A small proline-rich protein (SPRR) gene variant contributes to atopic eczema and eczema-associated asthma susceptibility.

Introduction: There is some evidence that genetic variants in the epidermal differentiation complex (EDC) genes on chromosome 1q21 may be involved in the pathogenesis of atopic eczema (AE) similar to the well-known filaggrin gene (FLG) mutations. Aim: To evaluate the association of SNP in the small proline-rich protein 2B (SPRR2B) gene with atopic eczema and other allergic phenotypes and to investigate its possible interaction with FLG mutations. Material and methods: One hundred and eighty-eight children less than 2 years old were screened for the variant of allele rs6693927 in the SPRR2B gene and for 4 most prevalent FLG mutations. The variant of allele rs6693927 and all FLG mutations were genotyped by real-time polymerase chain reaction assays with subsequent melting curve analysis using SimpleProbe® probes. Results: The allele rs6693927[A] was associated with a significantly increased risk of AE (OR = 3.02; 95% CI: 1.17-8.00; p = 0.011) and the effect was independent of FLG risk alleles. The largest effects were observed in patients with a combined asthma-plus-eczema phenotype (OR = 5.44; 95% CI: 1.17-25.16; p = 0.029). Finally, in eczema, we found interactions of rs6693927[A] with FLG mutations, the risk of eczema was the most increased in the subjects who combined both rs6693927[A] allele and FLG mutations. Conclusions: The SPRR2B risk variant may play an important role in the development of atopic eczema and the particular eczema-associated asthma phenotype in young children. The effect seems to be independent of, and supplementary to, the well-known FLG mutations and may be modulated by gene-gene interactions.

Genetic Variants in Epidermal Differentiation Complex Genes as Predictive Biomarkers for Atopic Eczema, Allergic Sensitization, and Eczema-Associated Asthma in a 6-Year Follow-Up Case-Control Study in Children.

Atopic eczema is the most common chronic inflammatory skin disease of early childhood and is often the first manifestation of atopic march. Therefore, one challenge is to identify the risk factors associated with atopic eczema that may also be predictors of atopic disease progression. The aim of this study was to investigate the association of SNPs in hornerin (HRNR) and filaggrin-2 (FLG2) genes with childhood atopic eczema, as well as other atopic phenotypes. Genotyping for HRNR and FLG2 was performed in 188 children younger than 2 years of age, previously screened for the FLG null mutations, and followed at yearly intervals until the age of 6. We demonstrated that risk variants of HRNR rs877776[C] and FLG2 rs12568784[T] were associated with atopic eczema, allergic sensitization, and susceptibility to the complex phenotype-asthma plus eczema. These effects seem to be supplementary to the well-known associations for FLG mutations and may be modulated by gene-gene interactions. Additionally, in children with eczema, these genetic variants may also be considered, along with FLG mutations, as predictive biomarkers for eczema-associated asthma. In conclusion, our results indicate that genetic variants in the epidermal differentiation complex gene could contribute to the pathogenesis of atopic eczema and progression to subsequent allergic disease.

Breastfeeding and Allergy Effect Modified by Genetic, Environmental, Dietary, and Immunological Factors.

Breastfeeding (BF) is the most natural mode of nutrition. Its beneficial effect has been revealed in terms of both the neonatal period and those of lifelong effects. However, as for protection against allergy, there is not enough data. In the current narrative review, the literature within the last five years from clinical trials and population-based studies on breastfeeding and allergy from different aspects was explored. The aim of this review was to explain how different factors could contribute to the overall effect of BF. Special consideration was given to accompanying exposure to cow milk, supplement use, the introduction of solid foods, microbiota changes, and the epigenetic function of BF. Those factors seem to be modifying the impact of BF. We also identified studies regarding BF in atopic mothers, with SCFA as a main player explaining differences according to this status. Conclusion: Based on the population-based studies, breastfeeding could be protective against some allergic phenotypes, but the results differ within different study groups. According to the new research in that matter, the effect of BF could be modified by different genetic (HMO composition), environmental (cesarean section, allergen exposure), dietary (SCFA, introduction of solid food), and immunologic factors (IgG, IgE), thus partially explaining the variance.

