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

项目摘要 莱姆病，由于感染硬蜱传播的螺旋体伯氏疏螺旋体（Bb）是最 在美国，这是一种常见的病媒传播疾病，每年有超过30万例新病例。感染可能导致 在无症状IgG血清转化或引起临床疾病中表现为孤立性皮肤损害红斑 移行性（EM）或涉及皮肤、心脏、神经系统和/或关节的全身性疾病。所述感染是 早期发现时对抗生素最敏感，但那些患有播散性感染或 治疗延迟可能会经历使人衰弱的疾病，这种疾病可能对抗生素没有反应。的 与无症状IgG血清转换相关的对Bb的免疫应答，并引起特定器官 对系统参与知之甚少。这项提案将分析先天性和适应性免疫 在具有a）无症状IgG血清转换的充分表征的患者中Bb感染后出现的应答; B）孤立性EM; c）急性播散性感染伴多发性EM和/或神经系统疾病; d）晚期 关节炎的表现。我们将使用最先进的新技术来深入描述免疫系统 随着时间的推移，在表型和功能上对Bb产生反应，并将这些反应与临床相关 介绍和成果。将进行CyTOF等最先进的高分辨率技术 全血和滑液，以及表现出不同趋化因子受体的T细胞库群 将产生定义细胞因子分泌和组织迁移的结果以评价抗原特异性应答。的 通过CyTOF鉴定的血液和其他感兴趣的细胞群体中的应答T细胞亚群的转录组 将通过单细胞RNAseq进行分析。当细胞样品被分离时，将使用新的细胞技术。 限制，如在皮肤活检或CSF样品的情况下，在单细胞水平表征应答细胞 使用MuSIC（多光谱成像细胞术）进行表型分析。这些组织将被询问， 在血液中发现的免疫特征与这些部位特有的免疫特征，并将通过单细胞 RNAseq。沿着免疫反应，我们还将分析宿主代谢物组以确定代谢物 与不同临床结局相关的特征。使我们能够理解免疫反应 将采用系统生物学方法进行数据集成，包括临床状态、宿主代谢物和体外 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， 它们在莱姆病患者体内进化这种功能系统免疫学方法的输出将是 细胞外细菌感染后人体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，最终目标是确定未来 用于干预和预测对系统性疾病的易感性或抵抗力的靶点。

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.