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Objective:To establish a quantitative detection method for the main components of dust mite allergens Der p 1, Der p 2 specific immunoglobulin E (sIgE) by using the nano-magnetic particle chemiluminescence immunoassay.Methods:The performance indexes of the established method were evaluated after setting up and optimizing the chemiluminescence detection system and immune reaction conditions of sIgE for dust mite allergen. Serum sIgE levels of 50 suspected allergic patients with dust mite were determined by this chemiluminescence method. At the same time, this method was compared with the Phadia kit and the consistency was analyzed by Kappa test. Results:The optimal amount of magnetic beads was 25 μg, the optimal reaction buffer (pH=7.4) contained 0.1 mol/L Tris-HCl and 0.25%( W/ W) casein, the optimal coating solution contatined 20 mmol/L phosphate buffer (PB) and 1%( W/ W) bovine serum albumin (BSA), and the luminescence enhancement solution contained 0.05%( V/ V) Triton X-100. The two-step immunoreaction was adopted, and the detection could be completed with 20 μl sample at the optimal reaction temperature of 37℃. The limit of detection (LOD) of the established nano-magnetic particle chemiluminescence system in detecting Der p 1 and Der p 2 sIgE antibodies were both less than 0.01 kU/L, with the linear range of 0.2-100.0 kU/L, the precision of less than 7%, and the cross contamination rate of 0.19% and 0.21%. Compared with the Phadia system, the positive and negative coincidence rate of Der p 1 were 78.0%(32/41) and 9/9 with good consistency ( Kappa=0.65, P=0.008), and the positive and negative coincidence rate of Der P 2 were 93.3%(28/30) and 85.0%(17/20) with good consistency ( Kappa=0.79, P=0.003). Conclusion:The nano-magnetic particle chemiluminescence immunoassay is successfully established for detecting dust mite allergen sIgE, which has good detection performance and good consistency with Phadia system.
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RESUMEN Fundamento: La inmunoterapia consiste en la administración de dosis crecientes del alérgeno para controlar la sensibilidad hacia este. Objetivo: Describir el comportamiento de la inmunoterapia por vía subcutánea o sublingual a ácaros. Metodología: Estudio retrospectivo realizado en Cabaiguán, periodo de 2010 a 2019, el universo de 267 pacientes con pruebas positivas a ácaros y la muestra de 53 pacientes. Los datos se obtuvieron del registro de datos. Resultados: Usó la vía sublingual el 60.4 %, predominó el grupo de edad entre 5-18 años (41.5 %), la vía subcutánea se utilizó en un 39.6 %, fue más frecuente en el sexo masculino (58.5 %). La vía subcutánea se indicó mayoritariamente en la rinitis alérgica (20.7 %) y la sublingual en el asma (32.1 %); para ambas, fue el Dermatofagoide pteronyssinus el ácaro de mayor sensibilidad y utilización (43.4 %), el tiempo de administración dominante fue de 3 años (67.9 %). El control de los síntomas se alcanzó con la vía sublingual (37.7 %). En la vía sublingual no se presentaron eventos adversos en el 43.3 %, con la vía subcutánea 11.3 % de los eventos adversos fueron locales y 7.6 % sistémicos leves. Conclusiones: La vía sublingual fue la más utilizada, la de mayor control de los síntomas y menos eventos adversos en el estudio.
ABSTRACT Background: Immunotherapy consists of the administration of the allergen increasing doses to control sensitivity towards it. Objective: To describe the behavior of subcutaneous or sublingual immunotherapy to mites. Methodology: Retrospective study carried out in Cabaiguán, from 2010 to 2019, the universe of 267 patients with positive tests to mites and the sample 53 patients. Data were obtained from the data record. Results: 60.4 % used sublingual, the age group between 5-18 years prevailed (41.5 %), the subcutaneous was used in 39.6 %, and it was more frequent in males (58.5 %). The subcutaneous was indicated mainly in allergic rhinitis (20.7 %) and sublingual in asthma (32.1 %); for both, Dermatofagoide pteronyssinus was the highest sensitivity and use mite (43.4 %) the dominant administration time was 3 years (67.9 %). Symptom control was achieved by sublingual (37.7 %). In sublingual, there were no adverse events in 43.3 %, with the subcutaneous, 11.3 % of the adverse events were local and 7.6 % were mild systemic. Conclusions: Sublingual was the most used, the one with the highest control of symptoms and the fewest adverse events in the study.
