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The specific detection of serum IgE antibodies specific to allergens (sIgE Abs) that can crosslink the plural high-affinity IgE receptor (FcεRIα) molecules on the surface of mast cells or basophils with a multivalent allergen can reduce the false-positive diagnoses observed in chemiluminescent and fluorescence enzyme immunoassays for type-I allergic patients. In this study, we detected sIgE Abs to the egg-allergen ovalbumin (OVA) and the wheat-allergen gluten in the sera of rats sensitized with each allergen using an amplified luminescence proximity homogeneous assay by crosslinking (AlphaCL). OVA and gluten were reacted with each sIgE Ab in the sera. Then, acceptor and donor beads labeled with the human FcεRIα were added to the reacted solution. The luminescence intensity for anti-OVA IgE Abs in the sera with the removal of IgG Abs was observed in five of seven (71.4%) of the sensitized rats, whereas no signals were observed in any of the unsensitized rats. The AlphaCL could also detect anti-gluten sIgE Abs in the sera of sensitized rats, but not of unsensitized rats. In conclusion, we successfully detected sIgE Abs in the sera of rats sensitized to two allergens using the AlphaCL. This detection method has the potential to be used as a new diagnostic tool for type-I allergic patients. [ABSTRACT FROM AUTHOR]

Abstract Background Recent studies have found that total immunoglobulin E (IgE) and allergen-specific IgE were associated with some metabolic diseases. However, the role of IgE in metabolism among adolescents is still unclear. Herein, this study aims to investigate the associations of serum total IgE and allergen-specific IgE with insulin resistance (IR) in adolescents, in order to provide some reference for the prevention and treatment of metabolic diseases in a young age. Methods Data of 870 adolescents were extracted from the National Health and Nutrition Examination Survey (NHANES) database in 2005–2006 in this cross-sectional study. Weighted univariate and multivariate logistic regression analyses were utilized to screen covariates and explore the relationships of serum total IgE and allergen-specific IgE with IR. The evaluation indexes were odds ratios (ORs) and 95% confidence intervals (CIs). In addition, these relationships were also assessed in subgroups of allergy history, asthma history, and number of allergens. Results Among eligible adolescents, 168 had IR. No significant association between serum total IgE level and IR was found. However, adolescents with higher level of allergen-specific IgE to rye grass [OR = 0.47, 95%CI: (0.25–0.91)], white oak [OR = 0.57, 95%CI: (0.37–0.88)], or peanut [OR = 0.38, 95%CI: (0.15–0.97)] seemed to have lower odds of IR, whereas those had higher level of shrimp-specific IgE [OR = 2.65, 95%CI: (1.21–5.84)] have increased odds of IR. In addition, these associations between allergen-specific IgE and IR were also discovered in adolescents who had allergy history or asthma history, or had different numbers of allergens. Conclusion Paying attention to different allergens in adolescents may be important in the early identification of IR among this high-risk population. The study results relatively provided some reference for further exploration on IR prevention.

Background: Recent studies have found that total immunoglobulin E (IgE) and allergen-specific IgE were associated with some metabolic diseases. However, the role of IgE in metabolism among adolescents is still unclear. Herein, this study aims to investigate the associations of serum total IgE and allergen-specific IgE with insulin resistance (IR) in adolescents, in order to provide some reference for the prevention and treatment of metabolic diseases in a young age. Methods: Data of 870 adolescents were extracted from the National Health and Nutrition Examination Survey (NHANES) database in 2005–2006 in this cross-sectional study. Weighted univariate and multivariate logistic regression analyses were utilized to screen covariates and explore the relationships of serum total IgE and allergen-specific IgE with IR. The evaluation indexes were odds ratios (ORs) and 95% confidence intervals (CIs). In addition, these relationships were also assessed in subgroups of allergy history, asthma history, and number of allergens. Results: Among eligible adolescents, 168 had IR. No significant association between serum total IgE level and IR was found. However, adolescents with higher level of allergen-specific IgE to rye grass [OR = 0.47, 95%CI: (0.25–0.91)], white oak [OR = 0.57, 95%CI: (0.37–0.88)], or peanut [OR = 0.38, 95%CI: (0.15–0.97)] seemed to have lower odds of IR, whereas those had higher level of shrimp-specific IgE [OR = 2.65, 95%CI: (1.21–5.84)] have increased odds of IR. In addition, these associations between allergen-specific IgE and IR were also discovered in adolescents who had allergy history or asthma history, or had different numbers of allergens. Conclusion: Paying attention to different allergens in adolescents may be important in the early identification of IR among this high-risk population. The study results relatively provided some reference for further exploration on IR prevention. [ABSTRACT FROM AUTHOR]

The specific detection of serum IgE antibodies specific to allergens (sIgE Abs) that can crosslink the plural high-affinity IgE receptor (FcεRIα) molecules on the surface of mast cells or basophils with a multivalent allergen can reduce the false-positive diagnoses observed in chemiluminescent and fluorescence enzyme immunoassays for type-I allergic patients. In this study, we detected sIgE Abs to the egg-allergen ovalbumin (OVA) and the wheat-allergen gluten in the sera of rats sensitized with each allergen using an amplified luminescence proximity homogeneous assay by crosslinking (AlphaCL). OVA and gluten were reacted with each sIgE Ab in the sera. Then, acceptor and donor beads labeled with the human FcεRIα were added to the reacted solution. The luminescence intensity for anti-OVA IgE Abs in the sera with the removal of IgG Abs was observed in five of seven (71.4%) of the sensitized rats, whereas no signals were observed in any of the unsensitized rats. The AlphaCL could also detect anti-gluten sIgE Abs in the sera of sensitized rats, but not of unsensitized rats. In conclusion, we successfully detected sIgE Abs in the sera of rats sensitized to two allergens using the AlphaCL. This detection method has the potential to be used as a new diagnostic tool for type-I allergic patients.

Background: The nature of epitopes on Bet v 1 recognized by natural IgG antibodies of birch pollen allergic patients and birch pollen‐exposed but non‐sensitized subjects has not been studied in detail. Objective: To investigate IgE and IgG recognition of Bet v 1 and to study the effects of natural Bet v 1‐specific IgG antibodies on IgE recognition of Bet v 1 and Bet v 1‐induced basophil activation. Methods: Sera from birch pollen allergic patients (BPA, n = 76), allergic patients without birch pollen allergy (NBPA, n = 40) and non‐allergic individuals (NA, n = 48) were tested for IgE, IgG as well as IgG1 and IgG4 reactivity to folded recombinant Bet v 1, two unfolded recombinant Bet v 1 fragments comprising the N‐terminal (F1) and C‐terminal half of Bet v 1 (F2) and unfolded peptides spanning the corresponding sequences of Bet v 1 and the apple allergen Mal d 1 by ELISA or micro‐array analysis. The ability of Bet v 1‐specific serum antibodies from non‐allergic subjects to inhibit allergic patients IgE or IgG binding to rBet v 1 or to unfolded Bet v 1‐derivatives was assessed by competition ELISAs. Furthermore, the ability of serum antibodies from allergic and non‐allergic subjects to modulate Bet v 1‐induced basophil activation was investigated using rat basophilic leukaemia cells expressing the human FcεRI which had been loaded with IgE from BPA patients. Results: IgE antibodies from BPA patients react almost exclusively with conformational epitopes whereas IgG, IgG1 and IgG4 antibodies from BPA, NBPA and NA subjects recognize mainly unfolded and sequential epitopes. IgG competition studies show that IgG specific for unfolded/sequential Bet v 1 epitopes is not inhibited by folded Bet v 1 and hence the latter seem to represent cryptic epitopes. IgG reactivity to Bet v 1 peptides did not correlate with IgG reactivity to the corresponding Mal d 1 peptides and therefore does not seem to be a result of primary sensitization to PR10 allergen‐containing food. Natural Bet v 1‐specific IgG antibodies inhibited IgE binding to Bet v 1 only poorly and could even enhance Bet v 1‐specific basophil activation. Conclusion: IgE and IgG antibodies from BPA patients and birch pollen‐exposed non‐sensitized subjects recognize different epitopes. These findings explain why natural allergen‐specific IgG do not protect against allergic symptoms and suggest that allergen‐specific IgE and IgG have different clonal origin. [ABSTRACT FROM AUTHOR]

