Dermatophagoides farinae induces conjunctival epithelial cell damage to promote neutrophil migration and neutrophil extracellular traps formation / 中国血吸虫病防治杂志

OBJECTIVE@#To investigate the mechanisms underlying allergic conjunctivitis caused by conjunctival epithelial cell damage, neutrophil migration and neutrophil extracellular traps (NETs) formation induced by crude extracts of Dermatophagoides farinae mite (CDM).@*METHODS@#Human conjunctival epithelial cells were stimulated with 500, 1 000, 2 000, 4 000 ng/mL, and the expression levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and IL-8 were detected using quantitative real-time PCR (qPCR) assay and enzyme-linked immunosorbent assay (ELISA). The culture supernatant of human conjunctival epithelial cells was collected and co-cultured with neutrophils. Neutrophil migration was measured using Transwell migration assay, and the expression of NETs markers myeloperoxidase (MPO) and citrullinated histone H3 (CitH3) was quantified using immunofluorescence staining. Neutrophils were stimulated with phorbol 12-myristate 13-acetate (PMA), and then NETs were collected for treatment of human conjunctival epithelial cells. Cell apoptosis was detected using flow cytometry, and the levels of IL-6, TNF-α, IFN-γ and IL-8 were measured in the cell culture supernatant using ELISA.@*RESULTS@#Treatment with CDM at concentrations of 2 000 ng/mL and 4 000 ng/mL up-regulated IL-6, TNF-α, IFN-γ and IL-8 expression in human conjunctival epithelial cells. Following treatment with CDM at concentrations of 2 000 ng/mL and 4 000 ng/mL, the culture supernatant of human conjunctival epithelial cells promoted neutrophil migration and induced increases in the staining intensity of MPO and CitH3. In addition, increased NETs triggered the apoptosis of human conjunctival epithelial cells and IL-6, TNF-α, IFN-γ and IL-8 secretion in the culture supernatant of human conjunctival epithelial cells.@*CONCLUSIONS@#CDM induces human conjunctival epithelial cell damages, thereby promoting neutrophil migration and NETs formation, while the release of NETs further aggravates human conjunctival epithelial cell damages.

Evaluation of the action of precipitating agents to obtain allergenic proteins from crude mite extracts / Avaliação da ação de agentes precipitantes na obtenção de proteínas alergênicas presentes em extratos brutos de ácaros

Introduction: Allergen proteins found in dust mite extracts, such as Dermatophagoides farinae (DF), Dermatophagoides pteronyssinus (DP) and Tyrophagus putrescentiae (TP), are relevant for scientific studies in the allergy and immunotherapy fields. The precipitation/concentration of protein extracts may favor the aggregation of the allergens in homogenates. Objective and method: This paper investigates the precipitation process by submitting crude mite extracts to compounds such as ammonium sulfate (NH4)2SO4, trichloroacetic acid (TCA) and acetone. Results: The best results were obtained by fractionation with (NH4)2SO4 at 80% (w/v) saturation (~0° C), observing the protein markings on the electrophoresis gel. Major allergens were identified by immunoblot at 25 kDa (cysteine protease) for Der f and Der p; and 25 kDa, 30 kDa (tropomyosin) and Try p 3, near 26 kDa. For this percentage the total protein contents were 12.83 mg mL-1 for Der f, 24.78 mg mL-1 for Der p and 27.35 mg mL-1 for Try. Conclusion: An advantage of precipitation with (NH4)2SO4 over precipitation with acetone was the possibility of gradually obtaining protein fractions, which does not happen when using the latter. The addition of 80% (v/v) acetone to the mite extracts favored total protein precipitation in the concentrations 16.42 mg mL-1, 28.47 mg mL-1 and 13.41 mg mL-1. The use of TCA in concentrations above 20% (w/v) forms peptides that are not retained in the gel under the established experimental conditions, and dilute solutions of this acid are more efficient.

