Milk Allergen Micro-Array (MAMA) for Refined Detection of Cow's-Milk-Specific IgE Sensitization.

BACKGROUND: Immunoglobulin-E(IgE)-mediated hypersensitivity to cow's milk allergens is a frequent cause of severe and life-threatening anaphylactic reactions. Besides case histories and controlled food challenges, the detection of the IgE antibodies specific to cow's milk allergens is important for the diagnosis of cow-milk-specific IgE sensitization. Cow´s milk allergen molecules provide useful information for the refined detection of cow-milk-specific IgE sensitization. METHODS: A micro-array based on ImmunoCAP ISAC technology was developed and designated milk allergen micro-array (MAMA), containing a complete panel of purified natural and recombinant cow's milk allergens (caseins, α-lactalbumin, ß-lactoglobulin, bovine serum albumin-BSA and lactoferrin), recombinant BSA fragments, and α-casein-, α-lactalbumin- and ß-lactoglobulin-derived synthetic peptides. Sera from 80 children with confirmed symptoms related to cow's milk intake (without anaphylaxis: n = 39; anaphylaxis with a Sampson grade of 1-3: n = 21; and anaphylaxis with a Sampson grade of 4-5: n = 20) were studied. The alterations in the specific IgE levels were analyzed in a subgroup of eleven patients, i.e., five who did not and six who did acquire natural tolerance. RESULTS: The use of MAMA allowed a component-resolved diagnosis of IgE sensitization in each of the children suffering from cow's-milk-related anaphylaxis according to Sampson grades 1-5 requiring only 20-30 microliters of serum. IgE sensitization to caseins and casein-derived peptides was found in each of the children with Sampson grades of 4-5. Among the grade 1-3 patients, nine patients showed negative reactivity to caseins but showed IgE reactivity to alpha-lactalbumin (n = 7) or beta-lactoglobulin (n = 2). For certain children, an IgE sensitization to cryptic peptide epitopes without detectable allergen-specific IgE was found. Twenty-four children with cow-milk-specific anaphylaxis showed additional IgE sensitizations to BSA, but they were all sensitized to either caseins, alpha-lactalbumin, or beta-lactoglobulin. A total of 17 of the 39 children without anaphylaxis lacked specific IgE reactivity to any of the tested components. The children developing tolerance showed a reduction in allergen and/or peptide-specific IgE levels, whereas those remaining sensitive did not. CONCLUSIONS: The use of MAMA allows for the detection, using only a few microliters of serum, of IgE sensitization to multiple cow's milk allergens and allergen-derived peptides in cow-milk-allergic children with cow-milk-related anaphylaxis.

Extensively Hydrolyzed Hypoallergenic Infant Formula with Retained T Cell Reactivity.

BACKGROUND: Immunoglobulin E (IgE)-mediated cow's milk allergy (CMA) can be life-threatening and affects up to 3% of children. Hypoallergenic infant formulas based on hydrolyzed cow's milk protein are increasingly considered for therapy and prevention of cow's milk allergy. The aim of this study was to investigate the allergenic activity and ability to induce T cell and cytokine responses of an infant formula based on extensively hydrolyzed cow's milk protein (whey) (eHF, extensively hydrolyzed formula) supplemented with Galactooligosaccharides (GOS) and Limosilactobacillus fermentum CECT5716 (LF) to determine its suitability for treatment and prevention of CMA. METHODS: eHF and standard protein formula based on intact cow's milk proteins (iPF) with or without Galactooligosaccharide (GOS) and Limosilactobacillus fermentum CECT5716 (LF) were investigated with allergen-specific antibodies and tested for IgE reactivity and allergenic activity in basophil degranulation assays with sera from cow's milk (CM)-allergic infants/children. Their ability to stimulate T cell proliferation and cytokine secretion in cultured peripheral blood mononuclear cells (PBMC) from CM-allergic infants and children was studied with a FACS-based carboxyfluorescein diacetate succinimidyl ester (CFSE) dilution assay and xMAP Luminex fluorescent bead-based technology, respectively. RESULTS: An eHF supplemented with GOS and LF exhibiting almost no IgE reactivity and allergenic activity was identified. This eHF induced significantly lower inflammatory cytokine secretion as compared to an intact protein-based infant formula but retained T cell reactivity. CONCLUSIONS: Due to strongly reduced allergenic activity and induction of inflammatory cytokine secretion but retained T cell reactivity, the identified eHF may be used for treatment and prevention of CMA by induction of specific T cell tolerance.

Primary Immunodeficiencies in Russia: Data From the National Registry.

