Dermatophagoides Pteronyssinus Allergen Research Articles

Severity Assessment and IL-5 Evaluation after Immunotherapy in Atopic Dermatitis Mice

Background: The effectiveness of immunotherapy in atopic dermatitis (AD) has been debated for many years. Immunotherapy suppresses Th2 cytokines, such as IL-5, which play a crucial role in the pathogenesis of AD. This study assessed the effect of immunotherapy on severity and IL-5 expression in AD mice.Methods: Male BALB/c mice were separated into three groups. The mice were sensitized with Dermatophagoides pteronyssinus allergen for seven days, except the mice in control group. The house dust mite (HDM) immunotherapy was injected subcutaneously every 3 days for 1.5 months with increasing doses (0.1, 1, 10, 100 μg in 100 μL PBS) every 4 injections. The mice in AD model and control group received placebo injections. Following immunotherapy, the mice were exposed to HDM allergen patch two times with 2 weeks interval in between. The mice were evaluated for severity score as the clinical marker and IL-5 expression with semiquantitative method as the histological marker.Results: The evaluation of severity score from two independent researchers showed a substantial agreement (Cohen’s kappa 0.613, p <0.001). The severity score of the immunotherapy group was significantly lower than the AD model group, while both immunotherapy and AD model group had significantly higher score than control group. IL-5 expression in the immunotherapy group was lower than the AD model group and slightly higher than control group. The mean difference between groups was not significant.Conclusion: The severity of skin lesion and IL-5 expression in AD mice receiving immunotherapy were lower than AD model group.Keywords: Atopic dermatitis; BALB/c mice; House dust mites; IL-5, Skin severity score; Tropical disease

Effectiveness of allergen-specific immunotherapy in children with bronchial asthma sensitized to house dust mites

Background. The only method of treatment that can change the natural course of allergies is allergen-specific immunotherapy, which allows reducing the sensitivity of the child’s body to the causative allergen and decreasing the activity of chronic inflammation. The evidence base for the effectiveness of allergen-specific immunotherapy in the treatment of bronchial asthma (BA) has significantly expanded recently. However, the truly important results remain too limited to make clinically useful conclusions about the efficacy of sublingual allergen-specific immunotherapy (SLIT) in people with BA and further evidence is needed. The purpose of the study is to evaluate the effectiveness of SLIT as an additional treatment for atopic BA of varying severity in children sensitized to house dust mites. Materials and methods. The main group consisted of 80 children with BA aged from 5 to 17 years, they were divided into three groups according to the severity of the disease: group 1 — 20 children with mild intermittent asthma, group 2 — 30 children with mild persistent asthma, group 3 — 30 children with moderately severe persistent asthma. Thirty practically healthy children without BA were also examined as a control group. For SLIT, a sublingual ORALTEK spray produced by Inmunotek (Spain) was used, a mixture of house dust mite allergens Dermatophagoides pteronyssinus 50 %/Dermatophagoides farinae 50 %, 1 ml of the solution contains 30,000 IU of natural allergen extract. To assess the effectiveness of the treatment, we conducted an analysis of the total score of clinical manifestations of asthma and allergic rhinitis and the degree of asthma control according to the Asthma Control Test. We also analyzed the state of cellular and humoral immunity in the dynamics of therapy. These indicators were evaluated within 1 month before the beginning and a month after the end of the one-year course of therapy. To assess the validity and reliability of the obtained results, statistical data processing was carried out using Microsoft Excel 2016 and Statistica for Windows 10 programs. The Student’s t-test was applied to assess the reliability of the results. The difference between the compared values was considered statistically significant at p < 0.05. Results. The results of the statistical analysis confirmed a significant improvement in the control indicators of patients who received SLIT: reduction in the total score of clinical manifestations, the frequency of clinical symptoms and the use of emergency drugs (p < 0.05). By the end of the study, the dynamics of a decrease in these indicators compared to the data before the start of treatment (according to the above list) was as follows: in patients of group 1 — 82.5, 91.8, 94.5 %, in patients of group 2 — 80.5, 88.4, 92.7 %, in patients of group 3 — 76.7, 85.3, 91.2 %. After treatment with the addition of SLIT to the basic therapy, a reliable increase in IgE and sIgA was observed, an increase in the spontaneous proliferative response of T-lymphocytes to the mitogen in all three groups to the level of the controls, a probable decrease in eosinophilic inflammation and an improvement in indicators of the cellular and humoral immunity (p < 0.001). Conclusions. The prescription of SLIT to children with mild intermittent BA, mild persistent BA and perennial allergic rhinitis, moderate persistent BA and perennial allergic rhinitis provides an opportunity to obtain stable control of the disease, reduces the number of visits to the doctor and justifies the wide use of this method in clinical allergy practice.

