Probing basophil function in microfluidic systems for allergic disease diagnosis

探测微流体系统中的嗜碱性粒细胞功能以诊断过敏性疾病

• 批准号: 10302059
• 负责人: Sindy Kam-Yan Tang
• 金额: $ 22.07万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10302059
• 关键词: Address; Adoption; Adverse reactions; Allergens; Allergic; Allergic Disease; Allergic Reaction; Anaphylaxis; Basophils; Behavior; Biological Assay; Blood; Blood specimen; Cells; Charge; Chemotaxis; Clinical; Color; Complement; Coupled; Cytoplasmic Granules; Data; Development; Diagnosis; Diagnostic; Diagnostic tests; Disease; Disease model; Dose; Epidemic; Flow Cytometry; Food; Food Hypersensitivity; Frequencies; Goals; Gold; Grant; Healthcare; Human; Hypersensitivity; Immune; Immunology; Immunotherapy; In Vitro; Label; Leukocytes; Life; Measurement; Measures; Mediator of activation protein; Medical Care Costs; Methods; Microfluidics; Monitor; National Institute of Biomedical Imaging and Bioengineering; Oral; Outcome; Patient risk; Patients; Pilot Projects; Play; Predictive Value; Proteoglycan; Reaction; Recording of previous events; Risk; Role; Sensitivity and Specificity; Severities; Signaling Molecule; Source; Stratification; Surface; System; Testing; Time; Tissues; Validation; Whole Blood; Work; base; biochip; companion diagnostics; cost; design; disease diagnosis; electric impedance; food allergen; food challenge; food standard; high risk; improved; innovation; microfluidic technology; migration; mouse model; patient stratification; rapid test; tool

PROJECT SUMMARY Basophils play important roles in allergic diseases: their activation is increasingly used for food allergy diagnosis, and their migration into tissues has been correlated with the onset of anaphylaxis. However, since basophils are rare, they are less studied and utilized for diagnostics than other leukocytes, especially in ex vivo microfluidic systems. The overall goal of this project is to fill this gap by developing enabling microfluidic technologies to better quantify basophil function, and by using food allergy as a model disease. Food allergy has reached epidemic proportions and has become a significant source of healthcare burden. Common in vitro allergy tests lack sufficient accuracy. Oral food challenge, the gold standard for food allergy assessment, is often not performed as it places the patient at risk of anaphylaxis. As such, food allergy is often identified only after an adverse reaction that could be life-threatening. In addition, there is no reliable correlation between the onset of anaphylaxis and a minimum offending dose of allergen. Metrics to predict anaphylaxis risk in a patient are essentially non-existent. Our long-term goal is to address these challenges by developing 1) allergy diagnostic tests that are accurate, safe, rapid, and accessible, and 2) metrics to better stratify patients at high risk of anaphylaxis. The overall objective of this pilot grant is to test the hypotheses that: 1) impedance changes in basophils upon activation, which exposes negatively-charged exteriorized granule proteoglycans, is an accurate and simple label-free indicator for basophil activation, and 2) basophil migration and secretion profile correlates with reaction severity. Our approach includes using whole blood samples from clinically-proven and well-characterized peanut allergic subjects and healthy controls, and leveraging our interdisciplinary team's expertise in innovative microfluidic technology and human immunology. We will: (Aim 1) design and validate a label-free impedance-based basophil activation test using whole blood in the presence of food allergen ex vivo, and (Aim 2) design and validate a coupled basophil migration and multiplexed secretion assay ex vivo to determine the correlation with patient history of anaphylaxis.

项目概要 嗜碱性粒细胞在过敏性疾病中发挥重要作用：它们的激活越来越多地用于食品 过敏诊断及其向组织的迁移与过敏的发生相关 过敏反应。然而，由于嗜碱性粒细胞很少见，因此对它们的研究和诊断用途较少 比其他白细胞，特别是在离体微流体系统中。本项目的总体目标 是通过开发微流体技术来更好地量化嗜碱性粒细胞来填补这一空白 功能，并使用食物过敏作为模型疾病。 食物过敏已达到流行病的程度，并已成为一个重要的来源 医疗负担。常见的体外过敏测试缺乏足够的准确性。口服食物挑战， 食物过敏评估的黄金标准通常不会被执行，因为它将患者置于 过敏反应的风险。因此，食物过敏通常只有在出现以下不良反应后才能发现： 可能会危及生命。此外，发病与发病之间没有可靠的相关性。 过敏反应和过敏原的最小致病剂量。预测过敏反应风险的指标 病人基本上不存在。 我们的长期目标是通过开发 1) 过敏诊断测试来应对这些挑战 准确、安全、快速且易于访问，以及 2) 更好地对高风险患者进行分层的指标 过敏反应。该试点拨款的总体目标是测试以下假设：1) 阻抗 激活后嗜碱性粒细胞发生变化，暴露出带负电的外化颗粒 蛋白聚糖，是一种准确且简单的无标记嗜碱性粒细胞激活指示剂，2) 嗜碱性粒细胞迁移和分泌特征与反应严重程度相关。我们的方法包括 使用来自经临床证明和充分表征的花生过敏受试者的全血样本 和健康的控制，并利用我们的跨学科团队在创新方面的专业知识 微流控技术和人体免疫学。我们将：（目标 1）设计并验证无标签 在存在食物过敏原的情况下使用全血进行基于阻抗的嗜碱性粒细胞活化测试 体内，以及（目标 2）设计并验证偶联的嗜碱性粒细胞迁移和多重分泌 离体测定以确定与患者过敏反应史的相关性。