Serum Specific IgE Antibodies Research Articles

The Hydrogel Based Allergen-Coated Gold Nanoparticles for Topical Administration: A Possible Epicutaneous Immunotherapy in Pollen-Sensitized Mice?

ABSTRACT Background The rapid uptake of antigens by antigen-presenting cells (APCs) and their migration to draining lymph nodes in the initial hours after antigen administration in epicutaneous allergen specific immunotherapy (EPIT) prompted us to investigate whether the topical administration of allergens without patch application could alleviate allergy in pollen-sensitized mice. We evaluated the immunotherapeutic effect of topically administering hydrogel-based Gold nanoparticles (AuNPs) loaded with a total extract of Platanus orientalis pollen (Pla. ext (50 μg)-AuNPs) on intact skin. Methods Mice sensitized to P. orientalis pollen were divided into three groups and treated with Pla. ext (50 μg)-AuNPs: 1) patch with Pla. ext (50 μg)-AuNPs, 2) patch with Pla. ext (50 μg)-AuNPs in combination with hydrogel, and 3) topical application of Pla. ext (50 μg)-AuNPs in combination with hydrogel. The immunotherapeutic effects were evaluated by measuring serum specific and total IgE antibodies, total cell and eosinophil count in nasopharyngeal lavage fluid, cytokines in the supernatants of re-stimulated splenocytes by the total extract, and histological examination of lung and nasal mucosa. Results Topical administration of Pla. ext (50 μg)-AuNPs, like patch-based administration, significantly downregulated specific and total IgE and IL-4 production, promoted secretion of IFN-γ and IL-10, markedly reduced the number of inflammatory cells, particularly eosinophils, in nasopharyngeal lavage fluid (p < .05), and inhibited inflammation and pathological damage in lung and nasal mucosa. Conclusion Our results suggest that topical administration of AuNPs loaded with P. orientalis total pollen extract on intact skin could be a potential application for EPIT in the P. orientalis pollen -sensitized mice.

Prediction of Food Sensitization in Children with Atopic Dermatitis Based on Disease Severity and Epidermal Layer Impairment

Plain Language SummaryThe study has revealed notable associations between food sensitizations (FSs) and the disease severity as well as epidermal barrier impairment in children with atopic dermatitis (AD). Specifically, the findings indicate that children with AD who were sensitized to higher numbers of food allergens (FAs) had higher SCORAD scores and greater epidermal barrier impairment. In addition, SCORAD score and stratum corneum hydration (SCH) levels at lesional skin could be predictive factors for FS in children with AD. The findings of this study suggest that clinicians should consider screening for food sensitization in children with moderate-severe AD and/or low SCH levels at lesional skins.

Preanalytical temperature and storage stability of specific IgE antibodies in serum

Allergen-specific serum immunoglobulin E (sIgE) levels are a cornerstone in allergy diagnostics. While immunoglobulins are known to be relatively stable molecules, the influence of preanalytical factors on the stability of sIgE is not thoroughly investigated. We studied the effect of several preanalytical factors: (1) delayed centrifugation of serum samples in tubes with separating gel for 10, 24 and 48 h, (2) prolonged storage at 5 °C for 3, 7, 10 and 14 days, (3) storage tube type (primary tube with separating gel or secondary tube), (4) repeated freeze-thawing cycles, and (5) prolonged storage at −20 °C for 4 and 8 weeks. We found that sIgE is stable at room temperature for 48 h before centrifugation and for 10 days at 5 °C after centrifugation. There was no effect of the separating gel after storing serum for 1 week in the freezer. However, storage for 4–8 weeks, and introducing more than one freeze-thaw cycle resulted in a larger variation of sIgE levels. In conclusion, we found that sIgEs are stable under various preanalytical conditions, which allows for flexible handling of samples for a comprehensive portfolio of sIgE analyses.

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.

