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.
Animals ; Humans ; Extracellular Traps ; Neutrophils ; Interleukin-8/metabolism* ; Dermatophagoides farinae ; Interleukin-6/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Epithelial Cells ; Interferon-gamma/metabolism* ; Tetradecanoylphorbol Acetate/pharmacology*

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.
Allergens ; Caseins ; Dermatophagoides farinae ; Dermatophagoides pteronyssinus ; Diagnosis ; Gliadin ; Humans ; Immunoassay ; Immunoglobulin E ; Immunoglobulins ; Pyroglyphidae

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.
Allergens ; Dermatitis, Atopic ; Dermatophagoides farinae ; Dermatophagoides pteronyssinus ; Diagnosis ; Horses ; Hypersensitivity ; Immunoglobulin E ; In Vitro Techniques ; Intradermal Tests ; Mites ; ROC Curve ; Sensitivity and Specificity

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.
Blister ; Busan ; Child ; Dermatitis, Atopic ; Dermatophagoides farinae ; Eosinophils ; Exfoliatins ; Humans ; Immunoglobulin E ; Immunoglobulins ; Incidence ; Medical Records ; Retrospective Studies ; Seasons ; Skin ; Skin Diseases ; Staphylococcal Scalded Skin Syndrome ; Staphylococcus aureus

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.
Adolescent* ; Alternaria ; Ambrosia ; Animals ; Artemisia ; Cats ; Child* ; Cynodon ; Dactylis ; Dermatophagoides farinae ; Dogs ; Eosinophilia* ; Eosinophils ; Hospitals, Teaching ; Humans ; Hypersensitivity ; Immunoglobulin E ; Korea ; Leukocytes ; Lolium ; Male ; Phleum ; Pyroglyphidae ; Retrospective Studies ; Rhinitis* ; Seoul

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.
Allergens ; Allergy and Immunology ; Ambrosia ; Animals ; Artemisia ; Cats ; Dermatophagoides farinae ; Dermatophagoides pteronyssinus ; Diagnosis ; Dogs ; Electrophoresis ; Electrophoresis, Polyacrylamide Gel ; Humans ; Humulus ; Hypersensitivity ; Immunoblotting ; Immunoglobulin E ; Immunoglobulins ; In Vitro Techniques* ; Indicators and Reagents* ; Korea ; Methods* ; Pollen ; Skin* ; Sodium

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.
Air Pollutants* ; Allergens ; Animals ; Asthma ; Cats ; Child ; Child, Preschool* ; Cockroaches ; Dermatophagoides farinae ; Dermatophagoides pteronyssinus ; Dogs ; Dust ; Fungi ; Hair ; Humans ; Hypersensitivity ; Mites ; Poaceae ; Pollen ; Prevalence ; Rhinitis, Allergic* ; Seasons ; Skin ; Trees ; Urbanization

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.
Aged* ; Antibodies ; Asthma ; Basophils ; Demography ; Dermatophagoides farinae ; Dermatophagoides pteronyssinus ; Dust* ; Hospitals, University ; Humans ; Immunoglobulin A ; Immunoglobulin E ; Immunoglobulins ; Immunomodulation ; Immunotherapy ; Pyroglyphidae* ; Quality of Life ; Rhinitis* ; Rhinitis, Allergic ; Skin ; Sublingual Immunotherapy*

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.
Allergens ; Anaphylaxis* ; Dermatophagoides farinae ; Flow Cytometry ; Fluorescence ; Immunoglobulin G ; In Vitro Techniques ; Leukocytes ; Neutrophils* ; Respiratory Burst* ; Suspensions

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.
Adult ; Allergens ; Animals ; Asthma ; Bermuda ; Cats ; Cockroaches ; Dander ; Dermatitis, Atopic ; Dermatophagoides farinae ; Dermatophagoides pteronyssinus ; Dogs ; Dust ; Humans ; Hypersensitivity ; Medical Records ; Mites ; Poaceae ; Pollen ; Prevalence ; Retrospective Studies ; Rhinitis ; Rhinitis, Allergic ; Risk Factors ; Skin Tests ; Skin ; Thailand ; Urticaria