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: 7139194
• 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/7139194
• 关键词: Rodentias; autoantigens; biological signal transduction; biotechnology; 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）是美国大约10％的严重视觉障碍的原因。当前的疗法基于局部或全身性皮质类固醇，有或没有二线药物，例如环孢素A或抗代谢物。这些药物的严重不利影响是开发毒性毒性较小和更具体疗法的动力。实验性自身免疫性葡萄膜炎（EAU）是人类葡萄膜炎的特征良好动物模型，对猴子和啮齿动物EAU的研究已导致视网膜蛋白，S抗原和IRBP鉴定为葡萄膜炎的假定自身抗原。人们对开发针对葡萄膜炎的治疗疫苗有很大的兴趣，特别是对S抗原，IRBP和/或其他推定的葡萄蛋白的耐受性诱导（Recoverin，Opsin）。我们先前表明，STAT途径的激活在发育中受到调控，并在树突细胞（DC）分化和成熟中起作用。例如，在前体DC（PDC）和未成熟DC（IDC）中，STAT6信号通路被组成式激活，但随着这些DC分化为成熟DC（MDC），STAT6激活相关性的下降随着SOCS蛋白上调（JImmunol。172：2307-15）。相反，STAT1信号促进了直流成熟，并且在MDC中最强大。但是，与STAT6途径不同，STAT1信号传导不受SOC蛋白的反馈调节，这表明STAT1和STAT6途径在发育和成熟的DC中明显受到调节。因此，STAT1和STAT6似乎分别是MDC和IDC的谱系标记。在这个财政年度，我们工作的目的是利用树突状细胞种群中Stat蛋白的差异利用来通过在DC制剂中特异性删除STAT1基因来生成高度纯化的IDC。因此，我们已经生成了几种siRNA构建体，并正在分析siRNA技术对STAT1表达或通过沉默SOCS基因上调STAT6的效率和可行性。我们的最终目标是用IRBP或S抗原的免疫病表位加载纯化的IDC，以用作针对葡萄膜炎的治疗疫苗。