Expression of immune response genes in human corneal epithelial cells interacting with Asperrgillus flavus conidia
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PROVIDER: PRJNA733997 | ENA |
REPOSITORIES: ENA
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Project description:Human corneal epithelial cells internalize Aspergillus flavus conidia. But the biochemical response of corneal epithelial cells in response to A.flavus is not clear. In this study, NanoString analysis was used to study the immune genes expression in primary corneal epithelial cells and cell line infected with A. flavus conidia. The study revealed the differential expression of immune genes associated with TNF, Th17, NF-kB signaling and B cell receptor signaling in corneal epithelial cells. There is also difference in the response between cell line and primary cells to A. flavus implies the use of both cells to understand the host-pathogen interaction.
2021-06-01 | GSE175824 | GEO
Project description:Mycotic leratitis is the corneal inflammation predominantly caused by Fusarium and Aspergillus species. Corneal epithelium is the earliest cell type encounter the invading pathogen. The innate immune responses of human corneal epithelial cells against Aspergillus flavus is not known. Here we studied the role of human corneal epithelial cells against Aspergillus flavus infection. The results showed that corneal epithelial cells internalized Aspergillus flavus conidia through actin mediated polymerization surrounding the conidia. Further the actin inhibitor cytochalasin D treatment reduced the formation actin ring around the conidia. The engulfed conidia acquired endosomal proteins as revealed by immunofluorescence analsyisis. Mass spectromtery of phagosomal proteins confirmed the recruitment of endosomal proteins and other proteins involved in phagocytosis. These results show the involvement of corneal epithelial cells in anti fungal defense.
2021-04-07 | PXD019512 | Pride
Project description:A. flavus exoproteome reference map as well as identification of the exoproteins from corneal isolates.
2015-01-07 | PXD001296 | Pride
Project description:BackgroundAspergillus flavus, one of the causative agents of human fungal keratitis, can be phagocytosed by human corneal epithelial (HCE) cells and the conidia containing phagosomes mature into phagolysosomes. But the immunological responses of human corneal epithelial cells interacting with A. flavus are not clear. In this study, we report the expression of immune response related genes of HCE cells exposed to A. flavus spores using targeted transcriptomics.MethodsHuman corneal epithelial cell line and primary cultures were grown in a six-well plate and used for coculture experiments. Internalization of the conidia was confirmed by immunofluorescence microscopy of the colocalized endosomal markers CD71 and LAMP1. Total RNA was isolated, and the quantity and quality of the isolated RNA were assessed using Qubit and Bioanalyzer. NanoString nCounter platform was used for the analysis of mRNA abundance using the Human Immunology panel. R-package and nSolver software were used for data analysis. KEGG and FunRich 3.1.3 tools were used to analyze the differentially expressed genes.ResultsDifferent morphotypes of conidia were observed after 6 h of coculture with human corneal epithelial cells and found to be internalized by epithelial cells. NanoString profiling showed more than 20 differentially expressed genes in immortalized human corneal epithelial cell line and more than ten differentially expressed genes in primary corneal epithelial cells. Distinct set of genes were altered in their expression in cell line and primary corneal epithelial cells. KEGG pathway analysis revealed that genes associated with TNF signaling, NF-KB signaling, and Th17 signaling were up-regulated, and genes associated with chemokine signaling and B cell receptor signaling were down regulated. FunRich pathway analysis showed that pathways such as CDC42 signaling, PI3K signaling, and Arf6 trafficking events were activated by the clinical isolates CI1123 and CI1698 in both type of cells.ConclusionsCombining the transcript analysis data from cell lines and primary cultures, we showed the up regulation of immune defense genes in A. flavus infected cells. At the same time, chemokine signaling and B cell signaling pathways are downregulated. The variability in the expression levels in the immortalized cell line and the primary cultures is likely due to the variable epigenetic reprogramming in the immortalized cells and primary cultures in the absence of any changes in the genome. It highlights the importance of using both cell types in host-pathogen interaction studies.
| S-EPMC8728928 | biostudies-literature
Project description:Dr. Panjwani's laboratory is focusing on the mechanism by which galectins-3 and 7 mediate corneal epithelial cell migration. We are currently performing studies to: (i) identify and characterize the corneal epithelial cell surface and extracellular matrix (ECM) molecules which serve as counterreceptors of galectin-3 and -7, to establish whether the lectins modulate corneal epithelial cell migration by binding to well known integrins, growth factor receptors, and/or ECM molecules and (ii) determine whether galectin-3 mediates corneal epithelial cell migration indirectly by modulating the expression of key adhesion and/or signal transduction molecules by using small interfering RNA, cDNA microarrays and glycogene arrays. We have prepared three independent preparations of total RNA of corneal epithelial cells from WT mice (total six samples) for analysis of glycogene expression. Samples are Normal Cornea (Left eye) and Laser ablation + 16-18 hours healing (right eye)
2010-12-11 | E-GEOD-26009 | biostudies-arrayexpress
Project description:Aspergillus flavus strain:corneal isolates Genome sequencing and assembly
| PRJNA445644 | ENA
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
2011-10-09 | E-GEOD-24981 | biostudies-arrayexpress
Project description:Dr. Panjwani's laboratory is focusing on the mechanism by which galectins-3 and 7 mediate corneal epithelial cell migration.
2010-12-11 | GSE26009 | GEO
Project description:We successfully induced corneal epithelial cells from human iPSCs. Then, we perfomed global expression analysis using microarray to compare the character of hiPSC-derived corneal epithelial cells with that of the other kinds of cells. Total RNA was obtained from human iPSCs (hiPSCs), human iPSC-derived corneal epithelial cells (hiCECs), human corneal limbal epithelial cells (HCECs), human oral keratinocytes (HOKs), human dermal fibroblasts (HDFs) and six weeks-differentiated hiPSCs (hiPSC-derived ocular surface ectoderm, OSE) using the QIAZol reagent. A microarray analysis using Sure Print G3 human 8x60K slides (Agilent technologies) was performed at Takara Bio (Shiga, Japan).
2016-03-09 | E-GEOD-73971 | biostudies-arrayexpress
Project description:Our experiment is to compare Glyco-gene expression by interferon-gamma/TNF-treated human corneal epithelial cells also treated either with cell communication glycoprotein 'lacritin' or negative control deletion construct 'C-25 lacritin'. In a newly developing story, lacritin but not C-25 protects human corneal epithelial cells from INFG/TNF-induced cell death. This expression array will help work out the mechanism of protection to complement phosphoblotting and shRNA approaches currently underway. Lacritin targets its coreceptor 'syndecan-1' in lacritin-dependent cell proliferation and secretion. Lacritin appears to be important in human 'dry eye' and in promoting corneal epithelial cell survival in the presence of inflammatory cytokines.
2011-09-05 | E-GEOD-31931 | biostudies-arrayexpress