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G. Wildner, C. von Toerne, M. Diedrichs-Möhring, P. J. Nelson; Differential Gene Expression in Autoantigen-Specific T Cell Lines Causing Monophasic or Recurrent EAE in Lewis Rats. Invest. Ophthalmol. Vis. Sci. 2009;50(13):2634.
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© ARVO (1962-2015); The Authors (2016-present)
We have recently described monophasic (S-Ag peptide PDSAg-induced) or relapsing (induced by IRBP-peptide R14) experimental autoimmune uveitis in Lewis rats, which depends on the antigen specificity of the autoimmune response. We investigated the differences between PDSAg- and R14-specific T cell lines with respect to gene expression by gene arrays analysis and quantitative PCR (qPCR).
Three different cDNA preparations from each, PDSAg- or R14-specific T cell lines from the 2nd to 4th in vitro restimulation were compared by Affymetrix Rat Genome Array analysis. Arrays were analyzed by a new more sensitive single probe-based software ChipInspector (CI) from Genomatix. Subsequently a network analysis was performed and Gene Ontology (GO) categories were investigated. Genes differentially regulated and several genes from overrepresented GO categories but not annotated to the array were analyzed by qPCR.
Of the 28,000 genes analyzed by the rat genome chip the CI identified 35 genes to be upregulated in R14-specific T cells. By qPCR of 106 genes we finally identified 22 genes that were regulated over the threshold of twofold expression in R14-specific cells vs. PDSAg-specific T cell lines. These genes were related to pathways of activation, antigen presentation, inflammation, migration and survival.
Compared to PDSAg-specific T cells, R14-specific T cell lines induce earlier onset disease, relapsing intraocular inflammation and show elongated survival in the eye, which could be explained by the enhanced expression of the genes we have identified.
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