Laboratorio de Investigación en Genética de enfermedades complejas, Hospital Clínico San Carlos, Instituto de Investigación Sanitaria del Hospital Clínico San Carlos (IdISSC), Madrid, Spain.
Laboratorio de Investigación en Genética de enfermedades complejas, Hospital Clínico San Carlos, Instituto de Investigación Sanitaria del Hospital Clínico San Carlos (IdISSC), Madrid, Spain.
Methods Cell Biol. 2023;179:113-126. doi: 10.1016/bs.mcb.2022.09.020. Epub 2022 Oct 31.
Accurate celiac disease (CD) diagnosis must be performed in individuals following a gluten containing diet. Diagnostic procedures for individuals already on a gluten-free diet (GFD) avoiding long gluten reintroductions are still challenging. To deal with this issue, we developed an accurate but simple method that requires only a 3-day gluten challenge and circumvents the main limitations of previously suggested proposals such as requirement of specific peptides and unusual specialized lab facilities or high cost. In an attempt to standardize this methodology to be used in daily clinical practice, we describe here an optimized protocol for assessing activated gut-homing CD8 T cells in blood combined with a short gluten challenge. Details about the amount and type of gluten antigen and the starting material are included, as well as the strategy to easily characterize and identify the cells of interest using flow cytometry. This methodology constitutes a diagnostic tool for CD diagnosis of high specificity and sensitivity for seropositive disease (>95%) as an alternative to long-term gluten challenge and open new possibilities to test the response to gluten in research and clinical trials.
在遵循含麸质饮食的个体中必须进行准确的乳糜泻 (CD) 诊断。对于已经遵循无麸质饮食 (GFD) 的个体，避免长期重新引入麸质的诊断程序仍然具有挑战性。为了解决这个问题，我们开发了一种准确但简单的方法，仅需进行 3 天的麸质挑战，规避了之前提出的建议的主要限制，例如需要特定的肽和不寻常的专门实验室设备或高成本。为了尝试将这种方法标准化，以便在日常临床实践中使用，我们在此描述了一种优化的方案，用于评估血液中活化的肠道归巢 CD8 T 细胞，并结合短期的麸质挑战。其中包括有关麸质抗原的量和类型以及起始材料的详细信息，以及使用流式细胞术轻松对感兴趣的细胞进行特征描述和鉴定的策略。该方法构成了一种用于 CD 诊断的工具，具有高特异性和敏感性（>95%），可作为长期麸质挑战的替代方法，并为研究和临床试验中测试对麸质的反应开辟了新的可能性。
