Basophil phenotype and activation in multi-food allergy and oral immunotherapy

项目总结 食物过敏(FA)是一种潜在的致命疾病，需要改进诊断、临床监测、 和治疗。嗜碱性粒细胞是血液中最少见的一种，已被认为是重要的 FA和食物过敏反应的致病因素。某些细胞表面表型特征可以 帮助鉴定在体内或体外被过敏机制激活的嗜碱性粒细胞。在这个项目中， 我们将确定血液嗜碱性粒细胞的表型和功能特征是否可以作为 预测多种食物过敏原对FA临床反应性严重程度的方法 食品(这里：多FA)，为了提高口服免疫疗法(OIT)方案治疗FA的安全性和有效性， 原则上，为每个对食物过敏的人定制免疫治疗方案。 在目标1中，我们将进行详细的免疫表型鉴定，以监测表型(S)和激活状态 多发FA患者血液中嗜碱性粒细胞及其亚群的体外基线测量 (这里的定义是没有过敏原或抗IgE的体外刺激)。血液将在手术前采集 患者参加多OIT(定义为同步)的先导性、机械性、2期临床试验 OIT具有多种食物过敏原)，使用或不使用奥马珠单抗或杜匹单抗，并以多个间隔进行 在免疫治疗(IT)过程中和结束后(见项目1)；我们还将分析血液 来自没有接受OIT治疗的多FA参与者、没有FA的特应性患者和健康的非特应性对照。这 工作将检验假设：1)活化和/或其他标志物在嗜碱性粒细胞中的表达 多FA患者在细胞体外刺激前的基线状态与患者的急性临床相关 在IT前和IT期间对食物过敏原的反应性，可用于监测 IT导致的脱敏和/或持续的无反应/“耐受性”。 在目标2中，我们将进行嗜碱性粒细胞表型和功能的体外研究(即，嗜碱性粒细胞激活试验 [蝙蝠])，在体外测量嗜碱性细胞对食物过敏原挑战的反应。这项工作将考验 假设，在多发性FA患者中，嗜碱性粒细胞表型变化的幅度(例如， 激活标记物)和/或功能(例如，介质的分泌) 食物过敏原体外与IT期间食物过敏原敏感性的临床证据减少相关 也可能与脱敏与持续无反应/“耐受性”的临床结果相关。 我们的主要长期目标之一是开发创新、快速和可靠的方法来监控嗜碱性细胞 临床上可用于预测一个人的临床敏感度的表型和功能 针对食物过敏原，预测OIT的临床结果，为个人定制OIT和其他IT方案 和/或监测免疫疗法的发展、有效性或持久性。

PROJECT SUMMARY Food allergy (FA) is a potentially fatal disorder in need of improved methods for diagnosis, clinical monitoring, and treatment. Basophils, the least common of the blood granulocytes, have been implicated as important contributors to the pathology of FA and food-induced anaphylaxis. Certain cell surface phenotypic features can help to identify basophils which have been activated by allergic mechanisms in vivo or in vitro. In this project, we will determine whether phenotypic and functional features of blood basophils can be used as part of an approach to predict the severity of clinical reactivity to food allergens in people with FA induced by multiple foods (herein: multi-FA), to improve the safety and efficacy of oral immunotherapy (OIT) protocols to treat FA, and, in principle, to customize immunotherapy protocols for each individual food allergic person. In Aim 1, we will perform detailed immunophenotyping to monitor the phenotype(s) and activation status of basophils and their subpopulations in the blood of participants with multi-FA, as measured ex vivo at baseline (defined herein as without ex vivo stimulation with allergens or anti-IgE). Blood will be obtained before the patients’ enrollment in a pilot, mechanistic-focused, phase 2 clinical trial of multi-OIT (defined as simultaneous OIT with multiple food allergens) conducted with or without omalizumab or dupilumab and at multiple intervals during the course of and after the end of such immunotherapy (IT) (see Project 1); we also will analyze blood from multi-FA participants not treated with OIT, atopic people without FA, and healthy non-atopic controls. This work will test the hypotheses: 1) that the expression of activation and/or other markers in blood basophils of multi-FA patients at baseline, before the cells’ ex vivo stimulation, correlates with the patients’ acute clinical reactivity to food allergens before and during IT and can be used to monitor the development and durability of desensitization and/or sustained unresponsiveness/“tolerance” induced by IT. In Aim 2, we will perform in vitro studies of basophil phenotype and function (i.e., basophil activation tests [BATs]), to measure basophil responses to challenge with food allergens ex vivo. This work will test the hypothesis that, in multi-FA patients, the magnitude of changes in basophil phenotype (e.g., expression of activation markers) and/or function (e.g., secretion of mediators) induced by challenge of their basophils with food allergens ex vivo correlates with the reduction in clinical evidence of food allergen sensitivity during IT and may also correlate with clinical outcomes of desensitization vs. sustained unresponsiveness/“tolerance”. One of our key long term goals is to develop innovative, rapid, and reliable methods to monitor basophil phenotype and function that can be used clinically for such purposes as predicting a person’s clinical sensitivity to food allergens, predicting the clinical outcome of OIT, customizing OIT and other IT protocols for individual people, and/or monitoring the development, effectiveness, or durability of immunotherapy.