Introdução: As proteínas alergênicas presentes nos extratos dos ácaros de poeira, tais como Dermatofagoides farinae (DF), Dermatofagoides pteronyssinus (DP) e Tyrophagus putrescentiae (TP) são relevantes para estudos científicos na área de alergias e aplicação em imunoterapias. A precipitação/concentração desses extratos proteicos pode favorecer a agregação de alérgenos nos homogenatos. Objetivo e método: O trabalho investiga o processo de precipitação, submetendo os extratos brutos de ácaros de poeira a compostos como sulfato de amônio (NH4)2SO4, ácido tricloroacético (ATC) e acetona. Resultados: Os melhores resultados foram obtidos por fracionamento com (NH4)2SO4 em 80% (m/v) de saturação (~ 0°C), observando as marcações proteicas no gel de eletroforese. Os alérgenos principais foram identificados por immunoblot em 25 kDa (cisteína protease) para Der f 1 e Der p 1; e 25 kDa, 33 kDa (tropomyosin), 11 kDa para Tyr. Para esse percentual, os teores de proteína total foram de 12.83 mg mL-1 para DF; 24,78 mg mL-1 para DP; e 27,35 mg mL-1 para TP. Conclusão: A vantagem da precipitação com (NH4)2SO4 frente à precipitação com acetona foi a possibilidade de gradativamente se obter frações proteicas, o que não acontece quando utilizado esse solvente. A adição de 80% (v/v) de acetona aos extratos de ácaros favoreceu a precipitação total de proteína nas concentrações 16,42 mg mL-1; 28,47 mg mL-1; e 13,41 mg mL-1. O uso de ATC em concentrações acima de 20% (m/v) forma peptídeos que não são retidos no gel nas condições experimentais estabelecidas, sendo eficiente soluções mais diluídas desse ácido.

Validation of PROTIA™ Allergy-Q 64 Atopy® as a Specific IgE Measurement Assay for 10 Major Allergen Components

PURPOSE: Component-resolved diagnostics (CRD) is expected to provide additional diagnostic information in allergic patients. PROTIA™ Allergy-Q 64 Atopy®, a recently developed CRD-based multiplex specific immunoglobulin E (sIgE) assay, can quantitatively measure sIgE to major allergen components. METHODS: The sIgE detection by PROTIA™ Allergy-Q 64 Atopy® and ImmunoCAP® assays was compared using the sera of 125 Korean allergic patients. Group 1 and 2 allergens of house dust mites (HDMs; Dermatophagoides farinae (Der f) 1 and Der f 2 in PROTIA™ Allergy-Q 64 Atopy®, Dermatophagoides pteronyssinus (Der p) 1 and Der p 2 in ImmunoCAP®), Bet v 1, Fel d 1, Que a 1, ω-5 gliadin, α-lactalbumin, β-lactoglobulin, casein and α-Gal were measured by both assays. RESULTS: Comparing the results from the 2 assays, the agreement rate for all the 10 allergens was > 88% (group 1 HDM allergen, 100%; group 2 HDM allergen, 94.6%; Bet v 1, 97.4%; Fel d 1, 90.5%; Que a 1, 89.2%; α-lactalbumin, 96%; β-lactoglobulin, 88%; casein, 88%; ω-5 gliadin, 96%; α-Gal, 100%). Correlation analysis indicated that, all the 10 allergen sIgEs showed more than moderate positive correlation (Pearson correlation coefficients > 0.640). Additionally, intra-class comparison showed more than high correlation for all the 10 allergens (Spearman's rank correlation coefficients > 0.743). CONCLUSIONS: PROTIA™ Allergy-Q 64 Atopy® is reliable and comparable to the ImmunoCAP® assay for component-resolved diagnosis.

Evaluation of multiple allergen simultaneous (sIgE) testing compared to intradermal testing in the etiological diagnosis of atopic dermatitis in horses

Although intradermal testing (IDT) is commonly used in the etiological diagnosis of allergies, in vitro testing for specific IgE (sIgE) is an attractive alternative. Currently, new laboratory techniques in veterinary allergological practice, including multiple allergen simultaneous tests (MASTs), gradually supersede in vivo tests. Both, serological (sIgE) and IDTs in fourteen atopic Malopolski horses were performed. Correlation and agreement between test results were evaluated. Receiver operating characteristic analysis showed that sIgE to Acarus siro had the best diagnostic performance (Area under the ROC curve [AUC] = 0.969), followed by Dermatophagoides pteronyssinus (AUC = 0.844), Dermatophagoides farinae (AUC = 0.813) and Tyrophagus putrescentiae (AUC = 0.803). A significant positive correlation between IDT and MAST was found for A. siro (r(S) = 0.870; p = 0.00005), and D. farinae (r(S) = 0.657; p = 0.011). There was significant moderate agreement for 2 of 5 allergens, A. siro (κ = 0.569) and D. farinae (κ = 0.485) in semiquantitative assessment and significant fair to substantial agreement for 3 of 5 allergens, D. pteronyssinus (κ = 0.689), A. siro (κ = 0.569), D. farinae (κ = 0.432) in dichotomic assessment. Sensitivity ranged from 44% to 89%, depending on the allergen, while specificity was significantly higher for all allergens in MAST (60%–100%); the mean accuracy was 73% (manufacturer cut-off) and 77.4% (optimal cut-off) based on the Youden index. Compared with IDT, serological MAST showed good detection performance for 60% allergen sIgE in dichotomic assessment with substantial diagnostic capability, but careful clinical interpretation is needed for some allergens.

