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Open Access 01-12-2015 | Research

Human severe sepsis cytokine mixture increases β2-integrin-dependent polymorphonuclear leukocyte adhesion to cerebral microvascular endothelial cells in vitro

Authors: Chris Blom, Brittany L Deller, Douglas D Fraser, Eric K Patterson, Claudio M Martin, Bryan Young, Patricia C Liaw, Payam Yazdan-Ashoori, Angelica Ortiz, Brian Webb, Greg Kilmer, David E Carter, Gediminas Cepinskas

Published in: Critical Care | Issue 1/2015

Abstract

Introduction

Sepsis-associated encephalopathy (SAE) is a state of acute brain dysfunction in response to a systemic infection. We propose that systemic inflammation during sepsis causes increased adhesion of leukocytes to the brain microvasculature, resulting in blood-brain barrier dysfunction. Thus, our objectives were to measure inflammatory analytes in plasma of severe sepsis patients to create an experimental cytokine mixture (CM), and to use this CM to investigate the activation and interactions of polymorphonuclear leukocytes (PMN) and human cerebrovascular endothelial cells (hCMEC/D3) in vitro.

Methods

The concentrations of 41 inflammatory analytes were quantified in plasma obtained from 20 severe sepsis patients and 20 age- and sex-matched healthy controls employing an antibody microarray. Two CMs were prepared to mimic severe sepsis (SSCM) and control (CCM), and these CMs were then used for PMN and hCMEC/D3 stimulation in vitro. PMN adhesion to hCMEC/D3 was assessed under conditions of flow (shear stress 0.7 dyn/cm²).

Results

Eight inflammatory analytes elevated in plasma obtained from severe sepsis patients were used to prepare SSCM and CCM. Stimulation of PMN with SSCM led to a marked increase in PMN adhesion to hCMEC/D3, as compared to CCM. PMN adhesion was abolished with neutralizing antibodies to either β2 (CD18), α_L/β₂ (CD11α/CD18; LFA-1) or α_M/β₂ (CD11β/CD18; Mac-1) integrins. In addition, immune-neutralization of the endothelial (hCMEC/D3) cell adhesion molecule, ICAM-1 (CD54) also suppressed PMN adhesion.

Conclusions

Human SSCM up-regulates PMN pro-adhesive phenotype and promotes PMN adhesion to cerebrovascular endothelial cells through a β2-integrin-ICAM-1-dependent mechanism. PMN adhesion to the brain microvasculature may contribute to SAE.

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Title: Human severe sepsis cytokine mixture increases β2-integrin-dependent polymorphonuclear leukocyte adhesion to cerebral microvascular endothelial cells in vitro
Authors: Chris Blom
Brittany L Deller
Douglas D Fraser
Eric K Patterson
Claudio M Martin
Bryan Young
Patricia C Liaw
Payam Yazdan-Ashoori
Angelica Ortiz
Brian Webb
Greg Kilmer
David E Carter
Gediminas Cepinskas
Publication date: 01-12-2015
Publisher: BioMed Central
Published in: Critical Care / Issue 1/2015
Electronic ISSN: 1364-8535
DOI: https://doi.org/10.1186/s13054-015-0883-z

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Human severe sepsis cytokine mixture increases β2-integrin-dependent polymorphonuclear leukocyte adhesion to cerebral microvascular endothelial cells in vitro

Abstract

Introduction

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Introduction

Methods

Results

Conclusions

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