A Fast Procedure for the Detection of Defects in Toll-like Receptor Signaling
A Fast Procedure for the Detection of Defects in Toll-like Receptor Signaling
复制标题
一种快速检测 Toll 样受体信号传导缺陷的方法
作者：
H. von Bernuth;Cheng;C. Rodríguez;Shen;B. Garty;L. Maródi;H. Chapel;M. Chrabieh;Richard L. Miller;C. Picard;A. Puel;J. Casanova
目标. Toll样受体信号传导的先天性缺陷最近被描述为原发性免疫缺陷，其使受影响的儿童易患危及生命的感染。白细胞介素-1受体相关激酶-4缺乏的患者易患侵袭性肺炎球菌病，而IL-93 B缺乏的患者易患单纯疱疹病毒性脑炎。这些遗传性疾病诊断不足，部分原因是诊断目前需要昂贵和耗时的技术，只有少数专业中心在世界各地。因此，我们的目标是开发一种廉价且快速的测试来检测Toll样受体信号传导中的缺陷。患者和方法。我们使用流式细胞术来评估38名健康对照和7名具有遗传定义的Toll样受体信号传导缺陷的患者（5名白细胞介素-1受体相关激酶-4缺乏症患者和2名IL-93 B缺乏症患者）的粒细胞上的膜结合L-选择素的裂解，这些患者在各种Toll样受体激动剂的激活下。结果所有白细胞介素-1受体相关激酶-4缺陷患者的粒细胞在Toll样受体1/2、2/6、4、7和8激动剂激活后，以及所有IL-93 B缺陷患者的粒细胞在Toll样受体7和8激动剂激活后，均观察到L-选择素脱落受损。所有的健康对照组都对这些刺激做出了反应。结论.通过流式细胞术对粒细胞上的膜结合L-选择素切割的评估可能被证明可用于检测Toll样受体途径中的原发性免疫缺陷，如白细胞介素-1受体相关激酶-4缺陷和IL-93 B缺陷。这个过程是廉价和快速的。因此，它可能适用于全球范围内侵袭性肺炎球菌疾病儿童和单纯疱疹病毒性脑炎患者的常规检测。
OBJECTIVES. Inborn defects in Toll-like receptor signaling are recently described primary immunodeficiencies that predispose affected children to life-threatening infections. Patients with interleukin-1 receptor-associated kinase-4 deficiency are prone to invasive pneumococcal disease, and patients with UNC-93B deficiency are prone to herpes simplex virus encephalitis. These genetic disorders are underdiagnosed, partly because diagnosis currently requires expensive and time-consuming techniques available at only a few specialized centers worldwide. We, therefore, aimed to develop a cheap and fast test for the detection of defects in Toll-like receptor signaling. PATIENTS AND METHODS. We used flow cytometry to evaluate the cleavage of membrane-bound L-selectin on granulocytes in 38 healthy controls and in 7 patients with genetically defined Toll-like receptor signaling defects (5 patients with interleukin-1 receptor-associated kinase-4 deficiency and 2 patients with UNC-93B deficiency), on activation with various Toll-like receptor agonists. RESULTS. Impaired L-selectin shedding was observed with granulocytes from all of the interleukin-1 receptor-associated kinase-4-deficient patients on activation with agonists of Toll-like receptors 1/2, 2/6, 4, 7, and 8 and with granulocytes from all of the UNC-93B-deficient patients on activation with agonists of Toll-like receptors 7 and 8. All of the healthy controls responded to these stimuli. CONCLUSIONS. The assessment of membrane-bound L-selectin cleavage on granulocytes by flow cytometry may prove useful for the detection of primary immunodeficiencies in the Toll-like receptor pathway, such as interleukin-1 receptor-associated kinase-4 deficiency and UNC-93B deficiency. This procedure is cheap and rapid. It may, therefore, be suitable for routine testing worldwide in children with invasive pneumococcal disease and in patients with herpes simplex encephalitis.