The Characteristics of Staphylococcal Scalded Skin Syndrome in Atopic Dermatitis / 고신대학교의과대학학술지

OBJECTIVES: Staphylococcal scalded skin syndrome (4S), a blistering dermatosis caused by exfoliative toxins from Staphylococcus aureus, occurs frequently in patients with atopic dermatitis (AD). However, association between 4S and AD has not rarely been reported. We investigated the characteristics of 4S according to AD status.METHODS: The study included 146 children with 4S who visited Busan St. Mary's Hospital from 2007–2018. Clinical features were analyzed from medical records and pictures, and 4S was classified as localized or generalized. We also retrospectively investigated the preceding conditions and test results related to AD.RESULTS: Among 146 patients with 4S, median age was 2.0 years, and 35 (24.0%) had AD. Since 2007, the incidence of both 4S and AD have increased, without obvious seasonal patterns. Generalized and localized disease occurred in 90 and 56 patients, respectively. Twenty-four of 35 patients with AD (68.6%) and 32 of 111 (28.8%) without AD had localized disease. Significant differences were observed between the groups (P = 0.000). Among those with AD, the most common preceding condition was skin infection or unknown (45.2%); however, respiratory disease was the most common (47.9%) among patients without AD. Eosinophil levels were higher in the AD group (P = 0.002), and there were no statistically significant differences in total immunoglobulin E (IgE), Dermatophagoides farinae (Df IgE), egg-white IgE, and culture results between the groups.CONCLUSIONS: Localized 4S frequently occurred without preceding conditions in children with AD and usually arose from skin infection compared to generalized 4S.

Immunological analysis of allergenic cross-reactivity between Cheyletus malaccensis and Dermatophagoides farinae, Dermatophagoides pteronyssinus and Blomia tropicalis / Análise imunológica da reatividade cruzada alergênica entre Cheyletus malaccensis e Dermatophagoides farinae, Dermatophagoides pteronyssinus e Blomia tropicalis

Objective: The mite Cheyletus malaccensis is cited in the literature as a predator of other mite species. Little is known about its protein composition, and few studies have evaluated its ability to trigger atopic respiratory allergic reactions. The present study aims to investigate the protein profile fingerprint present in Cheyletus malaccensis extract and to evaluate its immunologic reactivity in the presence of specific immunoglobulins (IgE) from the serum of individuals diagnosed with allergy to the mites Dermatophagoides farinae, Dermatophagoides pteronyssinus and Blomia tropicalis. These three species carry proteins responsible for the most cases of atopic respiratory allergies, hence the interest in comparing them to Cheyletus malaccensis. Methods: Samples of aspirated dust containing Cheyletus malaccensis were collected from households in the city of Rio de Janeiro, Brazil. From the collected mass of this mite, extracts were prepared for analysis. Proteins present in the extracts were identified by electrophoresis under denaturing conditions. Results: Proteins with a molecular mass of 24 kDa, 26 kDa, 12 kDa, 45 kDa and 70 kDa were visualized. The immunoblotting assay showed positive cross-reactivity for proteins of molecular mass ranging from 20 kDa to 45 kDa. These results indicate that specific links were established between IgE present in the serum of individuals allergic to the comparator mite and proteins from Cheyletus malaccensis. Conclusions: These findings are relevant for their potential clinical and immunotherapeutic applications, as well as information base for further studies.

Objetivo: O ácaro Cheyletus malaccensis é referido na literatura como um predador de outras espécies de ácaro. Pouco se sabe sobre sua composição proteica, e poucos estudos avaliaram sua habilidade de desencadear reações alérgicas respiratórias atópicas. O objetivo do presente estudo é investigar a impressão digital do perfil proteico presente em um extrato de Cheyletus malaccensis e avaliar sua reatividade imunológica na presença de imunoglobulinas (IgE) específicas do soro de indivíduos diagnosticados com alergia aos ácaros Dermatophagoides farinae, Dermatophagoides pteronyssinus e Blomia tropicalis. Essas três espécies carregam proteínas responsáveis pela maioria dos casos de alergias respiratórias atópicas, o que justifica o interesse em compará-las ao Cheyletus malaccensis. Métodos: Amostras de poeira aspirada contendo Cheyletus malaccensis foram coletadas de domicílios na cidade do Rio de Janeiro, no Brasil. A partir da massa coletada desse ácaro, extratos foram preparados para análise. As proteínas presentes nos extratos foram identificadas por eletroforese sob condições desnaturantes. Resultados: Proteínas com massa molecular de 24 kDa, 26 kDa, 12 kDa, 45 kDa e 70 kDa foram visualizadas. O ensaio imunoenzimático mostrou reatividade cruzada positiva para proteínas de massa molecular variando de 20 kDa a 45 kDa. Esses resultados indicam que ligações específicas foram estabelecidas entre a IgE presente no soro de indivíduos alérgicos ao ácaro usado como comparador e proteínas de Cheyletus malaccensis. Conclusões: Os achados são relevantes por seu potencial clínico e aplicações imunoterapêuticas, bem como sua base de informações para futuros estudos.