Background: Idiopathic anaphylaxis (IA) is an unresolved concern. Hidden allergens may be relevant in IA and in nonsteroidal anti-inflammatory drug hypersensitivity (NSAID-HS)., Objective: To identify hidden elicitors for IA and NSAID-HS by a skin prick test (SPT) (13 allergens) and allergen-specific IgE (sIgE) panel (12 allergens) and to determine the value of each tested allergen., Methods: We retrospectively included all patients from 2018 to 2021 referred with a suspicion of IA or NSAID-HS by history in whom SPT and/or sIgE to allergens of the IA panel were performed. Patient characteristics from patients' records included comorbidities, history and symptoms of anaphylaxis, serum baseline tryptase level, total IgE level, SPT, sIgE, challenge results, and final diagnoses., Results: A total of 134 patients (77 female, mean age 39.7 ± 14.6 years) were included. Median serum baseline tryptase and total IgE levels were 4.23 μg/L and 133.5 kU/L, respectively. Allergologic workup with the IA panel resulted in positive SPT and sIgE in 61 (47%) and 66 (60%) patients, respectively. In those, confirmation or exclusion of allergy, mostly by challenge, led to a definitive diagnosis in 61 of 134 patients (46%). Skin prick test was most frequently positive to gluten (22.4%) and sIgE to ω5-gliadin (21.6%), which correlated with the history (r = 0.310, P < .001; and r = 0.407, P < .001, respectively). In 28 of 134 patients (21%) with initially suspected IA or NSAID-HS, challenges confirmed occult food allergy in which wheat allergy dependent on augmentation factors was the most frequent cause of anaphylaxis (19%)., Conclusions: Wheat allergy dependent on augmentation factors should be considered in all patients with anaphylaxis of unknown cause or after NSAID intake., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The measurement of antigen-specific serum IgE is common in clinical assessments of type I allergies. However, the interaction between antigens and IgE won't invariably trigger mast cell activation. We previously developed the IgE crosslinking-induced luciferase expression (EXiLE) method using the RS-ATL8 mast cell line; however, the method may not be sensitive enough in some cases. In this study, we introduced an NF-AT-regulated luciferase reporter gene into the RBL-2H3 rat mast cell line and expressed a chimeric high-affinity IgE receptor (FcεRI) α chain gene, comprising an extracellular domain from humans and transmembrane/intracellular domains from rats. We generated multiple clones expressing the chimeric receptor. Based on their responsiveness and proliferation, we selected the HuRa-40 clone. This cell line exhibited significantly elevated human α chain expression compared to RS-ATL8 cells, demonstrating a 10-fold enhancement of antigen-specific reactivity. Reproducibility across different batches and operators was excellent. Moreover, we observed a detectable response inhibition by an anti-allergy drugs (omalizumab and cyclosporin A). HuRa-40 cells—which carry the human-rat chimeric IgE receptor—comprise a valuable reporter cell line for the EXiLE method. Their versatility extends to various applications and facilitates high-throughput screening of anti-allergy drugs. Crosslinking of IgE antibodies by multivalent antigens is a key event in allergic reactions. We established a novel cell line (HuRa-40) that expresses human-rat chimeric FcεRIα subunit and luciferase reporter gene. HuRa-40 cells enable highly sensitive measurements of IgE crosslinking-induced luciferase expression (EXiLE), compared with previously used RS-ATL8 cells. This is because HuRa-40 cells express abundant FcεRIα subunits. HuRa-40 cells are also well-suited for high-throughput screening of anti-allergic drugs due to their high reproducibility and wide dynamic range. [Display omitted] • A novel human-rat chimeric FcεRI α chain was designed and expressed in rat mast cells. • The HuRa-40 clone abundantly expressed the chimeric receptor, enabling highly sensitive and reproducible EXiLE responses. • HuRa-40 cells are an ideal tool for high-throughput screening of anti-allergy drugs. [ABSTRACT FROM AUTHOR]

The aim of the study was to study the range of substances that cause respiratory sensitization in adults and children living in Dnipro as of 2019. Materials and research methods. Enzyme-linked immunosorbent assay was used to study the serum of venous blood, which determined the specific IgE of up to 9 allergens that are most common in the home. Kits for quantification of allergen-specific IgE from Vitrotest Specific-IgE, Ukraine were used. Conducted allergy diagnosis for 380 people with certain features of allergy history of various types of allergies, who went to the laboratory to specify the etiological root cause of their disease. Research results. As a result of the analysis of the received data the distribution of allergens on their prevalence among the population of Dnipro was established. The degree of hypersensitivity to each specific allergen and their ability to cross-react with each other was determined. Among the allergens of the household panel, the most dangerous were allergens of molds, which caused a sensitization reaction in 292 people, which was 76.8 % of all surveyed. The top three (in prevalence) of household allergens included: cat epithelium, which caused sensitization in 125 people (32.9 %) and Derm mite. Farinae, sensitivity to which was found in 117 patients (30.7 %). The share of inadequately strong allergopathological reactions is one third of all examined and prevails in people with sensitization to allergic agents of the household panel such as cat epithelium, Derm mite. Farinae and Derm. Pteronyssinus. Hyperractivity to fungi of the genus Candida and mold has been found in 9-12 % of people. The lowest severity of sensitization was observed in the epithelium of the dog and house dust, and low sensitivity in the reactivity structure of allergopathological reactions was observed for allergens of the epidermal group (down and feathers of poultry) and cockroaches. The development of cross-reactions took place between allergens of epidermal origin: there was a hypersensitivity to the hair and epithelium of dogs and cats (r=0.94, P<0.01), poultry feathers and cockroaches (r=0.99, P<0.01). This association of cross - reactions is possible, given the theory of minor and major proteins, and is due to their similarity: the similarity of the inclusion in the allergen structures of specific forms of molecules inherent in both allergic agents that may have allergy-stimulating effects. Conclusions. Cross-linking between several types of related allergens is the best way to further investigate this issue. It makes sense to include in the laboratory study a molecular method for the determination of major and minor proteins in the case of the greatest relationship to determine not only a specific allergen as an etiological factor, but also a specific sensitizing protein that is part of them. This is of great importance for subsequent immunotherapy when the removal of only one (major) protein agent can reduce the risk of susceptibility reactions to several types of allergens [ABSTRACT FROM AUTHOR]

