Project description:We inoculated ARPE-19 human retinal pigment epithelial cells with EBOV, and followed course of infection by immunocytochemistry and measurement of titer in culture supernatant. To interrogate transcriptional responses of infected cells, we combined RNA sequencing with in silico pathway, gene ontology, transcription factor binding site and network analyses. Human retinal pigment epithelial cells were permissive to infection with EBOV, and supported viral replication and release of virus in high titer. Unexpectedly, 28% of 560 up-regulated transcripts in EBOV-infected cells were type I IFN responsive, indicating a robust type I IFN response.
Project description:Retinal pigment epithelial cells are critical for eye function and loss of cell function is linked to age-related blindness. Relatively little is known about the transcriptional regulatory networks in these cells. The datasets presented here are ChIP-seq experiments for RNA polymerase II , transcription factors and histone modifications in human retinal pigment epithelial cells. ChIP-Seq for transcription factors, RNA polymerase, histone modifications and CTCF in retinal pigment epithelial cells
Project description:In this study we examined the effect of T cell-derived cytokines on retinal pigment epithelial (RPE) cells with respect to expression of complement components. We used an in vitro co-culture system in which CD3/CD28-activated human T cells were separated from the human RPE cell line (ARPE-19) by a membrane. Differential gene expression in the RPE cells of complement factor genes was identified using gene arrays, and selected gene transcripts were validated by q-RT-PCR. Protein expression was determined by ELISA and immunoblotting. Co-culture with activated T cells increased RPE mRNA and/or protein expression of complement components C3, factors B, H, H-like 1, CD46, CD55, CD59, and clusterin, in a dose-dependent manner. Soluble factors derived from activated T cells are capable of increasing expression of complement components in RPE cells. This is important for the further understanding of inflammatory ocular diseases such as uveitis and age-related macular degeneration.
Project description:Comparitive analysis of the retinal stem/progentior cells derived from the ciliary and iris pigment epithelial cells and the subsequent differentiated cells derived from the stem/progenitor cells. Gene expression profiling has shown great promise in analysing the reprogramming pattern of the cells under various culture conditions, in this context we analysed the various differential gene expression pattern of the neurospheres derived from the ciliary and iris pigment epithelial cells, and their differentiated cells. This provide an insite for the stem cell transplantation studies. We analysed 3 biological cultures derived from each catogory like the primary cells of both ciliary and iris pigment epithelial cells, the stem/progenitor cells (Neurospheres), neurosphere derived differentiated cells. No technical replicates were performed.
Project description:T cells that encounter cultured ocular pigment epithelial cells in vitro are inhibited from undergoing T cell receptor-triggered activation. Because retinal pigment epithelial (RPE) cells are able to suppress T-cell activation, we studied whether RPE cells could suppress cytokine production by activated T helper (Th) cells. In this study we showed that primary cultured RPE cells greatly suppressed activation of bystander CD4+ T cells in vitro, especially the cytokine production by the target T helper cells (Th1 cells, Th2 cells, Th17 cells, but not Th3 cells). Cultured RPE cells and RPE-supernatants significantly suppressed IL-17 producing CD4+ T cells, and RPE cells fully suppressed polarized Th17 cell lines that induced by recombinant proteins, IL-6 and TGFb2. Moreover, RPE cells failed to suppress IL-17 producing T cells in the presence of rIL-6. In addition, Th17 cells exposed to RPE were suppressed via TGFb, which produce RPE cells. These results indicate that retinal PE cells have immunosuppressive capacity in order to inhibit Th17-type effector T cells. Thus, ocular resident cells play a role in establishing immune regulation in the eye. Retinal pigment epithelium suppresses Th17 cells
Project description:Human embryonic stem-cell derived retinal pigment epithelial cells were co-stimulated with TNF-α and TGF-β1 to model a retinal scarring complication, proliferative vitreoretinopathy. Thermogelling polymer prevented this scarring complication. To understand the polymer-cell interaction and ultimately the mechanism underlying it's anti-scarring activity, RNA-Seq was performed.
Project description:Comparitive analysis of the retinal stem/progentior cells derived from the ciliary and iris pigment epithelial cells and the subsequent differentiated cells derived from the stem/progenitor cells. Gene expression profiling has shown great promise in analysing the reprogramming pattern of the cells under various culture conditions, in this context we analysed the various differential gene expression pattern of the neurospheres derived from the ciliary and iris pigment epithelial cells, and their differentiated cells. This provide an insite for the stem cell transplantation studies.
Project description:Epithelial-mesenchymal transition (EMT) of retinal pigment epithelial (RPE) cells is a hallmark of PVR, which remains an unresolved clinical challenge leading to vision loss. Here, we identified disregulation of junctional adhesion molecule C (JAM-C) and transcriptional co-activator with PDZ-binding motif (TAZ) as key regulators of RPE EMT.
