Krystyna Wasowska-Królikowska

FOXP3, IL-10, and TGF-β genes expression in children with IgE-dependent food allergy.

BackgroundRegulatory T cells (Tregs) have an essential role in tolerance and immune regulation. However, few and controversial data have been published to date on the role and number of these cells in food allergic children. The forkhead/winged-helix transcription factor box protein 3 (FOXP3) is considered the most reliable marker for Tregs.ObjectiveThis study aims to investigate the FOXP3, interleukin (IL)-10, and transforming growth factor (TGF-β) genes expression in children with IgE-dependent food allergy.Material and MethodsThe study group consisted of 54 children with IgE-dependent food allergy (FA) and a control group of 26 non-atopic healthy children. The diagnosis of FA was established using questionnaires, clinical criteria, skin prick tests, serum sIgE antibodies (UniCAP 100 Pharmacia Upjohn), and a double-blind placebo control food challenge. In order to assess gene expression, the isolation of nucleated cells was performed using Histopaque-1077 (Sigma-Aldrich, Germany). The concentration of RNA obtained was measured using a super-sensitive NanoDrop ND1000 spectrophotometer (Thermo Scientific, USA). A reverse transcription reaction was performed using a commercially available set of High Capacity cDNA Archive Kit (Applied Biosystems, USA). Analysis have been carried out in the genetic analyzer 7900HT Real-Time PCR (Applied Biosystems, USA).ResultsThe average level of the FOXP3 gene expression in the studied group was 2.19 ± 1.16 and in the control group 2.88 ± 1.66 (p = 0.03). The average level of IL10 mRNA expression in the study group was 13.6 ± 1.07 and was significantly lower than corresponding values in the control group 14.3 ± 1.1 (p = 0.01). There were no significant differences in the average level of the TGF-β mRNA expression in the study group (3.4 ± 0.4) and controls (3.5 ± 0.3; p > 0.05). The FOXP3 gene expression was the highest in children who acquired tolerance to food (3.54 ± 0.75), lower in heated allergen-tolerant children (2.43 ± 0.81), and the lowest in heated allergen-reactive children (1.18 ± 0.5; p = 0.001 control vs heated allergen reactive; p = 0.005 heated allergen tolerant vs heated allergen reactive; p = 0.001 outgrown vs heated allergen reactive). The significant tendency toward lower total IgE levels with a higher FOXP3 mRNA expression was detected (n = 54; Pearson r = −0.4393; p = 0.001).ConclusionsChildren with FA showed statistically significant lower level of the FOXP3 and IL10 gene expression than healthy children. Children acquiring tolerance to the food show significantly higher levels of the FOXP3 gene expression than children with active FA. The correlation between the level of FOXP3 and total IgE was detected.

Bronchial reactivity in schoolchildren allergic to food

BACKGROUND Food allergy (FA) is associated with an increased risk of asthma. OBJECTIVE To evaluate whether bronchial hyperreactivity (BHR) occurs in children with FA depending on the presence of respiratory symptoms. METHODS Fifty-four children with FA and 62 without FA were studied for BHR, defined as a provocation concentration of methacholine that caused a decrease in forced expiratory volume in 1 second of 20% (PC20). The diagnosis of FA was established using questionnaires, clinical criteria, skin prick tests, serum specific IgE antibodies, and a double-blind placebo-controlled food challenge. RESULTS Among nonasthmatic children, BHR was diagnosed in 15 (47%) with FA and in 7 (17%) without FA (P < .005); BHR was demonstrated in 7 children (41%) with FA alone. All the patients with asthma with or without FA had BHR. The mean (SD) PC20 in children with FA alone was 2.8 [1.38] mg/mL and was significantly higher than that in children with asthma alone (0.88 [1.01] mg/mL) or with asthma and FA (0.96 [0.83] mg/mL) (P < .001). BHR was detected in 5 children (36%) with a mild anaphylactic reaction provoked by food, in 23 (74%) with a moderate reaction, and in all the children with a severe reaction. CONCLUSIONS Children with FA had increased BHR independent of respiratory symptoms. Although BHR occurs in asymptomatic children with FA, its course is milder than that in children with FA and asthma. Factors that determine BHR prevalence in children with FA are similar to those in children without FA.

Prevalence of EGG derangement in newly diagnosed type 1 diabetes in childhood.