The aim of the study was to study the range of substances that cause respiratory sensitization in adults and children living in Dnipro as of 2019. Materials and research methods. Enzyme-linked immunosorbent assay was used to study the serum of venous blood, which determined the specific IgE of up to 9 allergens that are most common in the home. Kits for quantification of allergen- specific IgE from Vitrotest Specific-IgE, Ukraine were used. Conducted allergy diagnosis for 380 people with certain features of allergy history of various types of allergies, who went to the laboratory to specify the etiological root cause of their disease. Research results. As a result of the analysis of the received data the distribution of allergens on their prevalence among the population of Dnipro was established. The degree of hypersensitivity to each specific allergen and their ability to cross-react with each other was determined. Among the allergens of the household panel, the most dangerous were allergens of molds, which caused a sensitization reaction in 292 people, which was 76.8 % of all surveyed. The top three (in prevalence) of household allergens included: cat epithelium, which caused sensitization in 125 people (32.9 %) and Derm mite. Farinae, sensitivity to which was found in 117 patients (30.7 %). The share of inadequately strong allergopathological reactions is one third of all examined and prevails in people with sensitization to allergic agents of the household panel such as cat epithelium, Derm mite. Farinae and Derm. Pteronyssinus. Hyperractivity to fungi of the genus Candida and mold has been found in 9-12 % of people. The lowest severity of sensitization was observed in the epithelium of the dog and house dust, and low sensitivity in the reactivity structure of allergopathological reactions was observed for allergens of the epidermal group (down and feathers of poultry) and cockroaches. The development of cross-reactions took place between allergens of epidermal origin: there was a hypersensitivity to the hair and epithelium of dogs and cats (r=0.94, P<0.01), poultry feathers and cockroaches (r=0.99, P<0.01). This association of cross-reactions is possible, given the theory of minor and major proteins, and is due to their similarity: the similarity of the inclusion in the allergen structures of specific forms of molecules inherent in both allergic agents that may have allergy-stimulating effects. Conclusions. Cross-linking between several types of related allergens is the best way to further investigate this issue. It makes sense to include in the laboratory study a molecular method for the determination of major and minor proteins in the case of the greatest relationship to determine not only a specific allergen as an etiological factor, but also a specific sensitizing protein that is part of them. This is of great importance for subsequent immunotherapy when the removal of only one (major) protein agent can reduce the risk of susceptibility reactions to several types of allergens [ABSTRACT FROM AUTHOR]

Introduction: In childhood, the so-called allergic march involves progression from IgE sensitization to allergy-related symptoms. Both IgE sensitization and relevant clinical symptoms are required for the diagnosis of allergy, but concordance between test results and clinical symptoms varies greatly, creating challenges for the diagnostics and for the prediction of outcomes. We assessed the prevalence of IgE sensitization and allergy symptoms, concordance between 2 IgE sensitization testing methods, and predictive value of these tests in relation to clinical symptoms in young Finnish children. Methods: The current study included 2 series of children: a birth cohort, in which the participants were followed prospectively from birth up to 3 years, and a young children cohort observed from 3 to 5 years of age. They were regularly monitored for sensitization by measuring serum allergen-specific IgEs (sIgEs) and performing skin prick tests (SPTs). The emergence of atopic dermatitis, wheezing, and symptoms associated with food allergies was recorded. Results: Over the first 5 years of life, the prevalence of sIgE sensitization was 46%, while it was 36% for positive SPTs. Disease prevalence was 26% for atopic dermatitis, 25% for wheezing, and 19% for symptoms associated with food allergies. Concordance between sIgE and SPT results was good for aeroallergens, but poor for dietary allergens. The association between clinical symptoms and sensitization was stronger at 5 years than at 3 years of age. The proportion of children with concordant combinations of allergy symptoms and sensitization markers in contrast to those with discordant combinations increased from 3 to 5 years. Conclusion: In early childhood, testing for IgE sensitization predicts allergy-related symptoms in an age-dependent manner, but not particularly well. Tests predict symptoms caused by aeroallergens clearly better than those caused by dietary allergens. The clinical relevance of sensitization testing in early life is therefore limited in the prediction of true allergy. [ABSTRACT FROM AUTHOR]

Background: Allergic rhinoconjunctivitis is a public health problem. Allergen Immunotherapy is an effective and safe treatment, that modifies the natural course of allergic disease and induces long‐term tolerance. Objective: To correlate basophil and antibody biomarkers of subcutaneous immunotherapy to clinical outcomes and cellular changes in target tissue. Methods: Adults suffering from allergic rhinoconjunctivitis due to grass pollen allergy were randomized to receive subcutaneous immunotherapy (n = 18) or to an open control group (n = 6). Patients reported daily symptom and medication scores and weekly rhinitis related quality of life scores during four pollen seasons. Biomarkers were measured every 3 months for three years treatment and every 6 months in the follow‐up year. Nasal and cutaneous allergen challenge tests were performed annually. Leukocyte subsets were assessed in nasal mucosa biopsies at baseline and after treatment. Results: Subcutaneous immunotherapy led to a 447‐fold decrease in basophil sensitivity during the first treatment year. This remained 100‐fold lower than baseline during the 3 year‐treatment period and 10‐fold lower during the follow‐up year (n = 18, P =.03). Decrease in basophil sensitivity after three weeks of treatment predicted long‐term improvement in seasonal combined symptom and medication scores (ῥ=−0.69, P =.0027) during three years of treatment. AUC of IgE‐blocking factor correlated to nasal allergen challenge (ῥ = 0.63, P =.0012) and SPT (ῥ = 0.45, P =.03). Plasma cell numbers in the nasal mucosa increased during treatment (P =.02). Conclusion: Decrease in basophil sensitivity after three weeks of subcutaneous allergen immunotherapy predicted the clinical outcome of this treatment. [ABSTRACT FROM AUTHOR]

(D) Taxa relative abundance at phylum level in non-sensitized (aIgE < 0.35 kUA/L) and sensitized (aIgE > 0.35 kUA/L) individuals gl Allergen-specific serum IgE measurements revealed that approximately half of the subjects were sensitized to airborne and food allergens (aIgE > 0.35 kU SB A sb /L, I n i = 30), and of these, 43% were sensitized to birch pollen (bIgE > 0.35 kU SB A sb /L, I n i = 13), and 50% to timothy pollen (tIgE > 0.35 kU SB A sb /L, I n i = 15) (Figure S1A-C). In conclusion, we show that high allergen-specific serum IgE levels in our study subjects are associated with potent pro-inflammatory, and attenuated anti-inflammatory responses to allergens. Keywords: 16S rRNA gene sequencing; allergen-specific IgE; amplicon sequence variant (ASV); host-microbe interactions; skin microbiota EN 16S rRNA gene sequencing allergen-specific IgE amplicon sequence variant (ASV) host-microbe interactions skin microbiota 1280 1284 5 04/14/21 20210401 NES 210401 The prevalence of allergic diseases continues to increase worldwide along with industrialization and urbanization. [Extracted from the article]

