Biomaterials to study tolerance immune induction kinetics

研究耐受免疫诱导动力学的生物材料

• 批准号: 10223121
• 负责人: Kristy M Ainslie
• 金额: $ 37.66万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-14 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10223121
• 关键词: Abbreviations; Acetals; Acids; Address; Adoptive Transfer; Affect; Aftercare; American; Animal Model; Anti-Inflammatory Agents; Antibodies; Antigen Presentation; Antigen-Presenting Cells; Antigens; Autoimmune; Autoimmune Diseases; Biocompatible Materials; Biological Models; C57BL/6 Mouse; CD80 gene; Cardiotoxicity; Cell Communication; Cell physiology; Cells; Clinical; Cohort Effect; Communication; Data; Delayed Hypersensitivity; Dendritic Cells; Development; Dexamethasone; Dextrans; Disease; Dose; Encapsulated; Engineering; Evaluation; Experimental Autoimmune Encephalomyelitis; Experimental Models; FOXP3 gene; Formulation; Generations; Glycolates; Goals; Hepatotoxicity; Hour; IL2RA gene; ITGAM gene; ITGAX gene; Immune; Immune Tolerance; Immunologics; Immunologist; Immunosuppression; In Vitro; Infection; Inflammation; Interferon Type II; Interferons; Interleukin-10; Kinetics; Lipopolysaccharides; Major Histocompatibility Complex; Malignant Neoplasms; Measures; Mediating; Methods; Modeling; Molecular Sieve Chromatography; Multiple Sclerosis; Mus; Myelin Basic Proteins; Onset of illness; Ovalbumin; Particle Size; Particulate; Pathogenesis; Peptides; Periodicity; Phagocytes; Phagolysosome; Phagosomes; Pharmaceutical Preparations; Polyesters; Polymers; Proteins; Proteolipids; Publishing; Reading; Regulatory T-Lymphocyte; Relapse; Research; Research Personnel; Risk; Role; Signal Transduction; Sirolimus; Spleen; Splenocyte; System; T cell response; T-Lymphocyte; T-Lymphocyte Subsets; Technology; Time; Transforming Growth Factors; Vaccines; Work; attenuation; biodegradable polymer; central nervous system demyelinating disorder; clinically relevant; cytokine; design; effector T cell; experimental study; immunological synapse; in vivo; macrophage; mouse model; multiple sclerosis treatment; nano; nanoparticle; oligodendrocyte-myelin glycoprotein; particle; response; side effect

ABSTRACT We plan to explore the kinetics of the immune synapse, as it relates to generation of tolerance, using nano/microparticles (Ps) fabricated from the polymer acetalated dextran (Ac-DEX). Previously, we have shown attenuation of clinical score with treatment of Ac-DEX particles encapsulating myelin basic protein (MBP) and dexamethasone (DXM), using a C57Bl/6 mouse model of experimental autoimmune encephalomyelitis (EAE). We continued this research and illustrated that Ac-DEX particles encapsulating proteolipid protein (PLP) and rapamycin (Rapa) in a SJL relapse and remitting model of EAE completely reduced clinical score to baseline when given after disease onset. The degree of reduction of clinical score for both of these EAE models was greater for the Ac-DEX particles systems than observed in other published antigen-specific EAE treatments that used particle systems. Our particle system is unique because it relies on the highly tunable polymer Ac-DEX. Ac-DEX is ideal for delivery of agents to phagocytic cells because it is acid-sensitive and has significantly increased degradation in the low acid (~pH 5) of the phagosome. In addition to this it has tunable degradation rates that can range from hours to months, which is a unique range from commonly used polyesters (e.g. poly(lactic-co-glycolic acid) (PLGA)) that have degradation on the order of months. Moreover, Ac-DEX is unique from polyesters because its degradation products are pH neutral, and do not have the potential to shift the local pH or damage sensitive payloads. We have shown that Ac-DEX particles have degradation rates that affect both antibody and cellular response for traditional vaccine and hypothesize similar effects for generation of tolerance. Therefore, we hypothesize that Ac-DEX particles promote antigen specific immune tolerance by inducing Tregs and that the cyclic acetal coverage of Ac-DEX impact degradation rate and modulate the immune synapse. We have three specific aims to address this hypothesis. Aim 1 is focused on formulation of the polymer and particles. Both a MS representative antigen as well as the model antigen OVA will be encapsulated. Particle parameters like size and loading will be determined. Ac-DEX polymer with various cyclic acetal coverages will be fabricated to degrade over a broad range of times. Aim 2 focuses on in vitro and in vivo studies to understand the immune synapse and how that relates to particle degradation times. The metrics for evaluation will be generation of inducible T-regulatory cells (iTregs). Furthermore, we will optimize systems using a delayed type hypersensitivity (DTH) model of inflammation. The relationship between particle degradation and generation of tolerance will be optimized. In Aim 3, the optimized formulation will be evaluated in a model of MS and expression of Tregs, as well as other immunological characterizations will be carried out. The overall goal of this work is to evaluate the Ac- DEX particles systems as an antigen-specific treatment for MS and to understand the influence of release of tolerance agents (e.g. antigen and Rapa) on the generation of antigen specific immune tolerance.

