Ocular inflammatory diseases, including uveitis, cause significant visual loss. Previous non-human investigations have identified several cell types, receptor systems and metabolic intermediates that have led to treatment approaches for human patients. However,information on the human genetic expression of these steps in defined inflammatory disease states is lacking. This non-intervention study proposes to obtain peripheral blood and tissue specimens from patients enrolled in other intramural trials for ocular inflammatory diseases and to apply contemporary cDNA microarray technologies for the analysis of differential gene expression. Test results will not be reported to participants or used for diagnostic or therapeutic purposes.The study?s primary objective is to identify unique gene expression profiles as well as disease relevant genes for patients with ocular inflammatory disease at defined clinical stages using cDNA microarray analysis. This will help provide further insight to understand the pathological mechanisms and potential targets for treatment. Some 3,000-5,000 genes will be examined starting with a selected set associated with interleukin (IL) proteins and their receptors, and with tumor necrosis factors (TNF). Purified peripheral blood mononuclear cells (or whole blood lysates using RNA isolation procedures) will be used to isolate total RNA from these samples. Samples will be taken during periods of active or recurring inflammatory disease and again during periods of quiescence after treatment. The microarray tools and methods for genetic analysis are now available at the NEI.In conjunction with these studies are functional assays and cell surface markers of negative regulatory cells, so called ?suppressor cells?. A correlation is being made between T cell activation and these negative regulatory signals and clinical activity.We continue to collect data from patients during the active phase of their uveitis and when their diesease became quiescent. To date, there have been 55 patients enrolled onto this protocol since approval in February 2002. We have recruited 55 patients until October 2005. Of the 55 patients enrolled in the protocol 31 are female, 24 are male, and 1 is considered a child. Among them, 33 are white, 16 are black, 3 are Asian, 2 are Hispanic and 1 is with unknown racial background. All the patients have been enrolled in another NEI protocol for evaluation or treatment prior to be enrolled into this protocol and signed consent form before samples were taken. For all 55 patients who have been enrolled either at a single stage of the disease or at multiple stages of the disease, 74 total RNA samples have been collected from purified PBMCs, 101 serum samples stored and 65 FACS analysis for multiple surface markers have been analyzed. In addition, 57 proliferation assays have also been performed to assess T cell responses to S-antigen, a self-antigen for non-infectious uveits. Of the 55 patients enrolled, 12 patients have undergone at least two stages of the disease (i.e., active to inactive or vice versa) and are considered completed for clinical sampling. We have done 48 microarrary analyses for 24 RNA samples from 10 patients using the NIA array membranes. We have also run 20 microarray using the SupperArray membranes for 11 patients and 1 normal pool. We have, thus far, identified 58 putative genes which may specifically up- or down-regulated in uveitis patients as compared to normal. Some of the genes are known to contribute to uveitis, such as interferon gamma. Some are new genes that have not been previously shown significant for uveitis. We are planning to confirm a selected panel of genes from that 58 candidate gene lists which we believe significant.
| Li, Zhuqing; Mahesh, Sankaranarayana P; Kim, Ben J et al. (2003) Expression of glucocorticoid induced TNF receptor family related protein (GITR) on peripheral T cells from normal human donors and patients with non-infectious uveitis. J Autoimmun 21:83-92 |