Skin prick test reactivity to aeroallergens in adult allergy clinic in Thailand: a 12-year retrospective study

BACKGROUND: The global prevalence of allergic rhinitis, asthma, and atopic dermatitis has risen significantly over the last 2 decades. Allergic sensitization to aeroallergen is a major risk factor in developing the allergic disease. The prevalence of aeroallergen sensitization varies in different regions and countries. OBJECTIVE: To determine the prevalence of common aeroallergen sensitization and the atopic status among adult patients. METHODS: A cross-sectional, retrospective study. The data were collected from medical records and database of the result of skin prick test of patients who had the allergic symptoms or chronic urticaria in adult allergy clinic, Ramathibodi hospital from January 2004 to December 2015. RESULTS: A total of 1,516 of patients (female, 1,118 [73.7%]) were enrolled. The mean ages of participants were 41.34 (standard deviation, ±16.5) years. Fifty-eight percent (58%) of patients were diagnosed with allergic rhinitis, 19.7%, 3.2%, and 9.2% with asthma, atopic dermatitis, and chronic urticaria respectively. In the chronic urticaria group, 57.4% underwent the positive skin prick test to common aeroallergens. Mites were responsible for the most common inhaled allergen sensitization in this study as 50.1% of Dermatophagoides pteronyssinus, 32% of Dermatophagoides farinae, and 31.5% of house dust. Cockroach was the second most common aeroallergen sensitization as 32.3% followed by grass pollen, Bermuda (21.1%) and timothy (13.6%). The animal dander, cat and dog, occupied 12.9 and 10% respectively. CONCLUSION: Mites were the most common cause of aeroallergen sensitization in all patients followed by cockroach, grass pollen, and animal dander. However, Bermuda sensitization has increased significantly in the last 6 years.

Aeroallergen sensitization and associated comorbid diseases of an adult Filipino population with allergic rhinitis

BACKGROUND: There is a large global variation in sensitization patterns to aeroallergens due to differences in climate, urbanization, and lifestyle. Knowledge of the most common inhalant allergens is important for appropriate prevention and management of allergic rhinitis (AR). OBJECTIVE: This study aims to provide data on aeroallergen sensitization patterns and associated comorbid diseases of adult Filipinos with AR. METHODS: Medical records of adult Filipinos seen in an Otolaryngology-Allergy Clinic from January 2011 to 2016 were reviewed. Inclusion criteria used was presence of clinically defined AR and positive skin test to at least one aeroallergen in the test panel. Demographics, comorbid conditions, and results of skin prick test were determined. Standard descriptive statistics were used for analysis. RESULTS: One hundred ninety-one adult patients were included in this study. Mean age was 38.8 years, and majority lived in an urban area (71.2%). Most patients exhibited polysensitization (97.4%). All exhibited sensitization to indoor and 86.9% to outdoor allergens. The most common indoor allergens were Dermatophagoides pteronyssinus (97.4%), Dermatophagoides farinae, (95.8%), cockroach (80.1%), and molds (72.8%). Bermuda (67%), Johnson grass (58.7%), and Acacia (58.2%) were the most common outdoor allergens. Urticaria (18.8%), dermatitis (16.8%), and asthma (11.5%) were the most common associated comorbid disease. Twelve percent of patients had more than one associated comorbid disease. Asthma + urticaria followed by asthma + dermatitis were the most common co-morbid combinations. One patient had three comorbid diseases: asthma + urticaria + rhinosinusitis. CONCLUSION: Compared to earlier studies, aeroallergen sensitization patterns of Filipinos remain unchanged. This study also identifies for the first time, the associated comorbid diseases of AR in this population. Understanding these factors can guide treatment strategies to reduce disease burden.