Objectives: The aim of this study was to evaluate the results of serum allergen-specific IgE testing in cats with a clinical diagnosis of asthma and to determine if the number of allergens with positive IgE reactivity and magnitude of positive IgE responses would be associated with the severity of clinical signs or airway eosinophilia. Methods: Medical records from 2008 to 2018 were retrospectively reviewed. Inclusion required a diagnosis of feline asthma based on consistent clinicopathologic features and bronchoalveolar lavage (BAL) cytology with >10% eosinophils; additionally, cats needed to have the results of serum allergen-specific IgE tests. Results: Eighteen cases satisfied the inclusion criteria. Median age was 5 years and the most common presenting clinical sign was cough (n = 10/18). Most cats lived exclusively indoors (n = 13/18). The median percentage of BAL eosinophils was 47%. Serum allergen-specific IgE testing supported an underlying allergic etiology in 14/18 (78%) cats, with all but one having polysensitization. The severity of clinical signs and magnitude of airway eosinophilia did not correlate with the degree of positive IgE reactivity. Conclusions and relevance: This study identified a strong association between the identification of allergen-specific IgE and cats with asthma, and the majority of these cats were polysensitized. However, larger numbers of allergens with positive IgE reactivity or magnitude of IgE reactivity were not significantly associated with clinical severity or airway eosinophilia. Knowledge of positive allergen-specific IgE results could guide allergen avoidance, regardless of the magnitude of IgE reactivity. [ABSTRACT FROM AUTHOR]

Abstract Background The pathogenesis of asthma, which is an allergic lung disease, is associated with a variety of allergens such as house dust mite, pollen, and mould, IgE containing serum IgE and allergen-specific-IgE, and inflammatory cytokines including thymus and activation-regulated chemokine (TARC)/CCL17. Because aging is an essential factor in the pathogenesis of asthma, we examined biomarkers related to asthmatic subjects depending on age. Results Physiological indices such as FEV1(forced expiratory capacity in 1 s), FEV1 (% predicted), and FEV1/FVC(forced vital capacity) (%) in asthmatic subjects were lower than those in normal subjects. Total IgE, Der p1 specific IgE, and Der f1 specific IgE were elevated in serum of asthmatics relative to normal individuals. Regulated on activation, normal T cell expressed and secreted (RANTES)/CCL5 in serum and interleukin 6 (IL-6), interleukin 8 (IL-8), monocyte chemoattractant protein (MCP)-1/CCL2, RANTES, and macrophage inflammatory protein (MIP)-1α/CCL3 in bronchoalveolar lavage fluid (BALF) of asthmatic subjects were higher than in normal individuals. Upon classification of experimental groups depending on age, physiological indices and Der p1-specific IgE (class) were decreased in middle aged adult and elderly adult groups relative to the young adult group. TARC levels in serum were strongly elevated in the elderly adult group relative to the young adult and the middle aged adult groups. TARC in serum was related to total IgE in serum in the elderly adult group. Conclusions Taken together, although TARC in serum and BALF is not different between normal and asthmatic individuals, TARC increases in serum of elderly asthmatic subjects. The level of TARC has a positive effect on the level of IgE in the elderly adult group. These findings may help us better understand the relationship of pathogenesis of allergic diseases and aging.

Mast cells drive the inappropriate immune response characteristic of allergic inflammatory disorders via release of pro‐inflammatory mediators in response to environmental cues detected by the IgE‐FcεRI complex. The role of TGF‐β‐activated kinase 1 (TAK1), a participant in related signaling in other contexts, remains unknown in allergy. We detect novel activation of TAK1 at Ser412 in response to IgE‐mediated activation under SCF‐c‐kit potentiation in a mast cell‐driven response characteristic of allergic inflammation, which is potently blocked by TAK1 inhibitor 5Z‐7‐oxozeaenol (OZ). We, therefore, interrogated the role of TAK1 in a series of mast cell‐mediated responses using IgE‐sensitized murine bone marrow‐derived mast cells, stimulated with allergen under several TAK1 inhibition strategies. TAK1 inhibition by OZ resulted in significant impairment in the phosphorylation of MAPKs p38, ERK, and JNK; and mediation of the NF‐κB pathway via IκBα. Impaired gene expression and near abrogation in release of pro‐inflammatory cytokines TNF, IL‐6, IL‐13, and chemokines CCL1, and CCL2 was detected. Finally, a significant inhibition of mast cell degranulation, accompanied by an impairment in calcium mobilization, was observed in TAK1‐inhibited cells. These results suggest that TAK1 acts as a signaling node, not only linking the MAPK and NF‐κB pathways in driving the late‐phase response, but also initiation of the degranulation mechanism of the mast cell early‐phase response following allergen recognition and may warrant consideration in future therapeutic development. [ABSTRACT FROM AUTHOR]

Introduction: The measurement of clinically important specific IgE (sIgE) antibody is pivotal for both diagnosis and management of allergy. Two methods may be distinguished depending on the number of antigens tested simultaneously: singleplex and multiplex. BioIC is a multiplex, advanced, automated microfluidic immunoassay system enabling simultaneous sIgE measurement against multiple allergens. ImmunoCAP is a singleplex assay for sIgE detection and gold standard method for diagnosis of allergy. Aim: To compare and validate the diagnostic capability of a multiplex sIgE assay - BioIC assay with a singleplex ImmunoCAP assay. Material and methods: Using both BioIC assay and ImmunoCAP assay, the sIgE level in serum samples from 20 allergic disease patients with respect to 33 allergens (16 inhalant allergens, 16 food allergens and 1 contact allergen) was measured. Receiver operating characteristic (ROC) area under the curve (AUC), qualitative and semiquantitative comparisons were performed to compare both sIgE measurement methods using statistical analyses. Results: ROC AUC analysis showed similar sensitivity and specificity of BioIC assay and ImmunoCAP assay. In qualitative analysis, the negative and positive agreements were 100% equal for each allergen. Spearman's rank correlation coefficients identified very high positive correlations between two assays for all tested allergens (p < 0.001). Conclusions: The BioIC showed agreement with ImmunoCAP assay. Sensitivity and specificity of both assays are similar, thus they showed similar diagnostic performance. However, careful interpretation of obtained results is necessary in clinical applications because of methodological differences between these two systems. [ABSTRACT FROM AUTHOR]

Background and Objectives: Different types of food introduced gradually in the diet will expose children to different food allergens, increasing the chance of developing allergic diseases. The aim of our study was to determine if allergen-specific IgE values can influence, depending on the diet, the prediction of remission of urticaria in children. Materials and Methods: This prospective study was conducted in 132 patients diagnosed over two years with urticaria, admitted to “Sf. Maria” Clinical Pediatric Hospital Iaşi. Total IgE assay was performed by ELISA, and determination of specific serum IgE by the CLA System Quanti Scan method (Innogenetics, Heiden, Germany). Data were gathered and statistical analysis was performed using statistical software SPSS, using descriptive and inferential statistics. Results: The determination of specific IgE to food allergens was performed on a total of 132 cases. The values of specific IgE were positive for one or more food allergens in 84 patients (63.64%). The most common allergens involved were: cow’s milk in 33.3% cases, egg white in 22.6% cases, and hazelnuts in 11.9% cases. The specific IgE values for the different types of food included in our study had a predictive value for disease remission. Conclusions: The determination of specific IgE confirms the presence of a particular food allergen and may have predictive value for the future development of an allergic manifestation.