摘要 我们计划探索免疫突触的动力学，因为它与耐受的产生有关，使用 由聚合物缩醛化葡聚糖（Ac-DEX）制造的纳米/微米颗粒（Ps）。此前，我们已经展示了 用包封髓磷脂碱性蛋白（MBP）的Ac-DEX颗粒治疗减轻临床评分， 地塞米松（DXM），使用实验性自身免疫性脑脊髓炎（EAE）的C57B1/6小鼠模型。 我们继续这项研究，并说明Ac-DEX颗粒包封蛋白脂质蛋白（PLP）和 雷帕霉素（Rapa）在EAE的SJL复发和缓解模型中完全降低临床评分至基线水平 在发病后服用。这两种EAE模型的临床评分降低程度均为 Ac-DEX颗粒系统比在其他公开的抗原特异性EAE治疗中观察到的更大， 使用粒子系统。我们的粒子系统是独一无二的，因为它依赖于高度可调的聚合物Ac-DEX。 Ac-DEX对于将药剂递送至吞噬细胞是理想的，因为它是酸敏感的并且具有显著的细胞毒性。 在吞噬体的低酸（~pH 5）中的降解增加。除此之外，它还具有可调的降解能力， 速率的范围可以从数小时到数月，这是与常用聚酯（例如， 聚（乳酸-共-乙醇酸）（PLGA）），其具有数月量级的降解。此外，Ac-DEX是独一无二的 因为其降解产物是pH中性的，并且不具有改变局部 pH值或损伤敏感有效载荷。我们已经表明，Ac-DEX颗粒的降解速率影响两者 传统疫苗的抗体和细胞应答，并假设产生耐受性的类似效果。 因此，我们假设Ac-DEX颗粒通过诱导TcR来促进抗原特异性免疫耐受， Ac-DEX的环缩醛覆盖影响降解速率并调节免疫突触。我们 有三个具体的目标来解决这个假设。目标1集中于聚合物和颗粒的配制。 MS代表性抗原以及模型抗原OVA都将被包封。粒子参数 将确定类似的尺寸和负载。将制备具有各种环状缩醛覆盖率的Ac-DEX聚合物 在很宽的时间范围内降解。目的2侧重于体外和体内研究，以了解免疫 以及它与粒子降解时间的关系。评估的指标将是 诱导型调节性T细胞（iT细胞）。此外，我们将使用迟发型超敏反应优化系统， (DTH)炎症模型。颗粒降解和公差产生之间的关系将是 优化在目标3中，还将在MS模型和TdR表达中评价优化的制剂， 因为将进行其它免疫学表征。这项工作的总体目标是评估AC- DEX颗粒系统作为MS的抗原特异性治疗，并了解 耐受剂（例如抗原和Rapa）对抗原特异性免疫耐受产生的影响。