No abstract

Background The peptide microarray is a novel assay which facilitates high-throughput screening of peptides with a small quantity of sample. Objective We sought to use overlapping peptides of milk allergenic proteins as a model system to establish a reliable and sensitive peptide microarray-based immunoassay for large scale epitope mapping of food allergens. Methods A milk peptide microarray was developed using commercially synthesized peptides (20-mers, 3 offset) covering the primary sequences of αs1-, αs2-, β-, and κ-caseins, and β-lactoglobulin. Conditions for printing and immunolabeling were optimized using a serum pool of five milk-allergic patients. Reproducibility of the milk peptide microarray was evaluated using replicate arrays immunolabeled with the serum pool, whereas specificity and sensitivity were assessed using serial dilution of the serum pool and a peptide inhibition assay. Results Our results show that epitopes identified by the peptide microarray were mostly consistent with those identified previously by SPOT membrane technology, but with specific binding to a few newly identified epitopes of milk allergens. Data from replicate arrays were reproducible (R≥0.92) regardless of printing lots, immunolabeling and serum pool batches. Using the serially diluted serum pool, we confirmed that IgE antibody binding detected in the array was specific. Peptide inhibition of IgE binding to the same peptide and overlapping peptides further confirmed the specificity of the array. Conclusions A reliable peptide microarray was established for large scale IgE epitope mapping of milk allergens and this robust technology could be applied for epitope mapping of other food allergens.

Our results suggest that supplementation with probiotic L. plantarum CJLP133 is beneficial in the treatment of pediatric AD.

IntroductionThe effects of weather and air pollution on the severity and persistence of atopic dermatitis (AD) are important issues that have not been investigated in detail. The objective of our study was to determine the short-term effects of meteorological variables and air pollution on AD symptoms in children.MethodsWe enrolled 177 AD patients with 5 years or younger from the Seoul Metropolitan Area, Korea, and followed for 17 months between August 2013 and December 2014. Symptoms records of 35,158 person-days, including itching, sleep disturbance, erythema, dry skin, oozing, and edema, were obtained. We estimated the effect of meteorological variables including daily mean temperature, relative humidity (RH), diurnal temperature range (DTR), rainfall and air pollutants including particulate matter with an aerodynamic diameter ≤10 μm (PM10), nitrogen dioxide (NO2), and tropospheric ozone (O3) on AD symptoms using a generalized linear mixed model with adjustment for related confounding factors.ResultsA 5°C increase in outdoor temperature and a 5% increase in outdoor RH was associated with 12.8% (95% confidence intervals (CI): 10.5, 15.2) and 3.3% (95% CI: 1.7, 4.7) decrease in AD symptoms, respectively, on the same day. An increase of rainfall by 5 mm increased AD symptoms by 7.3% (95% CI: 3.6, 11.1) for the days with <40 mm rainfall. The risk of AD symptoms increased by 284.9% (95% CI: 67.6, 784.2) according to a 5°C increase in DTR when it was >14°C. An increase in PM10, NO2, and O3 by 10 units increased the risk of AD symptoms on the same day by 3.2% (95% CI: 1.5, 4.9), 5.0% (95% CI: 1.4, 8.8), and 6.1% (95% CI: 3.2, 9.0), respectively.ConclusionExposure to meteorological variables and air pollutants are associated with AD symptoms in young children.