Objective: To evaluate gastric myoelectrical activity in children with newly diagnosed type 1 diabetes melliltus (T1DM) in relation to blood glucose control and visceral neuropathy. Methods: Percutaneous electrogastrograpy (EGG) was performed on 42 children (20 F; mean age 12.9 ± 3.1 years) with T1DM of <1 years duration and on 35 healthy controls (18 F; mean age 13.4 ± 3.6 years). After overnight fasting, a 30-minute EGG recording was followed by test meal consumption and then a 60-minute postprandial EGG aquisition. Fasting and postprandial periods were analyzed for gastric dysrhythmias, dominant frequency (DF) and additional parameters. In T1DM patients, HbA1c and blood glucose levels were measured and tests for visceral neuropathy were performed. Results: In 41 T1DM patients (98%), cardiovascular neuropathy tests were negative. In 12 of those patients (29%) and in 32 healthy controls (91%), electrogastrograms were normal. The percentages of fasting and postprandial gastric dysrhythmias were significantly higher in T1DM patients compared to controls (P < 0,05). In T1DM children after feeding, some normalization of gastric myoelectrical rhythm was observed: normogastria increased nearly 2-fold to 72.6 ± 22.9% and bradygastria decreased to 20.8 ± 20.4% from 52.3 ± 32.4% (P < 0.05). The percentages of fasting bradygastria and normogastria were correlated with glycemia level (r = −0.55 and r = 0.51, respectively; P < 0.05), as was postprandial DF (r = 0.41; P < 0.05). There was no correlation between HbA1c levels and EGG parameters. Conclusions: Derangement of the gastric myoelectrical activity is present in 71% of children with early stage T1DM. Glucose levels influence gastric myoelectrical activity, whereas long-term glucose control (HbA1c level) does not correlate with EGG parameters.

Electrogastrography in children and adolescents with type 1 diabetes: Weak correlation with metabolic control parameters

Aim: To evaluate gastric myoelectrical activity with respect to duration and metabolic control of type 1 diabetes mellitus (T1DM). Methods: 172 children and adolescents with T1DM (mean 14.4±3.7 y), divided into subgroups depending on diabetes duration (<5 and > 5 y), and 35 healthy controls (mean 13.93±3.59 y) were examined. All subjects underwent electrogastrography (EGG) performed after overnight fasting. In subjects with T1DM, haemoglobin A1c (HbA1c) and blood glucose levels during EGG records were measured. Results: 15.69% of T1DM patients and 91.42% of the controls fulfilled normal EGG criteria (p < 0.001). T1DM subjects had a lower percentage of fasting normogastria (34.56±27.35% vs 69.84±18.16%, p = 0.0001) and higher bradygastria (51.97±30.24% vs 19.11±15.01%, p = 0.0001) compared to controls. In diabetic patients, an increase in postprandial normogastria (60.37±23.96% vs 76.68±12.38, p < 0.05) and a decrease in bradygastria percentage (25.67±21.01% vs 9.58±7.13%, p < 0.05) was observed. In children with disease < 5 y, diabetes duration correlated with power ratio (r= ‐ 0.27, p = 0.01), postprandial normogastria (r= ‐ 0.24, p = 0.03) and tachygastria (r= 0.25, p = 0.02). Weak correlations between EGG parameters and glucose (preprandial dominant frequency r= ‐ 0.19, p < 0.05; postprandial normogastria r= 0.23, p < 0.01) and HbA1c levels (preprandial bradygastria r= 0.19, postprandial dominant power r= 0.23; p < 0.05) were observed.

Evaluation of Receptor Expression on Immune System Cells in the Peripheral Blood of Asthmatic Children Undergoing Food Challenges

Background: The role of food allergens in the induction of allergic reactions in the airways is not completely understood. The aim of the present study was to evaluate fluorocytometric assays of the peripheral blood during food challenge tests in children with asthma and food allergy. Patients and Methods: 22 children with asthma and concomitant food allergy and 18 children with asthma without food allergy participated in the study. Oral challenge tests were performed using double-blind, placebo-controlled food challenge. Blood samples were collected before and 4 and 24 h after the challenge. CD25 and CD23 antigen expression was determined with monoclonal antibodies using a FACSCalibur flow. Results: The evaluation of the CD25+ T subpopulation and CD19+CD23+ B lymphocytes revealed statistically significant differences between the study group and the control group. In children with asthma and food allergy, the cell pool consisted (on average) of 9 ± 2.8% of CD3+CD25+ cells before the challenge and of 10.3 ± 3.8% (mean delta: 1.623; p = 0.01) after the provocation. However, placebo challenge did not significantly change the number of this T-lymphocyte subpopulation (mean delta: –0.121; p > 0.05). The highest increase in the CD25+ T-subpopulation expression was found in patients with respiratory reactions during the positive food challenge (mean delta: 4.065; p < 0.004). Conclusions: An increase in CD25+ T-lymphocyte and CD23 B-lymphocyte populations after food allergen challenge may indicate their significant role in the pathogenesis of the active phase of the immunoinflammatory process in children with asthma and concomitant food allergy.