Development of dendritic cell vaccine against uveitis

抗葡萄膜炎树突状细胞疫苗的研制

• 批准号: 6968529
• 负责人: Charles E Egwuagu
• 金额: --
• 依托单位: NATIONAL EYE INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6968529
• 关键词: RNA interference; Rodentias; autoantigens; biological signal transduction; cell differentiation; cytokine; dendritic cells; gene targeting; small interfering RNA; transcription factor; uveitis; vaccine development

Uveitis (intraocular inflammatory diseases) is the cause of approximately 10% of severe visual handicap in the United States. Current therapy is based on topical or systemic corticosteroid, with or without second line agents such as cyclosporine A or anti-metabolites. Serious adverse effects of these drugs are the impetus for development of less toxic and more specific therapies for uveitis. Experimental autoimmune uveitis (EAU) is a well-characterized animal model of human uveitis and studies of monkey and rodent EAU have led to identification of the retinal proteins, S-Antigen and IRBP, as putative autoantigen of uveitis. There is significant interest in developing a therapeutic vaccine against uveitis, with particular emphasis on tolerance induction to S-Antigen, IRBP and/or other putative uveitogens (recoverin, opsin). We previously showed that activation of STAT pathways is developmentally regulated and plays a role in dendritic cell (DC) differentiation and maturation. For example, the STAT6 signaling pathway is constitutively activated in precursor DCs (pDCs) and immature DCs (iDCs) but declines as these DCs differentiate into mature DCs (mDCs) and the decline in STAT6 activation correlates with upregulation of SOCS proteins (J Immunol. 172:2307-15). In contrast, STAT1 signaling promotes DC maturation and is most robust in mDCs. However, unlike the STAT6 pathway, STAT1 signalling is not under feedback regulation by SOCS proteins, indicating that STAT1 and STAT6 pathways are distinctly regulated in developing and mature DC. Thus, STAT1 and STAT6 appear to be lineage markers of mDCs and iDCs, respectively. In this fiscal year the thrust of our work has been to exploit the differential utilization of STAT proteins in dendritic cell populations to generate highly purified iDC by specifically deleting STAT1 gene in DC preparations. We have therefore generated several siRNA constructs and are analyzing the efficiency and feasibility of siRNA technology to silence expression of STAT1 or upregulate STAT6 by silencing SOCS genes. Our ultimate goal is to load the purified iDCs with immunopathogenic epitopes of IRBP or S-Antigen for use as therapeutic vaccines against uveitis.

葡萄膜炎(眼内炎症性疾病)是美国约10%的严重视力障碍的原因。目前的治疗是基于局部或全身皮质类固醇，使用或不使用二线药物，如环孢素A或抗代谢药物。这些药物的严重不良反应是开发毒性较小、更具特异性的葡萄膜炎治疗方法的动力。实验性自身免疫性葡萄膜炎(EAU)是一种特征性的人葡萄膜炎动物模型，对猴和啮齿动物EAU的研究已导致视网膜蛋白S抗原和IRBP被确定为葡萄膜炎的可能自身抗原。人们对开发葡萄膜炎治疗性疫苗非常感兴趣，特别强调诱导对S抗原、红外线结合蛋白和/或其他可能的葡萄膜炎原(恢复素、视黄素)的耐受性。我们之前已经证明STAT通路的激活是发育调节的，并在树突状细胞(DC)的分化和成熟中发挥作用。例如，STAT6信号通路在前体DC(PDCs)和未成熟DC(IDCs)中被结构性激活，但随着这些DC分化为成熟DC(MDCs)而减弱，并且STAT6激活的下降与SOCS蛋白的上调相关(J免疫。172：2307-15)。相反，STAT1信号促进DC成熟，并且在MDCS中最强。然而，与STAT6通路不同的是，STAT1信号不受SOCS蛋白的反馈调节，这表明在发育和成熟的DC中，STAT1和STAT6通路受到明显的调控。因此，STAT1和STAT6似乎分别是MDCS和IDC的谱系标记。在本财年，我们的工作重点是利用树突状细胞群体中STAT蛋白的差异利用，通过在DC制剂中特异地删除STAT1基因来产生高纯度的IDC。因此，我们已经构建了几种siRNA结构，并正在分析siRNA技术通过沉默SOCS基因来沉默STAT1表达或上调STAT6表达的效率和可行性。我们的最终目标是将纯化的IDC负载IRBP或S抗原的免疫致病表位，作为葡萄膜炎的治疗性疫苗。