Site‐specific and endothelial‐mediated dysfunction of the alveolar‐capillary barrier in response to lipopolysaccharides

Infectious agents such as lipopolysaccharides (LPS) challenge the functional properties of the alveolar‐capillary barrier (ACB) in the lung. In this study, we analyse the site‐specific effects of LPS on the ACB and reveal the effects on the individual cell types and the ACB as a functional unit. Mon...

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Published inJournal of cellular and molecular medicine Vol. 22; no. 2; pp. 982 - 998
Main Authors Janga, Harshavardhan, Cassidy, Liam, Wang, Fanlu, Spengler, Dietmar, Oestern‐Fitschen, Stefanie, Krause, Martin F., Seekamp, Andreas, Tholey, Andreas, Fuchs, Sabine
Format Journal Article
LanguageEnglish
Published England John Wiley & Sons, Inc 01.02.2018
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Abstract Infectious agents such as lipopolysaccharides (LPS) challenge the functional properties of the alveolar‐capillary barrier (ACB) in the lung. In this study, we analyse the site‐specific effects of LPS on the ACB and reveal the effects on the individual cell types and the ACB as a functional unit. Monocultures of H441 epithelial cells and co‐cultures of H441 with endothelial cells cultured on Transwells® were treated with LPS from the apical or basolateral compartment. Barrier properties were analysed by the transepithelial electrical resistance (TEER), by transport assays, and immunostaining and assessment of tight junctional molecules at protein level. Furthermore, pro‐inflammatory cytokines and immune‐modulatory molecules were evaluated by ELISA and semiquantitative real‐time PCR. Liquid chromatography–mass spectrometry‐based proteomics (LS‐MS) was used to identify proteins and effector molecules secreted by endothelial cells in response to LPS. In co‐cultures treated with LPS from the basolateral compartment, we noticed a significant reduction of TEER, increased permeability and induction of pro‐inflammatory cytokines. Conversely, apical treatment did not affect the barrier. No changes were noticed in H441 monoculture upon LPS treatment. However, LPS resulted in an increased expression of pro‐inflammatory cytokines such as IL‐6 in OEC and in turn induced the reduction of TEER and an increase in SP‐A expression in H441 monoculture, and H441/OEC co‐cultures after LPS treatment from basolateral compartment. LS‐MS‐based proteomics revealed factors associated with LPS‐mediated lung injury such as ICAM‐1, VCAM‐1, Angiopoietin 2, complement factors and cathepsin S, emphasizing the role of epithelial–endothelial crosstalk in the ACB in ALI/ARDS.
AbstractList Infectious agents such as lipopolysaccharides (LPS) challenge the functional properties of the alveolar-capillary barrier (ACB) in the lung. In this study, we analyse the site-specific effects of LPS on the ACB and reveal the effects on the individual cell types and the ACB as a functional unit. Monocultures of H441 epithelial cells and co-cultures of H441 with endothelial cells cultured on Transwells® were treated with LPS from the apical or basolateral compartment. Barrier properties were analysed by the transepithelial electrical resistance (TEER), by transport assays, and immunostaining and assessment of tight junctional molecules at protein level. Furthermore, pro-inflammatory cytokines and immune-modulatory molecules were evaluated by ELISA and semiquantitative real-time PCR. Liquid chromatography-mass spectrometry-based proteomics (LS-MS) was used to identify proteins and effector molecules secreted by endothelial cells in response to LPS. In co-cultures treated with LPS from the basolateral compartment, we noticed a significant reduction of TEER, increased permeability and induction of pro-inflammatory cytokines. Conversely, apical treatment did not affect the barrier. No changes were noticed in H441 monoculture upon LPS treatment. However, LPS resulted in an increased expression of pro-inflammatory cytokines such as IL-6 in OEC and in turn induced the reduction of TEER and an increase in SP-A expression in H441 monoculture, and H441/OEC co-cultures after LPS treatment from basolateral compartment. LS-MS-based proteomics revealed factors associated with LPS-mediated lung injury such as ICAM-1, VCAM-1, Angiopoietin 2, complement factors and cathepsin S, emphasizing the role of epithelial-endothelial crosstalk in the ACB in ALI/ARDS.
Infectious agents such as lipopolysaccharides ( LPS ) challenge the functional properties of the alveolar‐capillary barrier ( ACB ) in the lung. In this study, we analyse the site‐specific effects of LPS on the ACB and reveal the effects on the individual cell types and the ACB as a functional unit. Monocultures of H441 epithelial cells and co‐cultures of H441 with endothelial cells cultured on Transwells ® were treated with LPS from the apical or basolateral compartment. Barrier properties were analysed by the transepithelial electrical resistance ( TEER ), by transport assays, and immunostaining and assessment of tight junctional molecules at protein level. Furthermore, pro‐inflammatory cytokines and immune‐modulatory molecules were evaluated by ELISA and semiquantitative real‐time PCR . Liquid chromatography–mass spectrometry‐based proteomics ( LS ‐ MS ) was used to identify proteins and effector molecules secreted by endothelial cells in response to LPS . In co‐cultures treated with LPS from the basolateral compartment, we noticed a significant reduction of TEER , increased permeability and induction of pro‐inflammatory cytokines. Conversely, apical treatment did not affect the barrier. No changes were noticed in H441 monoculture upon LPS treatment. However, LPS resulted in an increased expression of pro‐inflammatory cytokines such as IL ‐6 in OEC and in turn induced the reduction of TEER and an increase in SP ‐A expression in H441 monoculture, and H441/ OEC co‐cultures after LPS treatment from basolateral compartment. LS ‐ MS ‐based proteomics revealed factors associated with LPS ‐mediated lung injury such as ICAM ‐1, VCAM ‐1, Angiopoietin 2, complement factors and cathepsin S, emphasizing the role of epithelial–endothelial crosstalk in the ACB in ALI / ARDS .