Efficacy and Safety of Sublingual Immunotherapy in Elderly Rhinitis Patients Sensitized to House Dust Mites

PURPOSE: This study aims to determine the efficacy and safety of house dust mite (HDM)-sublingual immunotherapy (SLIT) in elderly patients with AR. METHODS: A total of 45 patients aged ≥ 60 years with HDM-induced AR who had ≥ 3 A/H ratio on skin prick test and/or ≥ 0.35 IU/L to both Dermatophagoides farinae and Dermatophagoides pteronyssinus by ImmunoCAP were enrolled in 4 university hospitals. To evaluate additional effects of HDM-SLIT, they were randomized to the SLIT-treated group (n = 30) or control group (n = 15). Rhinoconjunctivitis total symptom score (RTSS), rhinoscopy score, Korean rhinoconjunctivitis quality of life questionnaire, rhinitis control assessment test, asthma control test scores, and adverse reactions, were assessed at the first visit (V1) and after 1 year of treatment (V5); for immunological evaluation, serum levels of HDM-specific immunoglobulin A/IgE/IgG1/IgG4 antibodies and basophil response to HDMs were compared between V1 and V5 in both groups. RESULTS: There were no significant differences in demographics, RTSS, skin reactivity to HDMs, or serum total/specific IgE levels to HDMs (P < 0.05, respectively) between the 2 groups. Nasal symptom score and RTSS decreased significantly at year 1 in the 2 groups (P < 0.05). There were no significant differences in percent decrease in nasal symptom score and RTSS at year 1 between the 2 groups (P < 0.05); however, rhinoscopic nasal symptom score decreased significantly in the SLIT-treated group (P < 0.05). Immunological studies showed that serum specific IgA levels (not specific IgE/IgG) and CD203c expression on basophils decreased significantly at V5 in the SLIT-treated group (P = 0.011 and P = 0.001, respectively), not in the control group. The control group required more medications compared to the treatment group, but there were no differences in adverse reactions. CONCLUSIONS: It is suggested that HDM-SLIT for 1 year could induce symptom improvement and may induce immunomodulation in elderly rhinitis patients.

Neutrophil oxidative burst as a diagnostic indicator of IgG-mediated anaphylaxis

BACKGROUND: IgG-mediated anaphylaxis occurs after infusion of certain monoclonal antibody-based therapeutics. New in vitro tests are urgently needed to diagnose such reactions. We investigated whether allergens trigger neutrophil oxidative burst (OB) and if neutrophil OB occurs due to allergen-specific IgG (sIgG). METHODS: Neutrophil OB was measured by dihydrorhodamine 123 flow cytometry using a leukocyte suspension spiked with a very small patch of the allergen crude extract, Dermatophagoides farinae (Der f). The mean fluorescence intensity ratio of stimulated to unstimulated samples was calculated as the neutrophil oxidative index (NOI). RESULTS: The Der f-specific NOI (Der f-sNOI) showed a time-dependent increase after Der f extract addition. At 15 min activation, higher Der f-sIgG levels were associated with lower Der f-sNOI values in 31 subjects (P < 0.05). This inverse relationship occurs due to the initial blocking effect of free Der f-sIgG. Additionally, neutrophil OB was nearly absent (Der f-sNOI of −1) in two cases: a subject with undetectable Der f-sIgG levels and washed leukocyte suspensions deprived of Der f-sIgG. CONCLUSION: Allergens can trigger neutrophil OB via preexisting allergen-sIgG. Neutrophil OB can be easily measured in a leukocyte suspension spiked with the allergen. This assay can be used to diagnose IgG-mediated anaphylaxis.

Relation of allergic rhinitis, allergen sensitization, and air pollutants in preschool children

PURPOSE: We aimed to investigate the prevalence and allergen sensitization of allergic rhinitis (AR) in preschool children. We assessed the relationship of AR, allergen sensitization, and air pollutants. METHODS: Between 2012 and 2016 in Jincheon, skin prick testing and questionnaire survey were performed on 2,958 children. AR have diagnosed on the basis of current symptoms and sign of AR on the Korean International Study of Asthma and Allergies in Childhood questionnaire and 1 or more proven allergen sensitization. RESULTS: A total of 2,052 children completed survey and skin prick testing, and were included in the analysis. The prevalence of AR symptom within last 12 months and AR were 27.0% and 10.9%, respectively. During study, both prevalences were increased from 20.1% and 8.6%, respectively in 2012 to 29.1% and 12.8%, respectively in 2016. The total inhalant allergen sensitization rate was 32.6%. The most common inhalant allergen was Dermatophagoides farinae (23.1%), followed by Dermatophagoides pteronyssinus (22.5%), tree pollens (5.3%), weed pollens (5.2%), fungi (4.7%), dog hair (4.5%) cat fur (3.6%), grass pollens (1.1%), and cockroach (0.8%). Although there was no difference tree or weed pollen sensitization, pollen seasonal prevalence of current AR symptoms is highest in spring (80%) versus autumn (52.3%). Seasonal PM10 (particulate matter with a median aerodynamic diameter less than or equal to 10 µm in diameter) and SO2 (sulfur dioxide) levels were correlated with the prevalence of seasonal AR symptoms. Pollen seasonal current AR symptoms were significantly related to seasonal pollen sensitization, but not inhalant allergens including dust mites. CONCLUSION: There was a rapid increasing prevalence of AR in preschool children in the areas of urbanization and industrialization. Pollen seasonal current AR symptoms have a close relationship with PM10 and seasonal pollen sensitization.

