New Biomarkers of Hymenoptera Venom Allergy in a Group of Inflammation Factors.

Hymenoptera venom allergy significantly affects the quality of life. Due to the divergences in the results of the available test and clinical symptoms of patients, the current widely applied diagnostic methods are often insufficient to classify patients for venom immunotherapy (VIT). Therefore it is still needed to search for new, more precise, and accurate diagnostic methods. Hence, this research aimed to discover new biomarkers of Hymenoptera venom allergy in a group of inflammation factors using set of multi-marker Bioplex panel. The adoption of a novel methodology based on Luminex/xMAP enabled simultaneous determination of serum levels of 37 different inflammatory proteins in one experiment. The study involved 21 patients allergic to wasp and/or honey bee venom and 42 healthy participants. According to univariate and multivariate statistics, soluble CD30/tumor necrosis factor receptor superfamily, member 8 (sCD30/TNFRSF8), and the soluble tumor necrosis factor receptor 1 (sTNF-R1) may be considered as effective prognostic factors, their circulating levels were significantly decreased in the allergy group (p-value < 0.05; the Area Under the Curve (AUC) ~0.7; Variable Importance in Projection (VIP) scores >1.2). The obtained results shed new light on the allergic inflammatory response and may contribute to modification and improvement of the diagnostic and monitoring methods. Further, large-scale studies are still needed to explain mechanisms of action of studied compounds and to definitively prove their usefulness in clinical practice.

Factors causing oral and skin pathological features in the hyperimmunoglobulin E syndrome patient including the environmental component: a review of the literature and own experience.

The hyperimmunoglobulin E syndrome (HIES) is a rare multi-system disease with non-immunological as well as immunological abnormalities. The syndrome is characterized by a triad of the most distinctive symptoms, such as pneumonia with pneumatocele formation, recurring staphylococcal skin abscesses and a high serum concentration of IgE. Central mediators of immune responses such as STAT1 and STAT3 affect immune responses and contribute to changes of the skin microbiome which subsequently can amplify the defective immune response against microbial and fungal pathogens. Reactions related to an environmental factor, such as sun-induced skin changes, in individuals during long-term medication therapy have also been reported. The dermatological symptoms, oral status and other health problems of a hyperimmunoglobulin E syndrome paediatric patient are presented. HIES is of great importance to different professionals because sufferers require special preventive and therapeutic management from early infancy in order to avoid complications which can even prove to be life-saving for such patients.

MiRNA Expression Profile in the Airways is Altered during Pulmonary Exacerbation in Children with Cystic Fibrosis-A Preliminary Report.

MicroRNAs are small non-coding RNAs that regulate immune response and inflammation. We assumed that miRNAs may be involved in the immune response during cystic fibrosis pulmonary exacerbations (CFPE) and that altered expression profile in the airways and blood may underlie clinical outcomes in CF pediatric patients. METHODS: We included 30 pediatric patients diagnosed with cystic fibrosis. The biologic material (blood, sputum, exhaled breath condensate) was collected during pulmonary exacerbation and in stable condition. The miRNA expression profile from blood and sputum (n = 6) was done using the next-generation sequencing. For validation, selected four miRNAs were analyzed by qPCR in exosomes from sputum supernatant and exhaled breath condensate (n = 24). NGS analysis was done in Base Space, correlations of gene expression with clinical data were done in Statistica. RESULTS: The miRNA profiling showed that four miRNAs (miR-223, miR-451a, miR-27b-3p, miR-486-5p) were significantly altered during pulmonary exacerbation in CF patients in sputum but did not differ significantly in blood. MiRNA differently expressed in exhaled breath condensate (EBC) and sputum showed correlation with clinical parameters in CFPE. CONCLUSION: MiRNA expression profile changes in the airways during pulmonary exacerbation in CF pediatric patients. We suggest that miRNA alterations during CFPE are restricted to the airways and strongly correlate with clinical outcome.

Proteomic features characterization of Hymenoptera venom allergy.

