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 由于流行率的增加和地理范围的扩大，美国也出现了这种情况。据估计，有30万人 2017年，美国发生了LD病例。病程变化很大，可能包括 器官特异性和播散性受累，有急性和慢性表现。存在知识鸿沟 关于急性LD期间BB在血液中传播的免疫事件。我们的目标是 通过开发测量中性粒细胞-BB相互作用的新工具来弥合这一知识鸿沟 血。在前期工作中，我们论证了微流控技术用于寄主病原体研究的可行性。 血液中的相互作用。具体地说，我们发现(A)在加入AS后，中性粒细胞自发运动表型 血液中只有100个Bb，(B)在急性LD患者的血液样本中有类似的表型，以及(C)a 补体在与BB的早期相互作用中对血液中中性粒细胞的激活起关键作用。继续 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 活的、活动的BB在单细胞分辨率下(2)识别对BB的特征中性粒细胞反应，以及(3)探测继电器 中性粒细胞在少量BB激活血液中补体的作用。我们将开始申请 在特定抗体出现之前，测量急性LD患者中性粒细胞功能的新工具是 生产，并在治疗期间，以评估从血液中清除BB。总体而言，我们的研究将设计新的 用于诊断和监测LD的工具，并将扩大我们对宿主对LD的反应的知识。