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Retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach

BACKGROUND: Blood-retinal barrier cells are known to exhibit a massive phenotypic change during experimental autoimmune uveitis (EAU) development. In an attempt to investigate the mechanisms of blood-retinal barrier (BRB) breakdown at a global level, we studied the gene regulation of total retinal c...

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Autores principales: Lipski, Deborah A., Foucart, Vincent, Dewispelaere, Rémi, Caspers, Laure E., Defrance, Matthieu, Bruyns, Catherine, Willermain, François
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7076950/
https://www.ncbi.nlm.nih.gov/pubmed/32183784
http://dx.doi.org/10.1186/s12886-020-1333-5
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author Lipski, Deborah A.
Foucart, Vincent
Dewispelaere, Rémi
Caspers, Laure E.
Defrance, Matthieu
Bruyns, Catherine
Willermain, François
author_facet Lipski, Deborah A.
Foucart, Vincent
Dewispelaere, Rémi
Caspers, Laure E.
Defrance, Matthieu
Bruyns, Catherine
Willermain, François
author_sort Lipski, Deborah A.
collection PubMed
description BACKGROUND: Blood-retinal barrier cells are known to exhibit a massive phenotypic change during experimental autoimmune uveitis (EAU) development. In an attempt to investigate the mechanisms of blood-retinal barrier (BRB) breakdown at a global level, we studied the gene regulation of total retinal cells and retinal endothelial cells during non-infectious uveitis. METHODS: Retinal endothelial cells were isolated by flow cytometry either in Tie2-GFP mice (CD31(+) CD45(−) GFP(+) cells), or in wild type C57BL/6 mice (CD31(+) CD45(−) endoglin(+) cells). EAU was induced in C57BL/6 mice by adoptive transfer of IRBP1–20-specific T cells. Total retinal cells and retinal endothelial cells from naïve and EAU mice were sorted and their gene expression compared by RNA-Seq. Protein expression of selected genes was validated by immunofluorescence on retinal wholemounts and cryosections and by flow cytometry. RESULTS: Retinal endothelial cell sorting in wild type C57BL/6 mice was validated by comparative transcriptome analysis with retinal endothelial cells sorted from Tie2-GFP mice, which express GFP under the control of the endothelial-specific receptor tyrosine kinase promoter Tie2. RNA-Seq analysis of total retinal cells mainly brought to light upregulation of genes involved in antigen presentation and T cell activation during EAU. Specific transcriptome analysis of retinal endothelial cells allowed us to identify 82 genes modulated in retinal endothelial cells during EAU development. Protein expression of 5 of those genes (serpina3n, lcn2, ackr1, lrg1 and lamc3) was validated at the level of inner BRB cells. CONCLUSION: Those data not only confirm the involvement of known pathogenic molecules but further provide a list of new candidate genes and pathways possibly implicated in inner BRB breakdown during non-infectious posterior uveitis.
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spelling pubmed-70769502020-03-18 Retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach Lipski, Deborah A. Foucart, Vincent Dewispelaere, Rémi Caspers, Laure E. Defrance, Matthieu Bruyns, Catherine Willermain, François BMC Ophthalmol Research Article BACKGROUND: Blood-retinal barrier cells are known to exhibit a massive phenotypic change during experimental autoimmune uveitis (EAU) development. In an attempt to investigate the mechanisms of blood-retinal barrier (BRB) breakdown at a global level, we studied the gene regulation of total retinal cells and retinal endothelial cells during non-infectious uveitis. METHODS: Retinal endothelial cells were isolated by flow cytometry either in Tie2-GFP mice (CD31(+) CD45(−) GFP(+) cells), or in wild type C57BL/6 mice (CD31(+) CD45(−) endoglin(+) cells). EAU was induced in C57BL/6 mice by adoptive transfer of IRBP1–20-specific T cells. Total retinal cells and retinal endothelial cells from naïve and EAU mice were sorted and their gene expression compared by RNA-Seq. Protein expression of selected genes was validated by immunofluorescence on retinal wholemounts and cryosections and by flow cytometry. RESULTS: Retinal endothelial cell sorting in wild type C57BL/6 mice was validated by comparative transcriptome analysis with retinal endothelial cells sorted from Tie2-GFP mice, which express GFP under the control of the endothelial-specific receptor tyrosine kinase promoter Tie2. RNA-Seq analysis of total retinal cells mainly brought to light upregulation of genes involved in antigen presentation and T cell activation during EAU. Specific transcriptome analysis of retinal endothelial cells allowed us to identify 82 genes modulated in retinal endothelial cells during EAU development. Protein expression of 5 of those genes (serpina3n, lcn2, ackr1, lrg1 and lamc3) was validated at the level of inner BRB cells. CONCLUSION: Those data not only confirm the involvement of known pathogenic molecules but further provide a list of new candidate genes and pathways possibly implicated in inner BRB breakdown during non-infectious posterior uveitis. BioMed Central 2020-03-17 /pmc/articles/PMC7076950/ /pubmed/32183784 http://dx.doi.org/10.1186/s12886-020-1333-5 Text en © The Author(s). 2020 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 Article
Lipski, Deborah A.
Foucart, Vincent
Dewispelaere, Rémi
Caspers, Laure E.
Defrance, Matthieu
Bruyns, Catherine
Willermain, François
Retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach
title Retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach
title_full Retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach
title_fullStr Retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach
title_full_unstemmed Retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach
title_short Retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach
title_sort retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7076950/
https://www.ncbi.nlm.nih.gov/pubmed/32183784
http://dx.doi.org/10.1186/s12886-020-1333-5
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