Project 1: Molecular Profiling of the Immune Response in Lyme Disease

项目总结 莱姆病是由于感染硬蜱传播的伯氏疏螺旋体(BB)而引起的 美国常见的病媒传播疾病，每年新增病例超过30万例。感染可能导致 在无症状的免疫球蛋白血清转换或引起临床疾病时表现为孤立的皮损红斑 移行(EM)或涉及皮肤、心脏、神经系统和/或关节的全身性疾病。感染是 早期发现时对抗生素最敏感，但那些患有播散性感染或处于 治疗延误可能会经历衰弱的疾病，可能会对抗生素失去反应。这个 对BB的免疫反应与无症状的IgG血清转换相关，并引起特定的器官 人们对系统参与知之甚少。这项提议将描述先天免疫和获得性免疫 A)无症状的免疫球蛋白血清转换患者在BB感染后出现的反应； B)孤立的EM；c)伴有多种EM和/或神经系统疾病的急性播散性感染；以及d)晚期 关节炎的表现。我们将使用新的最先进的技术来深入描述免疫 随着时间的推移，对BB的反应，包括表型和功能，并将这些反应与临床相关 介绍和结果。将进行最先进的高分辨率技术，如CyTOF 全血和滑液以及显示不同趋化因子受体的T细胞库群体 将产生明确的细胞因子分泌和组织迁移，以评估抗原特异性反应。这个 CyTOF鉴定的血液和其他感兴趣细胞群中反应性T细胞亚群的转录 将通过单细胞RNAseq进行分析。当细胞样本被提取时，将使用新的纳米电池技术 限制，如在皮肤活检或脑脊液样本的情况下，在单细胞水平上表征反应细胞 表型使用MUSIC(多光谱成像细胞术)。这些相同的组织将被审问 血液中发现的免疫信号与这些部位独有的免疫信号，并将通过单细胞进一步评估 RNAseq.除了免疫反应，我们还将分析宿主代谢组以定义代谢物 签名与不同的临床结果相关。使得对免疫反应的理解成为可能 将采用系统生物学的方法进行数据集成，包括临床状态、宿主代谢物和体外数据 细胞反应，以支持对宿主代谢和免疫反应的深入分析和建模，如 它们在患有莱姆病的受试者中进化。这种功能系统免疫学方法的输出将是 感染细胞外细菌后人类代谢物和免疫特征的定义 病原体BB将与其他传染性病原体进行比较，最终目标是定义未来 干预和预测系统性疾病易感性或抵抗力的目标。

PROJECT SUMMARY Lyme disease, due to infection with the Ixodes tick-transmitted spirochete Borrelia burgdorferi (Bb) is the most common vector-borne disease in the United States, with over 300,000 new cases annually. Infection can result in asymptomatic IgG seroconversion or cause clinical disease manifesting as an isolated skin lesion erythema migrans (EM) or with systemic illness involving the skin, heart, nervous system and/or joints. The infection is most responsive to antibiotics when identified early, but those people with disseminated infection or in whom treatment is delayed can experience debilitating disease that can become unresponsive to antibiotics. The immune responses to Bb associated with asymptomatic IgG seroconversion and that give rise to specific organ system involvement are poorly understood. This proposal will profile the innate and adaptive immune responses that arise after Bb infection in well-characterized patients with a) asymptomatic IgG seroconversion; b) isolated EM; c) acute disseminated infection with multiple EM and/or neurologic disease; and d) the late manifestation of arthritis. We will use novel state-of-the-art technologies to deeply characterize the immune response to Bb over time, both phenotypically and functionally, and correlate these responses with clinical presentations and outcomes. State-of-the-art high-resolution technologies such as CyTOF will be conducted on whole blood and synovial fluid, and T cell library populations exhibiting different chemokine receptors defining cytokine secretion and tissue migration will be generated to evaluate antigen-specific responses. The transcriptomes of responding T cell subsets in blood and other cell populations of interest identified by CyTOF will be analyzed by single cell RNAseq. Novel nanowell technologies will be used when cell samples are limiting, as in the case of skin biopsies or CSF samples, to characterize responding cells at the single cell level phenotypically using MuSIC (MultiSpectral Imaging Cytometry). These same tissues will be interrogated for immune signatures found in blood vs those unique to these sites and will be further assessed by single cell RNAseq. Along with the immune responses, we will also profile the host metabolome to define metabolite signatures associated with divergent clinical outcomes. Enabling this understanding of the immune response will be a systems biology approach to data integration, including clinical status, host metabolites and in vitro cell responses, to support an in-depth analysis and modeling of the host metabolic and immune responses as they evolve in subjects with Lyme disease. The output of this functional systems immunology approach will be definitions of human metabolite and immune signatures following infection with an extracellular bacterial pathogen Bb that will be compared to other infectious pathogens, with the ultimate goal of defining future targets for intervention and predicting susceptibility or resistance to systemic disease.