Microfluidic Assays for Probing Neutrophil-Borrelia Interactions in Blood during Acute Lyme Disease

用于探测急性莱姆病期间血液中中性粒细胞与疏螺旋体相互作用的微流控分析

• 批准号: 10379279
• 负责人: Daniel Irimia
• 金额: $ 21万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10379279
• 关键词: Acute; Affect; Aftercare; American; Antibodies; Automobile Driving; Biological Assay; Blood; Blood Cells; Blood donor; Blood specimen; Borrelia; Borrelia burgdorferi; Cell Separation; Cells; Centers for Disease Control and Prevention (U.S.); Cessation of life; Chronic; Complement; Complement Activation; Development; Diagnosis; Diagnostic; Disease; Event; Flow Cytometry; Geographic Distribution; Geography; Health; Hour; Human Pathology; Immune; Immune response; Infection; Innate Immune Response; Knowledge; Lead; Leptospira; Leptospirosis; Leukocytes; Liver Failure; Lyme Disease; Measures; Meningitis; Microbe; Microfluidic Microchips; Microfluidics; Molecular; Monitor; Morphologic artifacts; Neutrophil Activation; Order Spirochaetales; Organ; Outcome; Pathologic Processes; Pathology; Patients; Phagocytosis; Phase; Phenotype; Prevalence; Reporting; Resolution; Respiratory distress; Rest; Role; Science; Serology test; Site; Symptoms; Syphilis; Ticks; Tissues; Treponema pallidum; United States; Work; cell motility; design; diagnostic tool; experience; genomic tools; neutrophil; novel; novel diagnostics; pathogen; renal damage; response; restoration; tick bite; tool

Lyme disease (LD) is caused by infection with Borrelia burgdorferi (Bb) and represents a growing problem in the United States due to increasing prevalence and expanding geographic range. It is estimated that 300,000 cases of LD occurred in the United States in 2017. The disease course is highly variable and can include organ-specific and disseminated involvement with acute and chronic manifestations. A knowledge gap exists regarding the immune events during the dissemination of Bb in blood during acute LD. Our objective is to bridge this knowledge gap through the development of novel tools that measure neutrophil-Bb interactions in blood. In preliminary work, we demonstrate the feasibility of microfluidic assays for the study of host-pathogen interactions in blood. Specifically, we found (A) a neutrophil spontaneous motility phenotype after spiking as few as one hundred Bb in blood, (B) a similar phenotype in blood samples from acute LD patients, and (C) a key role for complement in the activation on neutrophils in blood during early interactions with Bb. Continuing these efforts, we will optimize the neutrophil-assay-design to (1) study the interactions between neutrophils and live, motile Bb at single-cell resolution (2) identify signature neutrophil responses to Bb, and (3) probe the relay function of neutrophils during complement activation in blood by small numbers of Bb. We will begin applying the new tools to measuring neutrophil functions in patients during acute LD before specific antibodies are produced, and during treatment, to evaluate the Bb clearance from blood. Overall, our study will design new tools for the diagnostic and monitoring of LD and will expand our knowledge of host responses to LD.

莱姆病 (LD) 是由伯氏疏螺旋体 (Bb) 感染引起的，是一个日益严重的问题 美国由于患病率增加和地理范围扩大。估计有30万 2017 年美国发生了 LD 病例。病程变化很大，可包括 器官特异性和播散性受累，伴有急性和慢性表现。存在知识差距 关于急性 LD 期间 Bb 在血液中传播过程中的免疫事件。我们的目标是 通过开发测量中性粒细胞与 Bb 相互作用的新工具来弥补这一知识差距 血。在前期工作中，我们证明了微流体测定用于研究宿主病原体的可行性 血液中的相互作用。具体来说，我们发现 (A) 尖峰后中性粒细胞自发运动表型为 血液中只有 100 个 Bb，(B) 急性 LD 患者的血液样本中存在相似的表型，以及 (C) 在与 Bb 的早期相互作用过程中，补体在血液中中性粒细胞的激活中发挥着关键作用。继续 通过这些努力，我们将优化中性粒细胞检测设计，以 (1) 研究中性粒细胞和中性粒细胞之间的相互作用 单细胞分辨率下活的、活动的 Bb (2) 识别中性粒细胞对 Bb 的标志性反应，以及 (3) 探测中继 少量 Bb 激活血液中补体期间中性粒细胞的功能。我们将开始申请 在特异性抗体出现之前测量急性 LD 期间患者中性粒细胞功能的新工具 产生并在治疗期间评估血液中 Bb 的清除率。总的来说，我们的研究将设计新的 诊断和监测 LD 的工具，并将扩展我们对宿主对 LD 反应的了解。