Departamento de Microbiología y Parasitología, Facultad de Farmacia, Universidad de Sevilla, Seville, Spain.
Am J Clin Nutr. 2012 Mar;95(3):670-7. doi: 10.3945/ajcn.111.026708. Epub 2012 Jan 18.
Certain immunotoxic peptides from gluten are resistant to gastrointestinal digestion and can interact with celiac-patient factors to trigger an immunologic response. A gluten-free diet (GFD) is the only effective treatment for celiac disease (CD), and its compliance should be monitored to avoid cumulative damage. However, practical methods to monitor diet compliance and to detect the origin of an outbreak of celiac clinical symptoms are not available.
We assessed the capacity to determine the gluten ingestion and monitor GFD compliance in celiac patients by the detection of gluten and gliadin 33-mer equivalent peptidic epitopes (33EPs) in human feces.
Fecal samples were obtained from healthy subjects, celiac patients, and subjects with other intestinal pathologies with different diet conditions. Gluten and 33EPs were analyzed by using immunochromatography and competitive ELISA with a highly sensitive antigliadin 33-mer monoclonal antibody.
The resistance of a significant part of 33EPs to gastrointestinal digestion was shown in vitro and in vivo. We were able to detect gluten peptides in feces of healthy individuals after consumption of a normal gluten-containing diet, after consumption of a GFD combined with controlled ingestion of a fixed amount of gluten, and after ingestion of <100 mg gluten/d. These methods also allowed us to detect GFD infringement in CD patients.
Gluten-derived peptides could be sensitively detected in human feces in positive correlation with the amount of gluten intake. These techniques may serve to show GFD compliance or infringement and be used in clinical research in strategies to eliminate gluten immunotoxic peptides during digestion. This trial was registered at clinicaltrials.gov as NCT01478867.
来自 gluten 的某些免疫毒性肽可抵抗胃肠道消化,并且可与乳糜泻患者的因素相互作用,从而引发免疫反应。无麸质饮食(GFD)是乳糜泻(CD)的唯一有效治疗方法,应监测其依从性以避免累积性损伤。然而,目前尚无实用方法来监测饮食依从性和检测乳糜泻临床症状爆发的来源。
我们评估了通过检测人类粪便中的 gluten 和 gliadin 33-mer 等效肽段(33EPs)来确定乳糜泻患者的 gluten 摄入并监测 GFD 依从性的能力。
收集了健康受试者、乳糜泻患者和具有不同饮食条件的其他肠道病理受试者的粪便样本。使用免疫层析法和竞争性 ELISA 分析 gluten 和 33EPs,该 ELISA 使用高度敏感的抗 gliadin 33-mer 单克隆抗体。
体外和体内研究均表明 33EPs 的很大一部分具有抵抗胃肠道消化的能力。我们能够在健康个体摄入正常含 gluten 饮食、摄入 GFD 并同时受控摄入固定量 gluten 以及摄入 <100mg gluten/d 后在粪便中检测到 gluten 肽。这些方法还允许我们在 CD 患者中检测到 GFD 违规。
与 gluten 摄入量呈正相关,可在人类粪便中敏感地检测到 gluten 衍生肽。这些技术可用于显示 GFD 依从性或违规性,并在临床研究中用于在消化过程中消除 gluten 免疫毒性肽的策略。该试验在 clinicaltrials.gov 上注册为 NCT01478867。