Comparison of intradermal and serum testing for environmental allergen-specific immunoglobulin E determination in a laboratory colony of cats with naturally acquired atopic syndrome.

Allergen testing is used to select antigens included in the desensitisation vaccine. Intradermal skin test (IDT) is the gold standard in cats, yet allergen-specific immunoglobulin (Ig)E serological testing (ASIS) is often used. Feline data are lacking regarding the agreement between IDT and ASIS results. The first objective of the study was to establish a colony of cats with naturally acquired feline atopic syndrome (FAS). Further objectives were to define their hypersensitivity disorder to detail the allergen tests results, and to assess similarity between the allergen tests. Thirty-five cats with FAS and 10 control cats. Enrolled cats went through a five phase-screening and quarantine process before joining the colony. An elimination diet trial was performed on all FAS cats. ASIS and IDT were consecutively performed on all cats under sedation. Reactions to 34 allergens were compiled for the 45 cats. Global sensitivity and specificity of ASIS were 34.7% and 78.9%, respectively. Only flea (ICC = 0.26, p = 0.040) and Dermatophagoides pteronyssinus (ICC = 0.48, p < 0.001) allergens had a significant intraclass correlation (weak agreement). Two FAS cats had negative tests including one cat with a concomitant food allergy. This study depicts the first reported colony of cats with naturally acquired FAS. This is the first feline study to compare and show the poor agreement between allergen tests with a panel of 34 allergens. This colony also harbours two cats with FAS with negative allergen tests. These may represent the first described cats with an intrinsic form of atopic syndrome.

Oral administration of Lactobacillus delbrueckii subsp. lactis LDL557 attenuates airway inflammation and changes the gut microbiota in a Der p-sensitized mouse model of allergic asthma.

Atopic dermatitis (AD) is a chronic inflammatory skin disorder affecting up to 20% of children in developed countries. Although probiotics have shown promise as adjuvant treatments for AD, their mechanisms are not well understood. Building upon our previous studies, we investigated whether Lactobacillus gasseri and its moonlighting glyceraldehyde 3-phosphate dehydrogenase (GAPDH), namely LGp40, could be beneficial in AD management. In AD mouse models (SKH and C57BL/6J mice) with ovalbumin (OVA) and Dermatophagoides pteronyssinus (Der p) allergens, aligning with the "outside-in" and "inside-out" hypotheses, we administered L. gasseri orally and LGp40 intraperitoneally to investigate their protective effects. The evaluation involved measuring physiological, pathological, and immune function parameters. To delve deeper into the detailed mechanism of LGp40 protection in AD, additional assays were conducted using human skin keratinocytes (HaCaT) and monocytes (THP1) cell lines. L. gasseri and LGp40 enhanced skin barrier function and increased skin moisture retention. They also led to reduced infiltration of Langerhans cells in the dermis and mitigated skewed Th2 and Th17 immune responses. Moreover, LGp40 inhibited allergen-induced keratinocyte apoptosis through the blockade of the caspase-3 cascade and reduced the NLR family pyrin domain containing 3 (NLRP3) inflammasome in macrophages. These inhibitions were achieved through the activation of the peroxisome proliferator-activated receptor gamma (PPARγ) pathway. The results of this study provide a novel insight into the mechanism of action of probiotics in the prevention and treatment for allergic disorders through the moonlighting GAPDH protein.

A Case-Control Study Comparing the General Characteristics of Patients with Symptomatic Dermographism and Chronic Spontaneous Urticaria: Is Atopy a Risk Factor for Symptomatic Dermographism?

