The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur ) Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions

6 Fig 5 Expression of different antimicrobial peptides (AMPs) upon triggering with Toll-like receptor (TLR) ligands, interleukin (IL)-1β, and tumor necrosis factor (TNF)α was determined by polymerase chain reaction (PCR). AMP expression was largely independent of proinflammatory stimuli in the pleural epithelial normal pleura (NP)-1 cell culture. However, S100A7 production could be augmented by coincubation of NP-1 cells with TNFα (p = 0.029). (A) HBD1; (B) RNAse5; (C) RNAse7; (D) S100A7; (E) S100AB; (F) S100A9; and (G) cathelicidin. The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur ) Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions

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