Infectious agents such as lipopolysaccharides (LPS) challenge the functional properties of the alveolar-capillary barrier (ACB) in the lung. In this study, we analyse the site-specific effects of LPS on the ACB and reveal the effects on the individual cell types and the ACB as a functional unit. Monocultures of H441 epithelial cells and co-cultures of H441 with endothelial cells cultured on Transwells were treated with LPS from the apical or basolateral compartment. Barrier properties were analysed by the transepithelial electrical resistance (TEER), by transport assays, and immunostaining and assessment of tight junctional molecules at protein level. Furthermore, pro-inflammatory cytokines and immune-modulatory molecules were evaluated by ELISA and semiquantitative real-time PCR. Liquid chromatography-mass spectrometry-based proteomics (LS-MS) was used to identify proteins and effector molecules secreted by endothelial cells in response to LPS. In co-cultures treated with LPS from the basolateral compartment, we noticed a significant reduction of TEER, increased permeability and induction of pro-inflammatory cytokines. Conversely, apical treatment did not affect the barrier. No changes were noticed in H441 monoculture upon LPS treatment. However, LPS resulted in an increased expression of pro-inflammatory cytokines such as IL-6 in OEC and in turn induced the reduction of TEER and an increase in SP-A expression in H441 monoculture, and H441/OEC co-cultures after LPS treatment from basolateral compartment. LS-MS-based proteomics revealed factors associated with LPS-mediated lung injury such as ICAM-1, VCAM-1, Angiopoietin 2, complement factors and cathepsin S, emphasizing the role of epithelial-endothelial crosstalk in the ACB in ALI/ARDS.
Author Spengler, Dietmar
Cassidy, Liam
Fuchs, Sabine
Seekamp, Andreas
Wang, Fanlu
Tholey, Andreas
Janga, Harshavardhan
Oestern‐Fitschen, Stefanie
Krause, Martin F.
AuthorAffiliation 1 Department of Trauma Surgery and Orthopedics Experimental Trauma Surgery University Medical Center Schleswig‐Holstein Kiel Germany
2 Systematic Proteomics & Bioanalytics Institut für Experimentelle Medizin Christian‐Albrechts‐Universität zu Kiel Kiel Germany
3 Department of Pediatrics University Medical Center Schleswig‐ Holstein Kiel Germany
AuthorAffiliation_xml – name: 1 Department of Trauma Surgery and Orthopedics Experimental Trauma Surgery University Medical Center Schleswig‐Holstein Kiel Germany
– name: 2 Systematic Proteomics & Bioanalytics Institut für Experimentelle Medizin Christian‐Albrechts‐Universität zu Kiel Kiel Germany
– name: 3 Department of Pediatrics University Medical Center Schleswig‐ Holstein Kiel Germany
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ContentType Journal Article
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Keywords Liquid chromatography-mass spectrometry-based proteomics
lipopolysaccharides
alveolar-capillary barrier of the lung
epithelial-endothelial crosstalk
Language English
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SSID ssj0036139
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Snippet Infectious agents such as lipopolysaccharides (LPS) challenge the functional properties of the alveolar‐capillary barrier (ACB) in the lung. In this study, we...
Infectious agents such as lipopolysaccharides (LPS) challenge the functional properties of the alveolar-capillary barrier (ACB) in the lung. In this study, we...
Infectious agents such as lipopolysaccharides ( LPS ) challenge the functional properties of the alveolar‐capillary barrier ( ACB ) in the lung. In this study,...
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pubmed
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StartPage 982
SubjectTerms Adult
alveolar‐capillary barrier of the lung
Alveoli
Angiopoietin
Capillaries - drug effects
Capillaries - physiopathology
Cathepsin S
Caveolin 1 - metabolism
Cell Line
Cell Membrane Permeability - drug effects
Coculture Techniques
Cytokines
Electric Impedance
Endothelial cells
Endothelial Cells - drug effects
Endothelial Cells - metabolism
Endothelial Cells - pathology
Enzyme-linked immunosorbent assay
Epithelial cells
epithelial–endothelial crosstalk
Gene Expression Regulation - drug effects
Humans
Immunomodulation
Inflammation
Inflammation - pathology
Intercellular adhesion molecule 1
Interleukin 6
Lipopolysaccharides
Lipopolysaccharides - toxicity
Liquid chromatography
Liquid chromatography–mass spectrometry‐based proteomics
Lungs
Mass spectrometry
Mass spectroscopy
Monoculture
Original
Permeability
Phosphorylation - drug effects
Proteins
Proteomics
Pulmonary Alveoli - drug effects
Pulmonary Alveoli - physiopathology
Pulmonary Surfactant-Associated Proteins - metabolism
Tight Junction Proteins - metabolism
Vascular cell adhesion molecule 1
Zonula Occludens-1 Protein - metabolism
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Title Site‐specific and endothelial‐mediated dysfunction of the alveolar‐capillary barrier in response to lipopolysaccharides
URI https://onlinelibrary.wiley.com/doi/abs/10.1111%2Fjcmm.13421
https://www.ncbi.nlm.nih.gov/pubmed/29210175
https://www.proquest.com/docview/1990669085
https://search.proquest.com/docview/1973462232
https://pubmed.ncbi.nlm.nih.gov/PMC5783864
Volume 22
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