Novel Peripheral Blood Allergen Stimulation Test for Non-IgE-mediated Gastrointestinal Food Allergies in Neonates and Infants.
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE171660 | GEO | 2022/02/01
REPOSITORIES: GEO
Similar Datasets
Project description:Exclusively breast-fed infants can exhibit clear signs of IgE or non IgE-mediated cow’s milk allergy. The definite characterization of dietary cow’s milk proteins (CMP) that survive the maternal digestive tract to be absorbed into the bloodstream and secreted into breast milk remains missing. The aim of this study was to assess the occurrence of CMP-derived peptides in breast milk, using antibody-independent methods. Using high performance liquid chromatography-high resolution mass spectrometry in blinded assays, we identified 11 cow’s milk-derived peptides, including two β-lactoglobulin (2 out 6 samples) and one αs1-casein (1 out 6 samples) fragments, in breast milk from mothers receiving a cup of bovine milk daily. The β-lactoglobulin (β-Lg) fragments, namely f42-54 and f42-57, were absent in milk from mothers who observed a strict dairy-free diet (6 samples). In contrast, neither intact nor hydrolyzed β-Lg was detected by Western blot or competitive ELISA tests. CMP-derived peptides rather than intact CMP may sensitize or elicit allergic responses in the neonate through mother’s milk. Immunologically active peptides from the maternal diet could be involved in priming the newborn’s immune system to drive tolerogenic response in neonates and infants.
2016-07-19 | PXD003657 | Pride
Project description:Exclusively breast-fed infants can exhibit clear signs of IgE or non IgE-mediated cow’s milk allergy. The definite characterization of dietary cow’s milk proteins (CMP) that survive the maternal digestive tract to be absorbed into the bloodstream and secreted into breast milk remains missing. The aim of this study was to assess the occurrence of CMP-derived peptides in breast milk, using antibody-independent methods. Using high performance liquid chromatography-high resolution mass spectrometry in blinded assays, we identified 11 cow’s milk-derived peptides, including two ?-lactoglobulin (2 out 6 samples) and one ?s1-casein (1 out 6 samples) fragments, in breast milk from mothers receiving a cup of bovine milk daily. The ?-lactoglobulin (?-Lg) fragments, namely f42-54 and f42-57, were absent in milk from mothers who observed a strict dairy-free diet (6 samples). In contrast, neither intact nor hydrolyzed ?-Lg was detected by Western blot or competitive ELISA tests. CMP-derived peptides rather than intact CMP may sensitize or elicit allergic responses in the neonate through mother’s milk. Immunologically active peptides from the maternal diet could be involved in priming the newborn’s immune system to drive tolerogenic response in neonates and infants.
2017-04-02 | MSV000080880 | MassIVE
Project description:It is unclear to what degree protein degradation occurs in the infant stomach and whether peptides previously annotated for bioactivity are released. This study combined nanospray liquid chromatography separation with time of flight mass spectrometry, comprehensive structural libraries and informatics to interrogate milk of three human mothers and the gastric aspirates from their 4–12 day post-partum infants. Milk from the mothers contained almost two hundred distinct peptides, demonstrating enzymatic degradation of milk proteins beginning either during lactation or between milk collection and feeding. In the gastric samples, 649 milk peptides were identified, demonstrating that digestion continues in the infant stomach. The majority of peptides in both the intact milk and gastric samples were derived from β-casein. The numbers of peptides from β-casein, lactoferrin, α-lactalbumin, lactadherin, κ-casein, serum albumin, bile-salt associated lipase and xanthine dehydrogenase/oxidase were significantly higher in the gastric samples than the milk samples (p<0.05). Six hundred three peptides were significantly different in abundance between milk and gastric samples (p<0.05). Most of the identified peptides have previously identified biological activity. Gastric proteolysis occurs in the term infant in the first two weeks of life releasing biologically active milk peptides with immunomodulatory, antibacterial, and calcium-binding activity of clinical relevance to the proximal intestinal tract.
