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Monitoring of gluten-free diet compliance in celiac patients by assessment of gliadin 33-mer equivalent epitopes in feces.

Comino I, Real A, Vivas S, Síglez MÁ, Caminero A, Nistal E, Casqueiro J, Rodríguez-Herrera A, Cebolla A, Sousa C - Am. J. Clin. Nutr. (2012)

Bottom Line: The resistance of a significant part of 33EPs to gastrointestinal digestion was shown in vitro and in vivo.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.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Microbiología y Parasitología, Facultad de Farmacia, Universidad de Sevilla, Seville, Spain.

ABSTRACT

Background: 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.

Objective: 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.

Design: 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.

Results: 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.

Conclusions: 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.

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Related in: MedlinePlus

Analysis of mean (±SD) gluten amounts excreted in the feces of a celiac patient subjected to a gluten challenge for 6 d. Amounts of toxic peptide (ng/g feces) were determined by using a G12 competitive ELISA. Data were obtained from 3 independent experiments with samples run in triplicate. GFD, gluten-free diet; 33EPs, 33-mer equivalent peptidic epitopes.
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fig6: Analysis of mean (±SD) gluten amounts excreted in the feces of a celiac patient subjected to a gluten challenge for 6 d. Amounts of toxic peptide (ng/g feces) were determined by using a G12 competitive ELISA. Data were obtained from 3 independent experiments with samples run in triplicate. GFD, gluten-free diet; 33EPs, 33-mer equivalent peptidic epitopes.

Mentions: To assess the presence of gluten in feces of CD patients in remission, patients in remission after consumption of a GFD were given a gluten-containing diet for 6 d, and their stools were collected each day of the challenge period. The gluten challenge was carried out to confirm the diagnosis of CD and at the request of parents who were uncertain about the diagnosis. Out of the 3 patients who received the gluten challenge, 2 patients developed symptoms of vomiting, abdominal pain, and diarrhea within the first days of the administration of gluten and were excluded from the study. However, in the patient who tolerated the gluten challenge, fecal samples were collected for each of the 6 d that the challenge lasted, and the amount of gluten excreted was analyzed by using G12 antibody-based immunological methods to determine whether there was gluten excretion and the amounts of toxic peptides excreted during the period of gluten challenge. According to G12 competitive-ELISA results (Figure 6), excretion of gluten peptides started on the third day of the gluten challenge. Between days 3 and 6 of the challenge, fecal amounts of gluten reactive peptides ranged between 417 and 7037 ng 33EPs/g feces. These results were corroborated by using G12 immunochromatographic strips. In non-CD control subjects with other intestinal pathologies, the gluten content, although the subjects consumed a normal gluten-containing diet, ranged between 201 and 29,076 ng 33EPs/g feces. The intraindividual and interindividual differences seen in excretion could have been related to the diet, the type of gluten-containing food, and the variability in fermentative degradation.


Monitoring of gluten-free diet compliance in celiac patients by assessment of gliadin 33-mer equivalent epitopes in feces.

Comino I, Real A, Vivas S, Síglez MÁ, Caminero A, Nistal E, Casqueiro J, Rodríguez-Herrera A, Cebolla A, Sousa C - Am. J. Clin. Nutr. (2012)

Analysis of mean (±SD) gluten amounts excreted in the feces of a celiac patient subjected to a gluten challenge for 6 d. Amounts of toxic peptide (ng/g feces) were determined by using a G12 competitive ELISA. Data were obtained from 3 independent experiments with samples run in triplicate. GFD, gluten-free diet; 33EPs, 33-mer equivalent peptidic epitopes.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3278243&req=5

fig6: Analysis of mean (±SD) gluten amounts excreted in the feces of a celiac patient subjected to a gluten challenge for 6 d. Amounts of toxic peptide (ng/g feces) were determined by using a G12 competitive ELISA. Data were obtained from 3 independent experiments with samples run in triplicate. GFD, gluten-free diet; 33EPs, 33-mer equivalent peptidic epitopes.
Mentions: To assess the presence of gluten in feces of CD patients in remission, patients in remission after consumption of a GFD were given a gluten-containing diet for 6 d, and their stools were collected each day of the challenge period. The gluten challenge was carried out to confirm the diagnosis of CD and at the request of parents who were uncertain about the diagnosis. Out of the 3 patients who received the gluten challenge, 2 patients developed symptoms of vomiting, abdominal pain, and diarrhea within the first days of the administration of gluten and were excluded from the study. However, in the patient who tolerated the gluten challenge, fecal samples were collected for each of the 6 d that the challenge lasted, and the amount of gluten excreted was analyzed by using G12 antibody-based immunological methods to determine whether there was gluten excretion and the amounts of toxic peptides excreted during the period of gluten challenge. According to G12 competitive-ELISA results (Figure 6), excretion of gluten peptides started on the third day of the gluten challenge. Between days 3 and 6 of the challenge, fecal amounts of gluten reactive peptides ranged between 417 and 7037 ng 33EPs/g feces. These results were corroborated by using G12 immunochromatographic strips. In non-CD control subjects with other intestinal pathologies, the gluten content, although the subjects consumed a normal gluten-containing diet, ranged between 201 and 29,076 ng 33EPs/g feces. The intraindividual and interindividual differences seen in excretion could have been related to the diet, the type of gluten-containing food, and the variability in fermentative degradation.

Bottom Line: The resistance of a significant part of 33EPs to gastrointestinal digestion was shown in vitro and in vivo.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.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Microbiología y Parasitología, Facultad de Farmacia, Universidad de Sevilla, Seville, Spain.

ABSTRACT

Background: 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.

Objective: 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.

Design: 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.

Results: 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.

Conclusions: 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.

Show MeSH
Related in: MedlinePlus