BACKGROUND: Hymenoptera venom allergy is one of the most frequent causes of anaphylaxis. In its most severe form, the reaction to wasp and honey bee stings may be life-threatening. Therefore, immediate and proper diagnosis of venom allergy and implementation of suitable therapy are extremely important. Broadening the knowledge on the mechanism of the allergic reaction may contribute to the improvement of both diagnostic and treatment methods. Thus, this study aimed to discover changes in protein expression in serum of patients allergic to Hymenoptera (wasp and honeybee) venom and to point out proteins and peptides involved in the allergic inflammation. METHODS: Serum proteomic patterns typical to allergic patients and healthy volunteers were obtained with MALDI-TOF (matrix-assisted laser desorption/ionization-time of flight) mass spectrometer. The spectra were processed, analyzed and compared using advanced bioinformatics tools. The discriminative peaks were subjected to identification with liquid chromatography coupled with tandem mass spectrometry. RESULTS: This methodology allowed for the identification of four features differentiating between allergy and control groups. They were: fibrinogen alpha chain, coagulation factor XIII chain A, complement C4-A, and inter-alpha-trypsin inhibitor heavy chain H4. All of these proteins are involved in allergic inflammatory response. CONCLUSIONS: Extending the knowledge of the Hymenoptera venom sensitization will contribute to the development of novel, sensitive and specific methods for quick and unambiguous allergy diagnosis. Understanding the basis of the allergy at the proteomic level will support the improvement of preventive and therapeutic measures.

Interleukin 1ß polymorphism and serum level are associated with pediatric asthma.

BACKGROUND AND AIM: Interleukin-1 is a pro-inflammatory cytokine found in two forms (α and ß). The α form is mainly cell-bound, whereas IL-1ß is primarily secreted by macrophages in response to immune system stimulation. We hypothesized that polymorphic variants of interleukin 1 genes may play a role in childhood asthma risk. The aim of this study was to investigate if IL-1α and ß polymorphism is associated with asthma in a pediatric population and if the genotype affects its serum level. METHODS: The studied population included 310 children aged 6-18 years old (152 with asthma and 158 healthy children). Genotypes were determined with real-time PCR method using TaqMan Genotyping Assays. Serum level was measured with ELISA Set. Statistical analysis was done in Statistica v.12.0. Linkage disequilibrium and haplotype analysis was done in Haploview v. 4.2. RESULTS: We found that three IL-1ß polymorphisms rs1143634, rs1143633, and rs1143643 were associated with allergic asthma risk (P = 0.034; OR = 1.523; P = 0.024, OR = 1.477; 0.044, OR = 1.420, respectively). We also found a strong linkage disequilibrium between these polymorphisms and CAC haplotype was associated significantly with asthma risk (P = 0.023). For IL1α, we did not observe association with asthma. We then analyzed if IL-1ß expression was altered in serum and we found that asthmatic children showed significantly higher IL-1ß levels than healthy controls (P = 0.047). No association with asthma was observed for IL-1 α variants. CONCLUSIONS: This study indicates that IL-1ß gene polymorphism may affect allergic asthma risk in children.

Gastroesophageal reflux is not associated with short-term variability of parasympathetic activity in children.

PURPOSE: A lower parasympathetic activity was described in patients with the gastroesophageal reflux disease. We aimed to determine whether gastroesophageal reflux (GER) episodes are associated with a short-term parasympathetic tone variability in children. METHODS: In order to address this question we performed simultaneous 24-h esophageal multichannel intraluminal impedance-pH and electrocardiographic monitoring in 16 children (age range 6-18 years), including 8 with asthma and 2 with gastroesophageal reflux disease. After describing duration, height, and acidity of 483 GER episodes we also measured parasympathetic-related heart rate variability parameters in 4 time periods: preceding, containing, following GER, and in-between GERs (control). High frequency (HF) power and root-mean square differences of successive R-R intervals (r-MSSD) were assessed in 2.5-min and 1-min periods, respectively. RESULTS: We did not identify the searched short-term parasympathetic tone changes. CONCLUSIONS: In conclusion, GER episodes and their characteristics were not associated with short-term variability of parasympathetic activity in children.

Immune and clinical response to honeybee venom in beekeepers.

