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Cet abstract a été assigné à session FRO: Fund for Research in Ophthalmology
TitreFRO: Identification of the gene signature of retinal endothelial cells during classical experimental autoimmune uveitis, Th1- and Th17-dependent uveitis
Abstract Nr.1034
ButRetinal endothelial cell activation by autoreactive T cells plays a critical role in non infectious uveitis development. The aim of this work is to identify key genes regulated in endothelial cells during experimental autoimmune uveitis (EAU).
MéthodesTransgenic FVB/N-Tie2-GFP mice, in which vascular endothelial cells express GFP, are backcrossed with EAU-susceptible C57Bl/6 mice. The expression of GFP is characterized by immunofluorescence and FACS in order to validate the animal model. Retinal endothelial cells are isolated from dissociated retinas by FACSAria and their RNA purified for microarray analysis.
RésultatsFour generations of transgenic C57BL/6-Tie2-GFP mice have already been successfully generated. Immunofluorescence stainings confirm the expression of GFP on retinal endothelial cells, co-expressed with endoglin and CD31. FACS data indicate that up to 5% of dissociated retinal cells express CD31 but no GFP. Less than 1% of retinal cells are GFP+/CD31+. These cells can be efficiently sorted by FACS-Aria with an output of approximately 5.000 cells per animal. Quality RNA could be obtained from 10.000 cells in quantities compatible with microarray analysis.
ConclusionWe have validated our transgenic model, which seems to be more specific than retinal endothelial cell isolation based on CD31 expression. The next part of the project, which is the analysis of retinal endothelial cell mRNA modulation during EAU, is in progress.
Auteur 1
NomLIPSKI
InitialesD.
InstitutIRIBHM - Erasme Hospital
VilleBrussels
Auteur 2
NomDewispelaere
InitialesR.
InstitutIRIBHM - CHU Saint-Pierre
VilleBrussels
Auteur 3
NomFoucart
InitialesV.
InstitutIRIBHM - CHU Brugmann
VilleBrussels
Auteur 4
NomCaspers
InitialesL.
InstitutCHU Saint-Pierre
VilleBrussels
Auteur 5
NomBruyns
InitialesC.
InstitutIRIBHM
VilleBrussels
Auteur 6
NomWillermain
InitialesF.
InstitutIRIBHM - CHU Saint-Pierre
VilleBrussels
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