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Skin Tests , Antigens, Dermatophagoides , Sublingual Immunotherapy , Injections, Subcutaneous , MitesABSTRACT
Objective@#To explore the temporal and spatial characteristics of search volume (SV) of allergic rhinitis (AR) and related search terms on Baidu Index, as well as the correlation between SV from Baidu Index and epidemiology of AR.@*Methods@#SV of AR and related search terms in mainland area of China from January 2012 to December 2016 were extracted from Baidu Index. Firstly, we investigated the temporal and spatial characteristics of SV of AR and related search terms ("pollen allergy" and" dust mite allergy", etc.), and explored the correlation between SV of AR and SV of related search terms. Secondly, the SV of specific search terms in Beijing and Guangzhou were compared with the actual pollen count in Beijing from April to September in 2016 and the monthly outpatient volume of AR patients in the Otorhinolaryngology Hospital, The First Affiliated Hospital of Sun Yat-sen University from 2014 to 2016 respectively to analyze the consistency between SV from Baidu Index and the real-world data. Spearman correlation analysis was used to explore the correlation between different search terms according to the normality and homogeneity of variance of the data. P<0.05 was considered statistically significant.@*Results@#SV of AR showed repetitively seasonal pattern, with the first peak in May and the second peak from August to September. SV of AR in northeast China, north China and northwest China were correlated with both SV of "pollen allergy" and "dust mite allergy" (r value was 0.533, 0.638, 0.609, 0.791, 0.517, 0.682, respectively, all P<0.05). In east China, central China, south China and southwest China, SV of AR showed significant correlation only with SV of "dust mite allergy" (r value was 0.897, 0.884, 0.799, 0.937, respectively, all P<0.05), but not with SV of "pollen allergy" (r value was 0.110, 0.171, 0.020, 0.265, respectively, all P>0.05). The correlation between pollen count and the SV of AR on the day and SV of "pollen allergy" on the next day was the highest (r value was 0.692 and 0.713, respectively, all P<0.05). The SV of AR was correlated with the monthly outpatient volume of AR patients one month and two months later (r value was 0.523 and 0.503, respectively, all P<0.05).@*Conclusion@#The SV of AR and related search terms show different trends in different times and regions which are consistent with the AR epidemiological survey, and significantly correlate with pollen count and AR outpatient volume.
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Objective@#To explore the therapeutic effects of dendritic cells (DC) modified by the dust-mite-allergen(Der p1) gene on mouse model of allergic rhinitis (AR).@*Methods@#DC modified by the Der p1 gene (Der p1-DC) were prepared.Using random number table, 24 Balb\c mice were divided into four groups: immature DC (imDC)/AR group, dexamethasone/AR group, Der p1-DC/AR group and control group, with 6 mice in each group.AR mouse model was built with Der p1 and the mouse model of AR was established.The AR mice were respectively given by abdominal injection of Der p1-DC, imDC and dexamethasone.Normal control mice were treated with physiologic saline.ELISA method was used for determining the content of IgE, IgG1and histamine in blood.The relative expression of mRNA of IL-4 and IL-13 on nasal mucosa with protein was analyzed by RT-PCR and Westen blot methods.All the data were statistically analyzed by SPSS 19.0 statistical software, and the variance analysis was used in multiple groups of average samples.@*Results@#The contents of IgE, IgG1 and histamine in the mice of Der p1-DC/AR group were lower than those in imDC/AR group ((0.560±0.110) OD 450 nm vs (1.150±0.280) OD 450 nm, (0.690±0.054) OD 450 nm vs (0.920±0.125) OD 450 nm, (4 145±670) pg/ml vs (7 685±669) pg/ml, t value was 4.80, 4.14, 9.16, respectively, all P<0.05), and the expression of IL-4 and IL-13 on nasal mucosa in Der p1-DC/AR group was remarkedly lower than those in imDC/AR group (0.41±0.25 vs 1.59±1.02, 0.26±0.01 vs 1.10±0.09, t value was 2.75, 22.72, respectively, all P<0.05). There was no statistically significant difference between the mice treated with Der p1-DC and dexamethasone group.@*Conclusions@#The results showed that Der p1-DC could reduce inflammation in AR mice and decrease the expression of IL-4 and IL-13. It suggested that Der p1-DC can be used in the immunotherapy of AR mouse.