OBJECTIVE: The aim of the study was to assess immune response to honeybee venom in relation to the degree of exposure, time after a sting and clinical symptoms. MATERIALS AND METHOD: Fifty-four volunteers were divided into 2 groups: beekeepers and a control group. The serum levels of total IgE (tIgE), bee venom-specific IgE (venom sIgE), phospholipase A2-specific IgE (phospholipase A2 sIgE), tryptase and venom-specific IgG4 (venom sIgG4) were determined. In beekeepers, diagnostic tests were performed within 3 hours following a sting and were repeated after a minimum of 6 weeks from the last sting. In individuals from the control group, the tests were performed only once, without a sting. RESULTS: The tests showed significant differences in venom sIgE (beekeepers' median = 0.34 kUA/l, control group median = 0.29 kUA/l), baseline serum tryptase (beekeepers' median = 4.25 µg/l, control group median = 2.74 µg/l) and sIgG4 (beekeepers' median = 21.2 mgA/l, control group median = 0.14 mgA/l), confirming higher levels of the tested substances in the beekeepers than in the control group. A significant positive correlation was observed between phospholipase A2 sIgE concentration and severity of clinical symptoms after a sting in the group of beekeepers. It was also demonstrated that the clinical symptoms after a sting became less severe with increasing age of the beekeepers. CONCLUSIONS: The differences in the immune response to a bee sting between the beekeepers and individuals not exposed to bees were probably due to the high exposure of the beekeepers to honeybee venom allergens. This may suggest a different approach to the bee venom allergy diagnostic tests in this occupational group.

Neurotrophin serum concentrations and polymorphisms of neurotrophins and their receptors in children with asthma.

BACKGROUND: In the recent years numerous studies have analysed the effects of neurotrophins on allergic inflammation in airway diseases reporting increased neurotrophin levels locally in the airways as well as in serum of asthmatic patients. We aimed to investigate if levels of neurotrophins in serum of asthmatic children are influenced by the genotype of functional variants within genes encoding analysed neurotrophins and their specific receptors. METHODS: In the study we included 98 children diagnosed with asthma. Genotyping of 9 polymorphisms located in neurotrophins genes and their receptors genes was done with use of TaqMan SNP genotyping assays or PCR-RFLP. The serum levels of four neurotrophins (BDNF, NGF, NTF3, NTF4) were analysed during exacerbation of asthma symptoms with use of DuoSet ELISA Development Kit (R&D). RESULTS: The two patients with the genetic variant A/A of NTRK1 (rs6334) showed significantly higher NGF serum concentrations (113.4 and 218.1 pg/mL) as compared to the mean NGF serum concentrations in the total group of patients (34.8 pg/mL). We also observed a significant epistatic interactions between variants of NGF rs6330 and NTRK1 rs6334 that influenced NGF serum level (P = 0.0004). Analysis of four neurotrophins serum levels in relation to different genotypes of analysed neurotrophins genes showed no significant differences among analysed asthmatic children. CONCLUSIONS: Our results suggest that, among analysed neurotrophins, NGF serum levels may be influenced by the genotype of NTRK1 gene individually as well as in the interaction with NGF functional genetic variant suggesting their involvement in allergic inflammation in asthma. Serum levels of the other neurotrophins do not seem to be affected by the variants in the analysed genes.

Multilocus analysis of candidate genes involved in neurogenic inflammation in pediatric asthma and related phenotypes: a case-control study.

