Murine CD4+ T cells: naive vs. experimental autoimmune uveitis
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE213034 | GEO | 2022/09/10
REPOSITORIES: GEO
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Project 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, lipocalin 2, 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.
2020-01-28 | GSE144168 | GEO
Project description:Experimental autoimmune uveitis (EAU) in Lewis rats is a model for the clinical heterogeneity of human uveitis. The autoantigens inducing disease in the rat are also seen in human disease. Depending upon the specific autoantigen used, the experimental disease course can be either monophasic or relapsing/remitting and appears to be dictated by the T cell effector phenotype elicited. We investigated potential differences between monophasic and relapsing/remitting effector T cells using transcriptomic profiling and pathway analysis. RNA samples isolated from three independent T cell lines derived from each specificity where analyzed by microarrays. Microarray data was used to obtain transcriptomic changes reflecting signal transduction pathway dysregulation. Keywords: Two group comparison Comparison of two types of cell lines of two different antigen specificities.
2010-12-01 | E-GEOD-19652 | biostudies-arrayexpress
Project description:Cells were prepared from draining lymph nodes (dLN) from naïve C57BL6/N female mice, as well as from C57BL6/N female mice at day 1 and day 3 after subcutaneous immunization with MOG(35-55) emulsified in Complete Freund’s adjuvant (CFA). Some mice received the CpG-B-1826 (50 microg; TCCATgACgTTCCTgACgTT; TIB MOLBIOL Syntheselabor GmbH), which was added to the MOG(35-55)/CFA mixture before emulsification. Cells from dLN were directly lysed in 350 microlitres RLT buffer containing 1% beta-mercaptoethanol (Invitrogen). Mice were 6-10 weeks of age at the time of immunization. The goal was to identify genes differentially expressed between dLN cells from mice immunized with the classical EAE protocol versus mice immunized with the addition of CpG-B-1826 to the immunization cocktail in order to gain some insights into the immune-modulatory effect of this treatment using an unbiased approach.
2023-04-26 | GSE205280 | GEO
Project description:MOG-reactive CD4 T cells were isolated from draining lymph nodes (dLN) and spleen of C57BL6/N female mice on day 10 after subcutaneous immunization with MOG(35-55) emulsified in Complete Freund’s adjuvant (CFA). Some mice received the CpG-B-1826 (50 microg; TCCATgACgTTCCTgACgTT; TIB MOLBIOL Syntheselabor GmbH), which was added to the MOG(35-55)/CFA mixture before emulsification. To isolate MOG-reactive CD4 T cells, dLN and spleens were harvested from mice on day 10 post-immunization, and single cell suspensions were re-stimulated for 5 hours using a mixture of MOG(35-55) (20 microg/ml), anti-CD40, anti-CD28, and anti-CD40L-PE (all from a commercial kit – Miltenyi Biotec – cat number 130-093-129). After 5 hours, CD4-positive cells were enriched magnetically by MACS using anti-CD4 microbeads (Miltenyi Biotec GmbH, cat. 130-049-201, clone L3T4), and subsequently subjected to FACS sorting to isolate CD4+CD40L+ and CD4+CD40L- cells after staining with anti-CD4 (clone RM4-5, conjugated to Pacific Blue), including a bin channel for CD8-positive cells (clone 53-6.7) and a marker for staining dead cells (NHPO). Sorted CD4+CD40L+ and CD4+CD40L- cells were then quickly checked for purity and directly lysed in 350 microlitre RLT buffer containing 1% beta-mercaptoethanol (Invitrogen). Purity was >95%. Mice were 6-10 weeks of age at the time of immunization. The goal was to identify genes differentially expressed between MOG-reactive CD4 T cells in mice treated or not with CpG-B, and to characterize the transcriptome of autoreactive CD4 T cells in draining lymph nodes and spleen on day 10 after immunization.
