Allergic Subjects Research Articles

Detection of Allergen-IgE interaction in Allergic Children Through Combined Impedance and ROS Measurements

The aim of this paper is to provide an alternative and more efficient screening test for allergy with respect to traditional UNICAP and prick skin tests, which are commercially available nowadays. The proposed test is aimed at monitoring the interaction of the IgE protein with the allergen in an allergic subject. The idea relies on the fact that the interaction between the IgE and the corresponding allergen happens through binding mechanisms between the IgE and the allergen structure, changing the ionic content of the serum. This latter can be monitored by impedance measurements and confirmed by reactive oxygen species (ROS) evaluation in leukocyte subpopulations. Therefore, in this paper we propose a novel system for the detection of allergies based on impedance measurements and confirmed by ROS measurements. This type of measurement approach has never been presented in the literature for this purpose. A measurement protocol has been developed to guarantee reproducibility and repeatability of the assay. This paper describes both the instrument—specifically designed for impedance evaluation—and the measurement protocols for both methodologies, defining univocally, in this way, the overall measurement technique.

Toll-like receptor-induced expression of epithelial cytokine receptors on haemopoietic progenitors is altered in allergic asthma.

Haemopoietic progenitor cells (HPC) migrate to sites of allergic inflammation where, upon stimulation with epithelial cytokines, they produce Th2 cytokines and differentiate into mature eosinophils and basophils. They also express Toll-like receptors (TLR) involved in antimicrobial responses. The objective of this study was to compare TLR expression on peripheral blood HPC and TLR-induced responses, in particular changes in epithelial cytokine receptors, in healthy and asthmatic subjects at baseline and following allergen challenge. Ten healthy and 11 allergic asthmatic subjects were studied. HPC-enriched cell populations were stimulated with TLR-2, TLR-4 or TLR-9 ligands. TLR expression by circulating HPC and interleukin (IL)-25 (IL-17RB), IL-33 (ST2) and thymic stromal lymphopoietin receptor (TSLPR) expression after TLR ligation were examined by flow cytometry at baseline and, in asthmatics, following allergen challenge. The effects of dexamethasone (Dex) on TLR-induced responses were also assessed. Asthmatics had significantly lower circulating HPC expressing TLR-2 and TLR-9 with a similar trend for TLR-4. TLR-4 stimulation of HPC yielded higher numbers of TSLPR+ cells in asthmatics compared with healthy subjects. A similar trend was seen for TLR-9 ligation, an effect further augmented by allergen inhalation. Allergen challenge also enhanced TLR-induced ST2 expression on HPC. Treatment with Dex in vitro increased TLR-4-induced TSLPR expression but had no effect on other epithelial cytokine receptors. These data demonstrate an interaction between allergen and TLR ligand exposure in asthmatics. Allergen inhalation augments the TLR-induced inflammatory response by HPC, possibly leading to increased "in situ haemopoiesis" through up-regulation of TSLPR. These findings show that HPC may be a part of the pro-inflammatory cascade in pathogen-induced asthma exacerbation through their increased responsiveness to TLR stimulation.

Comparison of inflammatory cytokine release from nasal epithelial cells of non-atopic non-rhinitic, allergic rhinitic and polyp subjects and effects of diesel exhaust particles in vitro

BackgroundAlthough studies have reported an association between air pollutants and increased allergic airway diseases, such as allergic rhinitis and nasal polyposis, the underlying mechanisms are not fully understood. A limited number of studies have suggested that diesel exhaust particles (DEP) play a role in atopy and the pathogenesis of allergic upper airway diseases. The aim of this study was to investigate the effect of DEP on inflammatory cytokine release, and mRNA expression of transcription factors such as JNK and NF-β in primary nasal epithelial cells (NECs), in vitro. MethodsNECs from non-atopic, non-rhinitic subjects (controls) and patients with allergic rhinitis and nasal polyps were cultured and incubated with 0–100μg/ml DEP for 24h. ELISA and RT-PCR were used to assess the release of IL-8, GM-CSF, and RANTES, and mRNA expression for JNK and NF-κB, respectively. ResultsCompared to control cells, NECs from subjects with atopic polyps released significantly greater amounts of IL-8 (median=887 vs. 176.6pg/μg cellular protein; p<0.0001) and RANTES (median=0.191 vs. 0.02pg/μg cellular protein; p<0.001). While 50μg/ml DEP induced release of RANTES in NECs from patients with allergic rhinitis, 100μg/ml DEP decreased IL-8 levels in NECs from both control and allergic rhinitic subjects. DEP did not affect mRNA expression for JNK and NF-κB from NECs of subjects with polyps. ConclusionsNECs from subjects with various pathologies may respond differently to DEP.