Introduction: Symptomatic dermographism (SDerm) is the most common chronic inducible urticaria (CIndU) subtype. There is still limited information in the literature about clinical features, triggering factors, and accompanying comorbidities of SDerm. The aim of this study was to compare the clinical features and laboratory data of patients with SDerm and chronic spontaneous urticaria (CSU). Methods: The clinical features and laboratory data of patients with SDerm and CSU were compared retrospectively. The laboratory data and general characteristic features of the patients were obtained from the medical records. Results: The study included a total of 361 patients (CSU: 220, SDerm: 141). The rates of asthma (odds ratio [OR]: 1.79, p = 0.036), allergic rhinitis (OR: 6.03, p < 0.001), and thyroid disease (OR: 1.78, p = 0.039) were higher in patients with SDerm. The disease duration (median 12 months, p < 0.001) and regular antihistamine use (OR: 0.31, p < 0.001) were lower in patients with SDerm. Total IgE level (median: 193, p < 0.001), thyroid antibody positivity (OR: 1.93, p = 0.039), and atopy (OR: 8.81, p < 0.001) were higher in patients with SDerm. Dermatophagoides pteronyssinus (OR: 17.72, p < 0.001), Dermatophagoides farinae (OR: 17.20, p < 0.001), grass pollen (OR: 2.50, p < 0.026), cat epithelium (OR: 3.68, p < 0.023), and cockroach (OR: 4.93, p < 0.009) allergen positivity rates were higher in patients with SDerm. Conclusion: Atopic diseases such as asthma and allergic rhinitis and the sensitization rate to aeroallergens seem to be higher in patients with SDerm than in patients with CSU. The results of this study should be supported by multicenter studies of patients from different geographical regions.

Efficacy and Safety of Sublingual Immunotherapy using a Combination of Dermatophagoides Pteronyssinus and Blomia Tropicalis Extracts in Patients with Allergic Rhinitis: A Randomized, Double-Blind, Placebo-Controlled Trial

The aim of this study was to evaluate the safety and efficacy of sublingual immunotherapy (SLIT) with extracts of Dermatophagoides pteronyssinus and Blomia tropicalis in patients with moderate/severe persistent allergic rhinitis (AR). Methods: This was a double-blind, randomized, placebo-controlled trial of SLIT for the treatment of house dust mite-induced AR. A total of 65 patients, aged between 12 and 60 years, were treated for 12 months and randomized into two groups: SLIT and placebo. The SLIT group received a combination of extracts containing Dermatophagoides pteronyssinus and Blomia tropicalis allergens. All had their sensitization confirmed by skin prick test or serum-specific IgE. Total Nasal Symptom Score, RQLQ quality of life questionnaire, current treatment, and need for medication to control symptoms were recorded during the study. Total serum IgE, serum specific IgE, and IgG4 levels to Der p 1 and Blo t were assessed at baseline, 6 and 12 months after treatment. Results: There was no significant difference in the number of adverse events between groups. The SLIT group showed a significant reduction in the consumption of antihistamines to control symptoms (p &lt; 0.0001) when compared to placebo. There was no significant change in serum total IgE, serum specific IgE, and IgG4 to both allergens when comparing the SLIT and placebo groups. Conclusion: After one year, SLIT using a dose of 1 mcg of Der p 1/day and 753 UBE of Blo t/day proved to be effective and safe in controlling AR exacerbations.

Removal of N-Terminal Peptide Impacts Structural Aspects of an IgE-Reactive Recombinant Der p 5

(1) Background: Modification of the structural elements of allergens is widely used in the field of allergies. The goal of the present research was to express, purify, and characterize the shortened recombinant group 5 allergen of Dermatophagoides pteronyssinus (rDer p 5). (2) Methods: rDer p 5 storage stability and aggregation capacity were explored through in silico analysis, dynamic light scattering (DLS), and SDS-PAGE. Serum IgE reactivity and cytokine amount were investigated in sera or cell culture supernatants through ELISA, MULTIPLEX®, and Western blot analysis using sera from sensitized humans from Brazil, Colombia, and Ecuador. (3) Results: Dimeric rDer p 5 was detected through native PAGE, and this result was confirmed by data from DLS. The protein was thermically stable, as it did not degrade at 4 °C for 21 days. The shortened rDer p 5 was classified as a major IgE allergen in Brazil and Colombia, but minor in Ecuador. IL-13, IL-10, IL-1β, and IL-6 were significantly elevated in the sera of rDer p 5-reactive patients. The same cytokines plus IL-5 were more secreted by human cells upon rDer p 5 stimulation. (4) Conclusions: N-terminal peptide deletion led to a higher rDer p 5 folding stability, which, even though dimeric, was an IgE-reactive protein. Therefore, rDer p 5 could be used for molecular diagnostic applications or as backbone for hypoallergen design.

