PURPOSE: Japanese hop (Humulus spp.) and mugwort (Artemisia spp.) are notable causes of autumn pollinosis in East Asia. However, Japanese hop and mugwort pollen extracts, which are widely used for the diagnosis, have not been standardized. This study was performed to standardize Japanese hop and mugwort pollen extracts. MATERIALS AND METHODS: Allergen extracts were prepared in a standardized way using locally collected Humulus japonicus and purchased Artemisia vulgaris pollens. The immunoglobulin E (IgE) reactivities of prepared extracts were compared with commercial extracts via IgE immunoblotting and inhibition analyses. Intradermal skin tests were performed to determine the bioequivalent allergy unit (BAU). RESULTS: The IgE reactive components of the extracts via IgE immunoblotting were similar to those of commercial extracts. A 11-kDa allergen showed the strongest IgE reactivity in Japanese hop, as did a 28-kDa allergen in mugwort pollen extracts. Allergenic potencies of the investigatory Japanese hop and mugwort extracts were essentially indistinguishable from the commercial ones. Sums of erythema of 50 mm by the intradermal skin test (SigmaED50) were calculated to be 14.4th and 13.6th three-fold dilutions for Japanese hop and mugwort extracts, respectively. Therefore, the allergenic activity of the prepared extracts was 90827.4 BAU/mg for Japanese hop and 34412 BAU/mg for mugwort. CONCLUSION: We produced Japanese hop and mugwort pollen extracts using a standardized method. Standardized Japanese hop and mugwort pollen extracts will facilitate the production of improved diagnostic and immunotherapeutic reagents.
Allergens/*analysis/*immunology ; Antibody Specificity ; *Artemisia ; Bronchial Hyperreactivity/blood/immunology ; Cross Reactions ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoblotting ; Immunoglobulin E/blood/*immunology ; Pollen/*chemistry/*immunology ; Reference Standards ; Republic of Korea ; Rhinitis, Allergic, Seasonal

Allergen-specific immunotherapy (SIT) reduces allergen specific IgE (sIgE) levels and achieves clinical and immunological tolerance by modulating innate and adaptive immunological responses. Increased temperature and CO2 concentrations caused by climate changes contribute to an increase of pollen count and allergenicity that influences clinical SIT outcomes. In this study, we investigated the changes of IgE binding components to tree and weed pollens in pollinosis patients who showed a paradoxical increase of serum sIgE level during pollen-SIT. We enrolled nine patients who showed an increasing pattern of serum sIgE level to alder, birch, ragweed and mugwort pollens by enzyme-linked immunosorbant assay. IgE immunoblot analysis confirmed the intensification or new generation of major IgE binding components that could be induced by climate change. The findings suggest that the regular monitoring of sIgE levels and symptom changes is required to improve the clinical outcomes of SIT in patients undergoing SIT for tree and weed pollens.
Adult ; Climate Change ; *Desensitization, Immunologic ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunoglobulin E/*blood ; Male ; Middle Aged ; Pollen/immunology ; Rhinitis, Allergic, Seasonal/*therapy ; Skin Tests ; Young Adult

