The clinical effects of hospitalization in a low pollutant room on atopic dermatitis

BACKGROUND: Environmental pollutants are thought to be one of major triggers of atopic dermatitis (AD). OBJECTIVE: We attempted to evaluate the clinical effects of environment with low indoor pollutant levels on AD management. METHODS: Fifty-one children (mean age 1.7 years) with moderate to severe AD who failed to show improvement with conventional management were recruited. Disease severity was assessed by SCORAD (Scoring of AD) indices. They were admitted in a low pollutant oom for 3-4 days (mean 3.3 days) which was designed to keep low levels of dust, house dust mites, micro-organisms, and indoor air pollutants such as total volatile organic compounds (TVOCs), particulate matter (PM), and so on. Air pollutant levels in the low pollutant room were lower than primary standards defined by the Korean Ministry of Environment. we compared disease severity on admission and after discharge, and the pollutant levels of each patient's home and low pollutant room. RESULTS: The SCORAD was significantly reduced from 42.0 ± 11 .5 to 29.8 ± 8.9 (p < 0.001) by management in a low pollutant room. PM₂.₅, PM₁₀, formaldehyde, TVOCs, carbon dioxide, bacterial suspensions, and indoor molds were significantly higher in the patient's home than low pollutant room. Out of 29 patients who deteriorated after discharge to their home, 8 patients were admitted again, and their SCORAD was rapidly decreased from 53.1 ± 16.2 to 39.2 ± 9.8 (p = 0.036). CONCLUSION: Indoor air pollutants are likely to affect AD in susceptible individuals. Environmental control to lower indoor air pollutant levels might be necessary for better management of AD in some patients.

Assessment of Allergenicity in Genetically Modified Herbicide-Resistant Foods Using the Serum Screening Test / 소아알레르기및호흡기학회지

PURPOSE: This study aimed to evaluate the potential allergenicity of genetically modified (GM) herbicide-resistant food by using the serum screenning test. METHODS: Children with allergic disease were recruited, and those who were sensitized to soybean, corn or peanut were selected to obtain their sera. Sensitization to these food allergens was determined when the level of specific IgE was over 0.35 kU/L using ImmunoCAP (Pharmacia, Uppsala, Sweden). Immunoblot analyses were performed for soybean (n=50), corn (n=50) and peanut (n=20). Newly inserted gene was sequenced and cloned from GM soy (Roundup Ready Soybean, Monsanto), GM corn (Bt 11, Syngenta) and GM canola (MS8/RF3 canola, Bayer CropScience). These proteins, such as CP4 EPSPS, PAT, and BAR, were expressed and purified for the serum screening test. RESULTS: Immunoblot analysis using CP4 EPSPS and sera from soybean-sensitized children showed no bands. Likewise, sera from corn-sensitized children and PAT did not demonstrate IgE binding in immunoblot analysis. In addition, there were no reactions between BAR and sera from peanut-sensitized patients. CONCLUSION: The serum screening test using sera from allergic children and newly inserted protein (CP4 EPSPS, PAT and BAR) in GM soybean, GM corn and GM canola failed to show IgE binding in immunoblot analysis. The results of this study suggest that these newly inserted proteins may not cause allergic disease. Further studies using more sera from allergic children are needed to conclude the safety of herbicide-resistant GM food.

Evaluating the Allergenicity of Soybean by the Fermentation / 소아알레르기및호흡기학회지

PURPOSE: The aim of this study was to determine the allergenicity of soybean by fermentation. METHODS: Non-fermented soybean, fermented soybean by Bacillus subtilis KFCC 11293 and 3-step fermented soybean by Lactococcus lactis subsp. lactis IFO 12007 and Aspergillus oryzae and B. subtilis KFCC 11293 were extracted with phosphate buffered saline (PBS). In order to detect soybean-specific IgE, we performed SDS-PAGE and IgE immunoblot analysis by using 3 kinds of soybean extracts and sera from 9 patients with atopic dermatitis. All patients were sensitive to soybean, which were confirmed by CAP-FEIA. RESULTS: SDS-PAGE of non-fermented soybeans showed many bands, whereas only peptides of less than 15 kDa were found in fermented soybeans. IgE immunoblot analysis of fermented soybeans failed to detect specific IgE which were seen in non-fermented soybeans. CONCLUSION: Fermentation could reduce the allergenicity of soybeans by efficiently degrading antigenic proteins in soybeans. However, there was no significant difference between fermentation by B. subtilis and 3-step fermentation.

