Duffort Oscar, Palomares Oscar, Lombardero Manuel, Villalba Mayte, Barber Domingo, Rodríguez Rosalía, Polo Florentino
Departamento de Investigación y Desarrollo, ALK-ABELLO, Facultad de Ciencias Químicas, Universidad Complutense, Madrid, España.
Int Arch Allergy Immunol. 2006;140(2):131-8. doi: 10.1159/000092532. Epub 2006 Apr 3.
Clustered severe adverse reactions to immunotherapy with olive pollen extracts have been occasionally reported in areas where olive trees are extensively grown. Allergic patients from these areas, in addition to the major olive pollen allergen Ole e 1, frequently recognize a recently described allergen, Ole e 9.
We aimed to develop an immunoassay to measure Ole e 9 concentration and to study the variability of this allergen in olive pollen extracts.
Monoclonal antibodies (mAb) to Ole e 9 were produced from mice immunized with the pure allergen. One of these mAbs was used to develop a sandwich ELISA with an anti-olive pollen extract rabbit serum as the tracer. Olive pollen batches from several suppliers were analyzed using this method. These batches were also analyzed for Ole e 1 content and biological activity.
A 10-fold variation between the extreme values was found for the biological activity of the batches analyzed. Ole e 1 concentration showed a 25-fold variation. Variability of Ole e 9 concentration was extremely high, up to 161 times. The ratio Ole e 1/Ole e 9 varied in a range from 0.6 to 390.4.
The availability of a mAb-based ELISA for Ole e 9 made it possible for us to detect an important source of variability in olive pollen batches. This variability may be the cause of outbreaks of adverse reactions in the course of immunotherapy treatments, which have sometimes been observed among olive-allergic patients living in areas with very high levels of airborne olive pollen.
在橄榄树广泛种植的地区,偶尔会报告对橄榄花粉提取物免疫疗法的聚集性严重不良反应。来自这些地区的过敏患者,除了主要的橄榄花粉过敏原油橄榄孢粉素Ole e 1外,还经常识别出一种最近描述的过敏原Ole e 9。
我们旨在开发一种免疫测定法来测量Ole e 9的浓度,并研究这种过敏原在橄榄花粉提取物中的变异性。
用纯过敏原免疫小鼠,制备针对Ole e 9的单克隆抗体(mAb)。其中一种mAb用于开发一种夹心酶联免疫吸附测定(ELISA),以抗橄榄花粉提取物兔血清作为示踪剂。使用该方法分析了来自几个供应商的橄榄花粉批次。还对这些批次的Ole e 1含量和生物活性进行了分析。
在所分析批次的生物活性方面,发现极值之间存在10倍的差异。Ole e 1浓度显示出25倍的差异。Ole e 9浓度的变异性极高,高达161倍。Ole e 1/Ole e 9的比值在0.6至390.4的范围内变化。
基于mAb的Ole e 9 ELISA的可用性使我们能够检测橄榄花粉批次中一个重要的变异性来源。这种变异性可能是免疫治疗过程中不良反应爆发的原因,在生活在空气中橄榄花粉水平非常高的地区的橄榄过敏患者中有时会观察到这种情况。