Epigenetic Effects in Children With Cow's Milk Allergy Treated With Different Formulas (EPICMA II)

Epigenetic Effects Involved in Children With Cow's Milk Allergy: A Possible Effect of Hypoallergenic Formulas

Lactobacillus GG (LGG) is able to exert long lasting effects in children with atopic disorders. Nutramigen LGG accelerates tolerance acquisition in infants with cow's milk allergy. The mechanisms of these effects are still largely undefined. The effect of LGG could be related at least in part by the immunoregulatory role played by LGG. This probiotic can balance the generation of cytokines possibly involved in IgE- or non-IgE-mediated cow's milk allergy Interleulkin (IL)-4, IL-5, IL-10, IFN-γ , TGF-β, and TNF-Υ), which can contribute to modulation of inflammatory processes. The investigators have demonstrated that children with IgE-mediated CMA produce significantly higher level of IL-4 and IL-13 in response to cow's milk protein, and that tolerance is associated with a marked reduction of IL-13 production and a concomitant increased frequency of IFN-γ releasing cells. Epigenetics studies the heritable (and potentially reversible) changes of the genome inherited from one cell generation to the next which alter gene expression but do not involve changes in primary DNA sequences, highlighting the complexity of the inter-relationship between genetics and nutrition. There are three distinct, but closely interacting, epigenetic mechanisms (histone acetylation, DNA methylation, and non-coding microRNAs) that are responsible for modifying the expression of critical genes associated with physiologic and pathologic processes. The profile of epigenetic modifications associated with Th lineage commitment, coupled with the sensitivity of the early developmental period, has led to speculation that factors that disrupt these pathways may increase the risk of allergic diseases. Specifically, effects on DNA methylation and endogenous histone deacetylase inhibitors acting on specific pathways (Th1 and T regulatory cell differentiation) may favour Th2-associated allergic differentiation. MicroRNAs are another structural components of an epigenetic mechanism of post-transcriptional regulation of messenger RNA translation. It has been recently identified a specific Th2-associated microRNA (miR-21) that is critical for the regulation of Th cell polarization. It has been previously demonstrated an inverse DNA methylation pattern of cytokines involved in Th2 response (IL-4, IL-5) compared with cytokines involved in Th1 response (IL-10, INF- y) in children with CMA acquiring oral tolerance, with the most pronounced effects in those treated with Nutramigen LGG.

Study Overview

• Status: Completed
• Conditions: Cow Milk Allergy
• Intervention / Treatment: Dietary supplement: EHCF+LGG; Dietary supplement: RHF; Dietary supplement: EHWF; Dietary supplement: AAF; Dietary supplement: SF

• Study Type: Interventional
• Enrollment (Actual): 90
• Phase: Not Applicable

Contacts and Locations

Study Locations

• Italy
  • Naples, Italy, 80100
    • Pediatric Office

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 6 months to 1 year (Child)
• Accepts Healthy Volunteers: No

Description

Inclusion Criteria:

• children with cow's milk allergy

Exclusion Criteria:

• Concomitant chronic systemic diseases,
• congenital cardiac defects,
• active tuberculosis,
• autoimmune diseases,
• immunodeficiency,
• chronic inflammatory bowel diseases,
• celiac disease,
• cystic fibrosis,
• metabolic diseases,
• malignancy,
• chronic pulmonary diseases,
• malformations of the gastrointestinal tract,
• suspected eosinophilic esophagitis or eosinophilic enterocolitis,
• suspected food-protein-induced enterocolitis syndrome,
• suspected cow's milk protein-induced anaphylaxis,
• still on exclusion diet with one of the study formulas or with another dietary regimen because of cow's milk allergy,
• other food allergies.

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: Randomized
• Interventional Model: Parallel Assignment
• Masking: Double

Number of Arms

5

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: EHCF + LGG; Extensively hydrolyzed casein formula plus Lactobacillus rhamnosus GG	Dietary supplement: EHCF+LGG; Extensively hydrolyzed casein formula containing Lactobacillus rhamnosus GG
Active Comparator: RHF; Extensively hydrolyzed rice formula	Dietary supplement: RHF; Rice hydrolysed formula
Active Comparator: EHWF; Extensively hydrolyzed protein formula	Dietary supplement: EHWF; extensively hydrolysed whey formula
Active Comparator: AAF; Amino acid based formula	Dietary supplement: AAF; amino acid based formula
Active Comparator: SF; Soy formula	Dietary supplement: SF; Soy formula

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Epigenetic modifications in cytokines genes	TSDR demethylation (prop.) of FOXP3 in children with cow's milk allergy. The primary outcome was the time-averaged FoxP3 Treg-Specific-Demethylation-Region (TSDR) demethylation (prop.), which took into account three repeated measures (0,6,12 months).	12 months

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Concentration of metagenomic characteristics of the intestinal microbiota	evaluation of composition of gut microbiota	12 months
Concentration of metabolomic characteristics of the intestinal microbiota	evaluation of function of gut microbiota	12 months
The rate of patients with immune tolerance acquisition to cow's milk protein	Rate of patients with acquisition of tolerance to cow milk protein (rate of patients with outgrown cow's milk allergy)	12 months
microRNAs modifications	Expression of miR-125b miR-146a miR-155 miR-29a/b miR-27a miR-128 miR-21 miR-126 miR-145 miR-106a miR-193a5p miR-128	12 months
Epigenetic modifications in cytokines genes	Methylation rate (%) of interleukin IL-4, IL-5, IL-10, IFN-γ, in children with cow's milk allergy	6 and 12 months
Epigenetic modifications in cytokines genes	Serum levels (pg/ml) of interleukin IL-4, IL-5, IL-10, IFN-γ in children with cow's milk allergy	6 and 12 months
Epigenetic modifications in cytokines genes	Demethtylation rate and expression (fc, fold change, ΔΔCt method)) of FOXP3 in children with cow's milk allergy	6 and 12 months

Collaborators and Investigators

• Sponsor: Federico II University

Study record dates

Study Major Dates

• Study Start (Actual): December 10, 2019
• Primary Completion (Actual): December 31, 2023
• Study Completion (Actual): August 31, 2024

Study Registration Dates

• First Submitted: November 29, 2019
• First Submitted That Met QC Criteria: November 29, 2019
• First Posted (Actual): December 3, 2019

Study Record Updates

• Last Update Posted (Actual): April 14, 2026
• Last Update Submitted That Met QC Criteria: April 9, 2026
• Last Verified: December 1, 2025

More Information

Terms related to this study

• Additional Relevant MeSH Terms: Immune System Diseases; Hypersensitivity, Immediate; Hypersensitivity; Food Hypersensitivity; Milk Hypersensitivity
• Other Study ID Numbers: 28n15

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No