Objectives: The objective of this study was to examine the performance of AdvanSure AlloScreen Max with 92 different allergens compared to Polycheck Allergy and ImmunoCAP. The relationship of serum IgE concentration with the number and the highest class/level of positive allergen-specific IgEs was also examined.Methods: A total of 406 serum samples were included in this study. Discrepant cases between AdvanSure AlloScreen Max and Polycheck Allergy underwent ImmunoCAP testing for allergen-specific IgE.Results: Total agreement of the two multiple allergen simultaneous tests (MAST) was 92.5%. Compared to ImmunoCAP, total agreement rate was higher with AdvanSure AlloScreen Max (60.8%) than that with Polycheck Allergy (39.2%). Serum IgE concentration showed a significant and positive correlation with the number and the highest class/level of positive allergen-specific IgEs.Conclusions: A MAST assay panel containing as many allergens as possible would be more helpful in the allergen screening for patients with high serum IgE concentration. [ABSTRACT FROM AUTHOR]

Allergic conditions are prevalent equine diseases that can be diagnosed by clinical examination alone, but definitive diagnosis is more likely with laboratory testing. The ELISA Allercept© test was used to analyse the serum samples of 73 horses with allergic diseases. Sixty-one horses (83.5%) had allergen-specific IgE levels ≥ 150 ELISA Units (EU), the cut-off defined by the assay. Fifty-four horses had allergic dermatitis (AD) with high IgE levels to Tyrophagus putrescentiae (51.9%), Rumex crispus (48.1%), Tabanus (46.3%) and Dermatophagoides farinae/ D. pteronyssinus (40.7%). Seven horses with recurrent airway obstruction (RAO) had a high prevalence of T. putrescentiae (85.7%), followed by that of Acarus siro (57.1%) and D. farinae/D. pteronyssinus (57.1%). Horses affected with RAO had more positive reactions to mites (2.22 ± 0.84) than did horses with AD (1.51 ± 0.61, P < 0.05). A strong correlation of serum allergen-specific IgE level was found between Culex tarsalis and Stomoxys (r = 0.943) and between Dactylis glomerata and both Secale cereale (r = 0.79) and R. crispus (r = 0.696). These results indicate that among horses with allergic diseases in Spain, ELISA tests demonstrated a high prevalence of serum allergen-specific IgE in response to mites. Our study emphasises the importance of laboratory testing and updating allergy panels to improve the likelihood of a definitive diagnosis and the identification of allergens that should be included in allergic disease treatment. [ABSTRACT FROM AUTHOR]

Coeliac disease and HLA-conferred susceptibility to autoimmunity are associated with IgE sensitization in young children Keywords: aeroallergens; allergen-specific IgE; coeliac disease; dietary allergens; HLA At 3 y of age, children in the BC, who all carried HLA-conferred risk to type 1 diabetes and coeliac disease, were more frequently sensitized to (A) any allergen (P < .001), (B) dietary allergen (P < .001), and (C) aeroallergen (P = .001) compared to children in the YCC who represented the general population. Aeroallergens, allergen-specific IgE, coeliac disease, dietary allergens, HLA. [Extracted from the article]

Introduction and objective The association between airborne pollen counts or duration of pollen season and allergy symptoms is not always distinguished. The purpose of this study was to examine the correlation between pollen exposure (annual total pollen quantity and main pollen season length) of selected allergenic plants in the atmosphere of Bratislava, and concentration of allergen-specific immunoglobulin E (sIgE) in serum of patients with seasonal allergy during 2002–2003. Material and Methods The concentration of pollen was monitored by a Burkard volumetric pollen trap. At the same time, 198 pollen allergic patients were testing to determine the values of sIgE antibodies against selected pollen allergens; a panel of 8 purified allergens was used. Results The highest percentages of sensitization were detected for Poaceae and Ambrosia pollen allergens. The most abundant airborne pollen types were Urticaceae, Betula, Populus, Fraxinus, Pinus and Poaceae. The length of the pollen season varied. The longest pollen season was that of the Plantago – 105 days, and the shortest, Corylus – 20 days. A significant correlation was found between annual total pollen quantity and median sIgE values, especially in 2002. Conclusions A strong and significant positive correlation was observed between pollen counts, excluding Betula, and sIgE levels in both analysed years. The correlation was weaker and negative in the case of length of pollen season and sIgE values.

Purpose: To describe the profile of patients with allergic conjunctivitis (AC) regarding their demographics, symptomatology and specific allergen sensitization, in a Lebanese tertiary hospital.Methods: Cross-sectional study conducted at the Hôtel-Dieu de France hospital (Beirut, Lebanon) during a period of 18 months. Patients with seasonal or perennial AC presenting for ophthalmic consultation had measurements of total and specific IgE. A matching group of patients with AC seen at the allergist office during the same period underwent skin prick tests (SPTs).Results: Forty-four patients were enrolled for blood work by their ophthalmologists. Seasonal and perennial forms were almost equivalent. In total, 56.8% had positive specific IgE, with higher prevalence in patients with seasonal AC (p = 0.002), other associated allergies particularly allergic rhinitis (p = 0.002) or a family history of allergy (p = 0.005). Ocular surface severity scales were not shown as predictors. High levels of total IgE were commonly detected in those with positive specific IgE. Thirty-eight patients were assessed with SPT, and all had a positive result for at least one allergen. Dust mites were found to be the most frequent allergens based upon both specific IgE (72%) and SPT (92%), followed by Parietaria and other pollens.Conclusion: In our study, dust mites mono- or co-sensitization is present in the majority of patients with AC, with odds of positivity being higher using SPT than specific IgE. The latter are found more readily in seasonal AC and in the presence of personal and family history of allergy. [ABSTRACT FROM AUTHOR]

Background Several methods have been developed to detect allergen-specific IgE in sera. The passive IgE sensitization assay using human IgE receptor-expressing rat cell line RBL-2H3 is a powerful tool to detect biologically active allergen-specific IgE in serum samples. However, one disadvantage is that RBL-2H3 cells are vulnerable to high concentrations of human sera. Only a few human cultured cell lines are easily applicable to the passive IgE sensitization assay. However, the use of human induced pluripotent stem cells (iPSCs) to generate human mast cells (MCs) has not yet been reported. Methods The nuclear factor-kappa B (NF-κB)-responsive luciferase reporter gene was stably introduced into a human iPSC line 201B7, and the transfectants were induced to differentiate into MCs (iPSC-MCs). The iPSC-MCs were sensitized overnight with sera from subjects who were allergic to cedar pollen, ragweed pollen, mites, or house dust, and then stimulated with an extract of corresponding allergens. Activation of iPSC-MCs was evaluated by β-hexosaminidase release, histamine release, or luciferase intensity. Results iPSCs-MCs stably expressed high-affinity IgE receptor and functionally responded to various allergens when sensitized with human sera from relevant allergic subjects. This passive IgE sensitization system, which we termed the induced mast cell activation test (iMAT), worked well even with undiluted human sera. Conclusions iMAT may serve as a novel determining system for IgE/allergens in the clinical and research settings. [ABSTRACT FROM AUTHOR]

