Syngenta Central Toxicology Laboratory, Cheshire, United Kingdom.
J Immunotoxicol. 2005 Jul 1;1(3):189-99. doi: 10.1080/15476910490919140.
Allergen-specific IgE production is the central event in the pathogenesis of atopic disorders and increases in specific IgE serum antibodies are an indicator of immediate hypersensitivity responses in humans and in animal models of allergy. Consequently, accurate and user-friendly methods are needed to measure serum levels of allergen-specific IgE. This review examines historical and recent developments in in vivo and in vitro methods for the detection of allergen-specific IgE in humans and in animal models. Routinely, in vitro methods such as enzyme-linked immunosorbant assays or radioallergosorbant tests and in vivo methods such as the skin prick test (SPT) for humans and the passive cutaneous anaphylaxis assay (PCA) used in animals are utilized to detect allergen-specific IgE. While in vivo assays are usually more accurate than in vitro assays since they provide a functional readout of IgE activity, they are relatively costly and require considerable expertise. On the other hand in vitro assays are limited by the fact that the amount of allergen-specific serum IgG exceeds IgE antibody by several orders of magnitude, resulting in competition for allergen binding. Consequently, methods that use allergen as a direct capture step are limited by the availability of free allergen binding sites for IgE. In order to circumvent this problem, in vitro methods usually require prior depletion of IgG or use high amounts of allergen in order to facilitate availability of free binding sites for IgE detection. Clearly, these approaches are limited for small sample volumes and allergens that are in short supply. New methods such as protein microarray could potentially overcome this problem by providing high allergen concentrations in a relatively small reaction volume. Currently, in vitro methods are rarely used in isolation for prognosis but are used primarily to complement the information obtained from in vivo assays. With the emergence of new technologies it is conceivable that in vitro assays may in the future replace in vivo assays, however until then in vivo assays remain the gold standard of allergen-specific IgE detection.
过敏原特异性 IgE 的产生是特应性疾病发病机制中的核心事件,特异性 IgE 血清抗体的增加是人类和过敏动物模型中即刻超敏反应的指标。因此,需要准确且易于使用的方法来测量过敏原特异性 IgE 的血清水平。本文综述了人类和动物模型中检测过敏原特异性 IgE 的体内和体外方法的历史和最新进展。通常,体外方法如酶联免疫吸附测定或放射过敏原吸附试验,以及体内方法如皮肤点刺试验(SPT)用于人类和被动皮肤过敏反应测定(PCA)用于动物,用于检测过敏原特异性 IgE。虽然体内检测通常比体外检测更准确,因为它们提供了 IgE 活性的功能读数,但它们相对昂贵且需要相当多的专业知识。另一方面,体外检测受到限制,因为过敏原特异性血清 IgG 的量比 IgE 抗体高几个数量级,导致过敏原结合竞争。因此,使用过敏原作为直接捕获步骤的方法受到 IgE 结合的游离过敏原结合位点可用性的限制。为了克服这个问题,体外方法通常需要预先耗尽 IgG 或使用大量过敏原,以便为 IgE 检测提供游离结合位点。显然,这些方法对于小样本量和供应短缺的过敏原有限。新方法,如蛋白质微阵列,可以通过在相对较小的反应体积中提供高过敏原浓度来潜在地克服这个问题。目前,体外方法很少单独用于预后,但主要用于补充体内检测获得的信息。随着新技术的出现,可以想象,体外检测在未来可能会取代体内检测,然而,在此之前,体内检测仍然是过敏原特异性 IgE 检测的金标准。