Bachmeier-Zbären Noemi, Celik Alper, van Brummelen Robin, Roos Nadine, Steinmann Melanie, Hoang Jennifer A, Yin Xiaojun, Ditlof Christina M, Duan Lucy, Upton Julia E M, Kaufmann Thomas, Eggel Alexander, Eiwegger Thomas
Department of Rheumatology and Immunology, University Hospital Bern, Bern, Switzerland.
Centre for Computational Medicine, Hospital for Sick Children, Toronto, Ontario, Canada.
Allergy. 2025 Jan;80(1):215-226. doi: 10.1111/all.16341. Epub 2024 Sep 28.
Peanut allergy is among the most severe and common food allergies. The diagnosis has a significant impact on the quality of life for patients and their families. An effective management approach depends on accurate, safe, and easily implementable diagnostic methods. We previously developed a cell-based assay using Hoxb8 mast cells (Hoxb8 MCs) aimed at improving clinical allergy diagnosis. In this study, we assessed its diagnostic performance by measuring blinded sera from a prospectively enrolled and pre-validated peanut allergy cohort.
Hoxb8 MCs were passively sensitized with sera from peanut-allergic and peanut tolerant children and adolescents (n = 112). Degranulation of Hoxb8 MCs was quantified upon stimulation with dose-titrated peanut extract by means of flow cytometry, using CD107a as activation marker. The results from the Hoxb8 mast cell activation test (Hoxb8 MAT) were compared to established diagnostic assays such as the skin prick test (SPT), specific IgE (sIgE) levels, and the basophil activation test (BAT). Additionally, serum samples from BAT nonresponders were assessed with the Hoxb8 MAT.
Hoxb8 MAT displayed a robust dose-dependent activation to peanut extract, with a cutoff value of ≤5.2% CD107a positive cells. The diagnostic accuracy was highest at allergen concentrations ≥100 ng/mL, with an area under the receiver operating characteristic curve (AUROC) of 0.97, 93% sensitivity, and 96% specificity, outperforming traditional SPT and sIgE tests. When compared to BAT, Hoxb8 MAT exhibited comparable diagnostic efficacy. Moreover, sera from BAT nonresponders were accurately classified into allergics and nonallergics by the Hoxb8 MAT.
The Hoxb8 MAT demonstrated a very good diagnostic precision in patients prospectively assessed for peanut allergy comparable to the fresh whole blood-based BAT. Additionally, it demonstrated its value for accurate classification of BAT nonresponders into allergic and nonallergic individuals. Further investigations into its utility in the routine clinical setting are warranted.
花生过敏是最严重且常见的食物过敏之一。该诊断对患者及其家庭的生活质量有重大影响。有效的管理方法依赖于准确、安全且易于实施的诊断方法。我们之前开发了一种基于细胞的检测方法，使用Hoxb8肥大细胞（Hoxb8 MCs），旨在改善临床过敏诊断。在本研究中，我们通过检测来自前瞻性招募并预先验证的花生过敏队列的盲法血清来评估其诊断性能。
用花生过敏和花生耐受儿童及青少年（n = 112）的血清对Hoxb8 MCs进行被动致敏。通过流式细胞术，以CD107a作为激活标志物，在用剂量滴定的花生提取物刺激后，对Hoxb8 MCs的脱颗粒进行定量。将Hoxb8肥大细胞激活试验（Hoxb8 MAT）的结果与既定的诊断检测方法进行比较，如皮肤点刺试验（SPT）、特异性IgE（sIgE）水平和嗜碱性粒细胞激活试验（BAT）。此外，用Hoxb8 MAT评估BAT无反应者的血清样本。
Hoxb8 MAT对花生提取物表现出强烈的剂量依赖性激活，截断值为≤5.2% CD107a阳性细胞。在过敏原浓度≥100 ng/mL时诊断准确性最高，受试者操作特征曲线下面积（AUROC）为0.97，敏感性为93%，特异性为96%，优于传统的SPT和sIgE检测。与BAT相比，Hoxb8 MAT表现出相当的诊断效力。此外，Hoxb8 MAT将BAT无反应者的血清准确分类为过敏者和非过敏者。
Hoxb8 MAT在对花生过敏进行前瞻性评估的患者中显示出非常好的诊断精度，与基于新鲜全血的BAT相当。此外，它还证明了其在将BAT无反应者准确分类为过敏和非过敏个体方面的价值。有必要对其在常规临床环境中的效用进行进一步研究。
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