Pet ownership in pregnancy and methylation pattern in cord blood.

Having pets in the house during the first years of life has been shown to protect against allergies. However, the result of different studies is heterogeneous. The aim of this study was to evaluate the methylation pattern in cord blood in relation to pet ownership during pregnancy.We investigated the methylation patterns of 96 cord blood samples, participants of the Epigenetic Hallmark of Maternal Atopy and Diet-ELMA project, born to mothers who either owned pets (n = 32) or did not own pets (n = 64) during their pregnancy. DNA from cord blood was analysed using the Infinium methylation EPIC. For statistical analysis, RnBeads software was applied.We found 113 differentially methylated sites (DMs) in the covariate-adjusted analysis (FDR p < 0.05), with small methylation differences. The top DMs were associated with genes: UBA7, THRAP3, GTDC1, PDE8A and SBK2. In the regional analysis, two promoter regions presented with significance: RN7SL621P and RNU6-211P. Cis-regulatory element analysis revealed significant associations with several immune-related pathways, such as regulation of IL18, Toll signalling, IL6 and complement.We conclude that pet exposure during pregnancy causes subtle but significant changes in methylation patterns in cord blood, which are reflected in the biological processes governing both innate and adaptive immune responses.

Sensitisation patterns and allergy outcomes in pregnant women living in the urban area.

BACKGROUND: Worldwide, allergy affects more than one billion people, with particularly rising prevalence in industrialised areas. Specifically, young adults appear to be predominantly targeted for an allergy diagnosis. Allergic diseases in pregnancy are mainly pre-existing but could also occur de novo. The immunological changes while pregnant, with increased Th2 lymphocyte activity, can facilitate allergen sensitisation. OBJECTIVE: The aim of this study was to evaluate the pattern of specific IgE (sIgE) sensitisation to common inhalant and food allergens in pregnancy, and assess its relationship to self-reported allergic disease. METHODS: We assessed 200 pregnant women, aged 20-38 years (mean age = 29 years), participant of ELMA (Epigenetic Hallmark of Maternal Atopy and Diet) study, living in a metropolitan area, with no pregnancy associated metabolic complications, for total IgE and allergen specific IgE to 20 allergens. RESULTS: 48% of pregnant women were sensitised to at least one allergen, at a cut-off point of 0.35 kU/L and they were assigned as atopic. However 42% in atopic group were not reporting any allergic disease. The most common inhalant allergens were: pollen (24.5%) and animal dander (23.5%). The most common food allergens were: cow's milk (5.5%) and apples (4.5%). 7.5% of women reported asthma, 21.5% allergic rhinitis, 11.5% atopic dermatitis and 18.5% food allergy. 8.5% of were taking medication for asthma or allergies. Atopic dermatitis had the highest tendency to become more severe during pregnancy. Total IgE values were significantly higher in atopic women. CONCLUSIONS: Allergic sensitisation is a common phenomenon in pregnancy. Some sensitisations could be asymptomatic. Further studies should investigate if sensitisation in mothers confers risks for immune alterations in their children.

IL4RA gene expression in relation to I50V, Q551R and C-3223T polymorphisms.

BACKGROUND: Interleukin 4 (IL-4) and its receptor play important roles in the pathologies of asthma and atopy. The alpha subunit of the IL-4 receptor (IL-4RA) is included in 2 types of receptors which have different modulatory effects on immune responses. This distinct pattern reflects involvement in the immunopathology of both asthma and atopy. A number of studies have proven the association between IL4RA gene polymorphisms and asthma and atopy, but it is still an open question whether these variants are functional. OBJECTIVES: To analyze the data from IL4RA gene expression in PBMC in relation to specific polymorphisms - the most frequently studied I50V and Q551R and the less known C-3223T. MATERIAL AND METHODS: The analysis was performed for 36 subjects, both atopic and non-atopic. Real-time polymerase chain reaction (PCR) was used with specific primers for the quantification and genotyping. Delta Ct (ΔCT) and delta-delta Ct (ΔΔCT) values were used for the relative quantification of IL4RA expression in PBMC. RESULTS: We observed no significant differences in the IL4RA expression profile between the 3 genotypes. A trend toward higher relative expression was observed for homozygous minor I50V and C-3223T genotypes. CONCLUSIONS: We did not find a statistically significant relationship between the genetic polymorphisms and the relative expression of IL4RA. The effect of genetic polymorphism on IL4RA mRNA expression could interfere with other factors, such as environmental stimuli, and should be evaluated in future studies.