Tablet-lyophilizate containing a mixture of house dust mites usage for skin prick testing to confirm house dust mites sensitization

BACKGROUND: Since 2021, the production of water-salt allergens for skin allergy tests with household allergens has been suspended, which has led to the lack of an objective opportunity for skin prick tests in medical and preventive institutions in the Russian Federation. Quantitative determination of allergen-specific immunoglobulin class E in the blood serum by the ImmunoCAP method is currently difficult because of the lack of supplies in the laboratory.&#x0D; AIMS: To assess the possibility of using a drug solution of a lyophilizate mixture of house dust mites (Acarisax) containing house dust allergens Dermatophagoides pteronyssinus and Dermatophagoides farinae in equal amounts 12 SQ-HDM for the skin hood tests to confirm the sensitization of patients with mites of home dusts.&#x0D; MATERIALS AND METHODS: Skin prick tests were conducted with dilutions of one tablet per 2.5 and 5 mL of diluting liquid of the study drug with positive and negative controls. Statistical analysis was conducted using Statistica 13.3 or IBM SPSS 17.0. To analyze efficacy and safety, standard parametric tests were used: Students t-test for dependent/independent samples, analysis of variance for repeated measurements, MannWhitney U-test, and ShapiroWilk test. The analysis of AEs was based on an estimate of the frequency of undesirable phenomena/serious undesirable phenomena.&#x0D; RESULTS: The correlation was estimated between the total level of specific IgE to tick-borne allergens D. pteronyssinus and D. farinae measured during screening and the papule size 20 min after applying the drug in breeding with one tablet per 2.5 mL and 5 mL of a diluting liquid. The correlation coefficients (R) in both groups (total) were 0.735 (high correlation) and 0.762 (high correlation). The result was statistically significant (p 0.05) in both groups. The clinical test showed that the lyophilizate drug, a mixture of domestic dust mites containing dermal dust allergens D. pteronyssinus and D. farinae in equal quantities 12 SQ-HDM is a safe and informative method when used in skin hood tests to confirm the sensitization of patients to domestic dust ticks.&#x0D; CONCLUSIONS: For skin hood tests, to confirm the sensitization of patients to domestic dust ticks, the studied drug can be used in greater dilution one tablet per 5 mL of the diluting fluid. The lyophilizate mixture of dust mites containing D. pteronyssinus and D. farinae homemade allergens 12 SQ-HDM is safe and well tolerated by patients when used in skin test tests.

The importance of specific IgE antibodies in the epidemiology of allergic rhinitis and asthma (ECAP survey): part four. The relationship between the concentration of specific IgE antibodies in serum and types of asthma.

Specific immunoglobulins E (sIgE) are important parameters for the estimation of severity of allergic diseases. To determine the relationship between the concentration of specific IgE antibodies in serum and types of asthma. The concentration of sIgE antibodies against allergens Dermatophagoides pteronyssinus, cat dander, timothy grass, and Alternaria alternata were determined in the serum of 4077 respondents randomly selected from 8 regions (ECAP study). The positive results of sIgE (≥ 0.35 IU/ml or ≥ 0.7 IU/ml) were correlated to clinical diagnosis (types of asthma, skin-prick tests). sIgE antibodies against any allergen were detected in 9.9% (classes 1-6)/7.6% (classes 2-6) of healthy respondents. Comparing sIgE antibodies of respondents with intermittent asthma to sIgE antibodies of respondents with persistent asthma, no statistically significant differences were identified. Relating to allergens of D. pteronyssinus, cat dander, and A. alternata, sIgE antibodies were more frequently detected in respondents with atopic asthma and a negative skin-prick test as compared to healthy respondents with a negative skin-prick test (p < 0.005 to p < 0.001). Relating to allergens of D. pteronyssinus, cat dander, and timothy grass, sIgE antibodies were more frequently detected in respondents with atopic asthma and a weakly positive skin-prick test as compared to healthy respondents with weakly positive skin-prick test (p < 0.05 to p < 0.001). Regarding subjects with a negative or weakly positive skin test, when sIgE antibodies to the same allergen are detected, asthma is much more likely to occur.

The importance of specific IgE antibodies in the epidemiology of allergic rhinitis and asthma (ECAP survey): part five. The relationship between the concentration of specific IgE antibodies in serum and types of rhinitis.