Exhaled nitric oxide (eNO) has been proposed as a noninvasive marker of airway inflammation in asthma. In asthmatic patients, exhaled NO levels have been shown to relate with other markers of eosinophilic recruitment, which are detected in blood, sputum, bronchoalveolar lavage fluid and bronchial biopsy samples. The purpose of this study was to assess the possible relationship between eNO and allergic inflammation or sensitization in childhood asthma and allergic rhinitis. Subjects consisted of 118 asthmatic children, 79 patients with allergic rhinitis, and 74 controls. Their age ranged from 6 to 15 yr old. eNO level, peripheral blood eosinophil count, eosinophil cationic protein (ECP), serum total IgE level and specific IgE levels were measured. Methacholine challenge test and allergic skin prick test for common allergens were performed in all subjects. Atopic group (n = 206, 44.48 +/- 30.45 ppb) had higher eNO values than non-atopic group (n = 65, 20.54 +/- 16.57 ppb, P < 0.001). eNO level was significantly higher in patients with asthma (42.84 +/- 31.92 ppb) and in those with allergic rhinitis (43.59 +/- 29.84 ppb) than in healthy controls (27.01 +/- 21.34 ppb, P < 0.001) but there was no difference between asthma and allergic rhinitis group. eNO also had significant positive correlations with Dermatophagoides pteronyssinus IgE level (r = 0.348, P < 0.001), Dermatophagoides farinae IgE level (r = 0.376, P < 0.001), and the number of positive allergens in skin prick test (r = 0.329, P = 0.001). eNO had significant positive correlations with peripheral blood eosinophil count (r = 0.356, P < 0.001), serum total IgE level (r = 0.221, P < 0.001), and ECP (r = 0.436, P < 0.001). This study reveals that eNO level is associated with allergic inflammation and the degree of allergic sensitization.
Adolescent ; Allergens/immunology ; Animals ; Asthma/*immunology ; *Breath Tests ; Bronchial Provocation Tests ; Child ; Dermatophagoides pteronyssinus/immunology ; Eosinophil Cationic Protein/analysis/blood/immunology ; Eosinophils ; Exhalation ; Female ; Humans ; Hypersensitivity, Immediate/*immunology ; Immunoglobulin E/blood ; Leukocyte Count ; Male ; Nitric Oxide/*analysis ; Rhinitis, Allergic, Seasonal/*immunology

BACKGROUNDObjective evaluation of allergic rhinitis (AR) requires in vivo and in vitro tests. In vitro tests are important to assist or ensure the main allergens in multi-allergen-sensitive patients. The aim of this study was to evaluate the utility of serum specific IgE (sIgE) in the diagnosis of AR patients with multi-allergens in the Chinese population.

METHODSCombining a positive skin prick test (SPT) and clinical history as the diagnostic reference criteria of AR, we estimated concentrations of sIgE produced in response to the 7 most frequent allergens among 85 AR patients, using the UniCAP assay system.

RESULTSAmong 85 individuals with positive SPT results and allergen-specific nasal symptoms, sIgE concentration correlated well with SPT classes among all the tested allergens. Based on a clinical diagnosis and SPT results using a positive cut-off value of a class 1 score, the CAP test performed well and the sensitivity for different allergens ranged from 0.5 (giant ragweed) to 0.91 (Dermatophagoides pteronyssinus (Der p) and Dermatophagoides farinae (Der f), while specificity ranged from 0.93 (Der f) to 1.0 (animal hair, Der p and mugwort). When the cut-off score was adjusted to class 2, the sensitivity showed an increase overall while the remaining assessed items, including specificity, positive predictive value, negative predictive value and efficiency, showed an unacceptable decline.

CONCLUSIONSWell-established serum sIgE tests correlated well with SPTs. Setting a class 1 cut-off for positivity of SPT results was better than a class 2 setting for assessing the AR diagnostic value.

Adolescent ; Adult ; Allergens ; immunology ; Animals ; Child ; Female ; Humans ; Immunoglobulin E ; blood ; Male ; Middle Aged ; Rhinitis, Allergic, Perennial ; diagnosis ; Rhinitis, Allergic, Seasonal ; diagnosis ; Skin Tests

OBJECTIVE: To evaluate the diagnostic performance of the AllergyScreen system from Mediwiss An alytic GmbH and the Pharmacia CAP system from Pharmacia Diagnostics for the detection of four inhalant allergens in the diagnosis of allergic rhinitis. METHOD: All 35 serum samples were collected from patients who were referred to the allergist for a suspected allergic rhinitis between January to March in 2007. Patients were classified as study diagnosis-positive for inhalant allergy if they both had a positive clinical examination/history and a related positive skin test for the suspected inhalant allergen. RESULT: Comparing with the reference standard, the diagnostic indexes (sensitivity, specificity, accuracy, predictive value of positive, and predictive value of negative) of the CAP system and the AllergyScreen system were 0.96 vs 0.89, 0.84 vs 0.75, 0.89 vs 0.80, 0.78 vs 0.65 and 0.93 vs 0.93 respectively. The CAP system method had higher sensitivity, specificity and accuracy than AllergyScreen system method, but there have no statistical difference between two systems (P>0.05). CONCLUSION This data support the use of ImmunoCAP system and AllergyScreen system to identify potentially significant individual allergens in the diagnosis of allergic rhinitis. The diagnostic indexes between the two systems have no statistical difference. As a simple, rapid turnaround time and low-cost system, AllergyScreen system can test multi-allergens in one time, so it can be used as a complementary with the ImmunoCAP system.
Adolescent ; Adult ; Allergens ; blood ; immunology ; Child ; Female ; Humans ; Immunoglobulin E ; blood ; immunology ; Male ; Middle Aged ; Predictive Value of Tests ; Recoverin ; Rhinitis, Allergic, Seasonal ; blood ; diagnosis ; immunology ; Serologic Tests ; methods ; Skin Tests ; Young Adult