P501 Aspergillus fumigatus complicates one third of the patients with suspected bronchial asthma or pulmonary tuberculosis: Clinical validation of indigenously developed diagnostic kits

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PMObjectives Aspergillus fumigatus, an opportunistic fungus, causes complications in about 5%-20% of bronchial asthma and about 26% of pulmonary tuberculosis patients. Detection of Aspergillus fumigatus specific IgG and IgE antibodies in the patient serum is an excellent tool to screen for Aspergillus sensitization early on to employ anti-fungal drugs in the clinical management to stall the progression of lung fibrosis.MethodsNovel indigenous AfuPEPLISA assays were developed for the detection of specific IgG and IgE, based on the 12 amino acid long synthetic peptide epitope of Asp f1, an 18 kDa major allergen/antigen. The novel diagnostic kits were manufactured at a licensed GMP facility under a test license. Independent validation of the kits was pursued at PGIMER and VPCI hospitals in suspected bronchial asthma patients (n = 1307), and the diagnostic efficiency was compared with currently used ImmunoCAP assay.ResultsThe diagnostic specificity and sensitivity were found to be 95.7% and 89.8%, respectively, for IgG; and 94.2% and 70%, respectively for IgE AfuPEPLISA, and were not significantly different from ImmunoCAP assay. Screening of the suspected patients of pulmonary tuberculosis (PTB) at RBIPMT Hospital for the presence of A. fumigatus specific IgG and IgE antibodies was pursued using AfuPEPLISA kits. A total of 82 out of 254 suspected PTB patients (32.3%) were seropositive in agreement with the previous reports.See Figures 1 and 2 below.ConclusionThe study inferred that indigenously developed AfuPEPLISA kits are an economically viable option to integrate in the clinical management of patients with suspected bronchial asthma or PTB for efficient diagnosis of Aspergillus sensitization.

Estimating allergic sensitization to some mold fungi

The assessment of allergic sensitization of 894 patients (478 and 416 of them examined in 2018 and 2019, respectively) who were examined at the consultative and Diagnostic center of G.N. Gabrichevsky Scientific Research Institute for Epidemiology and Microbiology, Moscow, for some mold fungi was carried out by measuring serum specific IgE antibody level by using a highly specific and highly sensitive immunoblot method (RIDA AllergyScreen test systems, R-Biopharm AG, Germany). The prevalence of patients with elevated levels of specific IgE antibodies to mold fungi Alternaria alternata, Aspergillus fumigatus, Cladosporium herbarum and Penicillium notatum was found. All the examined patients at each time point was divided into two age groups: group 1 children aged 1 to 17 years, 548 children within the two-year period (303 of them girls and 245 boys), group 2 adults aged at least 18 years old, 346 subjects within the same period (231 of them women and 115 men). Results. In 2018, out of 478 serum samples, atopic sensitization to mold fungi was detected in 25.3% (n = 121) of the examined subjects, of which 17.9% (n = 86) and 7.4% (n = 35) was found in group 1 and group 2, respectively. In 2019, out of 416 serum samples, atopic sensitization to mold fungi was detected in 20.7% (n = 86) of the examined subjects, of which 13.5% (n = 56) and 7.2% (n = 30) was found in group 1 and group 2, respectively. In 2018, out of 121 positive samples, specific IgE antibodies to various mold fungi were slightly more often detected in females 53.7% (n = 65), of which 75.3% (n = 49) and 24.7% (n = 16) were girls and women, respectively. In contrast, in 2019, out of 86 positive samples, specific IgE antibodies to various mold fungi were slightly more often detected in males reaching 51.2% (n = 44), of which 79.5% (n = 35) and 20.5% (n = 9) were boys and men, respectively. Atopic sensitization to the mold fungus Alternaria alternata was most often detected: during the study period, out of 207 positive samples, it was found in 186 samples (89.9%). Serum IgE antibodies to mold fungi are more often detected in children under 17, in sex-independent manner.

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.