Specific immunoglobulins E (sIgE) are important parameters for the estimation of severity of allergic diseases. To determine the relationship between the concentration of specific IgE antibodies in serum and types of rhinitis. The concentration of sIgE antibodies against allergens Dermatophagoides pteronyssinus, cat dander, timothy grass, and Alternaria alternata were determined in the serum of 4077 respondents randomly selected from 8 regions (ECAP study). The positive results of sIgE (≥ 0.35 IU/ml or ≥ 0.7 IU/ml) were correlated to clinical diagnosis (types of rhinitis, skin-prick tests). sIgE antibodies are more frequently detected in respondents with intermittent/seasonal allergic rhinitis and a negative skin-prick test as compared to healthy respondents with a negative skin-prick test (p < 0.05 to p < 0.001). Relating to allergens of D. pteronyssinus and cat dander, sIgE antibodies are more frequently detected in respondents with persistent/perennial allergic rhinitis and a negative or weakly positive skin-prick test as compared to healthy respondents with a negative or weakly positive skin-prick test (p < 0.05 to p < 0.001). The occurrence of intermittent/seasonal allergic rhinitis is much more probable in respondents with a negative skin-prick test, when IgE antibodies against the same allergen are detected. And the occurrence of persistent/perennial allergic rhinitis is much more probable in respondents with a negative or weakly positive skin-prick test with allergens of D. pteronyssinus or cat dander, when IgE antibodies against the same allergen are detected.

Clinical and Epidemiological Features of Allergies in Schoolchildren of the Chechen Republic: Results of an Epidemiological Study

Background. One of the topical issues of modern medicine is the widespread increase in the incidence of allergic pathology. Objective. The aim of the study is to investigate clinical and epidemiological features of allergies in schoolchildren of the Chechen Republic. Methods. We performed clinical and diagnostic verification of allergies according to clinical guidelines in patients who positively answered the ISAAC questionnaire (n = 80) — 46 first-grade and 34 eighth-grade pupils. The first diagnostic step on studying the clinical manifestations in observed children was the history taking, complaints, physical examination. Laboratory and instrumental diagnosis included: IgE total, detection of skin sensitivity to allergens via standard pollen, domestic, epidermal, and food allergens (produced by I.M. Mechnikov Research Institute of Vaccines and Sera, Stavropol Research Institute of Vaccines and Sera), revealing of allergen-specific antibodies, component-divided allergen diagnosis via indirect immunofluorescence with ImmunoCAP analyzer, respiratory function evaluation with CareFusion, nitric oxide level detection in exhaled air via portable NObreath analyzer. If needed, specialised medical consultation were performed. Results. Asthma was detected in 37 (46.3%) children, allergic rhinitis (AR) — in 37 (46.3%), atopic dermatitis (AD) — in 5 (6.3%) according to the results of our study. The most common combination of medical conditions in examined patients was combination of asthma and AR. There were no statistically significant differences in age groups. No gender differences were reported in the study. The highest incidence of asthma and AR was revealed in city residents (p &lt; 0.05), while the AD incidence in the city and rural areas did not differ significantly. The sensibilization spectrum analysis in schoolchildren of the Chechen Republic revealed predominance of sensitivity to pollen (ragweed, herd grass) and domestic allergens (Dermatophagoides pteronyssinus, Dermatophagoides farinae). Sensitization to food allergens in children of the Chechen Republic did not have significant differences in the frequency of occurrence of individual allergens, that is, unlike pollen and household sensitization, a regional rating on the dominance of certain food allergens was not obtained. Sensibilization to epidermal allergens was also less common, the predominant one was the cat allergen. Conclusion. Thus, the first study on the territory of the Chechen Republic carried out according to the ISAAC program has revealed high prevalence of allergies in schoolchildren. Moreover, regional features of sensibilization structure have been identified. It will allow us to maintain economically optimal diagnosis of allergic diseases in children living in the Chechen Republic.

Effect of ozone on allergic airway inflammation.

