Palomares Oscar, Villalba Mayte, Quiralte Joaquín, Rodríguez Rosalía
Departamento de Bioquímica y Biología Molecular, Facultad de Química, Universidad Complutense de Madrid, Madrid, Spain.
Ann Allergy Asthma Immunol. 2006 Jul;97(1):61-5. doi: 10.1016/S1081-1206(10)61371-3.
Designing of methods for an accurate diagnosis is a main goal of allergy research. Olive pollen allergy is currently diagnosed using commercially available pollen extracts that do not allow identification of the molecules that elicit the disease.
To analyze the suitability of using the N- and C-terminal domains (NtD and CtD, respectively) of the 1,3-beta-glucanase Ole e 9, a major allergen from olive pollen, for in vitro diagnosis.
Serum samples from 55 olive-allergic patients were assayed using enzyme-linked immunosorbent assay to study hypersensitive patients with IgE reactivity to Ole e 9. The specific IgEs to NtD and CtD, obtained by recombinant technology, were determined by means of immunoblotting, enzyme-linked immunosorbent assay, and inhibition assays.
Thirty-one of 33 serum samples from Ole e 9-allergic patients were IgE reactive to recombinant NtD (rNtD) (n = 26 [79%]), recombinant CtD (rCtD) (n = 22 [67%]), or both (n = 17 [52%]). Nine patients (27%) were exclusively reactive to rNtD and 5 (15%) to rCtD. Inhibition assays of IgE binding to Ole e 9 with a mixture of both domains abolished 90% of the binding, whereas 44% and 45% were abolished when rNtD and rCtD were used, respectively.
Because sensitization to NtD or CtD of Ole e 9 could be correlated to vegetable food-latex-pollen cross-reactivity processes or to the exacerbation and persistence of asthma, respectively, these molecules could be used in vitro as markers of disease to classify patients and to design a patient-tailored immunotherapy approach.
设计准确的诊断方法是过敏研究的主要目标。目前,橄榄花粉过敏是使用市售花粉提取物进行诊断的,这些提取物无法识别引发该疾病的分子。
分析使用橄榄花粉主要过敏原1,3-β-葡聚糖酶Ole e 9的N端和C端结构域(分别为NtD和CtD)进行体外诊断的适用性。
采用酶联免疫吸附测定法检测55例对橄榄过敏患者的血清样本,以研究对Ole e 9具有IgE反应性的过敏患者。通过重组技术获得的针对NtD和CtD的特异性IgE,采用免疫印迹法、酶联免疫吸附测定法和抑制试验进行测定。
33例对Ole e 9过敏患者的血清样本中,有31例对重组NtD(rNtD)(n = 26 [79%])、重组CtD(rCtD)(n = 22 [67%])或两者(n = 17 [52%])呈IgE反应性。9例患者(27%)仅对rNtD有反应,5例(l5%)仅对rCtD有反应。用两个结构域的混合物对IgE与Ole e 9的结合进行抑制试验,可消除90%的结合,而分别使用rNtD和rCtD时,结合被消除的比例分别为44%和45%。
由于对Ole e 9的NtD或CtD致敏可能分别与植物性食物-乳胶-花粉交叉反应过程或哮喘的加重和持续有关,因此这些分子可在体外用作疾病标志物,用于对患者进行分类并设计针对患者的免疫治疗方法。