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Background: Epidemiological evidence indicates that the age at onset of asthma and allergen sensitization in early life is decreasing in people from Western countries. To explore latent trends, we conducted a retrospective examination of the age at onset of asthma and specific IgE antibodies against inhalant allergens in Japanese asthmatic children. Methods: We conducted a case series study of 103 consecutive children with atopic type of asthma (aged 2 years to 16 years, mean age 9.4 ± 3.4 years). Diagnoses of asthma and allergic rhinitis were defined according to Japanese guidelines. The onset of asthma and allergic rhinitis was also defined as any report of asthma and allergic rhinitis confirmed by a physician. Allergen sensitization was evaluated as specific serum IgE levels for 9 common inhalant allergens in peripheral blood. Atopic type of asthma was defined as a being positive for at least one aeroallergen. Results: Mean age at asthma onset was 2.3 ± 1.9 years, which is slightly lower than that of previous reports, including those published in Japan. A high prevalence rate of up to 80% was found for perennial antigens including Dermatophagoides spp. and house dust, as reported previously. Notably, some of the children aged at 1 year tested positive for these allergens. Conclusions: The age at onset of asthma seems to be decreasing in comparison with previous reports. Furthermore, the age at onset of allergen sensitization against inhalant allergens appears to follow this trend.

Background: : Japanese cedar pollen (JCP) is the most common allergen for seasonal allergic rhinitis in Japan. Little is known about local production of immunoglobulin (Ig)E in people with or without Japanese cedar pollinosis. The aims of this study were to measure levels of JCP-specific IgE in nasal secretions and determine correlations with levels in serum. Methods: : Forty-six subjects were enrolled in this study, comprising 24 symptomatic subjects, 9 asymptomatic subjects sensitized to JCP, and 13 subjects not sensitized to JCP. Nasal secretions were obtained during a period of Japanese cedar dispersal, and levels of JCP-specific IgE were measured with CAP-fluorescent enzyme immunoassay. Serum JCP-specific IgE and total IgE were also measured using the same method. Results: Among the 46 subjects enrolled, JCP-specific IgE in nasal secretions was measureable in 43 subjects. Irrespective of symptom development, sensitized subjects showed higher levels of JCP-specific IgE in nasal secretions than non-sensitized subjects. A significant moderate correlation was observed between JCP- specific IgE levels in nasal secretions and serum in all 43 subjects. With stratification by subject group, only symptomatic subjects showed a substantial correlation between JCP-specific IgE levels in nasal secretions and serum. Conclusions: : Our results imply a certain association between JCP-specific IgE in nasal secretions and sensitization of Japanese cedar pollinosis. Therefore, levels of allergen-specific IgE in nasal secretions can be used as an alternative diagnostic marker for allergic rhinitis patients.

Background: Allergen-specific IgE levels in human sera support the diagnosis of allergies. However, commonly used laboratory tests are time consuming. The new point-of-care test FastCheckPOC 20 (FCP20) allows the simultaneous and semi-quantitative evaluation of IgE antibodies for 20 aeroallergens and food allergens. Methods: Two hundred patients with allergic symptoms were included in a multicenter performance evaluation using both whole blood and plasma or serum samples. FCP20 results were compared with the skin prick test (SPT) and the ImmunoCAP system (Phadia) which was used as reference for specific IgE values. Results: The concordance between FCP20 and ImmunoCAP was 76% for positive results and 80% for negative results over 20 allergens. Overall agreement (accuracy) of FCP20 with the ImmunoCAP was 80% across all 20 allergens. The concordance between FCP20 and SPT over all 20 allergens was 66% for positive results and 71% for negative results. Overall agreement of FCP20 with the SPT was 70%. Semi-quantitative FCP20 results (Levels 1-5) revealed nearly identical values for whole blood and plasma/serum test results. A good concordance was also observed between FCP20 levels and ImmunoCAP classes. Conclusions: FastCheckPOC 20 is reliable to determine allergen-specific IgE in human whole blood and plasma/serum samples. [ABSTRACT FROM AUTHOR]

The present study tested the hypothesis that a pan-allergen sensitization may affect the sensitization pattern. For this reason, 22 sensitization pattern allergens (SPA), common in Genoa (Italy), were selected for analyses. Successively, five of them, such as Pru p 3 as representative for LTP family, Bet v 1 and Pru p 1 for PR-10, and Bet v 2 and Pru p 4 for Profilin, were used as target allergens (TA). This retrospective study included 1059 subjects, (396 males and 663 females, mean age 42.8 years). The current study showed that sensitization to a pan-allergen entails higher odds to have other sensitizations. In addition, the co-sensitization pattern depends on the basis of the sensitizing pan-allergen family. LTP-sensitization is strongly associated with peanut sensitization, PR10 and profiling sensitization with hazelnut positivity. This study shows that a pan-allergen sensitization is frequently associated with co-sensitizations and the sensitization pattern depends on the sensitizing pan-allergen. [ABSTRACT FROM AUTHOR]

Introduction Component-resolved diagnostics using specific IgE to 2 S albumins has shown to be a valuable new option in diagnostic procedure. Ana o 3 is a 2 S albumin from cashew. The aim of this study was to investigate the role of Ana o 3-specific serum IgE in the diagnosis of cashew allergy and to identify cut-off levels to replace oral food challenges. Moreover, the value of additional determination of total IgE has been investigated. Methods In a multicentre study, we analysed specific IgE to cashew extract and Ana o 3 as well as total IgE in children with suspected cashew allergy using the Immuno CAP- FEIA and a standardized diagnostic procedure including oral challenges where indicated. Results A total of 61 patients were included in the study. Forty-two were allergic to cashew, and 19 were tolerant. In receiver operating curves, Ana o 3 discriminates between allergic and tolerant children better than cashew-specific IgE with an area under the curve of 0.94 vs 0.78. The ratio of Ana o 3-specific IgE to total IgE did not further improve the diagnostic procedure. Probability curves for Ana o 3-specific IgE have been calculated, and a 95% probability could be estimated at 2.0 kU/l. Conclusion Specific IgE to Ana o 3 is a valuable tool for the diagnosis of cashew allergy. Considering its positive predictive value, it might allow to make a considerable number of oral challenges superfluous. [ABSTRACT FROM AUTHOR]