Peanut Allergen Threshold Study (PATS): Novel single-dose oral food challenge study to validate eliciting doses in children with peanut allergy

Eliciting doses (EDs) of allergenic foods can be defined by the distribution of threshold doses for subjects within a specific population. The ED05 is the dose that elicits a reaction in 5% of allergic subjects. The predicted ED05 for peanut is 1.5mg of peanut protein (6mg of whole peanut). We sought to validate the predicted peanut ED05 (1.5mg) with a novel single-dose challenge. Consecutive eligible children with peanut allergy in 3 centers were prospectively invited to participate, irrespective of previous reaction severity. Predetermined criteria for objective reactions were used to identify ED05 single-dose reactors. Five hundred eighteen children (mean age, 6.8years) were eligible. No significant demographic or clinical differences were identified between 381 (74%) participants and 137 (26%) nonparticipants or between subjects recruited at each center. Three hundred seventy-eight children (206 male) completed the study. Almost half the group reported ignoring precautionary allergen labeling. Two hundred forty-five (65%) children experienced no reaction to the single dose of peanut. Sixty-seven (18%) children reported a subjective reaction without objective findings. Fifty-eight (15%) children experienced signs of a mild and transient nature that did not meet the predetermined criteria. Only 8 (2.1%; 95% CI, 0.6%-3.4%) subjects met the predetermined criteria for an objective and likely related event. No child experienced more than a mild reaction, 4 of the 8 received oral antihistamines only, and none received epinephrine. Food allergy-related quality of life improved from baseline to 1month after challenge regardless of outcome (η2=0.2, P<.0001). Peanut skin prick test responses and peanut- and Ara h 2-specific IgE levels were not associated with objective reactivity to peanut ED05. A single administration of 1.5mg of peanut protein elicited objective reactions in fewer than the predicted 5% of patients with peanut allergy. The novel single-dose oral food challenge appears clinically safe and patient acceptable, regardless of the outcome. It identifies the most highly dose-sensitive population with food allergy not otherwise identifiable by using routinely available peanut skin prick test responses or specific IgE levels, but this single-dose approach has not yet been validated for risk assessment of individual patients.

The Impact on Allergy-Related Cells of a Birch Pollen Allergoid, with and without Monophosphoryl Lipid A, in Comparison with the Native Equivalent

Background: The clinical efficacy and safety of allergoid immunotherapy have been demonstrated in clinical trials. However, simultaneous monitoring of the immunological changes by allergoids versus allergens in the cells of the same individual has not been extensively performed, and the impact of concurrent Toll-like receptor 4 (TLR4) ligation has not been specified. Methods: Three types of birch allergen were utilized: glutaraldehyde-treated allergoid (extract A), the same allergoid plus monophosphoryl lipid A (MPL), i.e., TLR4 ligand (extract A*), and native allergen (extract B). Antigen-specific responses after the in vitro stimulation of blood cells with the extracts were assessed by studying costimulatory receptors on the B cell surface by flow cytometry, cytokine responses by ELISA, and CD63 and CD203c upregulation (basophil activation test) in allergic versus nonallergic subjects. Results: HLA-DR selectively increased upon allergen or allergoid treatment in the allergic group only. The extract types elicited similar cytokine responses, with IL-6 and IL-10 production detected only in certain atopic subjects. The allergoids revealed a strong reduction (100- to <10,000-fold) in basophil activation versus native allergen. Reactivity was undetectable in the basophils from nonallergic subjects. Conclusion: The allergenicity of the allergoid employed was sharply reduced when compared to the native allergen, while its immunogenicity was largely retained, especially in the presence of MPL. We also provide further evidence that allergic and nonallergic individuals show preexisting differences in their immune repertoires.