Comparison between Newly Developed and Commercial Inhalant Skin Prick Test Reagents Using In Vivo and In Vitro Methods

BACKGROUND: We developed skin prick test (SPT) reagents for common inhalant allergens that reflected the real exposure in Korea. The study aim was to evaluate diagnostic usefulness and allergen potency of our inhalant SPT reagents in comparison with commercial products. METHODS: We produced eight common inhalant allergen SPT reagents using total extract (Prolagen): Dermatophagoides farinae, Dermatophagoides pteronyssinus, oak, ragweed, mugwort, Humulus japonicus pollens, as well as cat and dog allergens. We compared the newly developed reagents with three commercially available SPT reagents (Allergopharma, Hollister-Stier, Lofarma). We measured total protein concentrations, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), major allergen concentration, and biological allergen potencies measured by immunoglobulin E (IgE) immunoblotting and ImmunoCAP inhibition test. RESULTS: Diagnostic values of these SPT reagents were expressed as positivity rate and concordance rate of the results from ImmunoCAP allergen-specific IgE test in 94 allergic patients. In vitro analysis showed marked differences in protein concentrations, SDS-PAGE features, major allergen concentrations, and biological allergen potencies of four different SPT reagents. In vivo analysis showed that positive rates and concordance rates of Prolagen® SPT reagents were similar compared to the three commercial SPT reagents. CONCLUSION: The newly developed Prolagen® inhalant SPT reagents are not inferior to the commercially available SPT reagents in allergy diagnosis.

The association between nasal eosinophilia and aeroallergen sensitization in children and adolescents with rhinitis

PURPOSE: To identify the correlation between nasal eosinophilia and aeroallergen sensitization in children and adolescents. METHODS: This is a retrospective study of patients below 18 years of age who had a history of rhinitis that lasted more than 2 weeks or had been repeated more than once a year, received nasal eosinophil examinations, and had serum specific IgE to aeroallergens measured at an Allergy Clinic in a single tertiary teaching hospital in Seoul, Korea. The percentage of nasal eosinophils was calculated by the number of eosinophils per total leukocytes in a high-power field of 1,000×. Data was analyzed to determine the association between nasal eosinophilia and 18 aeroallergens. RESULTS: Of the 245 patients included, 156 (63.7%) were male and the mean age (±standard deviation) was 7.9 years (±3.8). In total, 175 patients (71.4%) were sensitized to at least 1 of the 18 aeroallergens tested, and sensitization to house dust mite was most common. In addition, 118 (48.2%) and 69 patients (28.2%) had nasal eosinophilia of at least 1% and 5%, respectively. There were no significant correlations between serum total IgE or age and the percentage of nasal eosinophils. However, the percentage of nasal eosinophils in the group sensitized to any aeroallergens was significantly increased compared to the nonsensitized group (P=0.002). The percentage of nasal eosinophils was significantly higher in patients who were sensitized to Birch-Alder Mix, oak white, Bermuda grass, orchard grass, timothy grass, sweet vernal grass, rye, mugwort, short ragweed, Alternaria alternata, cats, dogs or Dermatophagoides farinae compared to those nonsensitized. CONCLUSION: Nasal eosinophilia was significantly associated with sensitization to aeroallergens.

Homology modeling and epitope prediction of Der f 33

Dermatophagoides farinae (Der f), one of the main species of house dust mites, produces more than 30 allergens. A recently identified allergen belonging to the alpha-tubulin protein family, Der f 33, has not been characterized in detail. In this study, we used bioinformatics tools to construct the secondary and tertiary structures and predict the B and T cell epitopes of Der f 33. First, protein attribution, protein patterns, and physicochemical properties were predicted. Then, a reasonable tertiary structure was constructed by homology modeling. In addition, six B cell epitopes (amino acid positions 34-45, 63-67, 103-108, 224-230, 308-316, and 365-377) and four T cell epitopes (positions 178-186, 241-249, 335-343, and 402-410) were predicted. These results established a theoretical basis for further studies and eventual epitope-based vaccine design against Der f 33.

