Instituto de Investigación en Ciencias de la Alimentación (CIAL), CSIC-UAM, Madrid, Spain.
Jaffe Food Allergy Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA.
Methods Mol Biol. 2021;2223:49-65. doi: 10.1007/978-1-0716-1001-5_4.
Egg allergy is one of the most common food allergies in children, being the most important allergenic proteins found in the egg white (EW). Allergy to EW shows a complex phenotype that involves a multifaceted reaction that can only be assessed in vivo. Although other routes of sensitization have been described, oral exposure to food antigens is one of the most suitable in humans. In mice, oral administration of allergenic proteins results in the development of tolerance, and the use of adjuvants, such as cholera toxin (CT), is required to promote Th2-biased immune responses over tolerogenic responses. In this regard, among the mouse strains that readily display Th2 responses, Balb/c has been widely used. Here, we describe a frequently used protocol of oral EW sensitization by using CT as an adjuvant and we explain in detail the methods that we have developed to analyze the sensitizing and eliciting capacity of EW proteins including evaluation of signs, measurement of serum levels of specific immunoglobulins, mast cell degranulation, cytokine secretion profile of allergen-reactive T cells, phenotyping of mesenteric lymph node- and spleen-derived dendritic and T cells by flow cytometry, and quantification of intestinal gene expression.
鸡蛋过敏是儿童最常见的食物过敏之一,其过敏原主要存在于蛋清(EW)中。EW 过敏表现出一种复杂的表型,涉及多方面的反应,只能在体内进行评估。尽管已经描述了其他致敏途径,但口服暴露于食物抗原是最适合人类的途径之一。在小鼠中,口服给予过敏原蛋白会导致耐受的发展,并且需要使用佐剂,如霍乱毒素(CT),以促进 Th2 偏向性免疫反应而不是耐受反应。在这方面,在容易显示 Th2 反应的小鼠品系中,Balb/c 已被广泛使用。在这里,我们描述了一种常用的通过 CT 作为佐剂进行 EW 口服致敏的方案,并详细说明了我们开发的用于分析 EW 蛋白致敏和引发能力的方法,包括评估症状、测量特异性免疫球蛋白的血清水平、肥大细胞脱颗粒、过敏原反应性 T 细胞的细胞因子分泌谱、通过流式细胞术对肠系膜淋巴结和脾脏来源的树突状细胞和 T 细胞进行表型分析以及定量肠道基因表达。