Komei Ito

Detection of IgE antibody against Candida albicans enolase and its crossreactivity to Saccharomyces cerevisiae enolase

Candida albicans 46 kDa protein, a glycolytic enolase enzyme, is an important allergen of the yeast. The purpose of the study was to detect circulating IgE and IgG antibodies against C. albicans enolase (CAE). We isolated CAE using sequential DEAE Sephacel and Pl 1 column chromatography from spheroptasts of C. albicans, and delected IgE and IgG antibody against CAE by immunoblotting. Crossreactivity of enolose of C. albicans and Saccharomyces cerevisiae was also examined by immunoblotting and immunoblot inhibition test. Among 54 sera with positive IgE RAST to C. albicans, IgE antibody against CAE was detected in 20 sera (37%) and IgG antibody in 27 sera (50%). The allergenic potency of CAE was confirmed using a skin‐prick test in three patients. Simultaneous IgE binding to S. cerevisiae enolase was only observed in four out of 20 sera reacting to CAE. Pre‐treatment of sera with CAE completely inhibited IgE binding to S. cerevisiae enolase. Whereas the latter only partially inhibited IgE binding to CAE. These results suggest that CAE shares some crossreacting epitopes with S. cerevisiae enolase, representing minor components of CAE but dominant segments of S. cerevisiae enolase.

IgE to Gly m 5 and Gly m 6 is associated with severe allergic reactions to soybean in Japanese children

IgE to Gly m 5 and Gly m 6 is associated with severe allergic reactions to soybean in Japanese children

Clinical Utility of IgE Antibodies to ω-5 Gliadin in the Diagnosis of Wheat Allergy: A Pediatric Multicenter Challenge Study

Background: There are contradictory results regarding the clinical usefulness of the determination of IgE antibodies to ω-5 gliadin in children with a suspicion of wheat allergy (WA). Methods: The study comprised 311 children and young adults with suspected wheat intolerance treated at three separate pediatric clinics and, with the exception of 25, were found to be positive in specific IgE antibody determinations to wheat. Their ages ranged from 6 months to 20.4 years (median age, 2.3 years). Possible relationships between IgE antibodies to ω-5 gliadin and a physician’s diagnosis of WA and challenge symptoms were studied. Results: The mean concentration of IgE antibodies to ω-5 gliadin was 1.2 kUA/l in WA patients and <0.35 kUA/l in patients without WA (p < 0.0001). Seventy-two percent of the WA patients had positive ω-5 gliadin levels and 75% of the patients without WA had negative levels. Logistic regression showed a significant relationship between the probability of WA and the concentration of IgE antibodies to ω-5-gliadin with a 2.6-fold (95% CI: 2.0–3.3) increased risk. Age was an important factor to consider as the risk of WA increased 5.4-fold (95% CI: 1.4–21) for children ≤1 year of age and 2.5-fold (95% CI: 2.0–3.2) for children >1 year of age with increasing levels of IgE. Conclusion: Detection of IgE to ω-5 gliadin seems to be associated with responsiveness to the challenge test and is particularly useful in infants with a suspicion of WA.

The usefulness of casein-specific IgE and IgG4 antibodies in cow's milk allergic children.

BackgroundCows milk allergy is one of the most common food allergies among younger children. We investigated IgE antibodies to milk, and IgE and IgG4 antibodies to casein, α-lactalbumin and β-lactoglobulin in cows milk allergic (CMA) and non-allergic (non-CMA) children in order to study their clinical usefulness.MethodsEighty-three children with suspected milk allergy (median age: 3.5 years, range: 0.8-15.8 years) were diagnosed as CMA (n = 61) or non-CMA (n = 22) based on an open milk challenge or convincing clinical history. Their serum concentrations of allergen-specific (s) IgE and IgG4 antibodies were measured using ImmunoCAP®. For the sIgG4 analysis, 28 atopic and 31 non-atopic control children were additionally included (all non-milk sensitized).ResultsThe CMA group had significantly higher levels of milk-, casein- and β-lactoglobulin-sIgE antibodies as compared to the non-CMA group. The casein test showed the best discriminating performance with a clinical decision point of 6.6 kUA/L corresponding to 100% specificity. All but one of the CMA children aged > 5 years had casein-sIgE levels > 6.6 kUA/L. The non-CMA group had significantly higher sIgG4 levels against all three milk allergens compared to the CMA group. This was most pronounced for casein-sIgG4 in non-CMA children without history of previous milk allergy. These children had significantly higher casein-sIgG4 levels compared to any other group, including the non-milk sensitized control children.ConclusionsHigh levels of casein-sIgE antibodies are strongly associated with milk allergy in children and might be associated with prolonged allergy. Elevated casein-sIgG4 levels in milk-sensitized individuals on normal diet indicate a modified Th2 response. However, the protective role of IgG4 antibodies in milk allergy is unclear.

Characterization of IgE‐binding epitopes on Candida albicans enolase

Candida albicans enolase is one of the important allergens in Candida allergy. We isolated and purified 46 kDa C. albicans enolase (CAE) from C. albicans and characterized epitopes for IgE antibody by lectin‐blotting and enzymatic digestion followed by sodium dodecyl sulfale polyacrylamide gel electrophoresis (SDS‐PAGE) and immunobiotting. Lectin blotting and deglycozilation indicated that this protein did not contain polysaccharide side chains. The purified CAE and recombinant fusion protein produced from CAE gene possessed common epitopes for IgE antibody. We estimated IgE binding epitopes on the basis of reported amino acid sequences from the analysis of cDNA encoding CAE. V8 protease digestion of CAE gave six polypeptide fragments (A‐F). The N‐termini of each fragment were confirmed by amino acid sequence and the C‐termini were estimated by molecular weights of each fragment and the specific cutting site of V8 protease. Fragment C (25.0 kDa; F‐171‐I‐399) reacted to 90% IgE antibodies examined, whereas fragments D (21.0 kDa; F‐171‐1‐360), E (16.2kDa: F‐171‐D‐317) and F (13.0kDa; A‐47‐E‐170) showed no IgE binding. Our results suggest that epitopes for IgE antibodies exist near the C‐terminal of the protein.