OBJECTIVES: Asthma is a heterogenous complex disorder caused by chronic inflammation of the airways. The key issue in genetic association studies of complex disorders is the identification of multiple low-risk genes that individually have little impact on the phenotype, but in combination account for the clinical manifestation of asthma. Since neurogenic inflammation is emerging as a candidate factor in the pathogenesis of asthma, the aim of the study was to investigate whether genetic variants of neurotrophin genes are associated with asthma disease severity or asthma-related phenotypes in a pediatric population. METHODS: We genotyped 27 polymorphisms located in neurotrophin genes, using TaqMan SNP genotyping assays or Polymerase Chain Reaction - Restriction Fragments Lengths Polymorphism (PCR-RFLP) in 200 children diagnosed with asthma and 226 controls. Interactions between 27 polymorphic loci and asthma-related phenotypes were determined using the Multifactor Dimensionality Reduction (MDR) method. RESULTS: In single marker analysis, we observed an association of MAP3K1 gene polymorphisms (rs702689 and rs889312) with asthma. We also observed that four Single Nucleotide Polymorphisms (SNPs) were associated with severe asthma. Analysis stratified by asthma-related phenotype revealed an association between atopy and NGFR (rs3785931), while BDNF (rs7124442), NTRK2 (rs1212171), NGFR (rs2072446), and FYN (rs3730353) variants were associated with increased exhaled nitric oxide (exNO). In addition, gene-gene interaction analysis revealed a significant epistatic interaction between MAPK (rs889312) and NGF (rs11102930) variants in asthma susceptibility. CONCLUSIONS: Our results suggest that genetic variants of MAP3K1 and NGF genes involved in the regulation of neurogenic inflammation may contribute to asthma, possibly via enhanced NGF expression and MAPK signaling pathway activation.

Health-related quality of life in Polish adolescents with Hymenoptera venom allergy treated with venom immunotherapy.

INTRODUCTION: Venom allergy, though rare, may seriously influence health-related quality of life (HRQoL). There is a paucity of research on HRQoL of adolescents and young adults with Hymenoptera venom allergy. The aim was to assess the level of HRQoL and to evaluate its independent predictors in Polish adolescents and young adults treated with venom immunotherapy. MATERIAL AND METHODS: A multicenter cross-sectional study based on the Vespid Allergy Quality of Life Questionnaire (VQLQ) adapted for Polish adolescents was used. The study sample included 87 patients (14-21 years) studied at different stages of venom immunotherapy (VIT). Statistical analysis was done with multivariate linear regression. RESULTS: Anxiety level was higher in patients with 4(th) grade of Mueller's classification (anaphylactic shock) than in those with 3(rd) grade (B = 0.84, 95% CI = 0.07-1.61, p = 0.03). Caution increased along with an increase of anxiety of adolescents treated with VIT (B = 0.54, 95% CI = 0.39-0.68, p < 0.01). Level of limitations increased with increasing caution of adolescents (B = 0.63, 95% CI = 0.35-0.91, p < 0.01). Discomfort increased along with a rise of caution of patients (B = 0.38, 95% CI = 0.22-0.55, p < 0.01). Similarly, it increased with an increase of their feeling of limitations (B = 0.37, 95% CI = 0.23-0.51, p < 0.01). The level of discomfort in adolescents treated with VIT was lower in those who were treated with conventional protocol in comparison to those treated with rush or ultra-rush ones (B = -0.47, 95% CI = -0.90 - -0.03, p = 0.04). CONCLUSIONS: Severity of anaphylactic reaction is an independent determinant of anxiety level in adolescents treated with VIT. The VIT protocol affects HRQoL of treated patients.

The hyperimmunoglobulin E syndrome--clinical manifestation diversity in primary immune deficiency.

The hyper-IgE syndromes are rare, complex primary immunodeficiencies characterized by clinical manifestation diversity, by particular susceptibility to staphylococcal and mycotic infections as well as by a heterogeneous genetic origin. Two distinct entities--the classical hyper-IgE syndrome which is inherited in an autosomal dominant pattern and the autosomal recessive hyper-IgE syndrome--have been recognized. The autosomal dominant hyper-IgE syndrome is associated with a cluster of facial, dental, skeletal, and connective tissue abnormalities which are not observable in the recessive type. In the majority of affected patients with autosomal dominant hyper-IgE syndrome a mutation in the signal transducer and the activator of the transcription 3 gene has been identified, leading to an impaired Th17 cells differentiation and to a downregulation of an antimicrobial response. A mutation in the dedicator of the cytokinesis 8 gene has been identified as the cause of many cases with autosomal recessive hyper-IgE syndrome and, in one patient, a mutation in tyrosine kinase 2 gene has been demonstrated. In this paper, the authors provide a review of the clinical manifestations in the hyper-IgE syndromes with particular emphasis on the diversity of their phenotypic expression and present current diagnostic guidelines for these diseases.

