Document Detail


Microarray analysis of gene expression in West Nile virus-infected human retinal pigment epithelium.
MedLine Citation:
PMID:  22509103     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
PURPOSE: To identify key genes differentially expressed in the human retinal pigment epithelium (hRPE) following low-level West Nile virus (WNV) infection.
METHODS: Primary hRPE and retinal pigment epithelium cell line (ARPE-19) cells were infected with WNV (multiplicity of infection 1). RNA extracted from mock-infected and WNV-infected cells was assessed for differential expression of genes using Affymetrix microarray. Quantitative real-time PCR analysis of 23 genes was used to validate the microarray results.
RESULTS: Functional annotation clustering of the microarray data showed that gene clusters involved in immune and antiviral responses ranked highly, involving genes such as chemokine (C-C motif) ligand 2 (CCL2), chemokine (C-C motif) ligand 5 (CCL5), chemokine (C-X-C motif) ligand 10 (CXCL10), and toll like receptor 3 (TLR3). In conjunction with the quantitative real-time PCR analysis, other novel genes regulated by WNV infection included indoleamine 2,3-dioxygenase (IDO1), genes involved in the transforming growth factor-β pathway (bone morphogenetic protein and activin membrane-bound inhibitor homolog [BAMBI] and activating transcription factor 3 [ATF3]), and genes involved in apoptosis (tumor necrosis factor receptor superfamily, member 10d [TNFRSF10D]). WNV-infected RPE did not produce any interferon-γ, suggesting that IDO1 is induced by other soluble factors, by the virus alone, or both.
CONCLUSIONS: Low-level WNV infection of hRPE cells induced expression of genes that are typically associated with the host cell response to virus infection. We also identified other genes, including IDO1 and BAMBI, that may influence the RPE and therefore outer blood-retinal barrier integrity during ocular infection and inflammation, or are associated with degeneration, as seen for example in aging.
Authors:
Luis Munoz-Erazo; Ricardo Natoli; Jan Marie Provis; Michelle Catherine Madigan; Nicholas Jonathan Cole King
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-03-27
Journal Detail:
Title:  Molecular vision     Volume:  18     ISSN:  1090-0535     ISO Abbreviation:  Mol. Vis.     Publication Date:  2012  
Date Detail:
Created Date:  2012-04-17     Completed Date:  2012-07-09     Revised Date:  2013-06-26    
Medline Journal Info:
Nlm Unique ID:  9605351     Medline TA:  Mol Vis     Country:  United States    
Other Details:
Languages:  eng     Pagination:  730-43     Citation Subset:  IM    
Affiliation:
Discipline of Pathology, Bosch Institute, School of Medical Sciences, Sydney Medical School, University of Sydney, Sydney, NSW, Australia.
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MeSH Terms
Descriptor/Qualifier:
Apoptosis / genetics,  immunology
Chemokines / genetics,  immunology
Epithelial Cells / cytology,  immunology*,  virology
Gene Expression / immunology*
Gene Expression Profiling
Humans
Membrane Proteins / genetics,  immunology
Multigene Family
Oligonucleotide Array Sequence Analysis
Primary Cell Culture
Proteomics
Retinal Pigment Epithelium / cytology,  immunology*,  virology
Toll-Like Receptors / genetics,  immunology
Transforming Growth Factor beta / genetics,  immunology
Viral Load
West Nile virus / physiology*
Chemical
Reg. No./Substance:
0/BAMBI protein, human; 0/Chemokines; 0/Membrane Proteins; 0/Toll-Like Receptors; 0/Transforming Growth Factor beta
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