Maternal atopy and offspring epigenome-wide methylation signature.

The increase in the prevalence of allergic diseases is believed to partially depend on environmental changes. DNA methylation is a major epigenetic mechanism, which is known to respond to environmental factors. A number of studies have revealed that patterns of DNA methylation may potentially predict allergic diseases.Here, we examined how maternal atopy is associated with methylation patterns in the cord blood of neonates.We conducted an epigenome-wide association study in a cohort of 96 mother-child pairs. Pregnant women aged not more than 35 years old, not currently smoking or exposed to environmental tobacco smoke, who did not report obesity before conception were considered eligible. They were further tested for atopy. Converted DNA from cord blood was analysed using Infinium MethylationEPIC; for statistical analysis, RnBeads software was applied. Gestational age and sex were included as covariates in the final analysis.83 DM sites were associated with maternal atopy. Within the top DM sites, there were CpG sites which mapped to genes SCD, ITM2C, NT5C3A and NPEPL1. Regional analysis revealed 25 tiling regions, 4 genes, 3 CpG islands and 5 gene promoters, (including PIGCP1, ADAM3A, ZSCAN12P1) associated with maternal atopy. Gene content analysis revealed pointwise enrichments in pathways related to purine-containing compound metabolism, the G1/S transition of the mitotic cell cycle, stem cell division and cellular glucose homoeostasis.These findings suggest that maternal atopy provides a unique intrauterine environment that may constitute the first environment in which exposure is associated with methylation patterns in newborn.

Chromosome 11q13.5 variant as a risk factor for atopic dermatitis in children.

INTRODUCTION: Atopic dermatitis is a chronic inflammatory skin disease with a strong genetic basis. Recent GWASs have identified a single nucleotide polymorphism on chromosome 11q13.5 (rs7927894) as novel susceptibility loci of atopic dermatitis. AIM: To evaluate the association of this genetic variant with atopic dermatitis and to investigate its possible interaction with filaggrin null mutations in children population. MATERIAL AND METHODS: One hundred eighty-eight children less than 2 years old were screened for the variant of allele of rs7927894 on chromosome 11q13.5 and for the 4 most prevalent filaggrin mutations. The variant of allele of rs7927894 and all filaggrin mutations were genotyped by real-time PCR assays with subsequent melting curve analysis using SimpleProbe® probes. RESULTS: The allele of rs7927894[T] was associated with a significantly increased risk of atopic dermatitis (OR = 2.21; 95% CI: 1.14-4.28; p = 0.015). Both allergic and non-allergic patient groups had rs7927894[T] allele significantly more frequently than the control group, however, the frequency of alleles did not differ in these two groups. Interestingly, when rs7927894 variant and filaggrin mutations were considered together, the risk of atopic dermatitis was the most increased in the subjects who combined both rs7927894[T] allele and filaggrin mutations (OR = 16.41; p = 0.003). CONCLUSIONS: Our results indicate that the rs7927894 variant on chromosome 11q13.5 may play a role in the development of atopic dermatitis, but this effect seems to be independent of allergic sensitization and of the well-established filaggrin risk alleles, but may be modulated by gene-gene interactions.

Genetic polymorphisms in pattern recognition receptors are associated with allergic diseases through gene-gene interactions.

