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Strain-dependent effects of clinical echovirus 30 outbreak isolates at the blood-CSF barrier

BACKGROUND: Echovirus (E) 30 (E-30) meningitis is characterized by neuroinflammation involving immune cell pleocytosis at the protective barriers of the central nervous system (CNS). In this context, infection of the blood-cerebrospinal fluid barrier (BCSFB), which has been demonstrated to be involv...

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Autores principales: Dahm, Tobias, Adams, Ortwin, Boettcher, Sindy, Diedrich, Sabine, Morozov, Vasily, Hansman, Grant, Fallier-Becker, Petra, Schädler, Sebastian, Burkhardt, Claus J., Weiss, Christel, Stump-Guthier, Carolin, Ishikawa, Hiroshi, Schroten, Horst, Schwerk, Christian, Tenenbaum, Tobias, Rudolph, Henriette
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5819246/
https://www.ncbi.nlm.nih.gov/pubmed/29463289
http://dx.doi.org/10.1186/s12974-018-1061-4
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author Dahm, Tobias
Adams, Ortwin
Boettcher, Sindy
Diedrich, Sabine
Morozov, Vasily
Hansman, Grant
Fallier-Becker, Petra
Schädler, Sebastian
Burkhardt, Claus J.
Weiss, Christel
Stump-Guthier, Carolin
Ishikawa, Hiroshi
Schroten, Horst
Schwerk, Christian
Tenenbaum, Tobias
Rudolph, Henriette
author_facet Dahm, Tobias
Adams, Ortwin
Boettcher, Sindy
Diedrich, Sabine
Morozov, Vasily
Hansman, Grant
Fallier-Becker, Petra
Schädler, Sebastian
Burkhardt, Claus J.
Weiss, Christel
Stump-Guthier, Carolin
Ishikawa, Hiroshi
Schroten, Horst
Schwerk, Christian
Tenenbaum, Tobias
Rudolph, Henriette
author_sort Dahm, Tobias
collection PubMed
description BACKGROUND: Echovirus (E) 30 (E-30) meningitis is characterized by neuroinflammation involving immune cell pleocytosis at the protective barriers of the central nervous system (CNS). In this context, infection of the blood-cerebrospinal fluid barrier (BCSFB), which has been demonstrated to be involved in enteroviral CNS pathogenesis, may affect the tight junction (TJ) and adherens junction (AJ) function and morphology. METHODS: We used an in vitro human choroid plexus epithelial (HIBCPP) cell model to investigate the effect of three clinical outbreak strains (13-311, 13-759, and 14-397) isolated in Germany in 2013, and compared them to E-30 Bastianni. Conducting transepithelial electrical resistance (TEER), paracellular dextran flux measurement, quantitative real-time polymerase chain reaction (qPCR), western blot, and immunofluorescence analysis, we investigated TJ and AJ function and morphology as well as strain-specific E-30 infection patterns. Additionally, transmission electron and focused ion beam microscopy electron microscopy (FIB-SEM) was used to evaluate the mode of leukocyte transmigration. Genome sequencing and phylogenetic analyses were performed to discriminate potential genetic differences among the outbreak strains. RESULTS: We observed a significant strain-dependent decrease in TEER with strains E-30 Bastianni and 13-311, whereas paracellular dextran flux was only affected by E-30 Bastianni. Despite strong similarities among the outbreak strains in replication characteristics and particle distribution, strain 13-311 was the only outbreak isolate revealing comparable disruptive effects on TJ (Zonula Occludens (ZO) 1 and occludin) and AJ (E-cadherin) morphology to E-30 Bastianni. Notwithstanding significant junctional alterations upon E-30 infection, we observed both para- and transcellular leukocyte migration across HIBCPP cells. Complete genome sequencing revealed differences between the strains analyzed, but no explicit correlation with the observed strain-dependent effects on HIBCPP cells was possible. CONCLUSION: The findings revealed distinct E-30 strain-specific effects on barrier integrity and junctional morphology. Despite E-30-induced barrier alterations leukocyte trafficking did not exclusively occur via the paracellular route. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12974-018-1061-4) contains supplementary material, which is available to authorized users.