Sensitization of Food Allergen in Breastfed Infant with Atopic Dermatitis / 대한지역사회영양학회지

Breastfeeding has been known as the best feeding practice to prevent allergies including atopic dermatitis (AD). However, the benefit on the prevention of allergic disease is still controversial. The objectives of this study were to examine the rate of sensitization to the protein of eggs, cow's milk and soy in exclusively breastfed infants and to evaluate antigen-antibody reaction between breast milk and serum of AD infant. Data on feeding and food hypersensitivity were obtained for 62 AD infants (32 male, 30 female) aged 0.7 kU/L by CAP assay (Pharmacia, Uppsala, Sweden) were considered positive. The rates of sensitization in breastfed infants were 41.9% (26/62) to egg, 30.6% (19/62) to milk and 18.0% (11/62) to soy. Immunoblotting analyses were performed using breast milk with the matched serum of seven AD infants (4 male/3 female). Binding patterns of AD infant's IgE to breast milk extract showed visible specific band for immunoglobulin, especially in case of a lactating mother who did not completely restricted ingestion of egg, milk and soy. These results indicate that sensitization to food allergen develops via breast milk feeding. Breast milk feeding should be recommended in infants at risk of developing allergic disease, but maternal intake of highly allergenic food might be restricted for prevention and treatment of food allergy among the babies with AD.

Sensitization of Food Allergen in Breastfed Infant with Atopic Dermatitis / 대한지역사회영양학회지

Breastfeeding has been known as the best feeding practice to prevent allergies including atopic dermatitis (AD). However, the benefit on the prevention of allergic disease is still controversial. The objectives of this study were to examine the rate of sensitization to the protein of eggs, cow's milk and soy in exclusively breastfed infants and to evaluate antigen-antibody reaction between breast milk and serum of AD infant. Data on feeding and food hypersensitivity were obtained for 62 AD infants (32 male, 30 female) aged 0.7 kU/L by CAP assay (Pharmacia, Uppsala, Sweden) were considered positive. The rates of sensitization in breastfed infants were 41.9% (26/62) to egg, 30.6% (19/62) to milk and 18.0% (11/62) to soy. Immunoblotting analyses were performed using breast milk with the matched serum of seven AD infants (4 male/3 female). Binding patterns of AD infant's IgE to breast milk extract showed visible specific band for immunoglobulin, especially in case of a lactating mother who did not completely restricted ingestion of egg, milk and soy. These results indicate that sensitization to food allergen develops via breast milk feeding. Breast milk feeding should be recommended in infants at risk of developing allergic disease, but maternal intake of highly allergenic food might be restricted for prevention and treatment of food allergy among the babies with AD.

Effect of Interleukin-9 on Cell Proliferation and Histamine Release of Cord Blood-derived Human Mast Cells / 소아알레르기및호흡기학회지

PURPOSE: Interleukin-9(IL-9), one of Th2-type cytokines, might be important in the pathophysiology of allergic diseases. We investigated the effect of IL-9 on human mast cells by assessing cell proliferation and histamine release. METHODS: Human umbilical cord blood cells were cultured in the presence of stem cell factor(SCF, 100 ng/mL) and IL-6(50 ng/mL) in liquid medium for 8 weeks. Then these cells were divided into 3 aliquots. Each aliquot was cultured for 4 more weeks in different conditions : SCF alone(100 ng/mL), IL-9 alone(50 ng/mL) and SCF+IL-9. Cell numbers were counted using hemocytometer. For evaluation of apoptosis, DNA fragmentation was determined by propidium iodide(PI) staining and flow cytometric analysis. Histamine concentration was measured by ELISA after stimulation with human IgE and anti-human IgE. RESULTS: Cell numbers increased significantly when they were cultured in the presence of SCF and IL-9 compared with SCF alone(P<0.05). Proliferation of mast cells was mediated by decreased apoptosis. Histamine release in activated mast cells was not different regardless of incubation with IL-9. CONCLUSION: IL-9 might be involved in allergic inflammation via proliferation of mast cells in target tissue.