Introduction: Primary immunodeficiencies (PID) are a group of rare genetic disorders with a multitude of clinical symptoms. Characterization of epidemiological and clinical data via national registries has proven to be a valuable tool of studying these diseases. Materials and Methods: The Russian PID registry was set up in 2017, by the National Association of Experts in PID (NAEPID). It is a secure, internet-based database that includes detailed clinical, laboratory, and therapeutic data on PID patients of all ages. Results: The registry contained information on 2,728 patients (60% males, 40% females), from all Federal Districts of the Russian Federation. 1,851/2,728 (68%) were alive, 1,426/1,851 (77%) were children and 425/1,851 (23%) were adults. PID was diagnosed before the age of 18 in 2,192 patients (88%). Antibody defects (699; 26%) and syndromic PID (591; 22%) were the most common groups of PID. The minimum overall PID prevalence in the Russian population was 1.3:100,000 people; the estimated PID birth rate is 5.7 per 100,000 live births. The number of newly diagnosed patients per year increased dramatically, reaching the maximum of 331 patients in 2018. The overall mortality rate was 9.8%. Genetic testing has been performed in 1,740 patients and genetic defects were identified in 1,344 of them (77.2%). The median diagnostic delay was 2 years; this varied from 4 months to 11 years, depending on the PID category. The shortest time to diagnosis was noted in the combined PIDs-in WAS, DGS, and CGD. The longest delay was observed in AT, NBS, and in the most prevalent adult PID: HAE and CVID. Of the patients, 1,622 had symptomatic treatment information: 843 (52%) received IG treatment, mainly IVIG (96%), and 414 (25%) patients were treated with biological drugs. HSCT has been performed in 342/2,728 (16%) patients, of whom 67% are currently alive, 17% deceased, and 16% lost to follow-up. Three patients underwent gene therapy for WAS; all are currently alive. Conclusions: Here, we describe our first analysis of the epidemiological features of PID in Russia, allowing us to highlight the main challenges around PID diagnosis and treatment.

Quality of life associated with maternal anxiety disorder in Russian children and adolescents with food allergy.

BACKGROUND: Food allergy negatively impacts the quality of life (QoL) and has been associated with increased maternal anxiety. There is currently a lack of data assessing QoL of food-allergic children and adolescents in Russia. We aimed to evaluate the performance of the Food Allergy Quality of Life Questionnaire (FAQLQ) measures in a Russian sample of children, adolescents, and mothers and to investigate association between child QoL and maternal general anxiety. METHODS: FAQLQ and Food Allergy Independent Measures (FAIM) were translated to Russian to be completed by participants. Mothers also filled the General Anxiety Disorder (GAD-7) questionnaire. Reliability, construct, and discriminant validity of the FAQLQs and association between FAQLQ scores and general anxiety were assessed. RESULTS: A total of 142 participants completed FAQLQ and FAIM. In total, 126 mothers completed GAD-7 questionnaire. All FAQLQs showed excellent internal consistency (α > 0.94). The means for the FAQLQ-PF, FAQLQ-CF, and FAQLQ-TF varied according to the number of foods avoided, severity of symptoms, and FAIM and GAD scores (ηp2 0.40 P < .001). One in 5 mothers had GAD-7 score indicative of moderate-to-severe anxiety. Association strength between GAD and FAQLQ increased with age. GAD score was associated with FAQLQ-PF score, controlling for multiple confounders. CONCLUSION: The FAQLQ showed good reliability and validity in Russian. This study identified number of foods avoided and reported reaction severity being associated with greater QoL impairment in Russian children and adolescents affected by food allergy. The significant association found between general anxiety in parents and QoL in children and adolescents has practice, screening, and resource implications.

A national survey of Russian physicians' knowledge of diagnosis and management of food-induced anaphylaxis.

OBJECTIVES: Food allergy is an increasing burden worldwide and is a common problem within paediatric populations, affecting 5%-8% of children. Anaphylaxis caused by food proteins is a potentially life-threatening condition and all healthcare practitioners should be aware of its recognition and management. Russia is the largest country in Europe but it is still unknown whether physicians are prepared to diagnose and manage food-induced anaphylaxis effectively. We aimed to examine physicians' knowledge of diagnosis and management of food-induced anaphylaxis. SETTING, POPULATION AND OUTCOMES: A survey was designed and published online at VrachiRF.ru website (for registered Russian-speaking practicing physicians). We obtained information on respondents' clinical settings, experience and specialty. Survey questions were based on a characteristic clinical scenario of anaphylaxis due to food ingestion. Outcome measures consisted of correct answers to the anaphylaxis diagnosis and management questions. RESULTS: From a total of 707 of physicians accessed in the survey, 315 (45%) responded to the clinical scenario. 16 respondents reported training in allergy-immunology and have been excluded from the analysis, leaving the final sample size of 299. Respondents were paediatricians (68%) and other specialties adult physicians (32%). Overall, 100 (33%) of respondents diagnosed anaphylaxis, but only 29% of those making the correct diagnosis administered adrenalin (1:1000) intramuscular. Respondents working in secondary/tertiary clinics diagnosed anaphylaxis significantly more often (p=0.04) when compared with primary care/private practice physicians. This difference was also apparent as the most important influence on responses in the multivariate analysis. CONCLUSIONS: In this national sample of Russian physicians, we found poor knowledge in both anaphylaxis diagnosis and management. Our data show that the chance of being properly diagnosed with anaphylaxis is 33% and being appropriately treated with adrenalin is 10%. These findings highlight lack of anaphylaxis knowledge among Russian physicians, both paediatricians and other specialists and illustrates the urgent need for allergy/anaphylaxis training.