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OBJECTIVE To explore the correlation of allergic rhinitis symptoms in preschool children with the household environment dust mites and house dust-mite-specific IgE, as to provide theoretical basis for pediatric allergic disease prevention and control work. METHODS The children with rhinitis who was born in the Second People's Hospital of Wuhu were selected in this study. When the children were 3 or 4 years old, they were followed up. The allergic rhinitis symptoms integral of the children were evaluated, the household environment dust samples of the children were collected. The content of dermatophagoides pteronyssinus allergen components 1(Der p1) and dermatophagoides farinae allergen components 1(Der f1) in dust samples were determined by the method of enzyme-linked immunosorbent(ELIEA). The children house dust-mite-specific serum IgE(sIgE) level was also examined. RESULTS Pediatric allergic rhinitis symptoms had a positive correlation with dermatophagoides pteronyssinus sIgE and dermatophagoides farinae sIgE(P0.05). CONCLUSION The allergic rhinitis symptoms of the children are positively correlated with household environment dust mites and dust mites specific IgE. That indicated the severity of allergic rhinitis symptom in children may be determined by detecting specific IgE levels. The prevention of allergic disease and improvement the quality of life of the allergic children can be done by strengthening the health education of dust mites and decreasing the content of bedroom environment dust mites.
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Objective To analyze results of atopy patch test (APT) with dust mite allergens at different concentrations in patients with atopic dermatitis (AD),and to evaluate the diagnostic value of APT.Methods Totally,85 patients with AD were enrolled into this study.All the patients underwent APT with 5 concentrations (3 000,5 000,7 000,10 000 and 12 000 pnu/g) of dust mite allergens,as well as skin prick test (SPT) with dust mite allergens.Dust mite allergens were obtained from two different manufacturers (group 1 and 2).Enzyme-linked immunosorbent assay (ELISA) was performed to detect allergen-specific immunoglobulin E (SIgE) in sera from these patients.The sensitivity,specificity and positive predictive value of APT,SPT and SIgE assay were compared,and the results of APT were compared among different concentrations of allergens and between allergens from different manufacturers.Results When SIgE assay served as the gold standard,the sensitivity,specificity and positive predictive value in the diagnosis of dust mite allergy were 79.41％,76.12％,and 64.29％ respectively for APT,73.53％,80.95％ and 67.57％ respectively for SPT with group 1 dust mite allergens,and 81.53％,77.78％ and 65.09％ respectively for APT,76.02％,79.85％ and 66.07％ respectively for SPT with group 2 allergens.When SPT was regarded as the gold standard,the sensitivity,specificity and positive predictive value in the diagnosis of dust mite allergy were 78.38％,77.42％,67.44％ respectively for APT,67.57％,84.21％,73.53％ respectively for SIgE assay with group 1 dust mite allergens,79.25％,80.63％ and 69.55％ respectively for APT,61.07％,82.54％ and 77.21％ respectively for SIgE assay with group 2 allergens.There were no significant differences in the sensitivity,specificity or positive predictive value of APT or SPT between the two groups of allergens.The positive rate of APT was 8.24％,22.35％,29.41％,44.71％ and 41.18％ respectively with group 1 allergens at 3 000,5 000,7 000,10 000 and 12 000 pnu/g,and 3.53％,23.53％,31.76％,34.12％ and 35.29％ respectively with group 2 allergens.No significant differences were observed in the positive rate of APT between group 1 and 2 allergens at same concentrations (all P ＞ 0.05),but a significant difference was observed in that between different concentrations of group 1 or 2 allergens (both P＜ 0.05).The positive rate of APT increased with the increase of allergen concentrations,but stopped rising when the concentrations of group 1 and 2 allergens reached 7 000 pnu/g and 5 000 pnu/g respectively.Conclusions The sensitivity of APT is relatively high for the diagnosis of dust mite allergy.The positive rate of APT increased with the increase in allergen concentrations,but stopped rising when the concentrations reached a certain level.
