Whole blood interleukin-2 release test to detect and characterize rare circulating gluten-specific T cell responses in coeliac disease
Whole blood interleukin-2 release test to detect and characterize rare circulating gluten-specific T cell responses in coeliac disease
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DOI：
10.1111/cei.13578
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发表时间：
2021-02-28
影响因子：
4.6
通讯作者：
Tye-Din, J. A.
中科院分区：
文献类型：
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作者：
Anderson, R. P.;Goel, G.;Tye-Din, J. A.
全血细胞因子释放试验(CRA)评估谷蛋白的细胞免疫功能可简化对乳糜泻(CD)的诊断和监测。我们的目的是确定电化学发光CRA检测免疫优势醇溶蛋白多肽反应的有效性。研究对象为HL A-DQ2中心点5(+)CD成人(队列1，n=6；队列2，n=12)和正常对照(队列3，n=9)。队列1接受为期3天的面筋挑战(GC)。在开始为期3天的GC后的3小时、6小时和6天，分别在基线和队列1采集血液。采用醇溶蛋白多肽刺激的细胞增殖、干扰素-γ-酶联免疫斑点(ELISPOT)和14-和3-复合体电化学发光CRA。泊松分布分析用于估计反应细胞频率。在队列1中，白细胞介素2主导了醇溶蛋白多肽刺激的全血细胞因子释放谱。GC可引起全身IL-2的急剧释放，并增加醇溶蛋白多肽刺激的干扰素-γELISPOT和全血CRA反应。GC后全血CRA以IL-2为主，但也包括干扰素-γ、C-X-C基序趋化因子配体10/干扰素-γ诱导蛋白10(CXCL10/IP-10)、干扰素-γ诱导的单核细胞因子(MIG)、IL-10、趋化因子(C-C基序)配体3/巨噬细胞炎症蛋白1-α(CCL3/MIP-1α)、肿瘤坏死因子-α和IL-8/CXCL8。在队列2和队列3中，醇溶蛋白多肽刺激的全血IL-2释放对无麸质饮食的CD患者具有100%的特异性和92%的敏感性；CD患者外周血中分泌IL-2到α-醇溶蛋白多肽的细胞频率估计为0中心点5~11个/ml。电化学发光法测定全血IL-2是检测CD中罕见的醇溶蛋白特异性T细胞的敏感方法，有助于CD的监测和诊断。有必要进行更大规模的研究，并用基于四聚体的分析进行验证。
Whole blood cytokine release assays (CRA) assessing cellular immunity to gluten could simplify the diagnosis and monitoring of coeliac disease (CD). We aimed to determine the effectiveness of electrochemiluminescence CRA to detect responses to immunodominant gliadin peptides. HLA-DQ2 center dot 5(+) CD adults (cohort 1, n = 6; cohort 2, n = 12) and unaffected controls (cohort 3, n = 9) were enrolled. Cohort 1 had 3-day gluten challenge (GC). Blood was collected at baseline, and for cohort 1 also at 3 h, 6 h and 6 days after commencing 3-day GC. Gliadin peptide-stimulated proliferation, interferon (IFN)-gamma enzyme-linked immunospot (ELISPOT) and 14- and 3-plex electrochemiluminescence CRA were performed. Poisson distribution analysis was used to estimate responding cell frequencies. In cohort 1, interleukin (IL)-2 dominated the gliadin peptide-stimulated cytokine release profile in whole blood. GC caused systemic IL-2 release acutely and increased gliadin peptide-stimulated IFN-gamma ELISPOT and whole blood CRA responses. Whole blood CRA after GC was dominated by IL-2, but also included IFN-gamma, C-X-C motif chemokine ligand 10/IFN-gamma-induced protein 10 (CXCL10/IP-10), CXCL9/monokine induced by IFN-gamma (MIG), IL-10, chemokine (C-C motif) ligand 3/macrophage inflammatory protein 1-alpha (CCL3/MIP-1 alpha), TNF-alpha and IL-8/CXCL8. In cohorts 2 and 3, gliadin peptide-stimulated whole blood IL-2 release was 100% specific and 92% sensitive for CD patients on a gluten-free diet; the estimated frequency of cells in CD blood secreting IL-2 to alpha-gliadin peptide was 0 center dot 5 to 11 per ml. Whole blood IL-2 release successfully mapped human leucocyte antigen (HLA)-DQ2 center dot 5-restricted epitopes in an alpha-gliadin peptide library using CD blood before and after GC. Whole blood IL-2 release assay using electrochemiluminescence is a sensitive test for rare gliadin-specific T cells in CD, and could aid in monitoring and diagnosis. Larger studies and validation with tetramer-based assays are warranted.