Comparative analysis of the allergenic characteristics and serodiagnostic potential of recombinant chitinase-like protein-5 and -12 from Sarcoptes scabiei

BackgroundScabies is caused by burrowing of the mite Sarcoptes scabiei into the stratum corneum. Currently, diagnosis via routine skin scraping is very difficult, and information on the allergenic identification of S. scabiei remains limited.MethodsWe performed comparative analysis of the serological diagnostic potential of recombinant S. scabiei chitinase-like protein-5 (rSsCLP5) and recombinant S. scabiei chitinase-like protein-12 (rSsCLP12) by measuring the levels of serum-specific IgG and IgE antibodies (Abs) as diagnostic markers. In addition, the allergenic characteristics of rSsCLP5 and rSsCLP12 were evaluated using IgE-binding experiments and skin tests.ResultsThe IgE Abs-based indirect enzyme-linked immunosorbent assay (ELISA) methods showed high sensitivity and specificity: the rSsCLP5-based assay had 93.5% sensitivity and 94.4% specificity; the rSsCLP12-based assay had 100% sensitivity and 98.1% specificity. The specific IgE Abs in infested mouse sera could bind rSsCLP5 and rSsCLP12. In skin tests, rabbits in the rSsCLP5 and rSsCLP12 groups and positive control (histamine) groups exhibited allergic reactions. Most test sites in the rSsCLP12 group had edema, bleeding spots, and even ulcers or scabs, but such allergy symptoms were rare in the rSsCLP5 group. Moreover, the allergic history rabbit group had more severe allergic reactions and lower levels of IgE Abs compared to the healthy rabbit group in the same protein group.ConclusionsThese findings validate the use of IgE Abs to rSsCLP5 and rSsCLP12 as potentially useful markers for diagnosing scabies. Moreover, both rSsCLP5 and rSsCLP12 have allergenic properties, and the potential allergen rSsCLP12 is a stronger allergen than rSsCLP5.Graphical

Пищевая и ингаляционная сенсибилизация при атопическом дерматите

Aim: to study the association of food and inhalation sensitization with the severity of atopic dermatitis (AtD) in order to improve its treatment and prevention methods. Patients and Methods: the study included 50 patients with AtD from 2 to 44 years old. The study included clinical and laboratory research methods. The survey analyzed the history and anamnesis vitae, estimated the age at the AtD onset, determined the nature of concomitant pathology, the possible causes of cutaneous exacerbation, the allergic severity in the family history, the severity score on the SCORAD (Scoring of Atopic Dermatitis) scale and the efficacy of previous therapy. The immunological examination consisted of the quantitative determination of specific IgE and IgG antibodies in the blood serum to inhaled and food allergens by immunochemiluminescence assay on an IMMULITE 2000 XPi analyzer. Results: according to the detection frequency of IgE to food allergens in patients with AtD: the leading positions were taken by the chicken egg (egg white in 26% of cases, egg yolk in 20%); IgE to banana was detected in 17% of cases; in 12% — to cow’s milk, more common — to the β-lactoglobulin fraction. When studying the activity of inhaled allergens, respiratory sensitization to the cat epithelium was detected in 26% of cases, and to black birch — in 24%. Hypersensitivity to herbs (wormwood, herbal mixture) and house dust allergens (8%, respectively) was found much less common. In severe AtD, IgE to respiratory allergens prevailed: at n=38, the IgE concentration to cat epithelium and black birch was 12.4±1.4 IU/mL each. Besides, there was a significant sensitization to food allergens, namely, the IgE concentration to egg white — 5.4±0.9 IU/mL. Conclusion: the study revealed the sensitization of patients with AtD to food (mainly egg white, egg yolk, banana and cow’s milk) and respiratory (mainly cat epithelium, black birch) allergens. Determination of the IgE concentration in the blood serum of patients showed that there was a significant concentration increase to egg white, cat and black birch in severe AtD. Therefore, it is advisable to test for specific allergens in all patients with moderate to severe AtD. KEYWORDS: food sensitization, inhalation sensitization, atopic dermatitis, allergy, allergens, antibodies, immunochemiluminescence assay, IgE, triggers. FOR CITATION: Vasilyeva E.A., Rusak Yu.E., Efanova E.N., Lebedeva L.N. Food and inhalation sensitization in atopic dermatitis. Russian Medical Inquiry. 2021;5(1):17–20. DOI: 10.32364/2587-6821-2021-5-1-17-20.

Validation study of a new chemiluminescent singleplex IgE assay in a set of Italian allergic rhinitis patients.