2014-04-15 | PXD000688 | Pride
Project description:<h4><strong>BACKGROUND: </strong>IgE-mediated cow's milk allergy (IgE-CMA) is one of the first allergies to arise in early childhood and may result from exposure to various milk allergens, of which β-lactoglobulin (BLG) and casein are the most important. Understanding the underlying mechanisms behind IgE-CMA is imperative for the discovery of novel biomarkers and the design of innovative treatment and prevention strategies.</h4><h4><strong>METHODS: </strong>We report a longitudinal <em>in vivo</em> murine model, in which two mice strains (BALB/c and C57Bl/6) were sensitized to BLG using either cholera toxin or an oil emulsion (n = 6 per group). After sensitization, mice were challenged orally, their clinical signs monitored, antibody (IgE and IgG1) and cytokine levels (IL-4 and IFN-γ) measured, and fecal samples subjected to metabolomics. The results of the murine models were further extrapolated to fecal microbiome-metabolome data from our population of IgE-CMA (n = 22) and healthy (n = 23) children (Trial: NCT04249973), on which polar metabolomics, lipidomics and 16S rRNA metasequencing were performed. In vitro gastrointestinal digestions and multi-omics corroborated the microbial origin of proposed metabolic changes.</h4><h4><strong>RESULTS: </strong>During mice sensitization, we observed multiple microbially derived metabolic alterations, most importantly bile acid, energy and tryptophan metabolites, that preceded allergic inflammation. We confirmed microbial dysbiosis, and its associated effect on metabolic alterations in our patient cohort, through <em>in vitro</em> digestions and multi-omics, which was accompanied by metabolic signatures of low-grade inflammation.</h4><h4><strong>CONCLUSION: </strong>Our results indicate that gut dysbiosis precedes allergic inflammation and nurtures a chronic low-grade inflammation in children on elimination diets, opening important new opportunities for future prevention and treatment strategies.</h4>
2024-02-15 | MTBLS6768 | MetaboLights
Project description:α-Lactalbumin is an abundant protein present in the milk of most mammals, and is associated with biological, nutritional and technological functionality. Its sequence presents N-glycosylation motifs, the occupancy of which is species-specific, ranging from no to full occupancy. Here, we investigated the N-glycosylation of bovine α-lactalbumin in colostrum and milk sampled from four individual cows, each at 9 time points starting from the day of calving up to one month post-partum. Using a glycopeptide-centric mass spectrometry-based glycoproteomics approach, we identified N-glycosylation at both Asn residues found in the canonical Asn-Xxx-Ser/Thr motif, i.e., Asn45 and Asn74 of the secreted protein. We found similar glycan profiles in all four cows, with partial site occupancies averaging at 35% and 4% for Asn45 and Asn74, respectively. No substantial changes in occupancy occurred over lactation at either site. Fucosylation, sialylation primarily with N-acetylneuraminic acid (NeuAc) and a high ratio of N,N'-diacetyllactosamine (LacdiNAc)/N-acetyllactosamine (LacNAc) antennae were characteristic biochemical features of the identified N-glycans. While no substantial changes occurred in site occupancy at either site during lactation, the glycoproteoform profile revealed lactational dynamics; the maturation of the α-lactalbumin glycoproteoform repertoire from colostrum to mature milk was marked by substantial increases in neutral glycans and the number of LacNAc antennae per glycan, at the expense of LacdiNAc antennae. While the implications of α-lactalbumin N-glycosylation on functionality are still unclear, we speculate that N-glycosylation at Asn74 results in a structurally and functionally different protein, due to competition with the formation of its two intra-molecular disulphide bridges.
2024-08-09 | PXD050331 | Pride
Project description:We have utilized the RNA isolated from breast milk fat globule (MFG) from lactating women from 6h to 42 days following delivery using the HumanHT-12 v4 Expression BeadChip (Illumina, Inc) to determine the temporal coordination of changes in gene expression in milk substrate synthesis processes. Of the 47,323 gene transcripts on the array, 16,623 transcripts were expressed. Major milk protein genes were among the most highly expressed and induced along with those involved in the metabolic and biosynthetic processes of carbohydrate and lipid. Milk fat synthesis mirrors expression of genes involved in fat synthesis, lipolysis and transport. In the lactose synthesis pathway, expression of α-lactalbumin mRNA was high already in the early milk and did not change. Lactose synthesis paralleled the induction of gene expression of proteins involved in UDP-galactose synthesis and transport. Serial milk samples were collected 6h following delivery, q12 h for the first 4 days and then weekly for 6 wks from 7 healthy, lean, exclusively breastfeeding women [7 x 15 = 105 samples]. RNA was isolated from the milk fat globules and utilized for microarray analyses.