N-glycan in cockroach allergen regulates human basophil function.

IntroductionCockroach allergen exposure elicits cockroach sensitization and poses an increased risk for asthma. However, the major components in cockroach allergen and the mechanisms underlying the induction of cockroach allergen‐induced allergy and asthma remain largely elusive. We sought to examine the role of cockroach‐associated glycan in regulating human basophil function.MethodsN‐linked glycans from naturally purified cockroach allergen Bla g 2 were characterized by MALDI‐TOF mass spectrometry. Binding of cockroach allergen to serum IgE from cockroach allergic subjects was determined by solid‐phase binding immunoassays. Role of cockroach associated glycan in histamine release and IL‐4 production from human basophils was examined. Expression of C‐type lectin receptors (CLRs) and their role in mediating glycan‐uptake in the basophils was also investigated.ResultsMALDI‐TOF mass spectrometric analysis of N‐glycan from Bla g 2 showed complex hybrid‐types of glycans that terminated with mannose, galactose, and/or N‐acetyl glucosamine (GlcNAc). Deglycosylated Bla g 2 showed reduced binding to IgE and was less capable of inducing histamine release from human basophils. In contrast, N‐glycan derived from Bla g 2 significantly inhibited histamine release and IL‐4 production from basophils passively sensitized with serum from cockroach allergic subjects. An analysis of CLRs revealed the expression of DC‐SIGN and DCIR, but not MRC1 and dectin‐1, in human basophils. Neutralizing antibody to DCIR, but not DC‐SIGN, significantly inhibited Bla g 2 uptake by human basophils. A dose‐dependent bindings of cockroach allergen to DCIR was also observed.ConclusionsThese observations indicate a previously unrecognized role for cockroach allergen‐associated glycans in allergen‐induced immune reactions, and DCIR may play a role in mediating the regulation of glycan on basophil function.

Periostin up-Regulates the Effector Functions of Eosinophils

Periostin, an extracellular matrix protein, is up-regulated in asthmatic airways, mainly by T helper (Th) 2 cytokines. Periostin functions as a matricellular protein in cell activation by binding to cell surface receptors. However, the role of periostin in the development of eosinophilic airway inflammation has not been fully clarified. Here, we examined whether periostin could modify eosinophil functions such as superoxide anion (O2−) generation, degranulation, and production of cytokine/chemokines. Eosinophils were isolated from the blood of healthy volunteers or allergic subjects, and their adhesion to recombinant human periostin was measured using eosinophil peroxidase assays. Eosinophil O2−generation was examined based on the superoxide dismutase-inhibitable reduction of cytochrome C. Eosinophil-derived neurotoxin (EDN) concentrations in cell media were measured by ELISA as a marker of degranulation, and concentrations of cytokine/chemokines were also measured. Periostin directly induces eosinophil adhesion, which was enhanced by IL-5. Periostin also activated other functions of eosinophils such as O2− generation and EDN release. Anti-αM or anti-β2 integrin monoclonal antibody suppressed the eosinophil adhesion, O2−generation, and EDN release induced by periostin. Finally, periostin increased the production of transforming growth factor (TGF-β)1, TGF-β2 and cysteinyl leukotrienes (cysLTs) from eosinophils. These findings suggested that periostin up-regulates eosinophil functions through αMβ2 integrin. These effects may be involved in the activation of eosinophils and in the development of remodeling in the airway of Th2-dominant asthma, and could possibly aggravate the disease.

Long-Term Treatment with 300IR 5-Grass Pollen Sublingual Tablet in Grass Pollen Allergic Subjects: Safety Outcome Year after Year

Long-Term Treatment with 300IR 5-Grass Pollen Sublingual Tablet in Grass Pollen Allergic Subjects: Safety Outcome Year after Year

Glucagon-like peptide-1 receptor expression on human eosinophils and its regulation of eosinophil activation.