The measurement of specific IgE to allergenic extracts and molecules in patients with allergic rhinitis (AR) is crucial for a precise diagnosis and further immunotherapy. Companies providing in vitro diagnostic methods in allergology continuously strive for the optimization and modernization of such methods. A new generation of automated allergy tests based on chemiluminescence detection and paramagnetic microparticles is now available, with possible advantages in sample volume, cost-effectiveness and avoidance of sample-related interference. To test whether sIgE antibody levels obtained with a new singleplex chemiluminescent method have a good agreement with the corresponding results obtained with a "gold standard" test. We tested sera from 368 AR patients. Specific IgE sera levels (kU/L) to a comprehensive panel of 15 allergen extracts and 6 molecules were tested with ImmunoCAP® (Thermo Fisher Scientific Inc, Phadia AB, Uppsala, Sweden) and NOVEOS™ (HYCOR® Biomedical, Garden Grove, CA, USA). We evaluated the qualitative and quantitative performance of the new NOVEOS system in matching the outcome of ImmunoCAP to each of the examined allergens. In relation to ImmunoCAP, the overall diagnostic sensitivity and specificity of sIgE tests with NOVEOS were 90.8% (95% CI=88.6-92.7) and 96.2% (95% CI=93.9-97.8), respectively. These values were higher when only molecules were considered (sensitivity=98.7% [95% CI=96.4%-99.7%]; specificity=94.2% [95% CI=88.4%-97.6%]) and lower when only extracts were considered (sensitivity=87.6% [95% CI=84.7%-90.2%]; specificity=97% [95% CI=94.4%-98.6%]). Spearman's correlation between the data set of both methods for a≥0.1kU/L cut-off was 0.84 (p<.001). The new singleplex NOVEOS system presented good results for qualitative and quantitative comparisons when testing specific serum IgE antibodies against a range of 21 allergens. This novel immunoassay system using only 4µl of sample per test appears to be robust and reliable and can, therefore, be used as an aid in allergy diagnosis.

Mannitol vs. methacholine in the evaluation of airway responsiveness in bakers' asthma.

Various indirect or direct airway challenge tests are used to measure nonspecific bronchial hyper-responsiveness (NSBHR). The evaluation of NSBHR in diagnosing occupational asthma (OA) is performed, e.g., to monitor the specific inhalation challenge test (SICT). The aim of this study was to preliminarily compare the results of methacholine and mannitol inhalation challenge tests in SICT monitoring in bakers with work-related airway symptoms. Four bakery workers with a suspicion of OA underwent single-blind placebo-controlled SICTs involving workplace allergens, accompanied by the evaluation of NSBHR with mannitol and methacholine, both before and after SICTs. Clinical examinations, spirometry tests, skin prick tests (SPTs) to common aeroallergens and occupational allergens, as well as tests to determine serum specific IgE antibodies to occupational aeroallergens were also performed. Positive SPTs results to occupational aeroallergens were found in all bakery workers, and specific IgE antibodies to flour were detected in 2 subjects. Three patients displayed positive SICT reactions. In all of these 3 patients, airway responsiveness to methacholine increased significantly. In 2 patients, airway reaction to mannitol was significant, whereas in 1 subject there was no increase in NSBHR after mannitol inhalation. The patient with a negative SICT result did not reveal any changes in NSBHR before and after the test, either to methacholine or mannitol. The data obtained by the authors show that there is no clear correlation between the methacholine and mannitol inhalation challenge tests in SICT monitoring. Preliminary results indicate the need for further investigations to evaluate the usefulness of the mannitol challenge test in the diagnostics of OA. Int J Occup Med Environ Health. 2020;33(2):235-9.

