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.

项目摘要 嗜碱性粒细胞在过敏性疾病中起重要作用：它们的激活越来越多地用于食物 过敏诊断及其迁移到组织已与开始的发作相关 过敏反应。但是，由于嗜碱性嗜嗜嗜性粒细胞很少 比其他白细胞，特别是在体内微流体系统中。该项目的总体目标 是通过开发启用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，并通过将食物过敏作为模型疾病。 食物过敏已达到流行病，并已成为重要的来源 医疗保健负担。常见的体外过敏测试缺乏足够的精度。口服食物挑战， 食物过敏评估的黄金标准通常不会进行，因为它将患者置于 过敏反应的风险。因此，通常只有在反应后才发现食物过敏 可能是威胁生命的。另外，在开始的发作之间没有可靠的相关性 过敏反应和最小违规剂量的过敏原。预测过敏反应风险的指标 患者本质上不存在。 我们的长期目标是通过开发1）过敏诊断测试来解决这些挑战 准确，安全，快速且可访问的，以及2）指标，以更好地对处于高风险的患者进行分层 过敏反应。该试点赠款的总体目标是检验以下假设：1）阻抗 激活后嗜碱性粒细胞的变化，暴露于负电荷的外部颗粒 蛋白聚糖是一种准确且简单的无标签指标，用于嗜碱性粒细胞，2） 嗜碱性迁移和分泌谱与反应严重程度相关。我们的方法包括 使用临床证实和特征良好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 微流体技术和人类免疫学。我们将：（目标1）设计和验证无标签 在食品过敏原的存在下，基于阻抗的嗜碱性嗜碱性激活测试使用全血 体内和（AIM 2）设计和验证耦合的嗜碱性粒细胞迁移和多重分泌物 测定离体以确定与过敏反应病史的相关性。