Epigenetic Effects in Children With Cow's Milk Allergy Treated With Different Formulas

Study Overview
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Brief Summary
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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.

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

Conditions
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Cow Milk Allergy

Intervention / Treatment
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* Dietary Supplement: EHCF+LGG
* Dietary Supplement: RHF
* Dietary Supplement: SF
* Dietary Supplement: EHWF
* Dietary Supplement: AAF


Participation Criteria
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Eligibility Criteria
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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.

Ages Eligible for Study
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Minimum Age: 6 Months
Maximum Age: 12 Months

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: Double


Arms and Interventions

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

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Epigenetic modifications in cytokines genes | Serum levels (pg/ml) of FOXP3 in children with cow's milk allergy | 12 months | 
| Epigenetic modifications in cytokines genes | Methylation rate (%) of FOXP3 in children with cow's milk allergy | 12 months | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| microRNAs modifications | Expression of miR-125b miR-146 miR-155 miR-29a/b miR-27 miR-128 let-7 family miR-21 miR-126 miR-145 miR-106a miR-375 miR-193a | 12 months | 
| 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 | 
| Epigenetic modifications in cytokines genes | Methylation rate (%) of interleukin IL-4, IL-5, IL-13, IL-10, IFN-γ, FoxP3, GATA3, T-bet, TGF-β, IL-17, IL-25, IL-12, IL-22, IL-6, IL-23 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-13, IL-10, IFN-γ, FoxP3, GATA3, T-bet, TGF-β, IL-17, IL-25, IL-12, IL-22, IL-6, IL-23 in children with cow's milk allergy | 6 and 12 months |