Specific Dog Allergen Immunoglobulin G Antibodies in Patients with Allergic Conjunctivitis

Purpose: The purpose of the study was to examine the role of total tear IgE, and specific serum dog IgG and IgE antibodies on the severity of allergic conjunctivitis. Methods: This study enrolled healthy subjects (control group, N=13), the patients with seasonal allergic conjunctivitis (seasonal group, N=13), and patients with perennial allergic conjunctivitis (perennial group, N=13). Skin prick test, tear IgE level, and serum specific dog IgE and IgG levels were examined. The severity of allergic conjunctivitis using a grading score (0-30) was also examined. Results: The levels of serum dog-specific IgE and IgG, and total tear IgE of the seasonal and perennial groups were higher as compared to those of the control group (all p&lt;0.05). The levels of serum dog-specific IgG of the perennial group were higher than those of the seasonal group (0.4 ± 0.6 vs. 0.0 ± 0.0). Multivariate analysis confirmed that the skin prick test result for dog allergen was related to the serum dog-specific IgG levels, but not IgE levels (p&lt;0.01). The severity of allergic conjunctivitis was related to the serum level of dog-specific IgG antibodies (p&lt;0.01). Conclusion: It was concluded that dog-specific IgG antibodies level may be associated with the severity of dog-related perennial allergic conjunctivitis.

Food-Specific IgE and IgG Antibodies in Patients With Chronic Rhinosinusitis With Nasal Polyps: A Case-Control Study.

EPOS 2012 states that investigation is needed to study a possible role for food allergy in the initiation and perpetuation of chronic rhinosinusitis with nasal polyps (CRSwNP). Our main goal was to compare serum levels of food-specific immunoglobulin G (IgG) and IgE antibodies in patients with CRSwNP and controls. A prospective case-control study with 33 patients with CRSwNP and 31 controls without CRS was carried out. Clinical data were gathered through a systematic interview and blood sample was collected. Enzyme-linked immunosorbent assay tests using OmegaDiagnostics kit with 40 food allergens for detection of specific IgG antibodies were performed and food-specific IgE antibodies were determined by immunoassay using ImmunoCAP. Immunoglobulin classes and IgG subclasses levels were also evaluated. Statistical analysis was performed using SPSS v.23. The overall sum of food IgG antibodies was significantly lower in CRSwNP compared to control group, and this difference was also observed for different specific IgG antibodies (corn, soya, grain legumes, pear and apple, berries, citric fruit). In controls, a positive correlation between IgG1 and the sum of food IgG antibodies was seen, but in CRSwNP group a negative correlation was found. In addition, a significant higher level of IgG1 and lower IgG2 and IgG3 was found among patients with CRSwNP. Levels of serum-specific IgE antibodies against multiallergen food mix (fx5) and against shrimp, strawberry, orange, rye, or egg yolk, as well as the sum of food IgE antibodies, did not differ significantly between the groups. These findings suggest that food allergy does not have an important role in CRSwNP etiopathogenesis, whether it is IgG or IgE mediated. Moreover, the observed suppression of specific IgG antibodies against food allergens, its negative correlation with IgG1 and the IgG1 switching in CRSwNP, can be related to deviated IgG responses against other targets (eg, airborne particles) and warrants future investigation.

Features of fungal sensitization in patients with asthma and cystic fibrosis: the results of a prospective study

Background. To estimate the frequency of fungal sensitization and allergic bronchopulmonary aspergillosis in patients with asthma or cystic fibrosis. Methods. The study included 205 patients with asthma and 190 patients with cystic fibrosis. For detection of fungal sensitization specific IgE antibodies in the blood serum with «AllergoELISA-specific IgE» test system and biotinylated fungal allergens were determined. Mycological examination included microscopy and cultural study of respiratory biomaterials. According to the indications, a CAT scan of the chest was performed. Results. The incidence of fungal sensitization in patients with asthma was 35,6%, CI [29,1%; 42,6%], in patients with cystic fibrosis - 56,8% [49,5%; 64%]. In patients with severe asthma the main fungal allergens were Alternaria spp. (70%) and Aspergillus spp. (60%); in patients with cystic fibrosis: Candida spp. - 73%, Alternaria spp. - 34%, Aspergillus spp. - 27%. The incidence of allergic bronchopulmonary aspergillosis (ABPA) in patients with asthma was 5,3%, in patients with cystic fibrosis - 5,7%. Conclusion Early detection of ABPA in patients with asthma and cystic fibrosis is very important as it changes the therapeutic measures of patients’ management and prevents progression of bronchiectasis and respiratory failure. Determination of specific Aspergillus spp. IgE antibodies in vitro is a necessary stage of diagnosis of ABPA in patients at risk.