2012-09-04 | E-GEOD-36936 | biostudies-arrayexpress
Project description:Human milk fat globules, by enveloping cell contents during their secretion into milk, are a rich source of mammary cell RNA. Here, we pair this non-invasive mRNA source with RNA sequencing technology to probe the milk fat layer transcriptome during three stages of lactation: colostral, transitional, and mature milk production. We find that transcriptional profiles cluster not by postpartum day, but by milk Na:K ratio, indicating that women sampled during the same postpartum time frame could be at markedly different stages of gene expression. Each stage of lactation is characterized by a dynamic range (105-fold) in transcript abundances not previously observed with microarray technology. We discovered that transcripts for isoferritins and cathepsins are strikingly abundant during colostrum production, highlighting the potential importance of these proteins for neonatal health. Two transcripts, encoding M-NM-2-casein (CSN2) and a-lactalbumin (LALBA), make up 45% of the total pool of mRNA in mature lactation. Genes significantly expressed across all stages of lactation are associated with making, modifying, transporting, and packaging milk proteins. Stage-specific transcripts are associated with immune defense during the colostral stage, up-regulation of the machinery needed for milk protein synthesis during the transitional stage, and the production of lipids during mature lactation. We observed strong modulation of key genes involved in lactose synthesis and insulin signaling. In particular, PTPRF may serve as a biomarker linking insulin resistance with insufficient milk supply. This study provides the methodology and reference data set to enable future targeted research on the physiological contributors to sub-optimal lactation in humans. Milk fat mRNA profiles were generated from Day 2 and mature milk samples obtained from lactating mothers
2013-07-22 | E-GEOD-45669 | biostudies-arrayexpress
Project description:Immunotherapy strategies have been studied for IgE-mediated food allergies but have not been studied for non-IgE mediated food allergy disorders, including eosinophilic esophagitis. We interrogated the transcriptional signatures of peripheral CD4+ cells isolated from pediatric eosinophilic esophagitis patients enrolled in a phase IIa clinical trial of milk-antigen epicutaneous immunotherapy. We analyzed differential expression in peripheral CD4+ cells before therapy in patients before and after milk-exclusion diet. RNA-sequencing analysis revealed that there were 244 differentially expressed genes in peripheral blood CD4+ cells of EoE patients consuming milk versus those eliminating it, and 129 DEGs in CD4+ cells isolated after EPIT versus after placebo (FDR<0.05). Gene set enrichment analysis identifies enrichment of hallmark interferon-a and -g response signatures in peripheral CD4+ T cells from EoE patients during active disease on a milk-containing diet. In post-EPIT therapy samples, our analysis identifies enrichment in T-cell receptor signaling, antigen presentation, costimulation, and cytokine signaling pathways. These data suggest a unique peripheral CD4+ T cell gene signature is present in EoE patients during active disease. We observe a distinct transcriptional profile post-EPIT therapy in EoE patients, suggesting that EPIT alters CD4+ responses in EoE patients.
2021-08-03 | GSE173895 | GEO
Project description:We have utilized the RNA isolated from breast milk fat globule (MFG) from lactating women from 6h to 42 days following delivery using the HumanHT-12 v4 Expression BeadChip (Illumina, Inc) to determine the temporal coordination of changes in gene expression in milk substrate synthesis processes. Of the 47,323 gene transcripts on the array, 16,623 transcripts were expressed. Major milk protein genes were among the most highly expressed and induced along with those involved in the metabolic and biosynthetic processes of carbohydrate and lipid. Milk fat synthesis mirrors expression of genes involved in fat synthesis, lipolysis and transport. In the lactose synthesis pathway, expression of α-lactalbumin mRNA was high already in the early milk and did not change. Lactose synthesis paralleled the induction of gene expression of proteins involved in UDP-galactose synthesis and transport.
2012-09-04 | GSE36936 | GEO
Project description:Whey proteins and caseins in breast milk provide bioactivities and also have different amino acid composition. Accurate determination of these two major protein classes will therefore provide a better understanding of human milk composition and function and also aid in developing improved infant formulas based on bovine whey proteins and caseins. In this study, we implemented a LC-MS/MS quantitative analysis based on iBAQ label free quantitation to estimate absolute concentrations of alpha-casein, beta-casein and kappa-casein in human milk samples (n=88) collected between day 1 and day 360 post partum. Total protein concentration measured by ninhydrin-based amino acid analysis ranged from 2.03 to 17.52 with a mean of 9.37�3.65 g/L. Casein subunits ranged from 0.04 to 1.68 g/L (alpha-), 0.04 to 4.42 g/L (beta-), and 0.10 to 1.72 g/L (kappa-), with beta-casein having the highest average concentration among the three subunits. Calculated whey/casein ratio ranged from 45:55 to 97:3. Linear regression analyses show significant decreases in total protein, beta-casein, kappa-casein, total casein, and whey/casein ratio during the course of lactation. Our study presents a novel and accurate analysis of human milk casein content, demonstrating a lower casein content than earlier believed, which has implications for improved infants formulas.
2017-06-23 | MSV000081195 | MassIVE