BACKGROUND: There is evidence that suggests variation in gene encoding pattern recognition receptors, the essential components of innate immunity, might be associated with atopic diseases. However, results have been inconclusive. OBJECTIVES: The aim of the study was to determine the individual associations and possible interactive effects of the CD14 (cluster of differentiation 14), TLR4 (toll-like receptor 4) and TLR2 (toll-like receptor 2) polymorphisms on allergic diseases. MATERIAL AND METHODS: The CD14 C-159T, TLR4 +896A/G and TLR2 A-16934T polymorphisms were identified in 115 children aged from 6 to 17 years. All subjects were selected using a detailed questionnaire which included questions on symptoms and each one underwent skin prick testing. All single-nucleotide polymorphisms (SNPs) were determined using real-time polymerase chain reaction (PCR) assays. RESULTS: There was no statistically significant correlation between the 3 polymorphisms (CD14 C-159T, TLR4 +896A/G and TLR2/-16934A/T) and either asthma, allergic rhinitis or atopy. We observed that children who were heterozygous or homozygous for both the CD14/-159T and TLR2/-16934A alleles had a 4-fold lower risk for asthma than children who were carriers of the T allele of CD14 but non-carriers of the A allele of TLR2, and an almost 3-fold lower risk for asthma when compared to all other groups. Concerning allergic rhinitis, a similar trend was observed. In addition, the presence of at least 1 A allele in the TLR2/-16934 polymorphism reduced the risk for asthma and allergic rhinitis, but only in children who were homozygous for the common A allele in the TLR4 +896 polymorphism. CONCLUSIONS: Our study supports the idea that the CD14, TLR2 and TLR4 polymorphisms may not be directly involved in the development of atopic diseases. However, our results suggest that their impact on the risk of asthma and allergic rhinitis might be modulated by gene-gene interactions.

IL-4RA gene expression in PBMC with regard to place of living and atopy status.

BACKGROUND: IL-4 and IL-4RA are key factors in allergic inflammation. IL-4 stimulates both IgE production and Th2 lymphocyte differentiation. Increased levels of IL-4 and IL-4RA have been shown in allergic patients. Genetic analyses have confirmed that polymorphisms within the IL-4RA gene influence the risk of allergy and can change the expression of the protein. Due to gene-environment interactions, this process is also likely to be modified by environmental exposure. OBJECTIVES: The aim of the study was to evaluate the IL-4RA gene expression in peripheral blood mononuclear cells (PBMC) from atopic and non-atopic subjects with regard to place of living (urban vs rural). MATERIAL AND METHODS: We enrolled 38 subjects into the study, 18 of whom were atopic. Atopy was estimated according to the results of a skin prick test. PBMC were isolated from whole blood, total RNA was extracted and reverse transcribed into cDNA. We performed real-time PCR to measure gene expression, the ACTB gene was chosen as a reference and the delta-delta Ct (ΔΔCT) method was applied for relative quantification. The Mann-Whitney U test was used for statistics. RESULTS: We did not observe any statistically significant differences in the gene expression profile between atopic and non-atopic subjects regardless of their place of living. However, a trend was observed for atopic rural inhabitants to have lower levels of IL-4RA gene expression than atopic subjects living in the town. CONCLUSIONS: The regulation of IL-4RA gene expression is complex and probably influenced by both genetic and environmental factors, such as farming exposures, which could provide the counterbalance to atopy.

Filaggrin loss-of-function mutations as a predictor for atopic eczema, allergic sensitization and eczema-associated asthma in Polish children population.

BACKGROUND: Loss-of-function mutations in the filaggrin (FLG) gene were identified as a major risk factor for atopic eczema. OBJECTIVES: The aim of the study was to investigate the importance of 4 common FLG null mutations in the susceptibility to atopic eczema and other allergic phenotypes in Polish children population. MATERIAL AND METHODS: The FLG mutations were determined in 158 children younger than 2 years of age. All subjects were selected using a detailed questionnaire and blood samples for total and specific IgE measurements were obtained. Cases of atopic eczema were diagnosed according to the criteria of Hanifin and Rajka and skin examination. All FLG mutations were genotyped by real-time PCR assays with a subsequent melting curve analysis using a SimpleProbe® probes. RESULTS: The combined genotype of all 4 mutations (carriage of ≥ 1 FLG mutation) was significantly associated with atopic eczema (p = 0.016). The odds ratio (OR) for individuals carrying 1 of these 4 null mutations was 5.52 (95% CI; 1.11 ÷ 37.12). The significant association between either the combined FLG genotype or 2282del14 deletion and eczema was seen only in the allergic group. The association with asthma was restricted to asthma occurring in the context of eczema (OR, 6.27; 95% CI, 0.89 ÷ 53.56; p = 0.042). CONCLUSIONS: Our study confirms the previous findings that FLG mutations are strongly associated with atopic eczema and confer a significant risk of allergic sensitization and asthma in the context of eczema. These results underline the role of the epidermal barrier and filaggrin insufficiency in the pathogenesis of atopic eczema and eczema-associated asthma.