Identification and characterization of the causative triatomine bugs of anaphylactic shock in Zhanjiang, China

BackgroundTwo health concerns primarily related to triatomine bugs are transmission of Trypanosoma cruzi through infective feces, and allergic reactions induced by triatomine bites. In the Southwestern United States, reduviid bugs bites commonly cause insect allergy. In South China, four cases of anaphylactic shock have been reported after this bite exposure. To further classify the species of these bugs and confirm the sensitization of the triatomine saliva, we caught triatomine bugs from the region where the bites occurred and performed phylogenetic and immunohistochemical (IHC) analysis.MethodsTriatomine bugs were collected in Donghai Island of Zhanjiang City in South China. The genomic DNA was extracted from three legs of the bugs. The fragments of mitochondrial 16S rRNA, cytochrome c oxidase subunit I (COI) gene and nuclear ribosomal 18S and 28S rRNA genes were obtained by PCR and sequenced. A phylogenetic tree was constructed based on the sequence of 16S rRNA gene using a maximum likelihood method with MEGA 7.0 software. Trypanosomal specific fragments and vertebrate COI genes were amplified from the fecal DNA to detect the infection of trypanosomes and analyze the blood feeding patterns, respectively. Paraffin-embedded sections were then prepared from adult triatomines and sent for IHC staining.ResultsWe collected two adult triatomine bugs in Donghai Island. Morphological and molecular analyses indicated that the triatomines were Triatoma rubrofasciata. No fragments of T. cruzi or other trypanosomes were detected from the fecal DNA. Mitochondrial gene segments of Homo sapiens and Mus musculus were successfully amplified. The allergens which induced specific IgE antibodies in human serum were localized in the triatomine saliva by IHC assay.ConclusionsThe two triatomine bugs from Donghai Island were T. rubrofasciata. They had bitten humans and mice. Their saliva should contain the allergens related to the allergic symptoms and even anaphylactic shock of exposed residents. Great consideration should be given to this triatomine bugs due to their considerable distribution and potential threat to public health in South China.

The basophil activation test differentiates between patients with alpha-gal syndrome and asymptomatic alpha-gal sensitization

Galactose-alpha-1,3-galactose (alpha-gal) syndrome is characterized by the presence of serum specific IgE antibodies to alpha-gal and delayed type I allergic reactions to the carbohydrate alpha-gal after consumption of mammalian (red) meat products and drugs of mammalian origin. Diagnostics currently rely on patient history, skin tests, determination of serum specific IgE antibodies, and oral food or drug challenges. We sought to assess the utility of different basophil parameters (basophil reactivity and sensitivity, the ratio of the percentage of CD63+ basophils induced by the alpha-gal-containing allergen to the percentage of CD63+ basophils after stimulation with anti-FcεRI antibody [%CD63+/anti-FcεRI], and area under the dose-response curve [AUC]) as biomarkers for the clinical outcome of patients with alpha-gal syndrome compared with subjects with asymptomatic alpha-gal sensitization. In addition to routine diagnostics, a basophil activation test (Flow CAST) with different concentrations of alpha-gal-containing allergens (eg, commercially available alpha-gal-carrying proteins and pork kidney extracts) was performed in 21 patients with alpha-gal syndrome, 12 alpha-gal-sensitized subjects, and 18 control subjects. Alpha-gal-containing allergens induced strong basophil activation in a dose-dependent manner in patients. Basophil reactivity at distinct allergen concentrations, the %CD63+/anti-FcεRI ratio across most allergen concentrations, the AUC of dose-response curves, and basophil allergen threshold sensitivity (CD-sens) with pork kidney extract were significantly higher in patients with alpha-gal syndrome compared with those in sensitized subjects. All parameters were negative in control subjects. The basophil activation test should be considered as an additional diagnostic test before performing time-consuming and potentially risky oral provocation tests. The %CD63+/anti-FcεRI ratio for all allergens and AUCs for pork kidney were the best parameters for distinguishing patients with alpha-gal syndrome from subjects with asymptomatic alpha-gal sensitization.