2023-04-26 | GSE205600 | GEO
Project description:Transcriptional profiling of mouse retina comparing naïve mouse and experimental autoimmune uveoretinitis (EAU)-developing mouse. EAU was induced by immunization of retinal autoantigens in adjuvants. Retinas were harvested 14 days after immunization.
2010-07-01 | GSE22617 | GEO
Project description:Experimental autoimmune uveitis (EAU) in Lewis rats is a model for the clinical heterogeneity of human uveitis. The autoantigens inducing disease in the rat are also seen in human disease. Depending upon the specific autoantigen used, the experimental disease course can be either monophasic or relapsing/remitting and appears to be dictated by the T cell effector phenotype elicited. We investigated potential differences between monophasic and relapsing/remitting effector T cells using transcriptomic profiling and pathway analysis. RNA samples isolated from three independent T cell lines derived from each specificity where analyzed by microarrays. Microarray data was used to obtain transcriptomic changes reflecting signal transduction pathway dysregulation. Keywords: Two group comparison
2010-12-01 | GSE19652 | GEO
Project description:Microbial sequencing revealed progressive reduction of gut microbiota that showed some differences in the two ABX groups compared to untreated controls. Interestingly, duration of ABX was associated with a gradual disappearance of the CD4+ and CD4+CD8+ subset of gut intraepithelial lymphocytes (IELs). This IEL subset is microbiota-dependent and is absent in germ-free mice. Relative abundance of Lactobacillus reuteri correlated with frequencies of CD4+CD8+ IELs and reduced EAU. Notably, IELs in culture suppressed antigen-specific activation of autoreactive T cells.
2023-11-17 | GSE236922 | GEO
Project description:Through this comparative proteomic study of equine CD4+ cells 376 membrane proteins could be identified. Several proteins showed changed abundance in horses affected by a spontaneous autoimmune disease (equine recurrent uveitis, ERU). Findings provide novel knowledge about changes in the CD4+ immune cell membrane proteome in a spontaneously and naturally occurring autoimmune disease in horses. The data is highly relevant for veterinary medicine and has proven translational quality for autoimmune uveitis in man.
2017-01-09 | PXD005384 | Pride
Project description:Uveitis is an immune-inflammatory disease that can cause blindness. However, little is known about a comprehensive atlas of the ocular cellular and inflammatory components in uveitis. Here, single-cell RNA sequencing was performed to construct a transcriptomic atlas of ocular cells from experimental autoimmune uveitis (EAU) and control mice to identify disease- associated immune cell subsets and molecules. We reveal 15 cell types in eyes and found an α-Synuclein positive microglia subset and plasma cell subset with high level of cytokines. The heterogeneous plasma cells subsets communicated with DC, T cells and retinal cells via various cytokines and molecular pairing. The G protein Rab1A participated in the inflammatory response of plasma cells. Altogether, we revealed the heterogeneous inflammatory genes, EAU-specific immune cell populations, and dysregulated communications of immune and retinal cells in EAU. This result provides a systematic view of the transcriptional map of uveitis and a foundation for studying the cellular and molecular mechanisms and pathogenesis of this disease.
2023-07-10 | GSE236389 | GEO
Project description:Dysregulation of Th17 differentiation was implicated in multiple inflammatory and autoimmune diseases including autoimmune uveitis. In the current study, we have provided evidence indicating that lactate-derived lactylation plays important roles in regulating Th17 differentiation. The lactylation level of CD4+ T cells was upregulated in EAU mice and inhibiting lactylation resulted in impaired EAU progression. We characterized the global lactylome of CD4+ T cells of normal and EAU mice. We found that the differentially lactylated proteins were enriched in pathways related to immune responses including leukocyte differentiation. Importantly, our results show that the lactylation level of Ikzf1 (K164) functions in regulating Th17 differentiation by differentially modulating gene expression patterns which are related to CD4+ T cell differentiation by CUT& Tag analysis. In view of the above mentioned well-documented evidence, Ikzf1 lactylation might represent an important regulator for Th17 differentiation in autoimmune uveitis.
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