Birmingham Neil, Payankaulam Sandhya, Thanesvorakul Sirinart, Stefura Bill, HayGlass Kent, Gangur Venu
Food Allergy and Immunology Laboratory, Department of Food Science and Human Nutrition, Michigan State University, East Lansing, MI 48823, USA.
J Immunol Methods. 2003 Apr 1;275(1-2):89-98. doi: 10.1016/s0022-1759(03)00008-5.
Passive cutaneous anaphylaxis (PCA) assay has been a gold standard method to measure allergen-specific IgE antibody (ASIgE Ab) levels in allergy mouse models. Many factors including stringent guidelines for laboratory animal use make PCA a difficult choice. Therefore, alternative methods are needed that can be readily applied for measurement of specific IgE antibody levels in mouse serum. Herein we describe a novel ELISA-based method that is more sensitive in comparison to PCA, IgE isotype-specific (because it has little cross-reactivity with IgG1 or IgG2a isotype) and highly reproducible (<10% inter- or intra-assay variation). Furthermore, we demonstrate the utility of this assay to measure specific IgE Ab against a variety of food extracts including chicken egg, peanut, almond, filbert/hazelnut and sweet potato. These findings are of particular interest to those who are seeking (i) to measure food-extract-specific IgE antibody in animal models and (ii) an alternative to the animal-based PCA method to measure mouse IgE antibodies.
被动皮肤过敏反应(PCA)试验一直是测量过敏小鼠模型中过敏原特异性IgE抗体(ASIgE Ab)水平的金标准方法。包括严格的实验动物使用指南在内的许多因素使得PCA成为一个困难的选择。因此,需要能够容易地用于测量小鼠血清中特异性IgE抗体水平的替代方法。在此,我们描述了一种基于ELISA的新方法,该方法与PCA相比更敏感,具有IgE同种型特异性(因为它与IgG1或IgG2a同种型几乎没有交叉反应)且具有高度可重复性(批间或批内变异<10%)。此外,我们证明了该试验在测量针对多种食物提取物(包括鸡蛋、花生、杏仁、榛子和红薯)的特异性IgE Ab方面的实用性。这些发现对于那些寻求(i)在动物模型中测量食物提取物特异性IgE抗体以及(ii)替代基于动物的PCA方法来测量小鼠IgE抗体的人来说特别有意义。