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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...
Autores principales: | , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
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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. |
format | Online Article Text |
id | pubmed-5819246 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
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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