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BACKGROUND AND OBJECTIVES: To evaluate IgE reactivity profiles to Der p 1, Der p 2, and Der p 10 in house dust mite (HDM) allergic rhinitis (AR) patients in Korea. Symptomatic and serologic changes after sublingual immunotherapy (SLIT) were analyzed according to IgE profiles. MATERIALS AND METHODS: Sixty AR patients diagnosed with an HDM allergy were tested for the presence of IgE antibodies against purified HDM allergen molecules (i.e., nDer p 1, rDer p 2, and rDer p 10) and native Dermatophagoides pteronyssinus (Dp) using ImmunoCAP 250®. Symptom scores and laboratory findings were analyzed after SLIT. RESULTS: Prevalence rates of IgE for Dp, Der p 1, Der p 2, and Der p 10 were 100%, 98.3%, 93.3%, and 8.3%, respectively. After one year of immunotherapy with SLITone○R (standardized to major allergens, but not Der p 10), symptom scores and laboratory findings improved in patients with and without Der p 10 IgE antibodies. CONCLUSION: In Korean AR patients, specific IgE antibodies to Der p 1 or Der p 2 are present in most Dp-allergic patients, while reactivity to Der p 10 is very low. Allergic symptoms improved in patients with Der p 10 IgE antibodies after SLIT.
Subject(s)
Humans , Allergens , Antibodies , Antigens, Dermatophagoides , Dermatophagoides pteronyssinus , Hypersensitivity , Immunoglobulin E , Immunotherapy , Korea , Prevalence , Pyroglyphidae , Rhinitis, Allergic , Sublingual ImmunotherapyABSTRACT
Objective To establish a cellular model for the expression of the C-type lectin dendritic cellspecific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN),and to provide a basis for the functional analysis of DC-SIGN.Methods The cDNA of DC-SIGN was obtained via PCR,and cloned into the eukaryotic expression vector porcine cytomegalovirus-enhanced green fluorescent protein (PCMV-EGFP) with EGFP at the N terminal of DC-SIGN.Then,the recombinant PCMV-EGFP-DC-SIGN plasmid was transfected into HEK293T cells followed by the detection of DC-SIGN expression using PCR,Western blot and flow cytometry.Confocal microscopy was performed to localize the expression of DC-SIGN-EGFP and visualize the recognization and internalization of the Derp2 allergen by DC-SIGN.Results The recombinant fluorescent fusion protein-expressing plasmid was successfully constructed.Both PCR and Western blot confirmed the expression of DC-SIGN.Flow cytometry showed that the expression of DC-SIGN was increased by approximately 50％ in HEK293T cells transfected with the recombinant expression plasmid compared with those untransfected.As confocal microscopy showed,the green fluorescence-labelled DC-SIGN was located on the cell membrane,which could bind to the red fluorescence-labelled antigen Derp2 and internalize it into the cells.Conclusions The recombinant DC-SIGN-EGFP fusion protein is characteristically located on the surface of 293T cells,which can be recognized by the DC-SIGN-specific antibody and is capable of internalizing the allergen Derp2,and may serve as an ideal cell model for further studies on molecular function of DC-SIGN.