Dexamethasone reduces lung eosinophilia, and VCAM-1 and ICAM-1 expression induced by Sephadex beads in rats

Airway eosinophilia is one of the key pathophysiologic features in asthma. The endothelial adhesion molecules, vascular cell adhesion molecule (VCAM-1) and intercellular adhesion molecule (ICAM-1), have previously been shown to play a crucial role in eosinophil recruitment into the inflamed airway. We have investigated the effects of dexamethasone on eosinophilia into the bronchoalveolar lavage fluid, and the upregulation of VCAM-1 and ICAM-1 expression, measured by immunoblotting, induced by i.v. injection of Sephadex beads into rats. The beads significantly increased the lung eosinophilia, and expression of VCAM-1 and ICAM-1 in the lung. Pretreatment with dexamethasone (0.1 to 2 mg/kg i.p.) strongly inhibited all the airway inflammatory events in a dose-dependent manner. In conclusion, glucocorticoids may be potent inhibitors of lung eosinophilia, at least in part, due to the prevention of the upregulation of VCAM-1 and ICAM-1 expression.

Effects of a Short-Term Parental Education Program on Childhood Atopic Dermatitis: A Randomized Controlled Trial

Parental education is important in managing childhood atopic dermatitis (AD). We evaluated the long‐term effects of a 2‐day parental education program (PEP) on childhood AD. In an investigator‐blinded, randomized controlled trial, 59 children age 6 months to 6 years with moderate to severe AD and their mothers were recruited in Japan. Participants were given a booklet about AD and received conventional treatment alone or in combination with a 2‐day PEP comprising three lectures, three practical sessions, and a group discussion. The primary outcome was evaluation of eczema severity using SCORing Atopic Dermatitis (SCORAD) at 6 months. Secondary outcomes included changes in symptom scores, amount of corticosteroid used, parental quality of life as determined according to the Dermatitis Family Impact questionnaire, and change in parental anxiety regarding the use of corticosteroids in their children. Participants in the PEP group had a significantly lower SCORAD score than those in the control group at 6 months (mean difference 10.0, 95% confidence interval [CI] = 2.3–17.7, p = 0.01) and objective SCORAD score (mean difference 7.1, 95% CI = 0.8–13.5, p = 0.03). The sleeplessness symptom score (mean difference 1.6, 95% CI = 0.0–3.1, p = 0.048) and corticosteroid anxiety score (p = 0.02) in the PEP group were significantly better than in the control group at 6 months. There was no significant difference between groups in the amount of corticosteroid used or quality of life. The PEP had positive long‐term effects on eczema severity and parental anxiety about corticosteroid usage.

Differential effects of topically applied formalin and aromatic compounds on neurogenic-mediated microvascular leakage in rat skin.

Various volatile organic compounds (VOCs) act as a causative agent of skin inflammation. We investigated the effect of topical application of several VOCs and formalin on microvascular leakage in rat skin. We tested capsaicin, which is a reagent that specifically causes the skin response via endogenously released tachykinins. Evans blue dye extravasation served as an index of the increase in skin vascular permeability. After shaving the abdomen, we applied formalin, m-xylene, toluene, styrene, benzene, ethylbenzene, acetone, diethyl ether, hexane, heptane, cyclohexane and capsaicin to the skin. At 40min after application, skin samples were collected. Among all of the VOCs tested, all of the aromatic compounds significantly produced skin microvascular leakage that was similar to formalin and capsaicin. We also investigated the skin responses seen after the intravenous administration of CP-99,994 (1.5 or 5mg/kg), which is a tachykinin NK1 receptor antagonist, ketotifen (1 or 3mg/kg), which is a histamine H1 receptor antagonist that stabilizes the mast cells, and the topical application of capsazepine (22.5 or 50mM), which is the transient receptor potential vanilloid 1 (TRPV1) antagonist. The response induced by formalin and capsaicin was completely inhibited by CP-99,994. On the other hand, the antagonist partially reduced the response induced by m-xylene, toluene and styrene by 39%, 50% and 46%, respectively. Capsazepine and ketotifen did not alter the response induced by formalin or any of the aromatic compounds. Like capsaicin, formalin and the aromatic compounds at least partially caused skin microvascular leakage, which was due to tachykinin NK1 receptor activation related to the release of tachykinins from the sensory nerve endings. However, it is unlikely that mast cells and TRPV1 play an important role in the skin response.

Measurement of specific IgE antibodies to Ses i 1 improves the diagnosis of sesame allergy

The number of reported cases of allergic reactions to sesame seeds (Sesamum indicum) has increased significantly. The specific IgE tests and skin prick tests presently available for diagnosis of sesame allergy are all based on crude sesame extract and are limited by their low clinical specificity. Thus, oral food challenge (OFC) is still the gold standard in the diagnosis.

The predictive relationship between peanut- and Ara h 2–specific serum IgE concentrations and peanut allergy

The predictive relationship between peanut- and Ara h 2-specific serum IgE concentrations and peanut allergy