Mechanisms of Pain and Immune Processes

概述：这个翻译研究计划涉及伤害性传递的分子和病理生理过程，以及研究慢性疼痛，自身免疫性疾病，传染病及其在人类患者中的交叉点的新方法。慢性神经性疼痛可影响身体的任何部位，并且可由于各种损伤、感染和自身免疫或代谢紊乱（例如，糖尿病周围神经病变）。我们正在测试的假设，在一些患者中，慢性疼痛是发起和/或维持相关的自身抗体对周围神经或雪旺细胞中的蛋白质产生的免疫病理过程。自身抗体是某些大纤维周围神经病变如副肿瘤性疾病的罪魁祸首。疼痛是一个组成部分，我们假设存在的自身抗体的蛋白质中发现的神经末梢产生的小直径疼痛感应（伤害性）C-或A-δ神经纤维。为了检验疼痛性神经病性病症具有自身免疫组分的假设，我们建立了一种灵敏的、定量的、液相发光测定法，该测定法使用在哺乳动物细胞中表达的重组蛋白抗原-荧光素酶融合物作为示踪剂。这种检测被称为荧光素酶免疫沉淀系统（LIPS），我们有多篇发表的论文表明LIPS可以稳健而灵敏地测量血清，血浆，脑脊液或唾液中的抗体。本研究的目标是了解（a）人类慢性疼痛疾病的分子和细胞生物学机制，以及（B）利用这些知识来设计新的治疗和诊断疼痛和其他疾病，它可以适应。 在这个项目中，自身免疫假说进行了探讨。我们建立的检测方法具有很大的通用性，由于其灵敏度，模块化，并使用重组DNA方法来产生蛋白质抗原示踪剂。我们的早期研究评估了抗体发挥作用的一系列疾病和病症，包括感染性疾病，有和没有神经系统受累，自身免疫性疾病，可能在患者亚群中具有神经系统病理学，以及其他类型的筛查和方法学变体。在许多神经自身免疫性疾病中，主要的自身抗原通常是质膜受体或离子通道。我们已经产生了几个这样的探针神经性疼痛疾病，特别是β-肾上腺素能受体和钾通道的基础上发表的报告，目前正在测试，以确定是否LIPS检测可以检测这些蛋白质的抗体在适当的患者。我们完成了一项关于带状疱疹或带状疱疹的研究，带状疱疹可以演变成一种疼痛的神经系统疾病，称为疱疹后神经痛（PHN）。 我们在一个亚群中检测到一些中和性抗细胞因子自身抗体，有趣的是，所有这些都有PHN。这一点很重要，因为它表明一些PHN患者可能需要额外的干预来控制这种疾病，而不仅仅是镇痛药物。 该项目最引人注目的方面之一是数据集及其用途的逐步分层和演变。随着我们增加测试抗原的数量，并对各种病症和疾病进行检测，我们对免疫和自身免疫反应进行了全面的评估。这是通过确定（a）对正向同源蛋白和蛋白片段的免疫应答的程度和特异性，（B）指示共同的抗原或一般机制的抗原谱的重叠，和（c）参与胞间或胞内通讯的整个信号传导途径内的抗原性来实现的。例子是干燥综合征中的TRIM蛋白家族和我们对红斑狼疮患者的基于自身抗原的分类。 公共卫生监测的一个新用途的例子是我们早期对唾液中自身抗体测量的演示，这突出了为许多类型的人类疾病建立非侵入性、基于唾液的测定或为大量或脆弱人群或儿科人群评估疫苗免疫状态的可行性。最终，可以实施完整的多种抗原分析，以更深入地了解许多复杂的人类疾病状态。

Overview: This translational research program addresses molecular and pathophysiological processes of nociceptive transmission and new ways to investigate chronic pain, autoimmune conditions, infectious diseases and their intersection in human patients. Chronic neuropathic pain can affect any part of the body and can occur due to a variety of insults, infections, and autoimmune or metabolic disorders (e.g., diabetic peripheral neuropathy). We are testing the hypothesis that, in some patients, chronic pain is initiated and/or maintained by immunopathological processes related to autoantibodies generated against proteins in peripheral nerve or Schwann cells. Autoantibodies are known culprits in certain large fiber peripheral neuropathies such as in paraneoplastic disorders. Where pain is a component, we hypothesize the presence of autoantibodies to proteins found in nerve endings arising from small diameter pain-sensing (nociceptive) C- or A-delta nerve fibers. To test the hypothesis that painful neuropathic conditions have an autoimmune component we established a sensitive, quantitative, liquid phase luminescence assay that uses recombinant protein antigen-luciferase fusions as tracers, which are expressed in mammalian cells. This assay is called luciferase immunoprecipitation systems (LIPS) and we have multiple published papers showing that LIPS can robustly and sensitively measure antibodies in serum, plasma, cerebrospinal fluid or saliva. The goals of this research are to understand (a) the molecular and cell biological mechanisms underlying human chronic pain disorders, and (b) to use this knowledge to devise new treatments and diagnostics for pain and other disorders to which it can be adapted. In this project an autoimmune hypothesis is explored. The assay methodology we established has great versatility due to its sensitivity, modularity, and use of recombinant DNA methodology to generate protein antigen tracers. Our early investigations evaluated a range of diseases and disorders that antibodies play a role in, including infectious diseases, with and without nervous system involvement, autoimmune disorders, which may have nervous system symptomology in subsets of patients, and other types of screening and methodological variants. In many neural autoimmune disorders the major autoantigens are frequently plasma membrane receptors or ion channels. We have generated several such probes for neuropathic pain disorders, notably the beta-adrenergic receptor and a potassium channel based on published reports, and are currently testing to determine if the LIPS assay can detect antibodies to these proteins in appropriate patients. We completed a study on shingles or herpes zoster that can evolve into a painful neurological disorder called post-herpetic neuralgia (PHN). We detected some neutralizing anti-cytokine autoantibodies in a subpopulation and, interestingly, all of these had PHN. This is important because it suggests that some patients with PHN may require additional intervention to control the disorder other than just analgesic medications. One of the most compelling aspects of this project is the progressive layering and evolution of the datasets and their uses. As we increase the number of test antigens, and assay across conditions and diseases, we assemble comprehensive evaluations of immune and autoimmune responses. This is accomplished by determination of (a) the extent and specificity of immune response to orthologous proteins and protein fragments, (b) overlap in antigen profiles indicative of common denominators or general mechanisms, and (c) antigenicity within an entire signaling pathway involved in inter- or intracellular communication. Examples are the TRIM family of proteins in Sjogren's Syndrome and our autoantigen-based classification of patients with Lupus Erythematosis. An example of a new use for public health monitoring is our earlier demonstration of autoantibody measurements in saliva, which highlights the feasibility of establishing non-invasive, saliva-based assays for many types of human diseases or assessment of vaccine immune status for large or vulnerable populations of people or the pediatric population. Eventually, full multiple antigen profiling can be implemented to obtain a deeper level of understanding of many complex human disease states.