运用多种实验室方法对各型遗传性血管性水肿患者的功能性C1酯酶抑制物进行评估。
Evaluating functional C1INH with multiple laboratory methods across Hereditary Angioedema types.
作者信息
Bardou Maine Luellah Demaret, Constantino-Silva Rosemeire Navickas, Alonso Maria Luiza Oliva, Teixeira Ana Júlia Ribeiro, Giavina-Bianchi Pedro Francisco, Mansour Eli, Pesquero João Bosco, Valle Solange Oliveira Rodrigues, Grumach Anete Sevciovic
机构信息
Clinical Immunology, Centro Universitario Faculdade de Medicina ABC, Santo Andre, SP, Brazil.
Department of Clinical Medicine, Immunology Service, Hospital Universitario Clementino Fraga Filho, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
出版信息
Front Immunol. 2025 Aug 26;16:1654078. doi: 10.3389/fimmu.2025.1654078. eCollection 2025.
INTRODUCTION
Hereditary Angioedema (HAE) is a rare genetic disease characterized by recurrent episodes of edema and classified into HAE with C1 inhibitor deficiency (HAE-C1INH types 1 and 2) and HAE with normal C1INH (HAE-nC1INH). This study evaluates the function of C1 inhibitor (fC1INH) in patients with suspected HAE using several laboratory methods: dried blood spot (DBS), chromogenic assay, and ELISA with FXIIa and PKa (plasma kallikrein). The comparative approach aims to improve early detection and understanding of C1INH dysfunction in all HAE subtypes to reflect real-world diagnostic scenarios.
METHODS
We assessed the diagnostic performance of four fC1INH assays in a cohort of 148 HAE patients: 84 with HAE-C1INH (72 type 1 and 12 type 2) and 64 with HAE-nC1INH (53 HAE-FXII and 11 HAE-UNK). The gold-standard chromogenic assay and the two substrate-specific ELISAs (PKa and FXIIa) were compared to a novel DBS-based LC-MS/MS assay using endogenous C1s activity. For all fC1INH assays, values >50% were considered within the normal range.
RESULTS
In HAE-C1INH, the DBS assay showed the highest specificity (type 1: 98.6%, type 2: 100%) and 100% sensitivity for both subtypes. ELISA-FXIIa also performed well (specificity: 97.2% and 91.7%). In contrast, ELISA-PKa and the chromogenic assay showed reduced specificity in type 2 (25% and 66.7%, respectively). Among patients with HAE-FXII, fC1INH levels were reduced by 36.5% by ELISA-FXIIa (19/52), 19.1% by DBS (9/47), and 3.8% by ELISA-PKa (2/52), and no alterations were detected by the chromogenic assay. Some of the changes seen in other tests may be partly related to pregnancy in a few patients. In the HAE-UNK group, all 11 patients had fC1INH >50% in all methods.
CONCLUSION
DBS-based LC-MS/MS and ELISA-FXIIa offer promising accuracy and broader applicability for early diagnosis of HAE types 1 and 2. The use of novel substrates and the inclusion of a clinically realistic cohort may enhance the translational relevance of these findings.
引言
遗传性血管性水肿（HAE）是一种罕见的遗传性疾病，其特征为反复发作的水肿，可分为C1抑制剂缺乏型HAE（HAE-C1INH 1型和2型）和C1INH正常型HAE（HAE-nC1INH）。本研究使用几种实验室方法评估疑似HAE患者中C1抑制剂（fC1INH）的功能：干血斑（DBS）、发色底物法和使用FXIIa和PKa（血浆激肽释放酶）的ELISA法。这种比较方法旨在改善所有HAE亚型中C1INH功能障碍的早期检测和理解，以反映现实世界中的诊断情况。
方法
我们评估了四种fC1INH检测方法在148例HAE患者队列中的诊断性能：84例HAE-C1INH患者（72例1型和12例2型）和64例HAE-nC1INH患者（53例HAE-FXII和11例HAE-UNK）。将金标准发色底物法和两种底物特异性ELISA法（PKa和FXIIa）与一种基于DBS的新型LC-MS/MS检测方法进行比较，该方法使用内源性C1s活性。对于所有fC1INH检测方法，值>50%被认为在正常范围内。
结果
在HAE-C1INH中，DBS检测显示出最高的特异性（1型：98.6%，2型：100%），且对两种亚型的敏感性均为100%。ELISA-FXIIa的表现也良好（特异性：97.2%和91.7%）。相比之下，ELISA-PKa和发色底物法在2型中显示出降低的特异性（分别为25%和66.7%）。在HAE-FXII患者中，ELISA-FXIIa使fC1INH水平降低了36.5%（19/52），DBS使fC1INH水平降低了19.1%（9/47），ELISA-PKa使fC1INH水平降低了3.8%（2/52），发色底物法未检测到改变。其他检测中观察到的一些变化可能部分与少数患者的妊娠有关。在HAE-UNK组中，所有11例患者在所有方法中fC1INH均>50%。
结论
基于DBS的LC-MS/MS和ELISA-FXIIa为1型和2型HAE的早期诊断提供了有前景的准确性和更广泛的适用性。使用新型底物和纳入临床实际队列可能会增强这些发现的转化相关性。
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