Glucagon-like peptide-1 (GLP-1) and its receptor are part of the incretin family of hormones that regulate glucose metabolism. GLP-1 also has immune modulatory roles. To measure the expression of the GLP-1 receptor (GLP-1R) on eosinophils and neutrophils in normal and asthmatic subjects and evaluate effects of a GLP-1 analog on eosinophil function. Peripheral blood samples were taken from 10 normal and 10 allergic asthmatic subjects. GLP-1R expression was measured on eosinophils and neutrophils. Subsequently, the asthmatic subjects underwent allergen and diluent inhalation challenges, and GLP-1R expression was measured. Purified eosinophils, collected from mild asthmatic subjects, were stimulated with lipopolysaccharide (LPS) and a GLP-1 analog to evaluate eosinophil cell activation markers CD11b and CD69 and cytokine (IL-4, IL-5, IL-8 and IL-13) production. Glucagon-like peptide-1 receptor is expressed on human eosinophils and neutrophils. Eosinophil, but not neutrophil, expression of GLP-1R is significantly higher in normal controls compared to allergic asthmatics. The expression of GLP-1R did not change on either eosinophils or neutrophils following allergen challenge. A GLP-1 analog significantly decreased the expression of eosinophil-surface activation markers following LPS stimulation and decreased eosinophil production of IL-4, IL-8 and IL-13, but not IL-5. Glucagon-like peptide-1 receptor is expressed on human eosinophils and neutrophils. A GLP-1 analog attenuates LPS-stimulated eosinophil activation. GLP-1 agonists may have additional adjunctive indications in treating persons with concomitant type 2 diabetes mellitus and asthma.

A procedure for grouping food consumption data for use in food allergen risk assessment

Food allergic subjects need to avoid the allergenic food that triggers their allergy. However, foods can also contain unintended allergens. Food manufacturers or authorities need to perform a risk assessment to be able to decide if unintended allergen presence constitutes a risk to food allergic consumers. One of the input parameters in risk assessment is the amount of a given food consumed in a meal. There has been little emphasis on how food consumption data can be used in food allergen risk assessment. The aim of the study was to organize the complex datasets from National Food Consumption Surveys from different countries (France, Netherlands and Denmark) to be manageable in food allergen risk assessment. To do this, a two-step method was developed. First, based on initial groups of similar food items, the homogeneity of consumption was evaluated using a customized clustering method. Then, the risk was calculated for each initial food group and its subgroups to verify if it also represents a relevant difference in risk. Forty-eight food groups were designated in Denmark (53 in the Netherlands, 54 in France). Finally, summary statistics and names for each food group for the Danish data illustrate the results when applying the procedure.

Update on Potential Therapies for IgE-Mediated Food Allergy

Food allergy is common, affecting up to 8% of children in the USA. Currently, therapy is limited to avoidance of the implicated allergen and availability of self-injectable epinephrine. However, several new approaches to food allergy are under investigation. This article reviews the published data on these new approaches. Oral immunotherapy, in which allergic subjects are exposed to increasing amounts of antigen, can be accomplished in the majority of allergic individuals. However, this approach is not a cure as most patients will react after cessation of regular intake. In addition, there is a high rate of side effects. Other approaches include epicutaneous immunotherapy, therapy with anti-IgE medications, and use of Chinese herbs. Several novel approaches on food allergy are under study. At the current time, these approaches show promise for preventing severe reactions to accidental exposures. However, there is little evidence that current approaches will represent a true cure for food allergy.

Component-resolved diagnostics in Thai children with cow's milk and egg allergy.