Background Although the mechanisms of the formation of anti-dense fine-speckled 70 (anti-DFS70) antibodies are not fully known, there is evidence in the literature that allergic reactions may play a role in their formation. Immunoglobulin E (IgE)-mediated immunopathological mechanisms are increasingly being elucidated in diseases such as atopic dermatitis and urticaria-related diseases. We aimed to reveal its relationship with anti-DFS70 in allergen-sensitive patients with positive specific IgE (sIgE) levels. Methodology The study included samples of 758 patients who underwent antinuclear antibody (ANA) screening and allergen-sIgE testing between January 2019 and January 2022. Patients' clinical diagnoses were retrospectively obtained from the hospital information management system. ANA was tested according to the instructions of the manufacturer by the indirect immunofluorescent antibody method using HEp-2 cell substrates (Euroimmun Luebeck, Germany). Allergen-sIgE was determined by chemiluminescence on the Immulite 2000 XPI system (Siemens Healthcare Diagnostics Products GmbH, Marburg, Germany) according to the instructions of the manufacturer. Results ANA pattern was detected in 74 samples included in the study. ANA-positive patients were divided into DFS70 (+) and DFS70 (-) groups. A statistically significant increase in the DFS70 pattern was observed in patients with a positive allergen-sIgE test (p < 0.0001). Both allergen-sIgE and DFS70 positivity were statistically significant in younger age groups (p < 0.05). The most common diagnosis was urticaria-related conditions in 23 (31%) patients with a positive allergy test. Conclusions Our study shows that the positivity of the DFS70 pattern is increased in allergen-sensitive patients. Therefore, the allergen-sIgE-mediated allergic disease should be considered in patients with isolated anti-DSF70. Studies with related disease groups are needed to determine whether there is a relationship between anti-DFS70 and allergy-related disease in these patients. If an immunopathological mechanism is not found, these false-positive results can be considered clinically insignificant, and unnecessary consultations can be avoided., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2023, Togay et al.)

Background: Allergic sensitisation is poorly documented in infants. This study aims to provide new insights into allergic sensitisation patterns and related factors in infancy.Methods: This study concerns 1860 infants involved in the Pollution and Asthma Risk: an Infant Study (PARIS) population-based birth cohort who had a standardised health examination when 18 months old, from 2004 to 2008. Sensitisation was assessed by measurements of serum specific IgE to 12 food and 4 inhalant allergens and defined by IgE≥0.35kUA/L. Information regarding lifestyle and environment were obtained from questionnaires prospectively administered.Results: Prevalence of allergic sensitisation to any allergen, to food allergens, and to aeroallergens was 13.8%, 12.3%, and 2.3%, respectively. Multiple sensitisation (to at least two allergens) concerned 6.2% of toddlers. Intrinsic factors such as male gender, family history of allergy, and high birth weight increased the risk of food allergen sensitisation and multiple sensitisation. Caesarean section was also positively associated with multiple sensitisation. Day-care attendance was negatively related to food allergen, aeroallergen, and multiple sensitisation. A cat entering the baby's room in early life was strongly associated with aeroallergen sensitisation (ORa 3.21, 95%CI: 1.29-8.01). An introduction of meat in infant's diet after 6 months of age was negatively related to food allergen sensitisation (ORa 0.46, 95%CI: 0.24-0.91).Conclusion: Our results suggest that intrinsic factors and indicators of exposure to microorganisms such as caesarean section and day-care attendance may be associated with inhalant as well as food allergen sensitisation in infancy. For example, male gender, family history of allergy, high birth weight, and caesarean section could be positively related whereas day-care attendance could be negatively related to both aeroallergen and food allergen sensitisation. Conversely, early life exposure to inhalant allergens or food allergens may be specifically linked to either aeroallergen sensitisation or food allergen sensitisation, respectively. [ABSTRACT FROM AUTHOR]

Background: Dry eye disease (DED) is a multifactorial disease of the tears and ocular surface. No epidemiologic reports have yet been published about the relationship between DED and allergic rhinitis or sensitization to allergens. Objective: To investigate the association between DED and allergic conditions in the general adult Korean population. Methods: This population-based cross-sectional study included 17,542 individuals (ages ≥ 19 years) who participated in the Korea National Health and Nutrition Examination Survey (KNHANES) from 2010 to 2012. Data for total serum immunoglobulin E (IgE) and allergen-specific IgE levels were available for 1857 subjects. Data were analyzed by using logistic regression to determine the association of DED with allergy or allergic conditions, including allergic rhinitis, atopic dermatitis, asthma, increased total serum IgE, and sensitization to house-dust mite (Dermatophagoides farinae), dog, and cockroach allergens. Results: The prevalence of DED, allergic rhinitis, atopic dermatitis, and asthma was 10.4, 13.1, 2.2, and 3.3%, respectively. The prevalence of the increased total serum IgE level and sensitization to D. farinae, dog, and cockroach allergens was 34.6, 15.7, 1.0, and 2.8%, respectively. After adjusting for general characteristics, DED was significantly associated with allergic rhinitis (odds ratio 1.58 [95% confidence interval, 1.38-1.81]) and D. farinae sensitization (odds ratio 1.80 [95% confidence interval, 1.17-2.79]). Conclusion: This study demonstrated a close association between DED and both allergic rhinitis and D. farinae sensitization in Korean adults. Further studies are warranted to establish temporal relationships between DED and allergic rhinitis. [ABSTRACT FROM AUTHOR]

Background: The prevalence of Japanese cedar (JC) pollinosis in Japanese children is increasing. However, few studies have reported the relationship between pollen count levels and the prevalence of pollinosis. To evaluate the relationship between JC pollen count levels and the prevalence of pollinosis in children, we investigated the sensitization and development of symptoms for JC pollen in two areas of Akita in northeast Japan with contrasting levels of exposure to JC pollen. Methods: The study population consisted of 339 elementary school students (10–11 years of age) from the coastal and mountainous areas of Akita in 2005–2006. A questionnaire about symptoms of allergic rhinitis was filled out by the students’ parents. A blood sample was taken to determine specific IgE antibodies against five common aeroallergens. Results: The mean pollen count in the mountainous areas was two times higher than that in the coastal areas in 1996–2006. The prevalence rates of nasal allergy symptoms and sensitization for mites were almost the same in both areas. On the other hand, the rates of nasal allergy symptoms and sensitization for JC pollen were significantly higher in the mountainous areas than in the coastal areas. The rate of the development of symptoms among children sensitized for JC pollen was almost the same in both areas. Conclusions: These results suggest that pollen count levels may correlate with the rate of sensitization for JC pollinosis, but may not affect the rate of onset among sensitized children in northeast Japan.

Background: An in vitro elicitation test employing human high-affinity IgE receptor-expressing rat mast cell lines appears to be a useful method for measuring mast cell activation using a patient's IgE and an allergen; however, such cell lines are sensitive to human complements in the serum. We have recently developed a new luciferase-reporting mast cell line (RS-ATL8) to detect IgE crosslinking-induced luciferase expression (EXiLE) with relatively low quantities of serum IgE. Methods: A total of 30 patients suspected of having egg white (EW) allergy were subjected to an oral food challenge (OFC) test; then, the performances of EW-specific serum IgE (CAP-FEIA), EW-induced degranulation, and EXiLE responses in RS-ATL8 cells were compared using receiver-operating characteristic (ROC) curve analysis. The patients' sera were diluted to 1:100, which causes no cytotoxicity when sensitizing the RS-ATL8 cells for the degranulation and EXiLE tests. Results: The area under the ROC curves was highest in the EXiLE test (0.977), followed by CAP-FEIA (0.926) and degranulation (0.810). At an optimal cutoff range (1.648-1.876) calculated from the ROC curve of the EXiLE test, sensitivity and specificity were 0.944 and 0.917, respectively. A 95% positive predictive value was given at a cutoff level of 2.054 (fold increase in luciferase expression) by logistic regression analysis. Conclusions: In contrast to in vivo tests, the EXiLE test appears to be a useful tool in diagnosing patients suspected of having IgE-dependent EW allergy without the risk of severe systemic reactions.

