Human pulmonary endothelial cell permeability after exposure to LPS-stimulated leukocyte supernatants derived from patients with early sepsis

American Journal of Physiology. Lung Cellular and Molecular Physiology
Aleksandra LeligdowiczMichael A Matthay

Abstract

Systemic immune activation is the hallmark of sepsis, which can result in endothelial injury and the acute respiratory distress syndrome (ARDS). The aim of this study was to investigate heterogeneity in sepsis-mediated endothelial permeability using primary human pulmonary microvascular endothelial cells (HPMECs) and the electric cell-substrate impedance sensing (ECIS) platform. After plasma removal, cellular component of whole blood from 35 intensive care unit (ICU) patients with early sepsis was diluted with media and stimulated with either lipopolysaccharide (LPS) or control media. Resulting supernatants were cocultured with HPMECs seeded on ECIS plates, and resistance was continually measured. A decrease in resistance signified increased permeability. After incubation, HPMECs were detached and cell adhesion proteins were quantified using flow cytometry and immunohistochemistry, and gene expression was analyzed with quantitative PCR. Significant heterogeneity in endothelial permeability after exposure to supernatants of LPS-stimulated leukocytes was identified. ICU patients with sepsis stratified into one of the following three groups: minimal (9/35, 26%), intermediate (18/35, 51%), and maximal (8/35, 23%) permeability. Maxi...Continue Reading

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Citations

Jan 10, 2020·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Pulin CheQiang Ding
Aug 7, 2020·American Journal of Physiology. Lung Cellular and Molecular Physiology·Julie A Bastarache
Jun 24, 2021·American Journal of Physiology. Cell Physiology·Khadeja-Tul Kubra, Nektarios Barabutis

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Methods Mentioned

BETA
PCR
flow cytometry

Software Mentioned

FlowJo
GraphPad
Excel
STATA
Prism

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