Component-resolved diagnostics (CRD) have garnered a lot of attention in recent years in the diagnosis of food allergies. We aimed to investigate sensitization against cow's milk and egg white components, and to study the clinical usefulness of serum food-specific immunoglobulin E (sIgE) to cow's milk and egg white components in Thai children with cow's milk and egg allergies. Children with IgE-mediated cow's milk and/or egg allergy were enrolled. Clinical reactions were determined. Specific IgE against cow's milk, egg white, alpha-lactalbumin (ALA), beta-lactoglobulin (BLG), casein, ovomucoid, ovalbumin and conalbumin were measured. Thirteen cow's milk allergic subjects and 32 egg allergic subjects were identified. The sensitization rate to BLG and casein was 91.7%, followed by ALA (66.7%) for cow's milk, and the sensitization rate to ovalbumin was 93.8%, followed by ovomucoid (81.3%) and conalbumin (37.5%) for egg. Patients in the urticaria group had a higher level of casein sIgE than the atopic dermatitis (AD) group but this difference was not significant (9.8 kUA/L vs. 0.9 kUA/L, p = 0.11). The level of ovomucoid sIgE was significantly higher in the non-AD group than in the AD group (3.8 kUA/L vs. 1.3 kUA/L, p = 0.048). BLG and casein for cow's milk and ovomucoid and ovalbumin for egg were the common components causing sensitization in cow's milk and egg allergic patients. Among the patients with cow's milk allergy, the level of casein sIgE in the urticaria group tended to be higher than the AD group, and in egg allergic patients, the non-AD group had a significantly higher ovomucoid sIgE level compared with the AD group.

T-Cell Proliferation Assay: Determination of Immunodominant T-Cell Epitopes of Food Allergens.

Characterization of allergen-specific T cells is critical to understand their contribution to disease pathogenesis. The identification of immunodominant T-cell epitopes is crucial for development of T-cell-based vaccines. Peptide-specific T-cell proliferation studies are usually performed in a library of short synthetic peptides (15mer or 20mer) with 3 or 5 offset spanning the entire length of the allergen. T-cell peptide epitopes lack the primary and tertiary structure of the native protein to cross-link IgE, but retain the ability to stimulate T cells. The peptides sequences can also be obtained either by in silico approaches and in vitro binding assays. The efficacy of T-cell epitope-based peptide immunotherapy has been proven in certain allergies. The present methodology describes T-cell proliferation assays using whole blood sample from allergic subjects.

Occurrence of COPD in Patients with Respiratory Allergy: A Clinico-Spirometric Evaluation in a Tertiary Hospital, Kolkata.

Smoking is established as the most important causative factor responsible for Chronic Obstructive Pulmonary Disease (COPD). Occurrence of allergy in COPD patients causes acute exacerbation of this disease, but role of allergy is not established in aetiopathogenesis of COPD. The present study was aimed at evaluation of occurrence of COPD in patients having symptoms suggestive of respiratory allergy. An observational cross-sectional study was conducted to evaluate occurrence of COPD in patients having respiratory allergic symptoms by routine spirometric screening. Five hundred and fifty urban patients aged 18-60 years (both gender) ailing from chronic respiratory symptoms like cough, wheeze and Shortness Of Breath (SOB), who were referred from OPDs of RGKMCH, Kolkata, were included in this study. After obtaining detailed clinical profile, patients were divided into two groups: subjects having additional clinical symptoms suggestive of respiratory allergy (n=260) like nasal catarrh, nasal stuffiness and sneezing and subjects with no symptoms suggestive of respiratory allergy (n=290). Thereafter, routine spirometry was carried out following recommendations of ATS/ERS (2005). Patients were then categorized based on FVC, FEV1, FEV1/FVC, FEF25-75 and PEFR percent predicted values. Study revealed that 18.97% of non-allergic population was suffering from COPD whereas only 7.69% of allergic subjects had COPD. This difference was statistically highly significant (p=0.0001). Although there was no significant difference in prevalence of respiratory symptoms between these two groups. Present study concludes that patients with respiratory allergy may have coexistent COPD but occurrence of COPD is much less than that in patients with no respiratory allergy.

