Regulation of Neuromyelitis Optica via Tolerance Induced by PLG Nanoparticles Encapsulating Aquaporin 4 Epitopes

项目摘要/摘要： 视神经脊髓炎(NMO)是以T细胞和抗体为特征的中枢神经系统自身免疫性疾病 对水通道蛋白水通道蛋白4(AQP4)和髓鞘少突胶质细胞蛋白(MOG)的反应。动校正 由于视神经和脊髓的慢性损伤，患者经常失明和瘫痪。 电源线。目前的治疗方法，如大剂量皮质类固醇、血浆置换和利妥昔单抗都很昂贵，非 治愈，未能提供持久的疾病控制。米勒实验室已经开发出一种使用羧化的方法 由FDA批准的生物聚合物聚乳酸-乙交酯(PLG)组成的可生物降解纳米颗粒 微囊化蛋白或多肽[PLG(Ag)]诱导T细胞耐受治疗自身免疫 多发性硬化症(EAE)模型以及其他自身免疫性和过敏性疾病。跟随静脉注射。输液， PLG(Ag)NPs被肝和脾边缘的耐受性抗原提呈细胞(APC)摄取 区域通过Marco清道夫受体。这些APC呈递被包裹的抗原并诱导 通过几种机制产生耐受性，如无能诱导和激活不同的Ag特异性亚群 树突状细胞。值得注意的是，包裹醇溶蛋白的PLG纳米粒目前正在进行第二阶段的临床测试 乳糜泻患者。NMO是一种可能受益于此的首选自身免疫性疾病 因大量证据表明该病主要由自身免疫反应驱动而进行的耐受性治疗 致AQP4。因此，我们建议检验用PLG纳米粒进行耐受治疗的假设。 包裹AQP4和/或其免疫优势T细胞表位将诱导特异性免疫治疗 NMO通过诱导无能和AQP4特异性Tregs。目标1将通过以下方式建立NMO的小鼠模型 用重组AQP4蛋白(RAQP4)启动的AQP4 KO小鼠活化T细胞的转移， 产生多个AQP4表位的特异性T细胞，或与NMO诱导的I-KO小鼠的T细胞反应。 AB/I-AS限制性AQP4201-220或I-Ab-限制性AQP4135-153与野生型B6或SJL受体的表位。作为NMO 患者经常表现出对MOG和PLP以及AQP4的自身抗体，我们将评估是否有表位 传播导致NMO小鼠对这些髓鞘蛋白的T细胞和抗体反应。目标2将决定 包被免疫优势基因AQP4201-220和/或AQP4135的PLG纳米粒诱导免疫耐受的实验研究 RAQP4和AQP4多肽诱导的NMO小鼠的153个T细胞表位目标3将确定 包裹rAQP4的PLG纳米粒对rAQP4转移引发的疾病的耐受性 人类NMO患者可能识别多个AQP4决定因素所致的多肽特异性T细胞 由不同的人类白细胞抗原II类分子呈递。耐受诱导将通过临床评分进行评估， 通过免疫标准(T细胞增殖和细胞因子)评估脱髓鞘和视神经炎 产生和抗体反应)。预计AQP4特异性耐受性的成功演示将 推动这一治疗NMO的新型免疫耐受诱导平台的快速临床测试。

PROJECT SUMMARY/ABSTRACT: Neuromyelitis optica (NMO) is an autoimmune disease of the CNS characterized by T cell and antibody responses to the water channel protein Aquaporin 4 (AQP4) and myelin oligodendrocyte protein (MOG). NMO patients often suffer from blindness and paralysis as a result of chronic damage to the optic nerves and spinal cord. Current treatments such as high dose corticosteroids, plasmapheresis and rituximab are expensive, non- curative and fail to provide durable disease control. The Miller lab has developed a method using carboxylated biodegradable nanoparticles composed of the FDA-approved biopolymer polylactide-co-glycolide (PLG) to encapsulate proteins or peptides [PLG(Ag)] for induction of T cell tolerance for treatment of autoimmune models of Multiple Sclerosis (EAE) as well as other autoimmune and allergic diseases. Following i.v. infusion, PLG(Ag) NPs are taken up by tolerogenic antigen presenting cells (APCs) in the liver and splenic marginal zone via the MARCO scavenger receptor. These APCs present the encapsulated antigen and induce tolerance by several mechanisms such as anergy induction and the activation of various subsets of Ag-specific Treg cells. Significantly, PLG NPs encapsulating gliadin are presently undergoing phase 2 clinical testing in celiac disease patients. NMO is a prime candidate autoimmune disease which may benefit from this tolerogenic therapy due to substantial evidence that the disease is primarily driven by autoimmune responses to AQP4. Therefore, we propose to test the hypothesis that tolerogenic treatment with PLG nanoparticles encapsulating AQP4 and/or its immunodominant T cell epitopes will induce specific immunotherapy of NMO via induction of anergy and AQP4-specific Tregs. Aim 1 will establish mouse models of NMO by transfer of activated T cells from AQP4 KO mice primed with either recombinant AQP4 protein (rAQP4), yielding T cells specific for multiple AQP4 epitopes, or with T cells from KO mice primed with NMO-inducing I- Ab/I-As-restricted AQP4201-220 or I-Ab-restricted AQP4135—153 epitopes to wildtype B6 or SJL recipients. As NMO patients often display autoantibodies to MOG and PLP as well as to AQP4 due, we will assess if epitope spreading leads to T cell and antibody responses to these myelin proteins in NMO mice. Aim 2 will determine the effects of tolerance induced by PLG NPs encapsulating the immunodominant AQP4201-220 and/or AQP4135- 153 T cell epitopes in rAQP4 and AQP4 peptide-induced NMO mice. Aim 3 will determine the effects of tolerance induced by PLG NPs encapsulating rAQP4 on disease initiated by transfer of rAQP4 vs. AQP4 peptide-specific T cells due to the possibility that human NMO patients recognize multiple AQP4 determinants presented by different HLA class II molecules. Tolerance induction will be assessed by clinical scoring, evaluation of demyelination and optic neuritis, and by immune criteria (T cell proliferation and cytokine production and Ab responses). It is anticipated that successful demonstration of AQP4-specific tolerance will propel the rapid clinical testing of this novel immune tolerance inducing platform for the treatment of NMO.