Immune Components in Human Milk Are Associated with Early Infant Immunological Health Outcomes: A Prospective Three-Country Analysis.

The role of breastfeeding in improving allergy outcomes in early childhood is still unclear. Evidence suggests that immune mediators in human milk (HM) play a critical role in infant immune maturation as well as protection against atopy/allergy development. We investigated relationships between levels of immune mediators in colostrum and mature milk and infant outcomes in the first year of life. In a large prospective study of 398 pregnant/lactating women in the United Kingdom, Russia and Italy, colostrum and mature human milk (HM) samples were analysed for immune active molecules. Statistical analyses used models adjusting for the site of collection, colostrum collection time, parity and maternal atopic status. Preliminary univariate analysis showed detectable interleukin (IL) 2 and IL13 in HM to be associated with less eczema. This finding was further confirmed in multivariate analysis, with detectable HM IL13 showing protective effect OR 0.18 (95% CI 0.04-0.92). In contrast, a higher risk of eczema was associated with higher HM concentrations of transforming growth factor ß (TGFß) 2 OR 1.04 (95% CI 1.01-1.06) per ng/mL. Parental-reported food allergy was reported less often when IL13 was detectable in colostrum OR 0.10 (95% CI 0.01-0.83). HM hepatocyte growth factor (HGF) was protective for common cold incidence at 12 months OR 0.19 (95% CI 0.04-0.92) per ng/mL. Data from this study suggests that differences in the individual immune composition of HM may have an influence on early life infant health outcomes. Increased TGFß2 levels in HM are associated with a higher incidence of reported eczema, with detectable IL13 in colostrum showing protective effects for food allergy and sensitization. HGF shows some protective effect on common cold incidence at one year of age. Future studies should be focused on maternal genotype, human milk microbiome and diet influence on human milk immune composition and both short- and long-term health outcomes in the infant.

Colostrum and Mature Human Milk of Women from London, Moscow, and Verona: Determinants of Immune Composition.

Cytokines and growth factors in colostrum and mature milk may play an important role in infant immune maturation, and may vary significantly between populations. We aimed to examine associations between environmental and maternal factors, and human milk (HM) cytokine and growth factor levels. We recruited 398 pregnant/lactating women in the United Kingdom, Russia, and Italy. Participants underwent skin prick testing, questionnaire interview, and colostrum and mature milk sampling. HM cytokine and growth factor levels were quantified by electro-chemiluminescence. We found significant geographical variation in growth factor levels, but no evidence of variation between sites in cytokine detectability. There was an inverse correlation between time of milk sampling and growth factor levels in colostrum for Hepatocyte Growth Factor (HGF) and TGFß1 and TGFß3, but not TGFß2, and levels were significantly higher in colostrum than mature milk for all growth factors. The kinetics of decline were different for each growth factor. Cytokines were present at much lower levels than growth factors, and the decline over time was less consistent. HM growth factors and cytokine levels vary between populations for unknown reasons. Levels of HM mediators decline at different rates postpartum, and these findings suggest specific biological roles for HM growth factors and cytokines in early postnatal development.

Prevalence of atopic diseases and the use of topical corticosteroids. Is there any connection?

The prevalence of atopic diseases (atopic dermatitis, bronchial asthma, allergic rhinitis) has considerably increased for the last 40 years. This tendency has coincided with the beginning of the epoch of the use of the topical corticosteroids, which have a potent immunomodulation action. This fact itself as well as a number of research results has allowed to formulate the following hypothesis: the use of topical corticosteroids in children of early age contributes to the increase of prevalence of atopic diseases in the developed countries. The offered hypothesis can explain lower prevalence of atopic diseases in rural areas and in children from families with the anthroposophic life style. In other side this hypothesis also capable to give explanation of increased level of atopic diseases among people with high socioeconomic level and in children from 1 child families. It is of a note that corticosteroids use in developing countries is limited due to economic reasons and active use of complementary medicine. If the proposed hypothesis is correct, a revision of the therapeutic approaches is necessary concerning the attitude towards application of topical corticosteroids in children suffering from various forms of eczema. The direct proof of this hypothesis can be found during prospective studies.