Effect of house dust mite on neutrophil apoptosis by cytokine secretion in lymphocytes

Although extract of Dermatophagoides farinae (DF) alone had no effect on neutrophil apoptosis, it inhibited neutrophil apoptosis in neutrophils cocultured with lymphocytes in both normal and allergic subjects. DF showed a stronger inhibition on the apoptosis of allergic neutrophils cocultured with allergic lymphocytes than that of normal neutrophils cocultured with normal lymphocytes. DF induced the secretion of IL-6, IL-8, MCP-1, and GM-CSF of the normal and allergic lymphocytes. The cytokine secretion due to DF in allergic lymphocytes is higher than in normal lymphocytes. The cytokine secretion of normal and allergic lymphocytes induced by DF was suppressed by PAR2i, a PAR2 inhibitor, LY294002, a PI3K inhibitor, AKTi, an Akt inhibitor, PD98059, an ER K inhibitor, and BAY-11-7085, an NF-κB inhibitor. Phosphorylation of ERK induced by DF was suppressed by PAR2i, LY294002 and AKTi, and NF-κB activity due to DF was inhibited by PAR2i, LY294002, AKTi, and PD98059.

Exploratory analysis of CD63 and CD203c expression in basophils from hazelnut sensitized and allergic individuals.

BackgroundSensitization to hazelnut (HN) is frequent and requires clarification to determine whether this sensitization is clinically relevant. The aim of this study was to investigate basophil activation profiles in HN-sensitized and allergic subjects.MethodsBasophil activation was determined by flow cytometric analyses of CD63 and CD203c expression using several HN allergen concentrations. Depending on their clinical reaction pattern, an oral allergy symptom group (OAS, n = 20), a systemic reaction group (n = 12) and a sensitized group without clinical symptoms (n = 20) were identified. Additionally, 10 non-allergic and non-sensitized individuals served as controls.ResultsCD63 and CD203c expression differed between allergic (OAS and systemic group) and sensitized subjects. The HN concentration required to activate 30% of CD203c+ basophils [effective concentration (EC)30] was significantly higher in sensitized versus the allergic group (p = 0.0089). This was more pronounced when the basophil allergen threshold sensitivity (CD-sens) was calculated (CD63: p = 0.018; CD203c: p = 0.009).ConclusionOur data indicate that the basophil activation test may provide information to better distinguish between sensitized and allergic subjects if several allergen concentrations are considered. CD203c expression displayed a better discrimination compared to CD63; therefore, its diagnostic value might be superior compared with CD63.Electronic supplementary materialThe online version of this article (doi:10.1186/s13601-016-0134-7) contains supplementary material, which is available to authorized users.

Old Apple (Malus domestica L. Borkh) Varieties with Hypoallergenic Properties: An Integrated Approach for Studying Apple Allergenicity.

Freshly consumed apples (Malus domestica L. Borkh) can cause allergic reactions because of the presence of four classes of allergens. Knowledge of the genetic factors affecting the allergenic potential of apples would provide important information for the selection of hypoallergenic genotypes, which can be combined with the adoption of new agronomical practices to produce fruits with a reduced amount of allergens. In the present research, a multiple analytical approach was adopted to characterize the allergenic potential of 24 apple varieties released at different ages (pre- and post-green revolution). A specific workflow was set up including protein quantification by means of polyclonal antibodies, immunological analyses with sera of allergic subjects, enzymatic assays, clinical assessments on allergic patients, and gene expression assays on fruit samples. Taken as a whole, the results indicate that most of the less allergenic genotypes were found among those deriving from selection processes carried out prior to the so-called "green revolution".

Cytokine production by PBMC and serum from allergic and non-allergic subjects following in vitro histamine stimulation to test fexofenadine and osthole anti-allergic properties

