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

BACKGROUNDGroup 2 innate lymphoid cells (ILC2s) are regarded as a novel population of lineage-negative cells that induce innate Type 2 responses by producing the critical Th2-type cytokines interleukin (IL)-5 and IL-13. ILC2s as key players in the development of allergic rhinitis (AR) have been proved, however, the effect of subcutaneous immunotherapy (SCIT) with dermatophagoides pteronyssinus extract (Der p-SCIT) on ILC2s in AR patients is not clear. This study aimed to investigate the response of ILC2s of peripheral blood in house dust mites (HDM)-sensitized Chinese patients with AR who received SCIT with Der P extract.

METHODSSeven healthy controls without symptoms of AR who had negative reactions to any of the allergens from skin-prick testing, nine patients diagnosed with persistent AR according to the Allergic Rhinitis and its Impact on Asthma (ARIA) guidelines, and 24 AR patients who received Der p-SCIT for 1.0-3.5 years were recruited for the study. ILC2s in the peripheral blood were evaluated using flow cytometry. The severity of their symptoms of all participants was rated based on the Total 5 symptom score.

RESULTSAmong 40 participants, 9 AR patients were assigned to the untreated group, 24 AR patients receiving Der p-SCIT were assigned to the immunotherapy group, and 7 healthy controls without symptoms of AR were assigned to healthy control group. The mean Total 5 symptom score of immunotherapy group was significantly lower than that of untreated group (4.3 ± 1.4 vs. 10.1 ± 2.5, P< 0.001). Similarly, the levels of ILC2s in the peripheral blood of immunotherapy group were significantly reduced compared with that in untreated group (P < 0.001), but were not significantly different from healthy controls (P = 0.775). Further subgroup analysis based on the duration of SCIT therapy (1.0-2.0 years [SCIT1-2], 2.0-3.0 years [SCIT2-3], and 3.0-3.5 years [SCIT3-3.5]) showed that the percentage of ILC2s was not significantly different between SCIT1-2, SCIT2-3, and SCIT3-3.5groups (SCIT1-2 vs. SCIT2-3: P = 0.268; SCIT1-2vs. SCIT3-3.5: P = 0.635; and SCIT2-3 vs. SCIT3-3.5: P = 0.787).

CONCLUSIONSThe present study highlighted the suppression of Der p-SCIT on ILC2s in HDM-AR patients. ILC2s identified in peripheral blood can be used as an effective biomarker for Der p-SCIT.

Adolescent ; Adult ; Animals ; Dermatophagoides pteronyssinus ; immunology ; Female ; Flow Cytometry ; Humans ; Interleukin-13 ; metabolism ; Interleukin-5 ; metabolism ; Lymphocytes ; immunology ; metabolism ; Male ; Rhinitis, Allergic ; immunology ; metabolism ; therapy ; Skin Tests ; Sublingual Immunotherapy ; Young Adult

OBJECTIVETo study the skin prick test (SPT) reactivity to house dust mite allergens in overweight and normal weight children with allergic asthma before and after standard subcutaneous specific immunotherapy.

METHODSTwo hundred and fifteen children with allergic asthma who had positive SPT responses to Dermatophagoides pteronyssinus (DP) and Dermatophagoides farinae (DF) were enrolled. According to the weight index, they were classified into overweight (n=63) and normal weight groups (n=152). Skin indices (SI) to DP and DF were compared between the two groups at 6 months and 1 year after standard subcutaneous specific immunotherapy.

RESULTSThe overweight group had a significantly larger histamine wheal diameter than the normal weight group after controlling the variation in testing time (P<0.05). After controlling the variation in weights, there were significant differences in the SIs to DP and DF before specific immunotherapy and at 6 months and 1 year after specific immunotherapy. At 6 months and 1 year after specific immunotherapy, the SIs to DP and DF were significantly reduced in both groups (P<0.05), and the overweight group had greater decreases in the SIs to DP and DF than the normal weight group.

CONCLUSIONSThe overweight children with allergic asthma have stronger responses to histamine than the normal weight patients. Specific immunotherapy can reduce the reactivity to dust mite allergens in children with allergic asthma. Within one year after specific immunotherapy, the overweight children with allergic asthma have a significantly greater decrease in the reactivity to dust mite allergens than the normal weight patients.

Adolescent ; Animals ; Asthma ; immunology ; therapy ; Child ; Child, Preschool ; Dermatophagoides farinae ; immunology ; Dermatophagoides pteronyssinus ; immunology ; Female ; Humans ; Immunotherapy ; Male ; Overweight ; immunology ; Pyroglyphidae ; immunology ; Skin Tests

OBJECTIVETo construct a vector encoding T-cell epitopes of major allergen group 1 of Dermatophagoides pteronyssinus as a vaccine delivered by MHC class II pathway.