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spelling pubmed-58192462018-02-21 Strain-dependent effects of clinical echovirus 30 outbreak isolates at the blood-CSF barrier Dahm, Tobias Adams, Ortwin Boettcher, Sindy Diedrich, Sabine Morozov, Vasily Hansman, Grant Fallier-Becker, Petra Schädler, Sebastian Burkhardt, Claus J. Weiss, Christel Stump-Guthier, Carolin Ishikawa, Hiroshi Schroten, Horst Schwerk, Christian Tenenbaum, Tobias Rudolph, Henriette J Neuroinflammation Research BACKGROUND: Echovirus (E) 30 (E-30) meningitis is characterized by neuroinflammation involving immune cell pleocytosis at the protective barriers of the central nervous system (CNS). In this context, infection of the blood-cerebrospinal fluid barrier (BCSFB), which has been demonstrated to be involved in enteroviral CNS pathogenesis, may affect the tight junction (TJ) and adherens junction (AJ) function and morphology. METHODS: We used an in vitro human choroid plexus epithelial (HIBCPP) cell model to investigate the effect of three clinical outbreak strains (13-311, 13-759, and 14-397) isolated in Germany in 2013, and compared them to E-30 Bastianni. Conducting transepithelial electrical resistance (TEER), paracellular dextran flux measurement, quantitative real-time polymerase chain reaction (qPCR), western blot, and immunofluorescence analysis, we investigated TJ and AJ function and morphology as well as strain-specific E-30 infection patterns. Additionally, transmission electron and focused ion beam microscopy electron microscopy (FIB-SEM) was used to evaluate the mode of leukocyte transmigration. Genome sequencing and phylogenetic analyses were performed to discriminate potential genetic differences among the outbreak strains. RESULTS: We observed a significant strain-dependent decrease in TEER with strains E-30 Bastianni and 13-311, whereas paracellular dextran flux was only affected by E-30 Bastianni. Despite strong similarities among the outbreak strains in replication characteristics and particle distribution, strain 13-311 was the only outbreak isolate revealing comparable disruptive effects on TJ (Zonula Occludens (ZO) 1 and occludin) and AJ (E-cadherin) morphology to E-30 Bastianni. Notwithstanding significant junctional alterations upon E-30 infection, we observed both para- and transcellular leukocyte migration across HIBCPP cells. Complete genome sequencing revealed differences between the strains analyzed, but no explicit correlation with the observed strain-dependent effects on HIBCPP cells was possible. CONCLUSION: The findings revealed distinct E-30 strain-specific effects on barrier integrity and junctional morphology. Despite E-30-induced barrier alterations leukocyte trafficking did not exclusively occur via the paracellular route. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12974-018-1061-4) contains supplementary material, which is available to authorized users. BioMed Central 2018-02-20 /pmc/articles/PMC5819246/ /pubmed/29463289 http://dx.doi.org/10.1186/s12974-018-1061-4 Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Dahm, Tobias
Adams, Ortwin
Boettcher, Sindy
Diedrich, Sabine
Morozov, Vasily
Hansman, Grant
Fallier-Becker, Petra
Schädler, Sebastian
Burkhardt, Claus J.
Weiss, Christel
Stump-Guthier, Carolin
Ishikawa, Hiroshi
Schroten, Horst
Schwerk, Christian
Tenenbaum, Tobias
Rudolph, Henriette
Strain-dependent effects of clinical echovirus 30 outbreak isolates at the blood-CSF barrier
title Strain-dependent effects of clinical echovirus 30 outbreak isolates at the blood-CSF barrier
title_full Strain-dependent effects of clinical echovirus 30 outbreak isolates at the blood-CSF barrier
title_fullStr Strain-dependent effects of clinical echovirus 30 outbreak isolates at the blood-CSF barrier
title_full_unstemmed Strain-dependent effects of clinical echovirus 30 outbreak isolates at the blood-CSF barrier
title_short Strain-dependent effects of clinical echovirus 30 outbreak isolates at the blood-CSF barrier
title_sort strain-dependent effects of clinical echovirus 30 outbreak isolates at the blood-csf barrier
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5819246/
https://www.ncbi.nlm.nih.gov/pubmed/29463289
http://dx.doi.org/10.1186/s12974-018-1061-4
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