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Objective To evaluate the efficacy and safety of specific immunotherapy (SIT) to allergic rhinitis in children.Methods Sixty-four patients with mite allergy allergic rhinitis in children,were divided into two groups by random digits table:treatment group and control group,each group with 32 cases.Treatment group was given SIT with standardized allergen vaccine for 3 years on the basis of symptomatic therapy,control group only received symptomatic therapy.Observation indexs included rhinitis symptoms score,drug score,skin index (SI),serum specificity IgE (sIgE),peripheral eosinophil (Eos) counting,development of asthma and the new sensitization.Results The Eos counting,SI after treatment 3 years in treatment group were significantly better than those before treatment and those in control group after treatment 3 years[(0.14 ± 0.12) × 109/L vs.(0.74 ± 0.18) × 109/L,(0.78 ± 0.36) × 109/L and 1.03 ± 0.13 vs.1.51 ± 0.32,1.51 ± 0.37] (P ＜ 0.01).There was no statistically significant difference in sIgE between two groups (P ＞0.05).The rhinitis symptoms score,drug score in two groups after treatment 1,2,3 years were significantly better than those before treatment (P ＜ 0.01).The rhinitis symptoms score,drug score in treatment group after treatment 1,2,3 years were significantly better than those in control group(P ＜ 0.01).The rate of new sensitization in treatment group was significantly lower than that in control group [3.1％ (1/32)vs.34.4％ (11/32),P ＜ 0.01].Conclusion Keeping long-term SIT is effective and safe for children's allergic rhinitis induced by mite,it also prevents new allergen appeared and allergic rhinitis development for asthma.
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La rinitis alérgica es una enfermedad respiratoria crónica de alta prevalencia y que tiene gran impacto en la calidad de vida y el desempeño laboral o escolar. Se relaciona estrechamentecon otras enfermedades alérgicas como el asma, la sinusitis y otras afecciones como la conjuntivitis y la otitis. Uno de los principales alérgenos que ocasionan la rinitis alérgica son los ácaros del género Dermatophagoides, en especial las especies D. pteronyssinus y D. farinae, aunque también se ha demostrado la importancia del ácaro Blomia tropicalis en estas afecciones.En este artículo de revisión se analiza la importancia de las reacciones alérgicas frente a estos ácaros, en especial del género Dermatophagoides, tomando en cuenta los estudios realizados en el contexto nacional y latinoamericano; también se explican los mecanismos inmunológicos de la hipersensibilidad tipo I en los pacientes sensibilizados con dichos alérgenos, incluyendo elproceso de sensibilización y de producción de IgE específica, así como el papel de las citocinas en este proceso. Así mismo, se discutirán los métodos diagnósticos y los esquemas terapéuticos que se utilizan, incluyendo la inmunoterapia.
Allergic rhinitis is a chronic respiratory disease of high prevalence that has great impact on the standard of living, and the performance at work or school. Allergic rhinitis is related with other allergic diseases, such as asthma, sinusitis, conjunctivitis, and otitis. One of the main allergens that cause it are the mites belonging to the Dermatophagoides genus, mostly the species D. pteronyssinus and D. farinae, although other studies have shown the importance of Blomia tropicalis in these allergic conditions. This review article aims to analyze the meaning of allergicreactions to such mites, especially those of the genus Dermatophagoides, with regards to studies performed in a national and Latin-American context. It also explains the immunological mechanisms linked to the processes of type I hypersensitivity in patients sensitized to these allergens, includingthe sensitization mechanism and specific lgE production, and the role of cytokines as part of the process. In addition, the diagnostic methods and treatments used, including immunotherapy, will be discussed.