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通过二维差异凝胶电泳(2D-DIGE)对小鼠卵类黏蛋白超敏反应进行蛋白质组学分析。

Proteomic analysis of ovomucoid hypersensitivity in mice by two-dimensional difference gel electrophoresis (2D-DIGE).

作者信息

Hobson D J, Rupa P, Diaz G J, Zhang H, Yang M, Mine Y, Turner P V, Kirby G M

机构信息

Department of Pathobiology, University of Guelph, Guelph, Ontario, Canada N1G 2W1.

出版信息

Food Chem Toxicol. 2007 Dec;45(12):2372-80. doi: 10.1016/j.fct.2007.06.039. Epub 2007 Aug 23.

Abstract

There is a need to develop reliable methods to assess the safety of genetically modified and other novel foods. The aim of this study was to identify protein biomarkers of food allergy in mice exposed to ovomucoid (OVM), a major food allergen found in chicken egg white. BALB/c mice were repeatedly sensitized by gavage with OVM and cholera toxin (CT) and control mice were exposed to a mixture of amino acids with CT. At the endpoint, all mice were challenged intraperitoneally with OVM and alum. Type-1 hypersensitivity was confirmed in OVM-sensitized mice by observation of clinical signs of anaphylaxis and elevated levels of plasma histamine, OVM-specific IgE and OVM-specific IgG by ELISA. Differential protein expression was assessed in albumin-depleted plasma as well as in mesenteric lymph node, liver, spleen, and ileum by two-dimensional difference gel electrophoresis (2D-DIGE). Differentially expressed proteins were identified by liquid chromatography with tandem mass spectrometry. Plasma proteins overexpressed in OVM-sensitized mice included haptoglobin (41-fold), serum amyloid A (19-fold) and peroxiredoxin-2 (1.9-fold). Further validation of these plasma proteins in other animal models of food allergy with different food allergens is required to assess their potential as candidate biomarkers for use in evaluating the allergenicity of novel foods.

摘要

需要开发可靠的方法来评估转基因食品和其他新型食品的安全性。本研究的目的是在暴露于卵类黏蛋白(OVM,一种存在于鸡蛋白中的主要食物过敏原)的小鼠中鉴定食物过敏的蛋白质生物标志物。通过向BALB/c小鼠灌胃OVM和霍乱毒素(CT)使其反复致敏,对照组小鼠则暴露于含有CT的氨基酸混合物中。在实验终点,所有小鼠均腹腔注射OVM和明矾进行激发。通过观察过敏反应的临床症状以及用ELISA检测血浆组胺、OVM特异性IgE和OVM特异性IgG水平升高,证实了OVM致敏小鼠存在1型超敏反应。通过二维差异凝胶电泳(2D-DIGE)评估了去除白蛋白的血浆以及肠系膜淋巴结、肝脏、脾脏和回肠中的差异蛋白表达。通过液相色谱-串联质谱鉴定差异表达的蛋白质。在OVM致敏小鼠中过表达的血浆蛋白包括触珠蛋白(41倍)、血清淀粉样蛋白A(19倍)和过氧化物酶2(1.9倍)。需要在其他使用不同食物过敏原的食物过敏动物模型中进一步验证这些血浆蛋白,以评估它们作为候选生物标志物用于评估新型食品致敏性的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6c9e/7126535/6681ee91b71b/gr1.jpg

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