METHODSThe nucleotide sequences of the 3 target genes were synthesized, including TAT, IhC and the recombinant fragment of Der p 1 encoding 3 T-cell epitopes. After amplification of the 3 target fragments by PCR and digestion with corresponding restriction endonucleases, the recombinant gene TAT-IhC-Der p 1-3T was ligated using T4 DNA ligase and inserted into the prokaryotic expression vector pET28a(+) to construct the recombinant plasmid pET-28a(+)-TAT-IhC-Der p 1-3T, which was confirmed by digestion with restriction endonucleases and sequencing. The recombinant vector was transformed into E. coli strain BL21 (DE3) and induced with IPTG, and the induced protein TAT-IhC-Der p 1-3T was detected by SDS-PAGE. After purification, the recombinant protein was confirmed by Western blotting and its allergenicity tested using IgE-binding assay.

RESULTSThe recombinant plasmid pET-28a-TAT-IhC-Der p 1-3T was successfully constructed as confirmed by restriction endonuclease digestion and sequencing and the expression of the recombinant protein TAT-IhC-Der p 1-3T was induced in E. coli. Western blotting verified successfull purification of the target protein, which showed a stronger IgE-binding ability than Der p 1.

CONCLUSIONWe successfully constructed a recombinant expression vector pET-28a-TAT-IhC-Der p 1-3T expressing a T-cell epitope vaccine delivered by MHC II pathway with strong IgE-binding ability, which provides a basis for further study on specific immunotherapy via MHC class II pathway.

Allergens ; immunology ; Animals ; Antigens, Dermatophagoides ; immunology ; Arthropod Proteins ; immunology ; Base Sequence ; Cloning, Molecular ; Cysteine Endopeptidases ; immunology ; Dermatophagoides pteronyssinus ; Epitopes, T-Lymphocyte ; Escherichia coli ; Gene Expression ; Genes, MHC Class II ; Genetic Vectors ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; immunology ; Vaccines ; immunology

OBJECTIVE: To study the therapeutic effects of the specific immunotherapy (SIT) on allergic rhinitis and allergic rhinitis combined bronchial asthma. METHED: All patients were classified into allergic rhinitis group (AR group) with 32 patients and allergic rhinitis combined bronchial asthma group (AR+BA group) with 32 patients. Another health control group with 32 cases was designed as well. The allergens,symptom scores and therapeutic effects of the former two-group patients were analysis, and the serums of all three-group cases were extracted to evaluate the specific Immunoglobin E(sIgE), Interleukin-4 (IL-4). The SPSS13. 0 package was applied to conduct t-test and chi-square test, and the difference of P<0. 05 was regarded as statistical significance. RESULT: The main allergens of 64 patients were dermatophagoides farinae and dermatophagoides pteronyssinus. The improvement of symptom scores before and after SIT was statistical significant with P<0. 05. Although total effective rate reached 100% , AR group was superior than AR+BA group in term of the efficacy comparison, and P<0. 05 indicated the statistical significance. The serum sIgE, IL-4 values of three groups were brought into comparison, and P<0. 05 indicated the statistical significance of the difference. CONCLUSION The SIT on the AR, AR+BA is a safe and effective treatment, but different disease responds diversely. The long-term treatment course is recommended.
Allergens ; immunology ; Animals ; Asthma ; therapy ; Dermatophagoides farinae ; immunology ; Dermatophagoides pteronyssinus ; immunology ; Humans ; Immunoglobulin E ; blood ; Immunotherapy ; Interleukin-4 ; blood ; Rhinitis, Allergic ; therapy ; Treatment Outcome

OBJECTIVE: We aimed to analyze the quality of life (QOL) in adults with allergic rhinitis according to the sensitization profile for relevant aeroallergens in Northern China, investigate the proportion of patients with coexisting asthma, and explore the correlation between QOL of rhinitis patients and the specific IgE level to the causative allergen. METHOD: One hundred and sixty-four allergic rhinitis patients participated in this study, whose clinical history, results of intradermal skin test and serum specific IgE levels to common aeroallergens in North China were collected. QOL was evaluated using the rhinoconjunctivitis quality of life questionnaire (RQLQ). RESULT: QOL of rhinitis patients was worse in those sensitized to tree pollens or weed pollens than those sensitized to house dust mites in Northern China. The proportion of patients with co-existing asthma was lower in tree pollen group than in house dust mite group or weed pollen group, and there was no significant difference between house dust mite group and weed pollen group. There was no significant correlation between QOL of rhinitis patients and the specific IgE level to the causative allergen. CONCLUSION In our study group, QOL of patients with allergic rhinitis varied with the allergen responsible for symptoms, but was not influenced by the specific IgE level to relevant allergen. The proportion of patients with co-existing asthma also varied with different pollen allergens. Rhinitis patients sensitized to weed pollens might be more likely to suffer from asthma than those sensitized to tree pollens.
Adult ; Allergens ; immunology ; Animals ; Asthma ; China ; Dermatophagoides pteronyssinus ; Humans ; Immunoglobulin E ; blood ; Pollen ; Pyroglyphidae ; Quality of Life ; Rhinitis, Allergic ; physiopathology