OBJECTIVE: To study the role of serum IL-10, 12, 13, 16 in patients with allergic rhinitis and vasomotor rhinitis. METHOD: The serum levels of IL-10, 12, 13, 16 were measured by ELISA in 30 cases of allergic rhinitis, 25 cases of vasomotor rhinitis and 20 healthy people. RESULT: The level of IL-12 in allergic rhinitis was (170.33 +/- 90.58) ng/L, which was significantly lower than that of normal controls [(376.69 +/- 140.70) ng/L, P < 0.01]. The levels of IL-13 and IL-16 in allergic rhinitis were (408.51 +/- 189.68) ng/L and (151.53 +/- 63.56) ng/L, which were significantly higher than those of normal controls [(151.92 +/- 85.08) ng/L, (60.65 +/- 32.45) ng/L, P < 0.01]. There were no significant difference of levels of IL-10, 13, 16 between vasomotor rhinitis and normal controls, while the level of IL-12 in vasomotor rhinitis was lower than that of normal controls [(196.03 +/- 96.31) ng/L vs. (376.69 +/- 140.70) ng/L, P < 0.01]. It was suggested that IL-10 had positive correlation with IL-12 (r = 0.73, P < 0.01), and IL-13 had positive correlation with IL-16 (r = 0.94, P < 0.01). CONCLUSION The imbalance of IL-12, IL-13 and IL-16 play crucial roles of regulation in the onset and developing of allergic rhinitis. Further research is needed on the role of IL-12 in vasomotor rhinitis.
Adolescent ; Adult ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Humans ; Interleukin-10 ; blood ; Interleukin-12 ; blood ; Interleukin-13 ; blood ; Interleukin-16 ; blood ; Male ; Middle Aged ; Rhinitis, Allergic, Perennial ; blood ; Rhinitis, Allergic, Seasonal ; blood ; Rhinitis, Vasomotor ; blood ; Young Adult

IL-28RA is one of the important candidate genes for complex trait of genetic diseases, but there is no published information of the genetic variation in this gene. We scanned the seven exons and their boundary introns sequence of IL-28RA including the promoter regions to analyze genetic variation sites, and identified eighteen single nucleotide polymorphisms (SNPs) and two variation sites. We chose seven SNPs (g.-1193 A>C, g.-30 C>T, g.17654 C>T, g.27798 A>G, g.31265 C>T, g.31911 C>T and g.32349 G>A) of them for large sample size genotyping, and assessed the association of genotype and allele frequencies of these SNPs between allergic rhinitis patients and non-allergic rhinitis controls. We also compared the genotype frequencies between Korean controls and Han Chinese control or Korean Chinese control. We investigated the frequencies of haplotype constructed by these SNPs between allergic rhinitis patients and non-allergic rhinitis controls. Our results suggested that the g.32349 G>A polymorphism of IL-28RA might be associated with susceptibility to allergic rhinitis (P=0.032), but seems to have no relationship with serum total IgE levels. The haplotype frequencies by these SNPs also show significant association between controls and allergic rhinitis patients.
Variation (Genetics) ; Rhinitis, Allergic, Seasonal/blood/*genetics ; Rhinitis, Allergic, Perennial/blood/*genetics ; Receptors, Cytokine/*genetics ; Promoter Regions (Genetics)/genetics ; Polymorphism, Single Nucleotide/*genetics ; Male ; Immunoglobulin E/blood ; Humans ; Haplotypes ; Genotype ; Genetic Predisposition to Disease/genetics ; Gene Frequency ; Female ; Exons/genetics ; Case-Control Studies ; Alleles ; Adult

