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乳糜泻患者和多发性骨髓瘤患者体内存在针对不同小麦品种麦醇溶蛋白的不同水平的体液免疫反应性。

Different levels of humoral immunoreactivity to different wheat cultivars gliadin are present in patients with celiac disease and in patients with multiple myeloma.

作者信息

Konic-Ristic Aleksandra, Dodig Dejan, Krstic Radmilo, Jelic Svetislav, Stankovic Ivan, Ninkovic Aleksandra, Radic Jelena, Besu Irina, Bonaci-Nikolic Branka, Jojic Njegica, Djordjevic Milica, Popovic Dragan, Juranic Zorica

机构信息

Institute for Oncology and Radiology of Serbia, Belgrade, Serbia.

出版信息

BMC Immunol. 2009 May 31;10:32. doi: 10.1186/1471-2172-10-32.

Abstract

BACKGROUND

Immunity to food antigens (gliadin, cow's milk proteins) is in the centre of the attention of modern medicine focused on the prevention of diseases, prevention which is based on the use of appropriate restriction diet. Detection of the enhanced levels of the immune reactions to antigen(s) present in food is from this point of view of great importance because there are reports that some of health disturbances, like celiac disease (CD) and some premalignant conditions, like monoclonal gammopathy of undetermined significance (MGUS), were vanished after the appropriate restriction diets. It is well known that gliadin is toxic to small bowel mucosa of relatively small population of genetically predisposed individuals, who under this toxic action develop celiac disease (CD). As the quantity of immunogenic gliadin could vary between different wheat species, the first aim of this work was to determine the percentage of immunogenic gliadin in ten bread wheat cultivars and in three commercially grown durum wheat cultivars. The second part of the study was initiated by results of previous publication, reporting that sera of some of multiple myeloma (MM) patients showed the presence of elevated levels of anti-gliadin IgA, without the enhanced levels of anti-gliadin IgG antibodies, determined with commercial ELISA test. It was designed to assess is it possible to reveal is there any hidden, especially anti-gliadin IgG immunoreactivity, in serum of mentioned group of patients. For this purpose we tested MM patients sera, as well as celiac disease (CD) patients sera for the immunoreaction with the native gliadin isolated from wheat species used for bread and pasta making in corresponding geographic region.

RESULTS

Gliadin was isolated from wheat flour by two step 60% ehanolic extraction. Its content was determined by commercial R5 Mendez Elisa using PWG gliadin as the standard. Results obtained showed that immunogenic gliadin content varies between 50.4 and 65.4 mg/g in bread wheat cultivars and between 20 and 25.6 mg/g in durum wheat cultivars. Anti-gliadin IgA and IgG immunoreactivity of patients' sera in (IU/ml) was firstly determined by commercial diagnostic Binding Site ELISA test, and then additionally by non-commercial ELISA tests, using standardized ethanol wheat extracts -gliadin as the antigen. In both patients groups IgA immunoreactivity to gliadin from different cultivars was almost homogenous and in correlation with results from commercial test (except for one patient with IgA(lambda) myeloma, they were more then five times higher). But, results for IgG immunoreactivity were more frequently inhomogeneous, and especially for few MM patients, they were more then five times higher and did not correlate with results obtained using Binding Site test.

CONCLUSION

Results obtained showed different content of immunogenic gliadin epitopes in various species of wheat. They also point for new effort to elucidate is there a need to develop new standard antigen, the representative mixture of gliadin isolated from local wheat species used for bread production in corresponding geographic region for ELISA diagnostic tests.

摘要

背景

对食物抗原(麦醇溶蛋白、牛奶蛋白)的免疫反应是现代医学关注的焦点,现代医学致力于疾病预防,这种预防基于采用适当的限制饮食。从这一角度来看,检测食物中存在的抗原免疫反应增强水平非常重要,因为有报告称,一些健康问题,如乳糜泻(CD)和一些癌前病变,如意义未明的单克隆丙种球蛋白病(MGUS),在采用适当的限制饮食后消失了。众所周知,麦醇溶蛋白对一小部分具有遗传易感性的个体的小肠黏膜有毒性,这些个体在这种毒性作用下会患上乳糜泻(CD)。由于免疫原性麦醇溶蛋白的含量在不同小麦品种之间可能有所不同,这项工作的首要目标是确定十个面包小麦品种和三个商业种植的硬粒小麦品种中免疫原性麦醇溶蛋白的百分比。该研究的第二部分是由先前的一篇出版物的结果引发的,该出版物报告称,一些多发性骨髓瘤(MM)患者的血清显示抗麦醇溶蛋白IgA水平升高,而用商业ELISA检测法测定的抗麦醇溶蛋白IgG抗体水平并未升高。其目的是评估在上述患者群体的血清中是否有可能揭示出任何隐藏的,尤其是抗麦醇溶蛋白IgG免疫反应性。为此,我们检测了MM患者血清以及乳糜泻(CD)患者血清与从相应地理区域用于制作面包和面食的小麦品种中分离出的天然麦醇溶蛋白的免疫反应。

结果

通过两步60%乙醇提取法从小麦粉中分离出麦醇溶蛋白。其含量通过使用PWG麦醇溶蛋白作为标准的商业R5门德斯ELISA法测定。获得的结果表明,面包小麦品种中免疫原性麦醇溶蛋白的含量在50.4至65.4毫克/克之间,硬粒小麦品种中在20至25.6毫克/克之间。患者血清的抗麦醇溶蛋白IgA和IgG免疫反应性(IU/ml)首先通过商业诊断结合位点ELISA检测法测定,然后另外通过非商业ELISA检测法测定,使用标准化的乙醇小麦提取物 - 麦醇溶蛋白作为抗原。在两个患者组中,对不同品种麦醇溶蛋白的IgA免疫反应性几乎是均匀的,并且与商业检测结果相关(除了一名患有IgA(λ)骨髓瘤的患者,其结果高出五倍以上)。但是,IgG免疫反应性的结果更频繁地不均匀,特别是对于少数MM患者,其结果高出五倍以上,并且与使用结合位点检测法获得的结果不相关。

结论

获得的结果表明不同小麦品种中免疫原性麦醇溶蛋白表位的含量不同。它们还指出需要做出新的努力来阐明是否有必要开发一种新的标准抗原,即从相应地理区域用于面包生产的当地小麦品种中分离出的麦醇溶蛋白的代表性混合物,用于ELISA诊断检测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a131/2696423/def7aaebb0cc/1471-2172-10-32-1.jpg

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