A Pilot Study of Intralymphatic Immunotherapy for House Dust Mite, Cat, and Dog Allergies

Several recent clinical trials reported that intralymphatic immunotherapy (ILIT) for some allergens, such as cat dander and pollen, induce tolerance more rapidly than conventional subcutaneous or sublingual immunotherapy, have a comparable duration of effect after only 3 injections, and do not provoke serious local or systemic reactions. However, the efficacy and safety of ILIT are using Dermatophagoides farinae (Df), Dermatophagoides pteronyssinus (Dp), and dog, which are indoor allergens that are commonly found globally, need to be evaluated. Furthermore, use of multiple allergens in ILIT should be investigated. We assessed the clinical efficacy and adverse effects of ILIT using aqueous Df, Dp, dog, and cat allergens or mixtures thereof in patients with allergic rhinitis. A total of 11 subjects with AR sensitized to Df, Dp, cat, and/or dog allergens received 3 intralymphatic inguinal injections of sensitized allergen extract (HollisterStier, New Orleans, LA, USA). Clinical parameters were assessed before ILIT, and 4 months and 1 year after the first injection. Rhinitis symptoms were alleviated and quality of life was improved 4 months after ILIT (P=0.012 and P=0.007, respectively), and these improvements lasted for 1 year after ILIT (P=0.047 and P=0.009, respectively). However, we observed 2 cases of anaphylaxis, one case of a moderate-to-severe systemic hypersensitivity reaction and the other case of a severe local reaction at the injection site after ILIT. In conclusion, ILIT can rapidly improve allergy symptoms and quality of life, and this effect lasts for 1 year. In hypersensitized patients, however, ILIT can provoke severe systemic and/or local hypersensitivity reactions when performed using aqueous allergen extracts.

Subcutaneous Immunotherapy for Allergic Asthma in a Single Center of Korea: Efficacy, Safety, and Clinical Response Predictors

Allergen-specific immunotherapy is the only causal treatment for allergic diseases. However, the efficacy of immunotherapy may vary around the world due to differences in climate, the nature of aero-allergens and their distribution. The aim of this study was to describe the effects of subcutaneous immunotherapy (SCIT) in Korean adults with allergic asthma (AA). As a retrospective cohort study, we reviewed medical records for 627 patients with AA in Korea who were sensitized to house dust mite (HDM) and/or pollens and who underwent SCIT with aluminum hydroxide adsorbed allergen extract from 2000 to 2012. Rates of remission, defined as no further requirement of maintenance medication, over time were determined by means of life tables and extension of survival analysis. Herein, 627 asthmatic patients achieved remission within a mean of 4.7 ± 0.2 years. The cumulative incidence rates of remission from AA were 86.9% upon treatment with SCIT. Baseline forced expiratory volume in the first second (FEV1) ≥ 80% (hazard ratio [HR], 3.10; 95% confidence interval [CI], 1.79–5.39; P < 0.001), and maintenance of immunotherapy for more than 3 years (HR, 1.82; 95% CI, 1.21–2.72; P = 0.004) were significant predictors of asthma remission during SCIT. In 284 patients on SCIT with HDM alone, initial specific immunoglobulin E (IgE) levels to Dermatophagoides pteronyssinus and Dermatophagoides farinae did not show significant difference between remission and non-remission group after adjusting demographic variables. In conclusion, SCIT was effective and safe treatment modality for patients with AA. Initial FEV1 ≥ 80% and immunotherapy more than 3 years were found to be associated with favorable clinical responses to SCIT.

Clinical Evaluation of Techniques for Measuring Nasal-Specific Immunoglobulin E in Pediatric Patients

Currently used methods for collecting nasal-specific immunoglobulin E (IgE) require administration of a large amount of saline into the nostrils, which is difficult in children. We designed two methods that are easy to use in children. We measured nasal-specific IgE and evaluated clinical characteristics of nasal-specific IgE in pediatric rhinitis. This study included 82 patients who visited Kyungpook National University Children's Hospital from December 2014 to July 2016 with rhinitis symptoms. Thirty patients used the spray method, and 52 patients used the swab method. We examined nasal-specific IgE to Dermatophagoides farinae, Dermatophagoides pteronyssinus, Alternaria, birch, and weed mix. Thirty patients had nasal-specific IgE concentrations of ≥ 0.35 kIU/L. There was a positive correlation between nasal-specific IgE and serum-specific IgE to D. farinae and D. pteronyssinus (r = 0.548, P < 0.001; r = 0.656, P < 0.001). If we regard positivity of skin prick test as standard, cutoff point of nasal-specific IgE to D. farinae was 0.11 kIU/L and to D. pteronyssinus was 0.12 kIU/L. Based on these cutoff points, five patients would be considered to have local allergic rhinitis. The methods showed relatively high positivity for nasal-specific IgE, which reflected the serum-specific IgE as well. These methods can be considered to diagnose local allergic rhinitis in children.

