Mousavi F, Kardar G A, Pourpak Z
Space Biology and Environment Center, Aerospace Research Institute, Ministry of Science Research and Technology, Tehran, Iran.
Immunology, Asthma and Allergy Research Institute, Tehran University of Medical Sciences, Tehran, Iran.
Allergol Immunopathol (Madr). 2019 May-Jun;47(3):272-276. doi: 10.1016/j.aller.2018.09.007. Epub 2018 Nov 28.
Murine models have been widely used in the study of allergy as sensitized mice can produce IgE and/or IgG1in response after the injection of an antigen/adjuvant combination. Ailanthus altissima pollen (AAP) has been recently reported as an emerging aeroallergen in Iran. So far, several AAP candidate allergens by the screening of allergen-specific IgE in the sera from AAP sensitized patients in Iran.
The aim of the present study was to detect and compare the allergens eliciting an IgE response in a mouse model, and in human, using pollen extract of A. altissima and an immunoproteomics based approach.
The pollen proteins were extracted in phosphate-buffered saline (PBS). Thirty male BALB/c mice were randomly divided into two groups of AP extract sensitized and sham that respectively received AAP PBS extract and a PBS control by intraperitoneal injections at regular intervals. The optimized AAP protein extracts were analyzed using 2D-gel electrophoresis and were subsequently confronted to pooled sera of sensitized mice.
Two-D gel electrophoresis of AAP extract allowed the separation of 125 protein spots distributed in a wide range of pI and molecular masses. Two-DE immunoblotting using pooled sera of sensitized mice led to the detection of 14 IgE reactive spots with molecular masses ranging from 12 to 40-42kDa.
The results do not correlate with our previous analyses using human AAP-sensitized sera. These findings reflect some differences in the sIgE reactivity to allergenic proteins in animal models.
小鼠模型已广泛应用于过敏研究，因为致敏小鼠在注射抗原/佐剂组合后可产生IgE和/或IgG1。臭椿花粉（AAP）最近被报道为伊朗一种新出现的气传过敏原。到目前为止，通过对伊朗AAP致敏患者血清中过敏原特异性IgE的筛选，已发现了几种AAP候选过敏原。
本研究旨在使用臭椿花粉提取物和基于免疫蛋白质组学的方法，检测并比较在小鼠模型和人类中引发IgE反应的过敏原。
花粉蛋白用磷酸盐缓冲盐水（PBS）提取。30只雄性BALB/c小鼠随机分为AAP提取物致敏组和假处理组，分别定期腹腔注射AAP PBS提取物和PBS对照。对优化后的AAP蛋白提取物进行二维凝胶电泳分析，随后与致敏小鼠的混合血清进行比对。
AAP提取物的双向凝胶电泳可分离出125个蛋白点，分布在较宽的等电点和分子量范围内。使用致敏小鼠混合血清进行的双向电泳免疫印迹检测到14个IgE反应性斑点，分子量范围为12至40 - 42 kDa。
结果与我们之前使用人类AAP致敏血清的分析不相关。这些发现反映了动物模型中sIgE对致敏蛋白反应性的一些差异。
