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Z. Li, S. P. Mahesh, B. Liu, S. Yeh, J. Lew, W. Lim, G. Levy Clarke, R. Buggage, R. B. Nussenblatt; Gene Expression Profiling of Non-Infectious Uveitis Patients Using Pathway Specific cDNA Microarray Analysis. Invest. Ophthalmol. Vis. Sci. 2007;48(13):1505. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
To study gene expression profiling of leukocytes from peripheral blood of patients with non-infectious uveitis.
Peripheral blood mononuclear cells (PBMCs) were isolated from 50 patients with clinically characterized non-infectious intermediate uveitis. Total RNA from PBMCs were extracted and quantified. Biotinylated probes were generated from total RNAs by incorporating biotinylated dUTP into synthesized cDNAs using a T7-polymerase linear amplification strategy. RNAs isolated from 20 healthy donors were pooled and served as normal control. Probes were hybridized to a pathway-specific cDNA microarray chip that contains some 400 autoimmune and inflammatory response related genes. Signals for specific binding were recorded by a CCD camera and gene expression profiling was analyzed using GEArray Suite software.
Despite tremendous heterogeneity of gene expression among patients and normal controls, there were clear differential gene expression patterns comparing those from patients to those from normal control. Although there was more than 80% of overlap of gene expression, some 15% of the genes were differentially expressed among patients compared to the normal donors. Several cytokine, chemokine or chemokine receptor genes were consistently more highly expressed among uveitis patients compared to the normal control. There seems less consistency in terms of down-regulated genes among patients when compared to normal control.
We show here that pathway specific cDNA microarray can be an efficient and economic strategy to profiling uveitis patients at the genomic level. Although preliminary and needing further confirmation, our data revealed several immune response genes that may be implicated in presumably autoimmune originated non-infectious uveitis.
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