Single-cell Immune Response to Controlled Gluten Ingestion in Pediatric Celiac Disease

NCT07362654 | Recruiting

• 1 other identifier: PI-0070-2024
• Study Type: interventional
• Target: 51
• Locations: 1 country
• Sites: 1

Brief Summary

This study investigates how the immune system of children with celiac disease responds to controlled, small amounts of gluten. Children on a strict gluten-free diet are randomly assigned to receive either placebo, 50 mg of gluten, or 5 g of gluten once daily for three days, simulating real-life accidental exposure or dietary transgression. Blood samples are collected on Day 1 (before gluten intake) and Day 8 (five days after the last dose). Stool and urine samples are also collected for complementary analyses. Using single-cell ribonucleic acid (RNA) sequencing, T-cell receptor sequencing, microRNA profiling, and exploratory metabolomics, the study aims to characterize changes in immune cell populations and gene expression after gluten exposure. The objective is to determine whether even very small amounts of gluten induce measurable systemic immune responses and whether these responses differ according to the dose administered. Understanding these mechanisms may support the development of new biomarkers and improve clinical management of pediatric celiac disease.

Conditions

Celiac Disease; Gluten Sensitivity; Autoimmune Diseases

Keywords

Celiac Disease; Gluten Exposure; Single-Cell RNA Sequencing; Gluten-Free Diet; Immune Response; Pediatrics; Omics

Outcome Measures

Primary Outcomes (2)

• Change in peripheral blood immune cell gene expression after controlled gluten ingestion

  Change from baseline (Day 1) to post-intervention (Day 8) in gene expression profiles of peripheral blood immune cells measured by single-cell RNA sequencing (scRNA-seq).

  Day 1 to Day 8

• Change in peripheral blood T-cell receptor repertoire after controlled gluten ingestion

  Change from baseline (Day 1) to post-intervention (Day 8) in T-cell receptor (TCR) diversity and clonality in peripheral blood T cells measured by TCR sequencing

  Day 1 to Day 8

Secondary Outcomes (6)

• Dose-dependent changes in immune gene expression after gluten exposure

  Day 1 to Day 8

• Dose-dependent changes in T-cell receptor repertoire clonality after gluten exposure

  Day 1 to Day 8

• Inter-individual variability in immune gene expression response to gluten exposure across participants

  Day 1 to Day 8

• Changes in serum exosomal microRNA expression after gluten exposure

  Day 1 to Day 8

• Changes in metabolomic profiles after gluten exposure

  Day 1 to Day 8

Study Arms (3)

Placebo Group

PLACEBO COMPARATOR

Participants receive a placebo preparation once daily on Days 1, 2, and 3. Blood samples are collected on Day 1 (baseline) and Day 8 (post-intervention). Participants and evaluators remain blinded to allocation.

Dietary Supplement: Placebo; Procedure: Collection of blood, stool and urine samples on Day 1 and Day 8.

Low-Dose Gluten (50 mg)

EXPERIMENTAL

Participants receive 50 mg of gluten once daily on Days 1, 2, and 3, simulating accidental low-level exposure. Blood samples are collected on Day 1 (baseline) and Day 8 (post-intervention). Participants and evaluators remain blinded to allocation.

Dietary Supplement: Gluten 50 mg; Procedure: Collection of blood, stool and urine samples on Day 1 and Day 8.

High-Dose Gluten (5 g)

EXPERIMENTAL

Participants receive 5 g of gluten once daily on Days 1, 2, and 3, simulating a dietary transgression. Blood samples are collected on Day 1 (baseline) and Day 8 (post-intervention). Participants and evaluators remain blinded to allocation.

Dietary Supplement: Gluten 5 g; Procedure: Collection of blood, stool and urine samples on Day 1 and Day 8.

Interventions

Placebo DIETARY_SUPPLEMENT

Placebo preparation identical in appearance and administration schedule to gluten doses. Administered once daily on Days 1, 2, and 3. Used as the comparator. Participants and evaluators remain blinded to allocation.

Placebo Group

Gluten 50 mg DIETARY_SUPPLEMENT

Participants receive 50 mg of gluten once daily on Days 1, 2, and 3. This dose simulates accidental low-level gluten exposure in children with celiac disease. Participants and evaluators remain blinded to allocation.

Low-Dose Gluten (50 mg)

Gluten 5 g DIETARY_SUPPLEMENT

Participants receive 5 g of gluten once daily on Days 1, 2, and 3. This dose simulates a dietary transgression in children with celiac disease. Participants and evaluators remain blinded to allocation.

