Ex-vivo whole blood secretion of interferon (IFN)-γ and IFN-γ-inducible protein-10 measured by enzyme- linked immunosorbent assay are as sensitive as IFN-γ enzyme- linked immunospot for the detection of gluten- reactive T cells in human leucocyte antigen (HLA)- DQ2.5+- associated coeliac disease
Ex-vivo whole blood secretion of interferon (IFN)-γ and IFN-γ-inducible protein-10 measured by enzyme- linked immunosorbent assay are as sensitive as IFN-γ enzyme- linked immunospot for the detection of gluten- reactive T cells in human leucocyte antigen (HLA)- DQ2.5+- associated coeliac disease
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DOI：
10.1111/cei.12232
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发表时间：
2014-02-01
影响因子：
4.6
通讯作者：
Anderson, R. P.
中科院分区：
文献类型：
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作者：
Ontiveros, N.;Tye-Din, J. A.;Anderson, R. P.
T细胞细胞因子释放测定用于诊断感染性疾病，但不是自身免疫性或过敏性疾病。乳糜泻（CD）是一种常见的T细胞介导的疾病，通过谷蛋白依赖性肠道炎症和血清学诊断。许多患者不能被诊断为CD，因为他们在医疗检查前减少了饮食中的麸质。在接受无麸质饮食（GFD）治疗的CD患者中，口服麸质激发可动员麸质反应性T细胞，可通过干扰素（IFN）-γ酶联免疫斑点（ELISPOT）或主要组织相容性复合物（MHC）II类四聚体进行测量。免疫优势肽在90%的具有HLA-DQ 2.5的患者中相当一致。我们的目的是开发全血检测面筋特异性T细胞。在谷蛋白攻击之前和之后，从确认患有CD的GFD供体（n=27，所有HLA-DQ 25（+））、确认不患有CD的GFD供体（n=6 HLA-DQ 2.5（+），11 HLA-DQ 2.5（-））和患有CD但不遵循GFD的供体（n=4，所有HLA-DQ 2.5（+））收集血液。在全血与肽或麦胶蛋白孵育后，通过酶联免疫吸附测定（ELISA）测量血浆IFN-γ和IFN-γ诱导蛋白-10（IP-10），并与IFN-γ ELISPOT相关联。在谷蛋白激发之前，没有T细胞测定可以区分CD患者和对照，但是在谷蛋白激发之后，全血IFN-γ ELISA和ELISPOT对于HLA-DQ 2.5（+）CD患者均为85%敏感性和100%特异性;全血IP-10 ELISA为94%敏感性和100%特异性。我们的结论是，全血细胞因子释放检测是敏感和特异性的检测谷蛋白反应性T细胞在CD，进一步的临床研究解决这些测试的效用在患者的CD诊断不确定是必要的。
T cell cytokine release assays are used to diagnose infectious diseases, but not autoimmune or allergic disease. Coeliac disease (CD) is a common T cell-mediated disease diagnosed by the presence of gluten-dependent intestinal inflammation and serology. Many patients cannot be diagnosed with CD because they reduce dietary gluten before medical workup. Oral gluten challenge in CD patients treated with gluten-free diet (GFD) mobilizes gluten-reactive T cells measurable by interferon (IFN)-gamma enzyme-linked immunospot (ELISPOT) or major histocompatibility complex (MHC) class II tetramers. Immunodominant peptides are quite consistent in the 90% of patients who possess HLA-DQ2.5. We aimed to develop whole blood assays to detect gluten-specific T cells. Blood was collected before and after gluten challenge from GFD donors confirmed to have CD (n=27, all HLA-DQ25(+)), GFD donors confirmed not to have CD (n=6 HLA-DQ2.5(+), 11 HLA-DQ2.5(-)) and donors with CD not following GFD (n=4, all HLA-DQ2.5(+)). Plasma IFN-gamma and IFN-gamma inducible protein-10 (IP-10) were measured by enzyme-linked immunosorbent assay (ELISA) after whole blood incubation with peptides or gliadin, and correlated with IFN-gamma ELISPOT. No T cell assay could distinguish between CD patients and controls prior to gluten challenge, but after gluten challenge the whole blood IFN-gamma ELISA and the ELISPOT were both 85% sensitive and 100% specific for HLA-DQ2.5(+) CD patients; the whole blood IP-10 ELISA was 94% sensitive and 100% specific. We conclude that whole blood cytokine release assays are sensitive and specific for detection of gluten-reactive T cells in CD; further clinical studies addressing the utility of these tests in patients with an uncertain diagnosis of CD is warranted.