Tunable Assembly of Regulatory Immune Signals to Promote Myelin-specific Tolerance

在自身免疫过程中，身体识别并攻击“自我”分子作为外来分子。目前的治疗采用广泛的 免疫抑制，这对患者有益，但可能使他们免疫功能低下。这种局限性， 沿着缺乏治愈方法，引发了人们对用免疫抑制剂控制自身免疫的策略的浓厚兴趣。 疫苗样的特异性，使免疫系统的其余部分完好无损。多份临床前报告和临床试验 我已经研究了这一理论，以对抗多发性硬化症（MS），一种神经退行性疾病，影响许多 退伍军人和发生时，髓鞘在中枢神经系统（CNS）的攻击，由免疫系统。一个 这些研究的重要发现是共同给予髓磷脂肽和耐受性免疫信号 可以促进调节性T细胞（Tregs）的发育，从而改善疾病。有趣的是， 阻碍髓鞘驱动的炎症控制的途径是Toll样受体（TLR）。在健康的个体中， 这些途径检测病原体相关模式以支持先天免疫。然而，最近的研究表明， Toll样受体信号传导--例如TLR 9--在人类MS和实验性自身免疫中升高 抑制TLR 9功能不仅减少炎症， 还促进TREGS并改善疾病。幼稚髓鞘反应性T细胞极化为炎性T细胞 细胞（例如，TH 17）或TREGS定位于脾和淋巴结（LN），协调免疫的组织。因此，在本发明中， 当髓磷脂存在于LN中时，帮助编程T细胞如何分化的策略-例如， 调节信号-可以产生大量髓鞘特异性TREGS，阻止病原性免疫细胞 没有广泛的压制。纳米技术在这一领域有很大的希望，通过增加控制， 靶向、释放动力学和多种信号的递送。然而，许多聚合物颗粒和其他材料 表现出激活炎症通路的内在特征，这可能会加剧自身免疫性疾病。 模仿生物材料吸引人的特征，同时消除炎症“载体”效应的策略可能是可行的。 对于MS或其他自身免疫性疾病的新疗法具有变革性意义。为了实现这一目标，拟议的研究 将使用聚离子免疫信号来创造完全由调节免疫构建的新型纳米结构胶囊， 信号和髓磷脂抗原。这些免疫荧光多层膜（“iPEM”）通过以下方式组装： 模板上的静电相互作用，然后将其移除，留下与髓鞘并列的疫苗胶囊 和TLR 9抑制性核酸（GpG）。由于没有载波，iPEM中信号的密度非常高 相对于具有包埋在基质中的货物的脂质或聚合物制剂。此外，颗粒使颗粒冷凝， 高密度信号，这是一种可以通过共定位促进向TREG分化的特征 髓鞘与GpG的关系。在小鼠中的生物分布研究表明，即使在贩运到LN时，iPEM 维持髓磷脂和调节信号的并置。在MS小鼠模型中的初步研究表明， 惊人的功效，iPEM治疗在100%的小鼠中停止疾病-临床评分为0，与之相比， 在91%的未治疗小鼠中出现几次瘫痪。拟议的工作将以这些调查结果为基础， 测试自身抗原和调节性免疫信号的组装在小鼠中产生耐受性的假设 MS模型和来自人类MS患者的样品，测试这种耐受性是否是髓鞘特异性的，并研究 功效的机制和持久性。具体目标是1）表征iPEM性质并在小鼠中筛选 来自巴尔的摩VA的MS患者组群的细胞和样品，2）在进行性小鼠模型中评估效力 的MS（EAE），并测试耐受性是否是髓鞘特异性的，3）阐明LN的结构和功能变化， 脾和导致耐受性的CNS，4）使用免疫组织化学方法测试耐受性是否可推广至其他自身抗原。 复发-缓解型MS模型（RR-EAE）。VA对该项目的支持可以使技术， 更具体和有效的治疗MS或其他自身免疫性疾病，影响许多退伍军人。

During autoimmunity, the body identifies and attacks “self” molecules as foreign. Current therapies employ broad immunosuppression, which is beneficial to patients, but can leave them immunocompromised. This limitation, along with the lack of cures, has sparked intense interest in strategies that could control autoimmunity with vaccine-like specificity, leaving the rest of the immune system intact. Several pre-clinical reports and clinical trials have investigated this theory to combat multiple sclerosis (MS), a neurodegenerative disease that impacts many Veterans and occurs when myelin in the central nervous system (CNS) is attacked by the immune system. An important finding from these studies is that co-administration of myelin peptide and tolerizing immune signals can promote the development of regulatory T cells (TREGS) that ameliorate disease. Interestingly, one set of pathways hindering control of myelin-driven inflammation are toll like receptors (TLRs). In healthy individuals, these pathways detect pathogen-associated patterns to support innate immunity. However, recent studies reveal that toll like receptor signaling – TLR9, for example – is elevated in human MS and in experimental autoimmune encephalomyelitis (EAE), a mouse model of MS. Suppressing TLR9 function not only reduces inflammation, but also promotes TREGS and improves disease. Polarization of naïve, myelin-reactive T cells into inflammatory T cells (e.g., TH17) or TREGS is localized to spleen and lymph nodes (LNs), tissues that coordinate immunity. Thus, strategies that help program how T cells differentiate when myelin is presented in LNs – for example, delivering regulatory cues – could generate large populations of myelin-specific TREGS that stop pathogenic immune cells without broad suppression. Nanotechnology holds great promise in this area through increased control over targeting, release kinetics, and delivery of multiple signals. However, many polymer particles and other materials exhibit intrinsic features that activate inflammatory pathways, which could exacerbate autoimmune disease. Strategies that mimic attractive features of biomaterials, while eliminating inflammatory “carrier” effects could be transformative for new therapies for MS or other autoimmune diseases. Toward this goal, the proposed research will use polyionic immune signals to create novel nanostructured capsules built entirely from regulatory immune signals and myelin antigens. These immune polyelectrolyte multilayers (“iPEMs”) are assembled through electrostatic interactions on a template, which is then removed to leave vaccines capsules that juxtapose myelin and TLR9-suppressive nucleic acids (GpG). Since there is no carrier, the density of signals in iPEMs is very high relative to lipid or polymer formulations with cargo embedded in a matrix. Further, the particles condense the signals at high densities, a characteristic that could promote differentiation toward TREG through co-localization of myelin with GpG in LNs. Biodistribution studies in mice reveal that even upon trafficking to LNs, iPEMs maintain juxtaposition of myelin and the regulatory signal. Pilot studies in a mouse model of MS demonstrate striking efficacy, with iPEM treatment stopping disease in 100% of mice – clinical score of 0, compared to development of several paralysis in 91% of untreated mice. The proposed work will build on these findings to test the hypotheses that assembly of self-antigen and regulatory immune signals generates tolerance in mouse models of MS and samples from human MS patients, test if this tolerance is myelin-specific, and investigate the mechanism and durability of efficacy. The specific aims are 1) characterize iPEM properties and screen in mouse cells and samples from the Baltimore VA’s MS patient cohort, 2) assess potency in a progressive mouse model of MS (EAE) and test if tolerance is myelin-specific, 3) elucidate the structural and functional changes in LNs, spleen, and the CNS that lead to tolerance, 4) test if tolerance is generalizable to other self-antigens using a relapsing-remitting model of MS (RR-EAE). VA support for this project could enable technology that creates more specific and effective treatments for MS or other autoimmune diseases that impact many Veterans.