Combining avidin with CD63 improves basophil activation test accuracy in classifying peanut allergy.
Combining avidin with CD63 improves basophil activation test accuracy in classifying peanut allergy.
复制标题
将抗生物素蛋白与 CD63 结合可提高花生过敏分类中嗜碱性粒细胞活化测试的准确性。
作者：
Castaño,Nicolas;Chua,Kaiser;Kaushik,Abhinav;Kim,Sungu;Cordts,SethC;Nafarzadegan,CeenaD;Hofmann,GradyH;Seastedt,Hana;Schuetz,JacksonP;Dunham,Diane;Parsons,EllaS;Tsai,Mindy;Cao,Shu;Desai,Manisha;Sindher,SayantaniB;Chinthr
传统的嗜碱性粒细胞活化试验(BATS)通过增加CD63的表达来检测嗜碱性粒细胞的激活。以前，荧光团标记的亲和素是一种带正电荷的分子，被发现与激活的嗜碱性细胞结合，在脱颗粒过程中往往会暴露出带负电荷的颗粒成分。本研究进一步比较了亲和素和CD63作为嗜碱性粒细胞激活生物标志物对花生过敏的分类。方法对70名花生过敏(N= 47)、花生以外的食物过敏(N= 6)或无食物过敏(N= 17)的受试者进行评估。我们对7种浓度的花生提取物(PE)和4种对照条件(无刺激剂、抗 E、fMLP和抗FCεRI)进行了BATS实验。用流式细胞仪检测嗜碱性粒细胞亲和素结合和CD63的表达。我们评估了Logistic回归和XGBoost模型用于花生过敏分类和特征识别。结果亲和素结合与CD63表达相关。这两个标记都区分了有花生过敏和没有花生过敏的受试者。虽然百分比很小，但在所有受试者中都发现了亲和素+/CD63−细胞亚群，这表明亲和素和CD63的结合可以更全面地鉴定活化的嗜碱性粒细胞。事实上，我们通过在七次PE剂量中结合亲和素和CD63获得了最好的分类准确性(97.8%的敏感性，96.7%的特异性)。联合PE剂量为10,000 ng/mL的亲和素和10,100 ng/mL的CD63的准确度相近。结论Avidin和CD63是与脱颗粒相关的可靠BAT激活标志物。它们的结合提高了对活化的嗜碱性粒细胞的鉴定，提高了花生过敏的分类准确率。
BackgroundConventional basophil activation tests (BATs) measure basophil activation by the increased expression of CD63. Previously, fluorophore‐labeled avidin, a positively‐charged molecule, was found to bind to activated basophils, which tend to expose negatively charged granule constituents during degranulation. This study further compares avidin versus CD63 as basophil activation biomarkers in classifying peanut allergy.MethodsSeventy subjects with either a peanut allergy (N= 47), a food allergy other than peanut (N= 6), or no food allergy (N= 17) were evaluated. We conducted BATs in response to seven peanut extract (PE) concentrations (0.01–10,000 ng/mL) and four control conditions (no stimulant, anti‐IgE, fMLP (N‐formylmethionine‐leucyl‐phenylalanine), and anti‐FcεRI). We measured avidin binding and CD63 expression on basophils with flow cytometry. We evaluated logistic regression and XGBoost models for peanut allergy classification and feature identification.ResultsAvidin binding was correlated with CD63 expression. Both markers discriminated between subjects with and without a peanut allergy. Although small by percentage, an avidin+/CD63−cell subset was found in all allergic subjects tested, indicating that the combination of avidin and CD63 could allow a more comprehensive identification of activated basophils. Indeed, we obtained the best classification accuracy (97.8% sensitivity, 96.7% specificity) by combining avidin and CD63 across seven PE doses. Similar accuracy was obtained by combining PE dose of 10,000 ng/mL for avidin and PE doses of 10 and 100 ng/mL for CD63.ConclusionsAvidin and CD63 are reliable BAT activation markers associated with degranulation. Their combination enhances the identification of activated basophils and improves the classification accuracy of peanut allergy.