Usefulness of House Dust Mite Nasal Provocation Test in Asthma

PURPOSE: We previously reported that the skin prick test was sensitive and the serum specific immunoglobulin E test was specific for predicting positive airway responses to house dust mites (HDMs) in patients with asthma. Because the nose and bronchus are one airway, the nasal provocation test would be more specific for predicting the bronchial responses to HDM than the skin test. METHODS: The allergy skin prick test and nasal and bronchial provocation tests using HDM (Dermatophagoides farinae) were performed in 41 young men (age, 19–28 years) who wanted military certification for asthma. The nasal responses to HDM was scored according to the severity of rhinorrhea, sneezing, and nose itching. RESULTS: The prevalence of a positive skin prick test to HDM did not significantly differ between patients with (n=24) and without (n=17) an early airway reaction (EAR; 79.2% vs 70.6%, P=0.534). However, the prevalence of a positive nasal test was significantly higher in the airway responders than in the others (37.5% vs 0%, P=0.005). The concordance of a positive response to the nasal test (κ=0.332, P=0.004) but not to the skin prick test (κ=0.091, P=0.529) was significant with an EAR. The diagnostic sensitivity of the nasal test (37.5%) was lower than that of the skin prick test (79.2%), but the specificity was higher (100% vs 29.4%). CONCLUSIONS: The skin prick test is more sensitive, whereas the nasal test is more specific and accurate, for predicting an EAR to HDM in patients with asthma.

Patterns of Inhalant Allergen Sensitization and Geographical Variation in Korean Adults: A Multicenter Retrospective Study

PURPOSE: Inhalant allergen sensitization is one of the major factors involved in the pathogenesis of allergic respiratory diseases. However, the sensitization is determined by interactions between genetic and environmental factors. Thus, testing panels of inhalant allergens may differ among geographical areas. Here we aimed to determine 10 common inhalant allergens in Korean adult patients with suspected respiratory allergies and to examine the variation between different geographical locations. METHODS: A total of 28,954 patient records were retrieved for retrospective analysis, from 12 referral allergy clinics located in 9 different areas. Inclusion criteria were Korean adults (≥18 years old) who underwent the inhalant allergen skin prick test for suspected history of respiratory allergy. The primary outcome was inhalant allergen skin prick response. Demographic and clinical information were also collected. Positive skin prick responses to allergens were defined as allergen-to-histamine wheal ratio ≥1. Based on skin test results, the most prevalent aeroallergens were determined. RESULTS: The overall prevalence of allergic sensitization was 45.3%. Dermatophagoides farinae and Dermatophagoides pteronyssinus were the most commonly sensitized allergens. Other common inhalant allergens were cat epithelium (8.1%), birch (7.7%), mugwort (6.9%), alder (6.7%), hazel (6.7%), beech (6.7%), oak (6.6%), and Tyrophagus putres (6.2%), in decreasing order frequency. These 10 inhalant allergens explained 90% of inhalant allergen sensitization in the study participants. However, distinct patterns of the 10 inhalant sensitization were observed in patients living in Chungnam and Jeju. American cockroach, Gernam cockroach, and Trichophyton metagrophytes were unique in Chungnam. Orchard, Japanese cedar, and Velvet were unique in Jeju. CONCLUSIONS: The present analysis suggests a panel of 10 most common inhalant allergens in Korean adult patients with suspected respiratory allergies, which explained 90% of inhalant allergen sensitization. This panel can be utilized as a practical and convenient tool for primary practice and epidemiological surveys of respiratory allergic diseases.

Regional Difference of Causative Pollen in Children with Allergic Rhinitis

The aim of this study was to investigate role of common pollen in Korean school-aged children with allergic rhinitis (AR) in 5 provinces (Incheon Metropolitan City-Gyeonggi Province, Chungcheongbuk-do, Gwangju Metropolitan City, Busan Metropolitan City, and Jeju Special Self-Governing Province), using a questionnaire and skin prick test, and to assess the differences among the residential regions. Among the enrolled 14,678 total children, 1,641 (22.0%) had AR. The sensitization rate to pollen (38.7%) was the second highest among examined allergens and significant differences were in the sensitization rates to trees, weeds, and grasses among the 5 provinces (P < 0.05). The sensitization to trees (25.2%) was the highest common among the pollen types and significant differences also were observed in the sensitization rates to alder, birch, Japanese cedar, oak, and elm among the 5 provinces. The sensitization rate to weeds (19.9%) was the second highest and significant differences were observed in the sensitization rate to Japanese hop, mugwort, and ragweed among the 5 provinces. The sensitization rate to house dust mite was 86.8%, the highest among examined allergens and that to Dermatophagoides farinae exhibited regional differences (P = 0.003) but not to D. farinae (P = 0.584). The sensitization rate to mold (13.5%) was the highest in Jeju and lowest in Busan, and a statistically significant difference was detected among the 5 provinces. These results support that examined pollen allergens are strongly associated with residential region due to regional causative pollen differences among children with AR within Korea to investigate the main pollen allergens.