MicroRNA expression data from human corneal epithelial cells exposed to media, human tear, media + P. aeruginosa antigens, tear + P. aeruginosa antigens
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ABSTRACT: Exposure to tear fluid can enhance corneal epithelial cells' ability to resist bacterial virulence mechanisms by upregulating epithelial-derived innate defense genes. The aim of this study was to further elucidate the mechanisms by which tear fluid modulates epithelial cell susceptibility to P. aeruginosa internalization and the relationship to known to be upregulated genes with tear fluid exposure. The hypothesis tested was that tear fluid effects on epithelial cells involve the induction of microRNA expression to modify innate defense gene responses to bacterial challenge. Human corneal epithelial cells were incubated in either 40 ul of fresh human tear fluid or high calcium KGM without antibiotics for 16 hours before extraction or before incubation with P. aeruginosa antigens for 3 h then extraction.
Project description:Exposure to tear fluid can enhance corneal epithelial cells' ability to resist bacterial virulence mechanisms by upregulating epithelial-derived innate defense genes. The aim of this study was to further elucidate the mechanisms by which tear fluid modulates epithelial cell susceptibility to P. aeruginosa internalization and the relationship to known to be upregulated genes with tear fluid exposure. The hypothesis tested was that tear fluid effects on epithelial cells involve the induction of microRNA expression to modify innate defense gene responses to bacterial challenge.
Project description:Recombinant protein of Pseudomonas aeruginosa hook protein FlgE was added to cultured human corneal epithelial cell line for 4 hours and the mRNA expression profiling was performed using Agilent 8*60K array and dual labeling. Three triplicates were included for FlgE treatment and PBS control respectively, thus producing three pairs of samples for array, namely E1PBS1, E2PBS2, E3PBS3.
Project description:We previously showed that pre-exposure of the cornea to TLR5 ligand flagellin induces profound mucosal innate protection against pathogenic microbes by reprogramming gene expression. To date, there was no genome-wide cDNA array to detect full scale of flagellin mediated reprogramming of gene expression in mucosal surface epithelial cells. Taking advantage of readily accessible, easily procurable epithelial cell population, this study is the first report to use genome-wide cDNA microarray approach to document genes associated with flagellin-induced protection against Pseudomonas aeruginosa infection in corneal epithelial cells (CECs). Total RNA obtained from isolated mouse corneal epithelial cells of the control (cells scrapped off from the corneas without infection), Pseudomonas aeruginosa infected (6 h post infection) and flagellin pretreated (24 h), followed by Pseudomonas aeruginosa infection (6 h).
Project description:This SuperSeries is composed of the following subset Series: GSE24979: MicroRNA-145 Regulates Human Corneal Epithelial Differentiation [Agilent-016436 array data] GSE24980: MicroRNA-145 Regulates Human Corneal Epithelial Differentiation [Agilent-014850 array data] Refer to individual Series
Project description:Purpose: Determination of inter- and intra-day variations in tear flow rate, tear fluid protein concentration as well as protein composition regarding their impact for future biomarker studies.Methods: Tear fluid was collected non-invasively from 18 healthy subjects performing Schirmer tests at four different time points repetitive in a period of two days. The tear flow rate with Schirmer test strips was measured. Proteins were extracted from strips and quantified using amino acid analysis. Protein composition was analyzed by data-independent (DIA) based mass spectrometry. To exclude any impairments to health, volunteers underwent a detailed neurological as well as an ophthalmological examination.Results: Whether tear fluid was collected from OS or OD did not affect the tear flow rate (p ≈ 0.63) or protein concentration (p ≈ 0.97) of individual subjects. Moreover, protein concentration was independent from the tear volume, so that a change in volume would only have an effect on total protein amount. When the examination days were compared, investigation of tear flow rate (p ≈ 0.001) and protein concentration (p ≈ 0.0003) indicated significant differences. Further mass spectrometric analysis of tear fluid revealed eleven differentially regulated proteins when comparing both examination days. Conclusion: Our findings provide evidence of inter-day variation in tear flow rate, tear proteome concentration and composition in healthy subjects, suggesting that inter-day variation need to be taken into consideration in biomarker research of tear fluid. Identified proteins were assigned to functions in the immune response, oxidative and reducing processes, as well as mannose metabolism.
Project description:Recombinant protein of Pseudomonas aeruginosa hook protein FlgE was added to cultured human corneal epithelial cell line for 4 hours and the mRNA expression profiling was performed using Agilent 8*60K array and dual labeling.
Project description:Human corneal endothelial cells (HCEC) form a monolayer by adhering tightly through their intercellular adhesion molecules. Located at the posterior corneal surface, they maintain corneal translucency by dehydrating the corneal stroma, mainly through the Na+- and K+-dependent ATPase (Na+/K+-ATPase). Because HCEC proliferative activity is low in vivo,we tried to activate proliferation of HCEC by inhibiting cyclin-dependent kinase inhibitors.We have here demonstrated microarray data of transduced human corneal endothelial cell lines. Affymetrix human U133 plus 2.0 array was used to transcriptionally profile to compare cultured human corneal endothelial cells and transduced human corneal endothelial cells.
Project description:Mass spectrometry-based proteomics by bottom-up approaches enables the unbiased and sensitive profiling of cellular proteomes and extracellular environments. Recent technological and bioinformatic advances permit identifying of dual biological systems in a single experiment, supporting investigation of infection from the perspective of both a host and pathogen. At the ocular surface, P. aeruginosa are commonly associated with biofilm formation and inflammation of the ocular tissues, causing damage to the eye. The interaction between P. aeruginosa and the immune system at the site of infection describes limitations in clearance of infection and enhanced pathogenesis. Here, we profile the extracellular environment (eye wash) of murine ocular surfaces infected with a clinical isolate of P. aeruginosa and detect neutrophil marker proteins, indicating neutrophil recruitment to the site of infection. In addition, we define the deepest murine corneal proteome to date and detect proteins, categories, and networks critical to the host response to infection. Moreover, we provide the first identification of bacterial-specific proteins in response to the host during bacterial biofilm formation of the eye. We validate our findings through in silico comparisons and enzymatic profiling. Overall, our work provides comprehensive profiling of the host-pathogen interface and uncovers differences between general and site-specific host responses to infection.
Project description:In order to test the hypothesis that fibroblasts from different tissues are phenotypically distinct from one another, we have subjected tendon, skin and corneal fibroblasts of fetal mouse to mechanical stimulation by fluid flow and analyzed the transcriptional responses of the cells using Affymetrix MOE430 chip set containing two arrays MOE430A and MOE430B. Experiment Overall Design: The experiment is done using Affymetrix MOE430 chip set containing two arrays MOE430A - platform GPL339 and MOE430B - platform MOE340. Three biological replicates for tendon, skin and corneal fibroblasts subjected to mechanical stimulation as well as not stimulated (control) were analyzed yielding 18 samples per chip (36 in total).
Project description:Ehf is a transcriptional regulator that is highly expressed and enriched in corneal epithelium. To gain insights into the role of Ehf in the corneal epithelium, we performed siRNA knockdown of Ehf in primary human corneal epithelial cells. Primary human corneal epithelial cells were transfected with siEhf or si controls, plated, and harvested at 72 hr.