The importance of specific IgE antibodies in epidemiology of allergic rhinitis and asthma - the Epidemiology of Allergic Diseases in Poland (ECAP) survey: part one. Influence of allergy risk factors on concentration of specific IgE antibodies in serum

The importance of specific IgE antibodies in epidemiology of allergic rhinitis and asthma - the Epidemiology of Allergic Diseases in Poland (ECAP) survey: part one. Influence of allergy risk factors on concentration of specific IgE antibodies in serum

The importance of specific IgE antibodies in epidemiology of allergic rhinitis and asthma - the Epidemiology of Allergic Diseases in Poland (ECAP) survey: part one. Influence of allergy risk factors on concentration of specific IgE antibodies in serum.

IntroductionSpecific immunoglobulins E (sIgE) are important parameters to estimate severity of allergic diseases.AimTo determine the influence of allergy risk factors on the concentration of specific IgE antibodies in serum.Material and methodsThe concentration of sIgE antibodies against allergens Dermatophagoides pteronyssinus, cat dander, timothy grass, Alternaria alternata were determined in serum of 4077 respondents randomly selected from 9 regions (ECAP study). The positive results of sIgE (≥ 0.35 IU/ml) were correlated to answers in questionnaires ECRHSII and ISAAC.ResultssIgE are more frequently detected in respondents declaring an allergic disease of a father than of a mother (D. pteronyssinus p < 0.05, A. alternata p < 0.01). An early beginning to attend school, kindergarten or nursery increases the frequency of sIgE detection (p < 0.05). If the number of children who slept in the same room as a respondent before the respondent was five years old, was lower, sIgE antibodies are more frequently detected (p < 0.05). Relating to: 1) all 4 allergens, sIgE are the most frequently detected in respondents declaring absence of a carpet/rug at home (p < 0.05), 2) cat dander – less frequently detected in respondents who have a cat at home at the age of 1–4 years (p < 0.005).ConclusionsCarpets/rugs capture a portion of mite, epidermal, mould allergens located at home, wherefore absence of a carpet or rug causes greater stimulation of an immune system by these allergens; as a consequence, IgE antibodies are the most frequently detected in respondents. Household contacts of the respondents, at the age of 1–4 years, with cats induced partial immune tolerance to cat dander.

Molecular and immunological characterisation of tropomyosin from Anisakis pegreffii.

Tropomyosin (TM) is a major allergen in shellfish, known to cross-react with mite, cockroach and/or some roundworm (nematode) TM. In this study, we aimed to express and purify TM from the parasitic nematode Anisakis pegreffii and also to characterise its cross-reactivity with TM from shellfish. A. pegreffii was isolated from the flathead tiger fish (Neoplatycephalus richardsoni) and characterised using single-strand conformation polymorphism (SSCP)-based sequencing of the first and second internal transcribed spacers (ITS-1 and ITS-2) of nuclear ribosomal DNA. The recombinant tropomyosin (rTM) of A. pegreffii was expressed, purified and confirmed by immunohistochemistry, sequencing and LC-MS/MS analyses. Immunohistochemistry showed the muscle and the base layer of the third-stage larvae (L3) of A. pegreffii as the location of TM in A. pegreffii. The molecular relationship of TM of A. pegreffii with homologs from other nematodes and crustaceans was inferred from phylogenetic analysis. Immunogenicity of TM from A. pegreffii was tested by immunoblotting, which showed that rTM from A. pegreffii binds to IgE from sera of patients with allergy to crustaceans. Immunoblotting also showed that the anti-TM monoclonal antibody (MAb) did not recognise rTM from A. pegreffii. The rTM from A. pegreffii was, however, recognised by anti-TM polyclonal antibodies (PAbs) as well as anti-crustacean polyclonal antibodies (PAbs). The detection of specific serum IgE antibody against parasite TM has been proposed as a useful approach for the diagnosis of parasite-induced allergy. The findings of this study merit further exploration of the cross-reactive allergenic proteins of Anisakis for improved, future diagnosis of allergenic diseases.