Background: In studies on allergies in mouse models, IgE production is an essential parameter to be evaluated. Here, we examine the effect of commercially available immunoreaction enhancer solutions and different blocking reagents in enzyme-linked immunosorbent assay (ELISA) for total or antigen-specific murine IgE in order to improve the assays. Methods: Sera from mice immunized with recombinant house dust mite major allergens, Der f 1 and Der p 1, were used for the assays. Total IgE was measured by sandwich ELISA using monoclonal antibodies against murine IgE. Antigen-specific IgE was assayed using allergen-coated plates. Sensitivity or signal intensity in ELISA was compared among conditions differing in the use of enhancer solutions, blocking reagents, or monoclonal antibodies, and incubation time. Results: Use of enhancer solutions improved the sensitivity of ELISA for total IgE by approximately 30-fold of that using a conventional buffer. A blocking reagent caused more unwanted enhancement of the background signal in blank wells in ELISA for total IgE compared with another blocking reagent, however, improved signal intensity in ELISA for antigen-specific ELISA without significant enhancement of the background signal. Optimal assay conditions were determined. Conclusions: Enhancer solutions are effective in improving ELISAs for total and antigen-specific murine IgE. Selection of blocking reagents was important to decrease unwanted enhancement of background signals and was effective in enhancing signals for positive samples. The ELISAs improved in this study are useful for the study of allergies in mouse models.

Summary: Background. In the diagnostic work up of allergy, determining allergen component-specific immunoglobulin E (IgE) is important for diagnosis, prognosis and choice of treatment. The purpose of this study was to evaluate the performance of the immunoblotting assay (Euroline) in detection of IgE antibodies against timothy grass and birch pollen allergen components compared to fluorescent enzyme assay (ImmunoCAP, Phadia 250).  Methods. A total of 128 serum samples from patients allergic to timothy grass and birch pollen were analysed. The levels of IgE antibodies to timothy grass and birch pollen were measured using Euroline DPA-Dx pollen 1 and ImmunoCAP assay. The two methods were then compared on binary (positive vs negative), semi-quantitative (IgE classes) and quantitative (concentration) levels. The two methods were also compared to results from skin prick testing. Results. The Euroline method showed a positive percentage agreement of 93% and negative percentage agreement of 94% with an overall accuracy of 94% when compared to ImmunoCAP. Kappa analysis showed moderate strength of agreement between the methods in determining IgE classes for 7/11 components tested. All components showed a positive correlation when analysed using Spearman's rank correlation. Conclusions. Overall, we found that there is good correlation between the Euroline and ImmunoCAP methods in measuring IgE sensitization.

Background The longitudinal pattern of allergen-specific IgE levels from the prenatal stage to early life has remained largely unexplored. Methods One-hundred and three mother-infant pairs, which were part of an ongoing population-based prospective birth cohort study of early childhood allergic diseases in Tainan, Taiwan, were included in this study. We examined the relationship of 20 allergen-specific IgE levels with blood samples of mothers, cord blood, and infants at 12 months of age using Spearman rank correlation, Kenal τ and McNemar test, respectively. Results Certain degree of IgE sensitization against most 20 examined specific allergens was observed in blood samples of mothers, cord blood, and infants at 12 months of age. When we further examined the association between allergy-related risk factors and atopy in infants at the first year of life, we found positive association between colic pain and atopy in infants at 12 months of age [adjusted odds ratios ( AOR) = 3.51; 95% confidence interval ( CI): 1.13-10.96; p = 0.03], and borderline significance between wheezing and atopy in infants at 12 months of age ( AOR = 4.58; 95% CI: 0.89-23.50; p = 0.07). Conclusion The findings from this study suggest that influence of maternal allergen-specific IgE levels on infant immune response might occur at birth and then wane in infants at 12 months of age. Positive association of colic pain and wheezing with atopy in infants at 12 months of age provides supportive evidence for the 'Allergy March' theory of allergy development in an Asian birth cohort. [ABSTRACT FROM AUTHOR]

We developed an in vitro system to diagnose allergy using an allergen microarray and photo-immobilization technique. Photo-immobilization is useful for preparing the allergen microarray because it does not require specific functional groups of the allergen and because any organic material can be immobilized by a radical reaction induced by photo-irradiation. To prepare the plates, allergen solutions were mixed with polymer and a bis- azidophenyl derivative, a photo-reactive cross-linker, the mixtures were micro-spotted on the plate, and the droplets were dried. The plate was irradiated with an ultraviolet lamp for immobilization. For the assay, human serum was added to the microarray plate. Allergen-specific immunoglobulin E (IgE) adsorbed on the micro- spotted allergen was detected by peroxidase-conjugated anti-IgE antibody. The chemiluminescence intensities of the substrate decomposed by the peroxidase were detected with a sensitive CCD camera. All allergens were immobilized by this method and used to screen allergen-specific IgE.

Background: An important portion of asthmatics do not respond to current therapies. Thus, the need for new therapeutic drugs is urgent. We have demonstrated a critical role for PARP in experimental asthma. Olaparib, a PARP inhibitor, was recently introduced in clinical trials against cancer. The objective of the present study was to examine the efficacy of olaparib in blocking established allergic airway inflammation and hyperresponsiveness similar to those observed in human asthma in animal models of the disease. Methods: We used ovalbumin (OVA)-based mouse models of asthma and primary CD4+ T cells. C57BL/6J WT or PARP-1-/- mice were subjected to OVA sensitization followed by a single or multiple challenges to aerosolized OVA or left unchallenged. WT mice were administered, i.p., 1 mg/kg, 5 or 10 mg/kg of olaparib or saline 30 min after each OVA challenge. Results: Administration of olaparib in mice 30 min post-challenge promoted a robust reduction in airway eosinophilia, mucus production and hyperresponsiveness even after repeated challenges with ovalbumin. The protective effects of olaparib were linked to a suppression of Th2 cytokines eotaxin, IL-4, IL-5, IL-6, IL-13, and M-CSF, and ovalbumin- specific IgE with an increase in the Th1 cytokine IFN-γ. These traits were associated with a decrease in splenic CD4+ T cells and concomitant increase in T-regulatory cells. The aforementioned traits conferred by olaparib administration were consistent with those observed in OVA-challenged PARP-1-/- mice. Adoptive transfer of Th2-skewed OTII- WT CD4+ T cells reversed the Th2 cytokines IL-4, IL-5, and IL-10, the chemokine GM-CSF, the Th1 cytokines IL-2 and IFN-γ, and ovalbumin-specific IgE production in ovalbumin-challenged PARP-1-/-mice suggesting a role for PARP-1 in CD4+ T but not B cells. In ex vivo studies, PARP inhibition by olaparib or PARP-1 gene knockout markedly reduced CD3/CD28-stimulated gata-3 and il4 expression in Th2-skewed CD4+ T cells while causing a moderate elevation in t-bet and IFN-γ expression in Th1-skewed CD4+ T cells. Conclusions: Our findings show the potential of PARP inhibition as a viable therapeutic strategy and olaparib as a likely candidate to be tested in human asthma clinical trials. [ABSTRACT FROM AUTHOR]