Improved T cell assay for identification of type 1 diabetes patients
Improved T cell assay for identification of type 1 diabetes patients
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DOI：
10.1016/j.jim.2009.03.004
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发表时间：
2009-05-15
影响因子：
2.2
通讯作者：
Palmer, Jerfy P.
中科院分区：
文献类型：
--
作者：
Brooks-Worrell, Barbara;Warsen, Adelaide;Palmer, Jerfy P.
糖尿病主要由两种临床独立的疾病组成：1型（T1D）和2型糖尿病（T2D）。T1D是一种针对β细胞的细胞介导的自身免疫性疾病，其特征是自身抗体（Ab）和T细胞对胰岛蛋白的反应性，而T2D是非自身免疫性疾病。尽管自身免疫性糖尿病的病理过程涉及T细胞，但糖尿病的免疫标志物主要集中在循环血清胰岛自身抗体的存在上。在两个蒙面NIH赞助的研讨会上，我们的细胞免疫印迹T细胞测定。该方法使用分离的人类胰岛分成18个分子量的部分，已被证实能够区分T1D患者和对照组，具有出色的特异性和敏感性。在这项研究中。我们利用第一次研讨会选择了T1D患者和对照组之间最具歧视性的人类胰岛的8个分子量部分。利用在第一个车间鉴定的这8个分子量分数，我们在第二个车间验证了这8个印迹切片的优先识别。然后，我们仅使用这8个印迹切片的数据重新计算了两个车间的细胞免疫印迹测定的敏感性和特异性。我们观察到，与两个车间的原始车间数据相比，敏感性和特异性都有所增加。使用8个而不是18个分子量区可以显著减少患者所需的血液量，从而允许对参与免疫调节研究的儿科患者进行细胞免疫印迹。这种改进的T细胞检测方法可以直接测量自身免疫性糖尿病患者的胰岛反应性T细胞反应，具有极好的敏感性和特异性，可能会改善干预研究期间患者的随访情况。(C) 2009 Elsevier B.V.版权所有
Diabetes mellitus is comprised primarily of two clinically separate diseases: type 1 (T1D) and type 2 diabetes (T2D). T1D is a cell-mediated autoimmune disease directed against the beta cells and characterized by autoantibody (Ab) and T cell reactivity to islet proteins whereas, T2D is non-autoimmune. Despite the fact that the pathological process in autoimmune diabetes involves T cells, immune markers of diabetes have primarily centered on the presence of circulating serum islet autoantibodies. In two masked NIH sponsored workshops, our cellular immunoblotting T cell assay. which uses isolated human islets separated into 18 molecular weight fractions, has been validated to be able to distinguish T1D patients from controls with excellent specificity and sensitivity. In this study. we utilized the first workshop to select eight molecular weight fractions of human islets that were the most discriminatory between T1D patients and controls. Using these eight molecular weight fractions identified in the first workshop, we validated the preferential recognition of these 8 blot sections in a second workshop. We then re-calculated the sensitivity and specificity of the cellular immunoblotting assay for both workshops using only the data from these 8 blot sections. We observed increases in both sensitivity and specificity compared to the original workshop data for both workshops. The use of 8 instead of 18 molecular weight regions allows for a significant reduction in the amount of blood needed from patients, thus allowing cellular immunoblotting to be performed on pediatric patients participating in immunomodulatory studies. This improved T cell assay, which directly measures islet reactive T cell responses in autoimmune diabetes patients with excellent sensitivity and specificity, will likely improve patient follow-up during intervention studies. (C) 2009 Elsevier B.V. All rights reserved.