Use Of Microarrays and Epigenetics In Gene Expression Of Uveitis & AMD Patients

眼部炎症性疾病，包括葡萄膜炎和黄斑变性，会导致严重的视力丧失。使用 各种免疫技术我们评估了这些疾病中免疫细胞的各种特征，包括信号通路，例如炎症和自身免疫通路。我们已经看到了葡萄膜炎的不同分子特征。特别令人感兴趣的是IL-22的鉴定。近年来，IL-22的表达与Th17细胞有关，而Th17细胞在AMD和葡萄膜炎患者的血清中升高。我们已经证明，IL-22导致原代培养的RPE细胞凋亡，可能是通过降低磷酸化的Bad水平。此外，我们还观察到老年性黄斑变性患者免疫细胞中IL-17活性升高。 我们已经报道了AMD患者免疫细胞的表观遗传学改变，但这一点仍有待证实。在葡萄膜炎的动物模型中，我们已经看到表观遗传改变药物对活动性疾病的表达产生了积极的影响，并在体外观察到了人类细胞的变化。此外，IL-17及其受体等免疫标志物的存在增加 在AMD的眼中是被注意到的，无论他们的机制是什么。此外，类固醇难治性葡萄膜炎患者外周血中有一种典型的类固醇难治性CD4T细胞亚群。先前的研究已经证明，这种类固醇耐药表型仅限于具有产生IL-17能力的中央记忆池的CD4细胞。因此，我们比较了Th1和Th17细胞对糖皮质激素的转录反应，以确定新的生物标志物和激素难治性疾病治疗干预的靶点。激素难治性患者比敏感患者有更大的倾向产生Th17细胞，并且两组患者的Th17细胞在接触地塞米松后的反应与Th1细胞不同。利用基因表达谱观察到对糖皮质激素的限制性反应。由于类固醇耐药性是一个重要的临床问题，这一信息强烈表明，针对Th17细胞或其来源的效应记忆T辅助细胞群体的新疗法将是药物开发的候选药物。有趣的是，在人类中，糖皮质激素治疗影响单核细胞亚型，诱导与T调节细胞诱导相关的亚型。 使用其他技术，我们发现葡萄膜炎患者的一个亚组的端粒长度明显缩短。此外，我们还注意到结节病患者循环中的IL-17与活动性疾病有关。结节病患者CD8细胞IL-17RC表达升高。此外，我们还注意到CD8 T细胞，现在被命名为Tc17细胞，产生IL-17，并以CD146表达为特征。我们还研究了眼炎性疾病患者中的树突状细胞。人们注意到的是CD1c MDCS与活性的关联，这似乎受到肿瘤坏死因子-α-p38 MAPK的调节。我们正在继续这一假设，即在疾病变得临床明显之前，这可能是一个活动性的生物标记物。

Ocular inflammatory diseases, including uveitis and AMD, cause significant visual loss. Using a variety of immune techniques we have evaluated various characteristics of immune cells in these diseases, including signaling pathways, e.g. inflammatory and autoimmune pathway. We have seen varying molecular signatures for uveitis. Of particular interest is the identification of IL-22. The expression of IL-22 has been recently associated with Th17 cells which is elevated in the serum of AMD and uveitis patients. We have shown that IL-22 resulted in apoptosis in cultured primary RPE cells, possibly by decreasing the phosphorylated-Bad level. In addition, we saw increased IL-17 activity in the immune cells of patients with age related macular degeneration. We have reported epigenetic alterations the immune cells of AMD patients but this remains to be verified. In an animal model for uveitis, we have seen that epigenetic altering medication affects positively the expression of active disease, and changes were noted in vitro on human cells. In addition an increase in the presence of immune markers such as IL-17 and its receptor in the eyes of AMD were noted, whatever their mechanism may be. In addition, patients with steroid refractory uveitis have a characteristic subpopulation of steroid refractory CD4+ T cells in their peripheral blood. Previously studies have demonstrated that this steroid refractory phenotype is restricted to the central memory pool of CD4+ cells which have the capacity to generate IL-17. We therefore compared transcriptomic responses of Th1 and Th17 cells to corticosteroids in order to identify novel biomarkers and targets for therapeutic intervention in steroid refractory disease. Steroid refractory patients have a greater propensity than sensitive patients to generate Th17 cells, and Th17 cells from either group of patients respond differently following exposure to Dex as compared with Th1 cells. Using gene expression profiling a restricted response to glucocorticoids was noticed. Of interest that there was a large genome shift in response to cyclosporine A. Since steroid resistance is an important clinical problem, this information would suggest strongly that new therapeutic which target either Th17 cells or the effector memory T helper cell population from which they are derived would be candidates for drug development. Of interest is the finding that in humans, glucocorticoid therapy affects the subtype of monocytes, inducing the subtype that is associated with the induction of T regulatory cells. Using additional techniques we have identified that a subgroup of uveitis patients have markedly shortened telomere length. In addition we have noted circulating IL-17 in sarcoidosis patients which is associated with active disease. Patients with sarcoidosis were noted to have an elevated IL-17RC expression on CD8+ cells. In addition we have noted that CD8+ T cells, which are now designated as Tc17 cells, produce IL-17 and are characterized by CD146 expression. We have also studies dendritic cells in patients with ocular inflammatory disease. What was noted was the association of CD1c+mDCs with activity, and this appears to be regulated by TNF-alpha-p38 MAPK. We are pursuing the hypothesis that this could be a biomarker for activity before the disease becomes clinically apparent.