BACKGROUNDCurrently anti-inflammatory therapy with steroids for allergic rhinitis need long-term repeated administration, although it is effective. Gene therapy is being suggested to substitute it. The aim of this study was to investigate nonviral vector mediated exogenous gene expression in COS-7 cells in vitro and the effect of intranasal mouse interleukin (mIL)-12 transgene expression on allergen induced eosinophil infiltration of nasal mucosa in a murine model of allergic rhinitis.

METHODSIn vitro COS-7 cells were infected with Epstein-Barr virus (EBV)/lipoplex. The expression of IL-12 p70 in cell culture supernatant was examined by enzyme-linked immunosorbent assay (ELISA). In mice with ovalbumin (OVA) induced allergic rhinitis, EBV/lipoplex was administered by nasal drops before OVA challenge once a day from day 1 to day 10. The expression of IL-12 mRNA and protein, the change of eosinophil count in nasal mucosa and serum total IgE were measured 24 hours after the last challenge.

RESULTSEBV/lipoplex could effectively transfect COS-7 cells. The expression of IL-12 p70 in cell culture supernatant was significantly more than in blank control. IL-12 via EBV plasmid vector transduction could be overexpressed in vivo. In pGEG.mIL-12 treated models, the nasal mucosa revealed a high level of widespread mIL-12 transduction by immunohistochemistry and in situ hybridization. Histological evaluation revealed marked suppression of eosinophil infiltration in nasal mucosa. The eosinophil count in allergic rhinitis group [(26.5 +/- 9.8)/high-power field (HPF)] was significantly increased over control group [(0.40 +/- 0.52)/HPF] (F = 56.94, P < 0.01), while the count in IL-12 gene therapy group [(4.60 +/- 2.63)/HPF] was significantly less than that of allergic group (F = 56.9, P < 0.01). Serum total IgE between in gene therapy mice [(88.83 +/- 6.71) ng/ml] and allergic rhinitis mice [(103.1 +/- 5.7) ng/ml] showed a significant difference (F = 1216, P < 0.05).

CONCLUSIONSNonviral EBV plasmid vector, pGEG.mIL-12 was able to overexpress exogenous gene both in vitro and in murine nasal mucosa in vivo. IL-12 overexpression via EBV/lipoplex could stem allergen induced eosinophil infiltration in nasal mucosa in murine models of allergic rhinitis, which may suggest a new cytokine immunogenetic therapy for allergic rhinitis.

Administration, Intranasal ; Animals ; COS Cells ; Cercopithecus aethiops ; Eosinophilia ; therapy ; Genetic Therapy ; Herpesvirus 4, Human ; genetics ; Immunoglobulin E ; blood ; Interleukin-12 ; genetics ; Lipids ; administration & dosage ; Male ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; metabolism ; Rhinitis, Allergic, Perennial ; therapy ; Rhinitis, Allergic, Seasonal ; therapy

OBJECTIVETo investigate the expression of T-bet mRNA in peripheral blood mononuclear cells (PBMC) as well as its relations with total IgE (TIgE), eosinophil cationic protein (ECP) in serum and nasal allergic symptoms in patients with allergic rhinitis (AR).

METHODSThe allergen, TIgE and ECP in serum of patients with AR were detected by Unicap CAP system. Blood sample was taken from 8 healthy individuals and 22 patients with allergic rhinitis. PBMC was isolated by density gradient centrifugation and one part of them was cultured with 50 microg/ml mite allergen. PBMC was subjected to analysis of T-bet mRNA expression using semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTSThe ratio of T-bet to beta-actin mRNA levels was 0.381 +/- 0.099 in patients and 0.750 +/- 0.067 in normal individuals, the difference was significantly (P <0.01). The expression intensity of T-bet mRNA had no relation to varying severity of allergic symptoms and concentration of ECP and the correlation coefficient was 0.187 and -0.165 (all P > 0.05). However, there was an inverse correlation between expression intensity of T-bet mRNA and TIgE concentration (r = -0.525, P < 0.05). Mean mRNA level (x +/- s) of T-bet expression before and after being stimulated by allergen was 0.381 +/- 0.099 and 0.365 +/- 0.104 respectively, which indicated no significant differences (P > 0.05).