We attempted to investigate the correlation between the severity of atopic dermatitis (AD) in children and the indoor level of house dust mite (HDM) allergens. Ninety-five patients (31.1 +/- 19.5 months of age) with AD were enrolled in this study, and serum specific IgE against Dermatophagoides pteronyssinus and D. farinae was measured. The severity of AD was assessed using the visual analogue scale on the same day of house dust collection. Living rooms and mattresses where the child usually slept were vacuumed for 2 minutes and concentrations of Der f 1 were measured by enzyme-linked immunosorbent assay. The skin symptoms were more severe in patients with Der f 1 concentrations in living room > 2 microg/g dust than < or = 2 microg/g dust (P = 0.018). This difference was noted in AD patients without sensitization to HDM (P = 0.004), but not in patients with sensitization. There was no difference in symptom severity according to Der f 1 concentrations in mattresses (P = 0.062). The severity of skin symptoms is associated with indoor concentrations of HDM in children with AD, and it is likely to act as nonspecific irritants as well as allergens in AD skin lesions.
Adolescent ; Animals ; Antigens, Dermatophagoides/*analysis ; Beds/parasitology ; Child ; Child, Preschool ; Dermatitis, Atopic/*diagnosis/pathology ; Dermatophagoides farinae/immunology ; Dermatophagoides pteronyssinus/immunology ; *Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunoglobulin E/blood ; Infant ; Male ; Severity of Illness Index

OBJECTIVE: To evaluate the efficacy and safety of subcutaneous immunotherapy with a standardized Dermatophagoides pteronyssinus extract for allergic rhinitis. METHOD: Sixty-eight patients with allergic rhinitis who received specific immunotherapy with a standardized dermatophagoides pteronyssinus extract were enrolled in this study. The clinical symptoms and signs were scored before and 2 years after treatment. Side effects were registered. Symptom and side effect scores were assessed to evaluate the clinical efficacy and safety. RESULT: There was a significant difference (P < 0.01) in symptom and medication scores between the two groups tow years after treatment. The patients had fewer symptoms and lower intake of medication. No severe systemic reactions were registered. CONCLUSION Immunotherapy with a standardized dermatophagoides pteronyssinus extract is effective and safe to treat patients clinically sensitive to this allergen.
Allergens ; therapeutic use ; Animals ; Dermatophagoides pteronyssinus ; immunology ; Female ; Humans ; Immunotherapy ; methods ; Male ; Rhinitis, Allergic ; therapy

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

OBJECTIVE: To study the pathogenesis of the patients with allergic rhinitis diagnosed by Skin Prick Test (SPT), especially about the epidemiologic data of the involved allergens. METHOD: The data was collected from 958 patients referred to the listed three allergy center and subsequently diagnosed as allergic rhinitis by SPT. RESULT: The intermittent mild type was more prevalent in male patients (40.5%); comparably the intermittent moderate type in female patients (70.0%); in the infant patients the moderate-severe type dominated (73.7%). The positive results of SPT comprised mainly of Dermatophagoides pteronyssinus (Der. p, 98.3%), Dermatophagoides farinae(Der. f, 96.8%), and Blomia tropicalis (Blot, 73.8%). Only 1.7% of the patients was allergic to single allergen, in contrary ,the majority of patients were allergic to multiple allergens. The positive rate to Der. p and Der. f was reversely increasing with age of the patients; and contrarily the number of positive allergens was increasing along with the age of the patients. The allergy to outdoor allergen was less common, and the positive rate in skin prick test was lower than the previously reported rate in North China ,such as timothy (3.9%), birch (2.7%), ragweed (2.0%), and mugwort (1.2%). CONCLUSION Dust Mite is the predominant allergen for patients with allergic rhinitis in Guangdong province; and the positive rate to outdoor allergen is lower than that in North China.
Adolescent ; Adult ; Allergens ; analysis ; immunology ; Animals ; Antigens, Dermatophagoides ; analysis ; immunology ; Child ; Child, Preschool ; China ; epidemiology ; Cohort Studies ; Dermatophagoides pteronyssinus ; immunology ; Female ; Humans ; Male ; Middle Aged ; Mites ; immunology ; Rhinitis, Allergic, Perennial ; diagnosis ; epidemiology ; Rhinitis, Allergic, Seasonal ; diagnosis ; epidemiology ; Skin Tests ; Young Adult