Bacterial microbiota of the upper respiratory tract and childhood asthma.

BACKGROUND: Patients with asthma and healthy controls differ in bacterial colonization of the respiratory tract. The upper airways have been shown to reflect colonization of the lower airways, the actual site of inflammation in asthma, which is hardly accessible in population studies. OBJECTIVE: We sought to characterize the bacterial communities at 2 sites of the upper respiratory tract obtained from children from a rural area and to relate these to asthma. METHODS: The microbiota of 327 throat and 68 nasal samples from school-age farm and nonfarm children were analyzed by 454-pyrosequencing of the bacterial 16S ribosomal RNA gene. RESULTS: Alterations in nasal microbiota but not of throat microbiota were associated with asthma. Children with asthma had lower α- and ß-diversity of the nasal microbiota as compared with healthy control children. Furthermore, asthma presence was positively associated with a specific operational taxonomic unit from the genus Moraxella in children not exposed to farming, whereas in farm children Moraxella colonization was unrelated to asthma. In nonfarm children, Moraxella colonization explained the association between bacterial diversity and asthma to a large extent. CONCLUSIONS: Asthma was mainly associated with an altered nasal microbiota characterized by lower diversity and Moraxella abundance. Children living on farms might not be susceptible to the disadvantageous effect of Moraxella. Prospective studies may clarify whether Moraxella outgrowth is a cause or a consequence of loss in diversity.

What the Genetic Background of Individuals with Asthma and Obesity Can Reveal: Is ß2-Adrenergic Receptor Gene Polymorphism Important?

The goal of this review was to evaluate the association of ß2-adrenergic receptor (ADRB2) gene polymorphisms with asthma and obesity. Asthma is the most common pediatric inflammatory disorder. The prevalence, severity, and hospitalization index for asthma have increased markedly in the last several decades. Interestingly, asthma is often diagnosed along with obesity. Genetic factors are essential for both conditions, and some of the candidate pleiotropic genes thought to be involved in the development of these diseases are ADRB2, vitamin D receptor (VDR), leptin (LEP), protein kinase C alpha (PRKCA), and tumor necrosis factor alpha (TNFα). The ADRB2 has been studied in multiple populations and more than 80 polymorphisms, mainly single-nucleotide polymorphisms, have been identified. For nonsynonymous Arg16Gly, Gln27Glu, and Thr164Ile, functional effects have been shown. In vivo, these polymorphisms have been evaluated to determine their association with both obesity and asthma, but the results are inconsistent and depend on the population studied or how the disease was defined. Currently, there are only few reports describing the genetic background for the comorbidity of asthma and obesity.

Health-related quality of life in rural children living in four European countries: the GABRIEL study.

OBJECTIVE: Measuring children's health-related quality of life (HRQOL) is of growing importance given increasing chronic diseases. By integrating HRQOL questions into the European GABRIEL study, we assessed differences in HRQOL between rural farm and non-farm children from Germany, Austria, Switzerland and Poland to relate it to common childhood health problems and to compare it to a representative, mostly urban German population sample (KIGGS). METHODS: The parents of 10,400 school-aged children answered comprehensive questionnaires including health-related questions and the KINDL-R questions assessing HRQOL. RESULTS: Austrian children reported highest KINDL-R scores (mean: 80.9; 95 % CI [80.4, 81.4]) and Polish children the lowest (74.5; [73.9, 75.0]). Farm children reported higher KINDL-R scores than non-farm children (p = 0.002). Significantly lower scores were observed in children with allergic diseases (p < 0.001), with sleeping difficulties (p < 0.001) and in overweight children (p = 0.04). The German GABRIEL sample reported higher mean scores (age 7-10 years: 80.1, [79.9, 80.4]; age 11-13 years: 77.1, [74.9, 79.2]) compared to the urban KIGGS study (age 7-10 years: 79.0, [78.7-79.3]; age 11-13 years: 75.1 [74.6-75.6]). Socio-demographic or health-related factors could not explain differences in HRQOL between countries. CONCLUSIONS: Future increases in chronic diseases may negatively impact children's HRQOL.

