Basophil phenotype & activation in peanut allergy and oral Immunotherapy

项目总结（见说明）：花生过敏（PA）是一种潜在的致命性疾病，需要改进诊断、临床监测和治疗方法。嗜碱性粒细胞，最常见的血液粒细胞，已被牵连的PA和花生引起的过敏反应的病理学的重要贡献者。某些细胞表面表型特征有助于识别体内或体外过敏机制激活的嗜碱性粒细胞。在这个项目中，我们试图确定血液嗜碱性粒细胞的表型和功能是否可以作为一种方法的一部分来预测PA个体受试者对花生的临床反应的严重程度，以提高口服免疫疗法（OIT）治疗PA的安全性和有效性，和/或为每个花生过敏个体定制OIT方案。在本项目的目标1中，我们将采用详细的免疫表型分析方法来监测PA儿童和成人受试者外周血中嗜碱性粒细胞的表型和活化状态，如在入组OIT治疗PA的II期临床试验（本U19申请的项目1）之前的基线和OIT过程中的多个时间间隔所测量的。这项工作将测试的假设，PA患者的血液嗜碱性粒细胞中的活化和细胞表面标志物的表达与他们的急性临床反应花生过敏原之前和期间OIT，可用于监测的发展，成功，和持久性的耐受性诱导OIT。在目标2中，我们将进行人嗜碱性粒细胞表型和功能的体外研究（即，嗜碱性粒细胞活化试验[BAT]），以测量这种嗜碱性粒细胞对攻击性（花生）与不相关的过敏原离体攻击的反应。这项工作将检验花生过敏原诱导的表型变化的幅度（例如，CD 20 Sc的表面表达）或功能（例如，PA患者的血液嗜碱性粒细胞的介质分泌）与OIT过程中花生过敏原敏感性的临床证据的降低相关。我们在这项工作中的关键目标之一是开发创新的，快速的，可靠的方法来监测嗜碱性粒细胞的表型和功能，可用于临床预测受试者的临床敏感性花生，预测OIT的临床结果，定制OIT方案最佳个体受试者，和/或监测OIT的发展，有效性或持久性。

PROJECT SUMMARY (See instructions): Peanut allergy (PA) is a potentially fatal disorder In need of improved methods for diagnosis, clinical monitoring, and treatment. Basophils, the least common blood granulocyte, have been implicated as Important contributors to the pathology of PA and peanut-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 seek to determine whether the phenotype and function of blood basophils can be used as part of an approach to predict the severity of clinical reactivity to peanut In individual subjects with PA, to improve the safety and efficacy of an oral Immunotherapy (OIT) protocol to treat PA, and/or to customize the OIT protocol to each individual peanut allergic subject. In Aim 1 of this project, we will employ detailed immuophenotyping approaches to monitor the phenotype and activation status of basophils in the peripheral blood of pediatric and adult subjects with PA, as measured at baseline before their enrollment in a phase 2 clinical trial of OIT for PA (Project 1 of this U19 application) and at multiple intervals during the course of OIT. This work will test the hypothesis that the expression of activation and cell surface markers in the blood basophils of PA patients correlates with their acute clinical reactivity to peanut allergen before and during OIT and can be used to monitor the development, success, and durability of tolerance induced by OIT. In Aim 2, we will perform in vitro studies of human basophil phenotype and function (i.e., basophil activation tests [BATs]), to measure the responses of such basophils to challenge with offending (peanut) vs. irrelevant allergens ex vivo. This work will test the hypothesis that the magnitude of peanut allergen-induced changes in the phenotype (e.g., surface expression ofCD20Sc) or function (e.g., secretion of mediators) of blood basophils of PA patients correlates with the reduction in clinical evidence of peanut allergen sensitivity over the course of OIT. One of our key goals in this work 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 subject's clinical sensitivity to peanut, predicting the clinical outcome of OIT, customizing the OIT protocol optimally for individual subjects, and/or monitoring the development, effectiveness, or durability of OIT.