Egg white-specific IgG western blotting in children with atopic dermatitis / 천식및알레르기

BACKGROUND AND OBJECTIVE: Specific IgE to food allergen is associated with atopic dermatitis, but it does not always show good clinical correlation. It has been suggested that IgG may be partly involved in allergen-induced reaction. This study was designed to investigate the clinical significance of specific IgG antibody to egg white (EW) in atopic dermatitis patients who showed improvement with egg elimination diet. METHOD: Eleven atopic dermatitis patients who responded to egg elimination diet were selected. They were classified into two groups; group I (n=5) with positive specific IgE to EW and group II (n=6) with normal levels of serum specific IgE. Two volunteers with no allergic diseases were enrolled in the control group. EW-specific IgG western blotting was performed with patient's serum and purified protein extracted from EW. RESULTS: IgG western blotting to EW in group I showed bands at 51.8 kD in two patients and bands at 35 kD in the others. In group II, two showed bands at 51.8 kD, and diffuse bands at 35 kD~51.8 kD were found in four patients. There were no bands in the control group. CONCLUSION: Regardless of the presence of specific IgE, specific IgG to EW was detected by western blotting in patients with egg-associated atopic dermatitis, suggesting that specific IgG may play a role in the pathogenesis of atopic dermatitis.

Sulfidoleukotrienes Production in Blood Leukocytes Stimulated by Mite Allergen / 소아알레르기및호흡기학회지

PURPOSE: Although asthma is the most common chronic disease in childhood, accurate diagnosis in infants and young children remains challenging clinicians. Allergen challenging tests in vitro have been used productively as an investigative tool in studies of the pathophysiology and diagnosis of asthma. Therefore, we compared the sulfidoleukotrien (sLT) concentration according to allergen leukocyte stimulation test, in normal versus asthmatic patients, to find better diagnostic tools. METHODS: From May through August, 1998, nine children were enrolled who presented positive skin reaction in Dermatophagoides pteronyssinus (D.p.), Dermatophagoides farinae (D.f.) as patient groups. We measured total eosinophil count, serum IgE, sLT concentration of three different allergen stimulation (100 ng/ml, 10 ng/ml, 1 ng/ml). RESULTS: sLT concentration in three different D.p., D.f. stimulation showed significant differences (P<0.01). Allergen concentration of 10 ng/ml was fit for stimulating peripheral leukocyte. The sLT concentration is correlated with IgE, total eosinophil count, but not with age. Actual concentrations of sLT was not measured in allergen stimulation test. Its interpretation of test results was complicated by the fact that several variants were involved in determining sampling time and appropriate sampling volume. Most importantly, the diagnostic sensitivity of the sLT concentration tests varies directly with the magnitude of IgE antibody and total eosinophil count. CONCLUSION: We emphasizes the role of allergen challenge in understanding the pathophysiology of young children asthma. It focuses on more accurate diagnosis with objective techniques for analyzing the leukocyte sLT release to antigen.

Characterization of IL-3, IL-5, GM-CSF-induced Eosinophils from Human CD34+ Cord Blood Cells / 소아알레르기및호흡기학회지

PURPOSE: Eosinophils are cells of the granulocyte lineage that participate in host defence against parasitic disease and mediate allergic inflammation. In this study by using the combination of cytokines IL-3, IL-5 and GM-CSF, we explore the characterization of cultured eosinophils from CD34+ CBCs. METHODS: Mononuclear cells were isolated heparinized umbilical cord blood by Ficoll-paque (1.077 g/ml) density gradient centrifugation method. The CD34-bearing hematopoietic progenitor cells were collected by elution after their adhesion to a magnetic cell sepatation (MACS) column. The CD34+ cells were incubated in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and were cultured in the presence of IL-3, IL-5, and GM-CSF in a 6-well plate bottomed tissue culture plate at 37 degrees C for 7-28 days in humidified 5% CO2 and 95% air atmosphere. For the identification of cultured eosinophils Wright's and Giemsa staining, RT-PCR, Southern blotting and FACS analysis are used. RESULTS: We analyzed the cultured eosinophils Wright's and Giemsa staining, the total cell number of cells increased 50-fold by days 28 of culture. Also, using the sensitive RT-PCR technique, we monitered the appearance of mRNA transcrips of EPO. Identity of each RT-PCR product was confirms by southern blotting with independent gene-specific oligonucleotide probes and we found increasing of hybridization signals for EPO at 7th culture days. In addtion, we identified eosinophils in cultured CD34+ CBCs by flow cytometry. As s results, we succeeded in developing of pure eosinophils efficiently from CD34+ of CBCs in the presence of IL-3, IL-5 and GM-CSF. CONCLUSION: The in vitro growth of CD34+ CBCs may provide a useful system to study growth factor and stage-dependent adhesion molecule expression, as well as function on developing eosinophils.