CONCLUSIONSAmong allergic patients whose allergen was mite, there was a down-regulated expression of T-bet mRNA, which had no relation to ECP concentration and allergic symptoms, but was one of important links in mechanisms of imbalance of Th1/Th2 in AR. There was no effect of specific allergen on T-bet mRNA in patients with AR T-bet was one of indirect factors that affected the level of IgE.

Adolescent ; Adult ; Case-Control Studies ; Child ; Child, Preschool ; Eosinophil Cationic Protein ; blood ; Female ; Humans ; Immunoglobulin E ; blood ; Male ; Rhinitis, Allergic, Seasonal ; blood ; T-Box Domain Proteins ; blood ; Young Adult

BACKGROUNDAtopy is a state of allergy to common antigens and is founded on an immune disturbance of exuberant Th2 activity and IgE production. There is also epidemiological and experimental evidence that exposure to mycobacteria has the potential to suppress the development of asthma or atopy. Since Th1 and Th2 immune mechanisms are significantly antagonistic, it is hypothesized that mycobacterial exposure may moderate atopic disease by modification of immune responses.

METHODSOne hundred and twenty mild to moderate persistent asthmatics accompanied with allergic rhinitis were randomly divided into four groups with one injection every other day for 18 times for group A with 1 ml of normal saline, B with 0.5 mg of Bacillus Calmette-Guérin polysaccharide nucleotide (BCG-PSN) and C with 1 mg of BCG-PSN, 36 times for group D with 0.5 mg of BCG-PSN. Markers for the severity of asthma and rhinitis including the amount of inhaled corticosteriod, bronchodilator and oral H1 blocker-loratidine being used to obtain optimal symptomatic control, symptom scores of asthma and allergic rhinitis, peak expiratory flow (PEF), histamine provocative dose that produces at least a 20% change in forced expiratory volume with in 1 second (PD20-FEV1), blood IgE levels as well as dermatophagoides pteronysinus (DP) and dermatophagoides farinae (DF) skin prick test were measured every visit for 6 months.

RESULTSThere were no differences for symptom scores of asthma, daily use of bronchodilator, PEF, PD20-FEV1, blood IgE as well as DF and DP skin prick test among the four groups. Score for allergic rhinitis decreased significantly in groups B, C and D on day 36 and 72 as compared with group A (P < 0.05). Score for allergic rhinitis increased after day 72 in group B and C while it was significantly lower in group D (P < 0.05). The patients in group D used less amount of inhaled beclomethosone than other groups (P < 0.05) from day 72 after the treatment to day 180. Oral loratadine consumption in groups B, C and D was significantly less on day 36 and 72 as compared with their baseline and group A after the treatment (P < 0.05). Group D maintained significantly lower dosage of oral loratadine until day 150 comparing with its baseline and group A.

CONCLUSIONSBCG-PSN has a symptomatic effect on allergic rhinitis. BCG-PSN may reduce the dosage of non-sedative H1 blocker loratadine as well as the dosage of inhaled beclomethosone in the treatment of mild to moderate asthma and allergic rhinitis.

Adult ; Asthma ; immunology ; therapy ; BCG Vaccine ; administration & dosage ; Double-Blind Method ; Female ; Humans ; Immunoglobulin E ; blood ; Injections, Intramuscular ; Interferon-gamma ; biosynthesis ; Male ; Middle Aged ; Nucleotides ; administration & dosage ; Polysaccharides, Bacterial ; administration & dosage ; Rhinitis, Allergic, Perennial ; immunology ; therapy ; Rhinitis, Allergic, Seasonal ; immunology ; therapy ; Skin Tests