Objective Initiation of a gluten-free diet without proper diagnostic work-up of coeliac disease is a frequent and demanding problem. Recent diagnostic guidelines suggest a gluten challenge of at least 14 days followed by duodenal biopsy in such patients. The rate of false-negative outcome of this approach remains unclear. We studied responses to 14-day gluten challenge in subjects with treated coeliac disease. Design We challenged 20 subjects with biopsy-verified coeliac disease, all in confirmed mucosal remission, for 14 days with 5.7 grams per oral gluten daily. Duodenal biopsies were collected. Blood was analysed by multiplex assay for cytokine detection, and by flow cytometry using HLA-DQ:gluten tetramers. Results Nineteen participants completed the challenge. Villous blunting appeared at end of challenge in 5 of 19 subjects. Villous height to crypt depth ratio reduced with at least 0.4 concomitantly with an increase in intraepithelial lymphocyte count of at least 50% in 9 of 19 subjects. Interleukin-8 plasma concentration increased by more than 100% after 4 hours in 7 of 19 subjects. Frequency of blood CD4+ effector-memory gut-homing HLA-DQ:gluten tetramer-binding T cells increased by more than 100% on day 6 in 12 of 15 evaluated participants. Conclusion A 14-day gluten challenge was not enough to establish significant mucosal architectural changes in majority of patients with coeliac disease (sensitivity ≈25%–50%). Increase in CD4+ effector-memory gut-homing HLA-DQ:gluten tetramer-binding T cells in blood 6 days after gluten challenge is a more sensitive and less invasive biomarker that should be validated in a larger study. Trial registration number NCT02464150

中文翻译：

---

HLA-DQ：在 14 天麸质挑战后，血液中的麸质四聚体测试比活检更能检测乳糜泻患者

目标 在没有适当的乳糜泻诊断检查的情况下开始无麸质饮食是一个常见且要求高的问题。最近的诊断指南建议对此类患者进行至少 14 天的麸质挑战，然后进行十二指肠活检。这种方法的假阴性结果率仍不清楚。我们研究了接受治疗的乳糜泻患者对 14 天麸质挑战的反应。设计 我们对 20 名经活检证实的乳糜泻患者进行了为期 14 天的试验，每天口服 5.7 克麸质。收集十二指肠活检。通过细胞因子检测的多重分析和使用 HLA-DQ:谷蛋白四聚体的流式细胞术分析血液。结果 19 名参与者完成了挑战。19 名受试者中有 5 名在攻击结束时出现绒毛变钝。19 名受试者中有 9 名的绒毛高度与隐窝深度之比降低了至少 0.4，同时上皮内淋巴细胞计数增加了至少 50%。4 小时后，19 名受试者中有 7 名的白细胞介素 8 血浆浓度增加了 100% 以上。在第 6 天，15 名接受评估的参与者中有 12 人的血液 CD4+ 效应记忆肠道归巢 HLA-DQ：谷蛋白四聚体结合 T 细胞的频率增加了 100% 以上。结论 14 天的麸质挑战不足以在大多数乳糜泻患者中建立显着的粘膜结构变化（敏感性 ≈25%–50%）。面筋挑战 6 天后，血液中 CD4+ 效应记忆肠道归巢 HLA-DQ：面筋四聚体结合 T 细胞的增加是一种更敏感、侵入性更小的生物标志物，应在更大的研究中得到验证。试验注册号 NCT02464150