Engineered DNA-particles to model immune events in systemic lupus erythematosus

摘要/摘要 系统性红斑狼疮(SLE或狼疮)是一种导致严重系统性红斑狼疮的典型自身免疫性疾病。 表现，其中肾炎是发病率和死亡率的主要原因。系统性红斑狼疮主要影响年轻人 女性，非洲裔美国人患者表现出比欧洲人更早和更严重的疾病 美国人。系统性红斑狼疮的一个重要特征是表达抗核分子抗体(抗核抗体 抗体(ANA))。ANA为诊断、分类和疾病活动性提供了血清标志物。而当 关于系统性红斑狼疮的发病机制已经有了广泛的研究，但对于其潜在的原因仍有许多未知之处。 促进炎症和肾脏损伤的机制。因此，系统性红斑狼疮的治疗是经验性的和频繁的。 效果不佳。目前的治疗方法会导致永久性器官损伤和严重的副作用，提供了一种强大的 为更有效和毒性更低的疗法所需的机制研究的理论基础。在患者中以及在 在系统性红斑狼疮的动物模型中，Anas形成免疫复合体是炎症和损伤的关键步骤。的 ANA可以形成免疫复合体，抗DNA抗体(抗DNA)具有突出的和得到充分验证的作用 在肾炎中，如从患者受影响的肾脏中分离抗DNA以及诱导 应用抗DNA单抗建立动物肾炎模型。虽然大多数机型都是免疫的 复合体是基于抗体与相对较低的可溶性蛋白质或核酸抗原的相互作用。 最近的研究表明，系统性红斑狼疮中的免疫复合体形成于细胞生成的颗粒上 称为细胞外小泡。我们认为DNA可以吸附在这些颗粒的表面形成一种 为抗DNA抗体提供靶标的“日冕”。这项研究的意义在于它的发展 控制良好的合成DNA粒子，以解决有关免疫学特性的基本问题 不能用自然产生的粒子寻址的粒子。这项研究的结果将是 对DNA与颗粒结合形成抗原底物的机制的理解；形成 颗粒对免疫复合体的作用；表面DNA对颗粒与免疫细胞相互作用的影响； 以及表面DNA在免疫刺激中的作用。目标1将确定吸附在其上的DNA的抗原性 颗粒作为颗粒直径、DNA长度和蛋白质电晕的函数，使用先前开发的 以抗体结合试验为读数。目标2将使用目标1中确定的抗原颗粒来阐明 DNA颗粒的体外免疫刺激活性、免疫应答和细胞内化的测定 这些DNA粒子。控制良好的合成粒子系统的开发提供了一种创新的 系统性红斑狼疮的研究进展。除了系统性红斑狼疮，自然产生的细胞生成的颗粒作为元素的作用 在疾病中，发病机制对其他自身免疫性和炎症性疾病具有重要意义。 0

Summary/Abstract Systemic lupus erythematosus (SLE or lupus) is a prototypic autoimmune disease that causes severe systemic manifestations, of which nephritis is the major cause of morbidity and mortality. SLE primarily affects young women, with African American patients showing much earlier and more severe disease than European Americans. An important feature of SLE is the expression of antibodies to nuclear molecules (anti-nuclear antibodies (ANAs)). ANAs provide serological markers for diagnosis, classification, and disease activity. While there has been extensive study of the pathogenesis of SLE, much remains unknown about the underlying mechanisms that promote inflammation and renal injury. As a result, SLE treatment is empiric and frequently ineffective. Current treatments can cause permanent organ damage and severe side effects, providing a strong rationale for the mechanistic studies necessary for more effective and less toxic therapies. In patients as well as animal models of SLE, the formation of immune complexes by ANAs is a key step in inflammation and injury. Of ANAs that can form immune complexes, antibodies to DNA (anti-DNA) have a prominent and well-validated role in nephritis, as shown by the isolation of anti-DNA from affected kidneys of patients, as well as the induction of nephritis in animal models by administration of monoclonal anti-DNA antibodies. While most models for immune complexes are based on antibody interaction with soluble protein or nucleic acid antigens of relatively low molecular weight, recent research suggests that immune complexes in SLE form on cell-generated particles known as extracellular vesicles. We propose that DNA can adsorb onto the surface of these particles to form a “corona” that provides a target for anti-DNA antibodies. The significance of this research is the development of well-controlled synthetic DNA-particles to address fundamental questions about the immunological properties of particles that are not addressable with naturally occurring particles. The outcome of this research will be the understanding of the mechanisms by which DNA binds to particles to form an antigenic substrate; the formation of immune complexes by particles; the effects of surface DNA on the interaction of particles with immune cells; and the role of surface DNA in immune stimulation. Aim 1 will determine the antigenicity of DNA adsorbed on particles as a function of particle diameter, DNA length, and a protein corona using a previously developed antibody binding assay as a readout. Aim 2 will use the antigenic particles identified in Aim 1 to elucidate the immunostimulatory activity of DNA-particles in vitro, measuring the immune response and cellular internalization of the DNA-particles. The development of a well-controlled synthetic particle system provides an innovative approach to the study of SLE. Beyond SLE, the role of naturally occurring, cell-generated particles as elements in disease pathogenesis has important implications for other autoimmune and inflammatory diseases. 0