Exposure to ozone (O3) is associated with increased risk of exacerbations of asthma, but the underlying mechanisms are not well studied. We sought to determine whether O3 exposure would enhance airway inflammatory responses to allergen and the GSTM1-null genotype would modulate this enhancement. In a crossover design, 10 asthmatic participants (5 with GSTM1-null genotype) who had specific sensitization to Dermatophagoides pteronyssinus (DP) were exposed to 160 ppb O3 or filtered air (FA) control for 4 hours on 2 separate days at least 3 weeks apart. At 20 hours after exposure, endobronchial challenge with DP allergen, and sham normal saline (NS) instillation, were performed in separate bronchi. Six hours later, a second bronchoscopy was performed to collect bronchoalveolar lavage (BAL) from the DP- and NS-challenged segments for analyses of inflammatory biomarkers. Linear regression compared cell and cytokine responses across the 4 exposure groups (FA-NS, O3-NS, FA-DP, O3-DP). Effect modification by GSTM1 genotype was assessed in stratified regressions. BAL eosinophil counts were increased in segments challenged with DP compared to sham-challenged segments (P < .01). DP challenge compared to sham also caused a significant increase in BAL concentrations of the TH2 cytokines IL-4, IL-5, IL-10, and IL-13 (P < .03 for all comparisons). O3 exposure did not significantly affect BAL cells or cytokine after DP challenge. Compared to GSTM1-present participants, GSTM1-null participants had significantly lower eosinophil (P < .041) and IL-4 (P < .014) responses to DP challenge after O3 exposure. While O3 did not cause a clear differential effect on airway inflammatory responses to allergen challenge, those responses did appear to be modulated by the antioxidant enzyme, GSTM1.

Specific IgE against the house dust mite allergens Der p 5, 20 and 21 influences the phenotype and severity of atopic diseases.

House dust mites (HDM) are among the most important sources for airborne allergens with high relevance for atopic diseases. Routine tests contain only 4 of 32 registered allergens of Dermatophagoides pteronyssinus. Clinical relevance and pathomechanistic properties of many allergens are not well understood. The association of several HDM allergens with allergic rhinitis, allergic asthma, and atopic dermatitis was investigated to identify allergens with biomarker potential and to transfer them into diagnostics. Eight out of nine D. pteronyssinus allergens (nDer p 1, rDer p 2, rDer p 5, rDer p 7, rDer p 10, rDer p 13, rDer p 20, rDer p 21, rDer p 23) were recombinantly expressed and purified. Sensitization patterns of 384 HDM-allergic individuals exhibiting different clinical phenotypes were analyzed with a serum-saving multiplex array. Sensitization to more than three mite allergens (sensitization count) was associated with allergic asthma and/or atopic dermatitis. Reactions to Der p 5 and Der p 21 were more frequent in allergic asthma compared to allergic rhinitis. Atopic dermatitis patients were more often sensitized to Der p 5, Der p 20, and Der p 21 among others. Der p 20-IgE > 80 kU/L was associated with severe atopic dermatitis in 75% of patients. This study demonstrates the clinical importance of the sensitization count and of certain allergens (Der p 5, Der p 20, and Der p 21) not available for routine diagnostics yet. Implementing them as well as the sensitization count in diagnostic measures will improve diagnosis and risk assessment of HDM-allergic patients.

Allergens from house dust and storage mites

SummaryHouse dust mites are among the most important allergy triggers worldwide. While mites of the genus Dermatophagoides occur almost worldwide, the tropical mite Blomia tropicalis and storage mites are only of importance for certain areas or groups of people. The most important allergens of Dermatophagoides pteronyssinus are Der p 1, Der p 2, and Der p 23 with immunoglobulin E (IgE)-binding frequencies of more than 70% and high allergenic activity. Also of importance are Der p 5, Der p 7, and Der p 21, which have IgE-binding frequencies of about 30%. According to the current state of knowledge, these six allergens are the allergens of clinical relevance which are also required for diagnosis and immunotherapy with individual components.

Allergy to Der p 23 influences the cytokine profile in patients with allergic asthma – a preliminary study

Objective Der p 23 is a major allergen of Dermatophagoides pteronyssinus, which could contribute to allergic asthma. The study compared the cytokine profile (Il-1beta, Il-4, Il-5, Il-6, Il-13, Il-17, TNF-alpha) in patients with allergic asthma, with confirmed allergy to D. pteronyssinus and with the presence or absence of allergy to Der p 23. Methods Among 173 included patients, the following combinations were analyzed: profile A – Der p 1 (+), Der p 2 (+), and Der p 23 (−) observed in 38 (22%) patients; profile B – Der p 1 (+), Der p 2 (+), and Der p 23 (+) in 87 (50.3%) patients; and profile C – Der p 1 (−), Der p 2 (−), and Der p 23 (+) in 15 (8.7%) patients. Results The mean concentration of Il-1beta was significantly lower in profile A than in profiles B and C: 10.51 ± 5.22 (pg/ml) vs. 21.92 ± 11.34 vs. 23.1 ± 8.56 (A vs. B for p = 0.03 and A vs. C for p = 0.019). Similar trends were observed for Il-5: 38.5 ± 10.45 (pg/ml) vs. 94.8 ± 54.11 vs. 103.61 ± 34.9 (A vs. B for p = 0.008 and A vs. C for p = 0.001). Conclusion The higher Il-1 and Il-5 activities observed in profiles B and C with Der p 23 (+) could be responsible for the more effective acceleration of allergic inflammation than in profile A with Der p 23.

