Development of Mass-Screening Method for the Identification of Patients at Risk of Developing Type 1 Diabetes Mellitus.

描述了用于识别具有发展为1型糖尿病（DM）风险的患者的大规模筛查方法的发展。研究结果如下：1.本文报道了一种检测牛奶蛋白伊加抗体的超灵敏酶免疫分析法（免疫复合物转移酶免疫分析法），并对糖尿病患者和健康人血清中的抗体水平进行了比较。然而，未观察到显著差异。如果血清中存在针对抗体的抗独特型抗体，则其可能干扰测定，因此采用适当的ELISA形式以尽量减少此类干扰。结果表明，无论是对牛奶蛋白质还是对其他食物蛋白质的伊加抗体水平，前者均显著高于后者. 50%的患者具有DQB 1 ^*0401-DRB 1 ^*0405单倍型。在单倍型患者中，牛奶蛋白伊加抗体、伊加和TGF-β的水平显著高于其他患者。这表明，在消化道中，通过单倍型加速了向Th 3的抗原呈递，随后，TGF-β产生增加，因此，伊加抗体产生增加。因此，伊加抗体应答，即粘膜免疫应答，成为鉴别DM患者危险性的可靠指标.已开发并验证了使用干燥尿液样本评估对牛乳蛋白的粘膜免疫应答的测定方法。此外，伊加抗体的水平在干燥的尿液样品和血清样品相关的结论，大规模筛选方法，其中粘膜免疫反应的牛奶蛋白是通过使用干燥的尿液进行评估，可以为我们提供一个可靠的方法，识别患者的风险发展为DM。

The study on the development of mass-screening method for the identification of patients at risk of developing type 1 diabetes mellitus (DM) is described. The results are as follows.1. An ultrasensitive enzyme immunoassay (immune-complex transfer enzyme immunoassay) for IgA antibodies to cow's milk proteins has been developed, and the antibody levels in sera from patients with DM and healthy subjects were compared. However, no significant difference was observed. If anti-idiotype antibodies to the antibodies were present in serum, they might interfere with the assay, so suitable ELISA format was employed to minimize such interference. As results, the levels of IgA antibodies not only to the cow's milk proteins but also to an other food protein were significantly higher in former than latter.2. 50% of the patients had DQB1^*0401-DRB1^*0405 haplotype. In the patients with the haplotype, the levels of IgA antibodies to the cow's milk proteins, IgA and TGF-beta were significantly higher than those in the other patients. This was suggested that, in the digestive tract, an antigen presentation to Th3 was accelerated by the haplotype, subsequently, TGF-beta production was elevated, consequently, IgA antibody production was enhanced. Therefore, the IgA antibody response, that is, mucosal immune response, to the food antigen become a reliable marker for identification of patients at risk of developing DM.3. The assay method to assess the mucosal immune response to the cow's milk proteins using dried urine samples has been developed and validated. In addition, the levels of IgA antibodies in the dried urine samples and serum samples were correlated.In conclusion, the mass-screening method, in which the mucosal immune response to the cow's milk proteins is assessed by using the dried urine, can provide us with a reliable method for identification of patients at risk of developing DM.