Protection from childhood asthma and allergy in Alpine farm environments-the GABRIEL Advanced Studies.

BACKGROUND: Studies on the association of farm environments with asthma and atopy have repeatedly observed a protective effect of farming. However, no single specific farm-related exposure explaining this protective farm effect has consistently been identified. OBJECTIVE: We sought to determine distinct farm exposures that account for the protective effect of farming on asthma and atopy. METHODS: In rural regions of Austria, Germany, and Switzerland, 79,888 school-aged children answered a recruiting questionnaire (phase I). In phase II a stratified random subsample of 8,419 children answered a detailed questionnaire on farming environment. Blood samples and specific IgE levels were available for 7,682 of these children. A broad asthma definition was used, comprising symptoms, diagnosis, or treatment ever. RESULTS: Children living on a farm were at significantly reduced risk of asthma (adjusted odds ratio [aOR], 0.68; 95% CI, 0.59-0.78; P< .001), hay fever (aOR, 0.43; 95% CI, 0.36-0.52; P< .001), atopic dermatitis (aOR, 0.80; 95% CI, 0.69-0.93; P= .004), and atopic sensitization (aOR, 0.54; 95% CI, 0.48-0.61; P< .001) compared with nonfarm children. Whereas this overall farm effect could be explained by specific exposures to cows, straw, and farm milk for asthma and exposure to fodder storage rooms and manure for atopic dermatitis, the farm effect on hay fever and atopic sensitization could not be completely explained by the questionnaire items themselves or their diversity. CONCLUSION: A specific type of farm typical for traditional farming (ie, with cows and cultivation) was protective against asthma, hay fever, and atopy. However, whereas the farm effect on asthma could be explained by specific farm characteristics, there is a link still missing for hay fever and atopy.

The protective effect of farm milk consumption on childhood asthma and atopy: the GABRIELA study.

BACKGROUND: Farm milk consumption has been identified as an exposure that might contribute to the protective effect of farm life on childhood asthma and allergies. The mechanism of action and the role of particular constituents of farm milk, however, are not yet clear. OBJECTIVE: We sought to investigate the farm milk effect and determine responsible milk constituents. METHODS: In rural regions of Germany, Austria, and Switzerland, a comprehensive questionnaire about farm milk consumption and other farm-related exposures was completed by parents of 8334 school-aged children, and 7606 of them provided serum samples to assess specific IgE levels. In 800 cow's milk samples collected at the participants' homes, viable bacterial counts, whey protein levels, and total fat content were analyzed. Asthma, atopy, and hay fever were associated to reported milk consumption and for the first time to objectively measured milk constituents by using multiple regression analyses. RESULTS: Reported raw milk consumption was inversely associated to asthma (adjusted odds ratio [aOR], 0.59; 95% CI, 0.46-0.74), atopy (aOR, 0.74; 95% CI, 0.61-0.90), and hay fever (aOR, 0.51; 95% CI, 0.37-0.69) independent of other farm exposures. Boiled farm milk did not show a protective effect. Total viable bacterial counts and total fat content of milk were not significantly related to asthma or atopy. Increased levels of the whey proteins BSA (aOR for highest vs lowest levels and asthma, 0.53; 95% CI, 0.30-0.97), α-lactalbumin (aOR for interquartile range and asthma, 0.71; 95% CI, 0.52-0.97), and ß-lactoglobulin (aOR for interquartile range and asthma, 0.62; 95% CI, 0.39-0.97), however, were inversely associated with asthma but not with atopy. CONCLUSIONS: The findings suggest that the protective effect of raw milk consumption on asthma might be associated with the whey protein fraction of milk.