Effect of Dermatophagoides pteronyssinus extract on the expression of genes involved in inflammation and tissue remodeling by peripheral blood mononuclear cells of allergic asthma patients

PurposeHouse dust mite allergy constitutes a risk factor for asthma development and is associated with a faster decline of lung function in allergic asthmatics (AAs).To evaluate the effect of Dermatophagoides pteronyssinus (Dp) allergens on the expression of genes involved in inflammation and tissue remodeling by peripheral blood mononuclear cells (PBMCs) isolated from the blood of AAs. Materials and methodsThe cells from AAs, allergic rhinitis without asthma patients (ARs), and healthy controls (HCs) were cultured in the presence of Dp, lipopolysaccharide (LPS), or without any stimulation. The expression of 84 genes was evaluated using a low-density microarray whereas, the quantitative expression analysis of selected genes was performed using a real-time polymerase chain reaction. The concentration of selected proteins in the cell culture supernatants was assessed using ELISA. ResultsStimulation of PBMCs with Dp and LPS resulted in a significant upregulation of 8 and 15 among 84 studied genes, respectively. The greatest upregulation was observed for CCL2 and CCL3 using Dp and LPS, respectively. In comparison with HCs, in AAs, significantly increased upregulation of CCL2 in response to Dp was found. The secretion of CCL2 and CCL3 by PBMCs reflected the pattern of gene expression at the mRNA level. The mean Dp-stimulated secretion of CCL2 by PBMCs of ARs was less than in AAs (p ​< ​0.01), both being notably greater than in the HCs (p ​< ​0.01). ConclusionRapid and potent upregulation of CCL2 expression by PBMCs in response to Dp may constitute an important contribution to the development of allergic asthma.

Identification of an immunodominant IgE epitope of Der p 39, a novel allergen of Dermatophagoides pteronyssinus

BackgroundHouse dust mites (HDMs) are the main source of indoor inhalatory allergens that cause IgE-mediated allergic diseases. The discovery and identification of HDM allergens are important for the diagnosis and treatment of allergic diseases. ObjectiveWe sought to identify a Group 39 Dermatophagoides pteronyssinus (Der p) allergen, namely Der p 39, and explore its immunodominant IgE epitopes. MethodsHomology analysis of amino acid (aa) sequences in HDM and human troponin C (TnC)-like protein was performed. Total RNA of Der p was extracted and used to amplify Der p 39 cDNA with specific primers. Recombinant Der p 39 protein was expressed with a pET-His prokaryotic expression system and purified with Ni-NTA resins. IgE binding was evaluated with western blot, dot blot, and enzyme-linked immunosorbent assay (ELISA) experiments. The IgE binding epitopes of Der p 39 were identified by observing HDM-allergic sera interactions with truncated and hybrid proteins formed from Der p 39 and human TnC-like proteins. ResultsThe Der p 39 open reading frame (ORF) cDNA was found to be 462 base pairs and registered in the NCBI library (GenBank no. MZ336019.1). Der p 39, which encoded 153 aa, was found to have 35.63% and 99.35% homology with human TnC and Dermatophagoides farina (Der f) 39, respectively. IgE-ELISA showed IgE binding with expressed and purified recombinant Der p 39 (18 kDa) in 5/87 (5.75%) HDM-allergic sera samples. Analyses of IgE binding with Der p 39-based truncated and hybrid proteins indicated that IgE binding epitopes are likely located in the C-terminal region and dependent on conformational structure. The data from this study were submitted to the World Health Organization and International Union of Immunological Societies (WHO/IUIS) Allergen Nomenclature database. ConclusionDer p 39 was identified as a minor HDM allergen with a conformational IgE binding epitope. These findings could have important theoretical implications in the development of HDM allergy diagnostics and therapeutics.

