Immunochemical detection of the occupational allergen, methylene diphenyl diisocyanate (MDI), in situ
Immunochemical detection of the occupational allergen, methylene diphenyl diisocyanate (MDI), in situ
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DOI：
10.1016/j.jim.2015.12.008
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发表时间：
2016-02-01
影响因子：
2.2
通讯作者：
Liu, Jian
中科院分区：
文献类型：
--
作者：
Wisnewski, Adam V.;Liu, Jian
聚氨酯生产中必不可少的二异氰酸酯化学品是公认的职业性哮喘的原因。异氰酸酯诱发哮喘的发病机制，包括该化学物质的吸收途径及其在暴露组织（尤其是肺部）中的分布，仍不清楚。我们开发了一种对二苯基甲烷二异氰酸酯（MDI）具有特异性的抗血清，并建立了其原位检测MDI的能力。通过用在完全弗氏佐剂中乳化的与钥孔血蓝蛋白（KLH）缀合的MDI免疫家兔，随后用不完全弗氏佐剂进行两次加强注射来诱导多克隆MDI特异性IgG。抗血清含有IgG，其通过斑点印迹分析识别各种不同的MDI缀合蛋白，但不识别未缀合或模拟暴露的蛋白。抗血清进一步证明了对与MDI缀合的蛋白质的特异性，但对其他常用的二异氰酸酯没有特异性。对鼻内暴露于MDI（作为可逆反应性谷胱甘肽结合物，如GSH-MDI）的小鼠的细胞离心法气道细胞和福尔马林固定石蜡包埋肺组织切片进行的免疫化学研究证明了抗血清检测组织样本中MDI的能力。数据表明MDI渗透到下呼吸道，局部沉积在呼吸道周围的上皮区域，并被肺泡巨噬细胞摄取。这种新的免疫化学试剂应该有助于进一步研究MDI的摄取和组织分布，特别是因为它与暴露的不良健康影响有关。(C)2015 Elsevier B. V.版权所有。
Diisocyanate chemicals essential to polyurethane production are a well-recognized cause of occupational asthma. The pathogenesis of diisocyanate-induced asthma, including the pathways by which the chemical is taken up and its distribution in exposed tissue, especially the lung, remains unclear. We developed an antiserum with specificity for methylene diphenyl diisocyanate (MDI) the most abundantly produced and utilized diisocyanate worldwide, and established its ability to detect MDI in situ. Polyclonal MDI-specific IgG was induced by immunizing rabbits with MDI-conjugated to keyhole limpet hemocyanin (KLH) emulsified in complete Freund's adjuvant, followed by two booster injections with incomplete Freund's adjuvant. The antiserum contains IgG that recognize a variety of different MDI conjugated proteins, but not unconjugated or mock exposed proteins by dot blot analysis. The antiserum further demonstrates specificity for proteins conjugated with MDI, but not other commonly used diisocyanates. Immunochemical studies with cytospun airway cells and formalin-fixed paraffin embedded lung tissue sections from mice intranasally exposed to MDI (as reversibly reactive glutathione conjugates, e.g. GSH-MDI) demonstrated the antiserum's ability to detect MDI in tissue samples. The data demonstrate penetration of MDI into the lower airways, localized deposition in the epithelial region surrounding airways, and uptake by alveolar macrophages. The new immunochemical reagent should be useful for further studies delineating the uptake and tissue distribution of MDI, especially as it relates to adverse health effects from exposure. (C) 2015 Elsevier B.V. All rights reserved.