High-Dose Gluten (5 g)

Collection of blood, stool and urine samples on Day 1 and Day 8. PROCEDURE

In addition to blood samples, stool and urine samples will be collected for complementary analyses (including fecal gluten immunogenic peptides (GIP) and exploratory metabolomic assays)

High-Dose Gluten (5 g); Low-Dose Gluten (50 mg); Placebo Group

Eligibility Criteria

• Age: 8 Years - 14 Years
• Sex: all
• Healthy Volunteers: No
• Age Groups: Child (0-17)

You may qualify if:

• Age 8 to 14 years at study entry.
• Diagnosis of celiac disease according to ESPGHAN 2020 criteria.
• At least 18 months on a strict gluten-free diet (GFD).
• Asymptomatic from a gastrointestinal perspective in the preceding weeks.
• Ability to swallow the gluten/placebo preparation.
• Written informed consent from parents/legal guardians and assent from the child.

You may not qualify if:

• Obesity defined as BMI ≥ 95th percentile according to WHO criteria.
• Diagnosed inflammatory bowel disease or diabetes mellitus.
• Chronic hepatic, pulmonary, renal, or rheumatologic disease.
• History of severe acute reactions to accidental gluten ingestion.
• Use of oral corticosteroids or immunosuppressive therapy in the previous 3 months.
• Any condition that, in the opinion of the investigators, may contraindicate participation or compromise study integrity.

Sponsors & Collaborators

• Hospital Regional de Malaga: collaborator
• Andaluz Health Service: collaborator
• Fundación Pública Andaluza para la Investigación Biomédica Andalucía Oriental: collaborator
• Universidad de Málaga: collaborator
• Biohealth Research Institute in Granada (ibs.GRANADA): collaborator
• Fundación Pública Andaluza para la Investigación de Málaga en Biomedicina y Salud: lead
• Consorcio Centro de Investigación Biomédica en Red (CIBER): collaborator

Study Sites (1)

Hospital Regional Universitario de Málaga

Málaga, 29010, Spain

RECRUITING

MeSH Terms

Conditions

Celiac Disease; Autoimmune Diseases

Interventions

Glutens; Defecation

Condition Hierarchy (Ancestors)

Malabsorption Syndromes; Intestinal Diseases; Gastrointestinal Diseases; Digestive System Diseases; Metabolic Diseases; Nutritional and Metabolic Diseases; Immune System Diseases

Intervention Hierarchy (Ancestors)

Prolamins; Grain Proteins; Plant Proteins; Proteins; Amino Acids, Peptides, and Proteins; Seed Storage Proteins; Digestive System Physiological Phenomena; Digestive System and Oral Physiological Phenomena

Study Officials

• Rafael Martín Masot, MD, PhD

  Hospital Regional Universitario de Málaga / IBIMA-BIONAND

  PRINCIPAL INVESTIGATOR

• Lara María Bossini Castillo, MD, PhD

  IBS Granada

  PRINCIPAL INVESTIGATOR

Central Study Contacts

Rafael Martín Masot, MD, PhD

CONTACT

+34 951290000; rafammgr@uma.es

Lara María Bossini Castillo, MD, PhD

CONTACT

+34 663148283; lbossinicastillo@ugr.es

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: RANDOMIZED
• Masking: QUADRUPLE
• Who Masked: PARTICIPANT, CARE PROVIDER, INVESTIGATOR, OUTCOMES ASSESSOR
• Masking Details: Quadruple-blind (participant, care provider, investigator, outcomes assessor). Only three designated team members not involved in routine clinical assessment or outcome evaluation have access to the randomization list in order to prepare the gluten/placebo doses and monitor safety. Data entry and statistical analyses are performed using a blinded database.
• Purpose: BASIC SCIENCE
• Intervention Model: PARALLEL
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Model Details: Participants with celiac disease are randomized into three parallel intervention groups receiving placebo, 50 mg gluten, or 5 g gluten.

Study Record Dates

• First Submitted: December 17, 2025
• First Posted: January 23, 2026
• Study Start: July 7, 2025
• Primary Completion: April 1, 2026
• Study Completion (Estimated): November 1, 2026
• Last Updated: April 27, 2026

Record last verified: 2025-11

Data Sharing

• IPD Sharing: Will not share

Locations

ES (1)