FXF is a third-generation antihistamine drug and osthole is assumed a natural antihistamine alternative. This paper compares peripheral blood mononuclear cell (PBMC) incubation with FXF and osthole, by studying FXF, osthole and histamine cytokine secretion in PBMC in vitro cultures. Mabtech kits determined the interleukins IL-1β, IL-4, IL-10, IL-13 and TNF-α. The influence of the above active substances on cytokine secretion in PBMC's and serum was assessed: cytokines were IL-1β, IL-4, IL-10, IL-13 and TNF-α; and cytokine levels secreted by untreated PBMCs in pure culture medium formed the absolute control (ctrl).We determined that osthole affects PBMC cytokine secretion to almost precisely the same extent as FXF (IL-1β, IL-4, IL-10 and TNF). In addition osthole had greater IL-13 blocking ability than FXF. Moreover, we observed significantly decreased IL-4 level in histamine/osthole theatment compared to histamine alone. Meanwhile, FXF not significantly decrease the level of IL-4 increased by histamine. This data indicates osthole's strong role in allergic inflamation.All results confirm our hypothesis that osthole is a natural histamine antagonist and therefore can be beneficially used in antihistamine treatment of conditions such as allergies.

Oral Food Desensitization in Children With IgE-Mediated Cow's Milk Allergy: Immunological Changes Underlying Desensitization.

PurposeThis study aimed to evaluate the safety and efficacy to induce clinical desensitization to cow's milk (CM) of an oral immunotherapy (OIT) protocol in a pediatric population with cow's milk allergy (CMA). In addition, the immune responses against β-casein, of peripheral blood mononuclear cells (PBMCs) from CMA patients, before and after the protocol were evaluated and compared to a nonallergic population.MethodsA group of 20 children with IgE-mediated CMA and 15 nonallergic children were recruited. Allergic subjects underwent an OIT protocol based on weekly doses of commercial semi-skimmed ultra-high temperature treated (UHT) CM, followed by a maintenance phase. Immune profiles and changes in all subjects were investigated by measuring Th1, Th2, and Treg cytokines, transcription factors, and specific IgE and IgG4 levels.ResultsThe CM-OIT protocol enabled to desensitize 70% of the allergic patients. Successful OIT was accompanied by significant increases in casein-specific IgG4 levels, together with a reduction in the concentration of antigen-specific IgE and in IL-5, IL-13, and IL-10 production by β-casein-stimulated PBMCs. Baseline significant differences observed between allergic and nonallergic children in IL-13 and IL-5 levels were no longer found once the protocol had finished.ConclusionsThe OIT protocol was safe and effective in inducing milk desensitization in 70% of the children with CMA, leading to alterations in their immune profiles toward a nonallergic phenotype.

Three peanut-allergic/sensitized phenotypes with gender difference.

Peanut-allergic reactions are heterogeneous ranging from mild symptoms to anaphylaxis. Identify peanut-allergic/sensitized phenotypes to personalize patient management. A combined factor and cluster analysis was used to study the phenotypes of 696 patients diagnosed with peanut sensitization and enrolled in the MIRABEL survey. The method was first applied to the 247 patients with an oral food challenge (OFC). It was then applied to the 449 patients without OFC to confirm the findings in an independent population. Three independent clusters emerged from the OFC subgroup. Cluster 1, 'Severe peanut allergy with little allergic multi-morbidity' (123 subjects), had the highest proportion of patients with positive OFC (92%), a medium level of peanut protein inducing a positive OFC (235mg), lower percentage of allergic multi-morbidity (2% asthma plus atopic dermatitis (A + AD), no cases of A + AD + multiple food allergies (MFA)). Cluster 2, 'Severe peanut allergy with frequent allergic multi-morbidity' (62 subjects), had a high proportion of patients with positive OFC (85%) with the lowest level of peanut protein inducing a positive OFC (112mg), 89% allergic subjects, 100% with allergic multi-morbidity (A + AD) and 84% with A + AD + MFA. Cluster 3, 'Mild peanut-allergic/sensitized phenotype' (62 subjects), had the lowest mean age, the lowest proportion of patients with positive OFC (53%) with a high level of peanut protein inducing a positive OFC (770mg), a low percentage of allergic multi-morbidity (48% A + AD + MFA). The two severe peanut-allergic phenotypes were more frequent in girls. The same clusters were found in the subgroup of patients without OFC. Besides the classic markers associated with lower threshold doses of OFC (such as SPT and rAra h 2), allergic multi-morbidity and female gender should also be taken into account to better adapt the progressive dosage of provocation tests.