Component-Resolved Diagnosis for Endotyping Patients with Chronic Rhinosinusitis with Nasal Polyps

Background: Chronic rhinosinusitis with nasal polyps (CRSwNP) is a predominantly type 2 inflammation-mediated disease of the nasal mucosa and paranasal sinuses that often coexists with asthma. The role of atopy in the development and severity of CRSwNP is still a controversial issue. Objective: The aim of our study was to propose a systematic allergy workup to identify atopic patients in the context of CRSwNP and to characterize their allergen sensitization profile (sources/molecules). Methods: Patients with a diagnosis of CRSwNP (n = 97) were studied in the otorhinolaryngologist and allergy settings. Demographic and clinical data were collected for each patient. Different allergen sensitization profiles (sources/molecules) were evaluated in atopic CRSwNP patients by using component-resolved diagnosis (CRD). Results: In our cohort of patients, the CRSwNP was frequently diagnosed during adulthood with significant impact on health-related quality of life. Asthma and atopy were the most common comorbidities with a prevalence of asthma in the atopic group. In CRSwNP patients sensitized to grass pollens and/or to house dust mites, the CRD analysis revealed a prevalence of sensitization to species-specific allergens of Phleum pratense (Phl p1, Phl p2, and Phl p5) or Dermatophagoides pteronyssinus (Der p1 and Der p2) rather than to cross-reactive ones. Conclusion: To define the allergen sensitization profile in atopic CRSwNP patients by CRD, it may be useful to better characterize type 2 inflammation, thus providing a personalized endotype-driven treatment.

Rationally designed hypoallergenic mutant variants of the house dust mite allergen Der p 21

BackgroundAllergic diseases figure among the most common immune-mediated diseases worldwide, affecting more than 25% of the world's population. Allergic reactions can be triggered by house dust mite (HDM) allergens, of which the so-called group 21 of allergens is considered as clinically relevant. MethodsHerein, we used a structural bioinformatics and immunoinformatics approach to design hypoallergenic mutant variants of the Der p 21 allergen of Dermatophagoides pteronyssinus, which were then recombinantly expressed in bacteria and tested for their IgE-reactivities. For this, we scanned the wild-type Der p 21 protein for all possible single amino acid substitutions in key IgE-binding regions that could render destabilization of the major epitope regions. ResultsFour main substitutions (D82P, K110G, E77G, and E87S) were selected to build mutant variants of the Der p 21 allergen, which were produced in their recombinant forms; two of these variants showed reduced reactivity with IgE. Molecular dynamic simulations and immune simulations demonstrated the overall effects of these mutations on the structural stability of the Der p 21 allergen and on the profile of immune response induced through immunotherapy. ConclusionsWhen produced in their recombinant forms, two of the Der p 21 mutant variants, namely proteins K110G and E87S, showed significantly reduced IgE reactivities against sera from HDM-allergic individuals (n = 20; p < 0.001). General significanceThis study successfully translated a rational in silico mutagenesis design into low IgE-binding mutant variants of the allergen rDer p 21. These novel hypoallergens are promising to compose next-generation allergen-immunotherapy formulations in near future.

Place of residence, gender, month of birth and age influence on the concentration of specific IgE antibodies in serum. The Epidemiology of Allergic Diseases in Poland (ECAP survey): part three.

IntroductionSpecific immunoglobulins E (sIgE) are important parameters to estimate the severity of allergic diseases.AimTo determine the influence of gender, age and place of residence on the concentration of specific IgE antibodies in serum.Material and methodsThe concentration of sIgE antibodies against allergens of Dermatophagoides pteronyssinus, cat dander, timothy grass, Alternaria alternata were determined in serum of 4077 respondents randomly selected from 8 regions (ECAP study). The positive results of sIgE (≥ 0.35 IU/ml) were correlated with answers to ECAP questionnaires.ResultsIgE antibodies are less frequently detected in respondents living in the country (p < 0.05 to p < 0.005); and they are less frequently detected in respondents living in cities of hardly industrialised regions than in respondents living in cities of industrialised regions (D. pteronyssinus p < 0.05, cat dander p < 0.01). IgE antibodies are more frequently detected in men (p < 0.005 to p < 0.001). IgE antibodies are more frequently detected in respondents born in May-July (timothy grass p < 0.005, Alternaria alternata p < 0.005) and less frequently detected in respondents born in August-October (timothy grass p < 0.005, Alternaria alternata p < 0.05).ConclusionsCharacteristics of the urban environment, non-specific factors and gender affect the concentration of specific IgE antibodies in serum. Influence of seasonal allergens within the first months of life and/or in the course of the foetal period alters the production of specific IgE antibodies.