Pulmonary lymphomatoid granulomatosis in Griscelli syndrome type 2.

Griscelli syndrome type 2 (GS2) is a rare autosomal-recessive disorder associated with a RAB27A gene mutation, and clinically manifesting as hypopigmentation, disseminated chronic encephalitis, and severe immunological disorders characterized by an accelerated hematological phase, also referred to as hemophagocytic syndrome (HS), or hemophagocytic lymphohistiocytosis (HLH). The authors report the diagnosis of GS2 in an 11-year-old girl with hypopigmentation, immunodeficiency, hepatosplenomegaly, severe neurological impairments, and fatal multiorgan failure. In this patient a diagnosis of pulmonary lymphomatoid granulomatosis (LG), an Epstein-Barr virus (EBV)-related lymphoproliferative disorder, was established from radiological and histological findings. Although EBV-related malignancies are common in immunocompromised patients, this is the first report of a diagnosis of pulmonary LG in a patient with GS2.

[Diagnosis and venom specific immunotherapy (VIT) in allergic children in Poland--how much the current practice follows the international guidelines?]. / Diagnostyka i immunoterapia u dzieci z alergia na jad owadów blonkoskrzydlych (VIT) w Polsce--jak wyglda w praktyce zgodnosc z miedzynarodowymi wytycznymi?

INTRODUCTION: Insect venom allergy requires a high level approach adequate to allergy intensity. In case of severe IgE-mediated sting reactions, in children older than five years, venom immunotherapy is a treatment of choice. AIM: Identification of current practices applied to venom allergic children in Poland and their adherence to the international guidelines. METHOD: Questionnaire survey concerning diagnostic and treatment rules was carried out in 8 centres of pediatric allergology, based on a similar audit conducted in the United Kingdom [Diwakar L. et al. Clin Exp Allergy 2008, 38: 1651]. RESULTS: In 5 centres both RAST and SPT tests were used as the first line of investigation. Subsequently 6 centres performed IDT. In three centres baseline serum tryptase levels were estimated. In case of sensitization to both bee and wasp venom in a child with the history of severe systemic reaction, but uncertain culprit insect, specific venom immunotherapy with both venoms was practised by 2 centres. In systemic reaction and not-detectable IgE in 6 centres child was followed-up in 6-12 months. Antihistamine premedication concerned all children in 7 centres. Six-week interval between booster doses was applied in half of centres. A target dose equal 100 mcg was used in 7 centres. Similarly all centres practiced 3-5 five year period of VIT. CONCLUSIONS: In Poland current practice with venom allergic children was conducted in congruence with most of the recommendations.

[Adaptation of Vespide Allergy Quality of Life Questionnaire for Polish children with venom allergy and for their parents]. / Adaptacja kwestionariuszy jakosci zycia dla dzieci z alergia na jad owadów oraz dla ich rodziców.

Aim of the study was to assess validity and reliability of adaptations of VQLQ for Polish children with Hymenoptera venom allergy and their parents. Sample under study consisted of 73 children aged up to 14 years, who were treated with specific venom immunotherapy (VIT) in 2008 in centres conducting this procedure in Poland, and their parents. Theoretical validity of the scales was assessed with exploratory factor analysis using principal component analysis method. Reliability of the scales was assessed in terms of internal consistency with Cronbach alpha coefficient. Results of analysis showed that both scales measure 4 dimension of quality of life and reliability of scales measuring particular dimensions is at least acceptable in case of scale for children, and high in case of scale for parents. Both adapted scales are valid and reliable tools measuring quality of life in children with Hymenoptera venom allergy and their parents' quality of life in the face of child's allergy.

[Health-related quality of life in children with Hymenoptera venom allergy treated with VIT and quality of life of their parents]. / Jakosc zycia warunkowana stanem zdrowia dzieci z alergia na jad owadów blonkoskrzydlych leczonych VIT oraz jakosc zycia ich rodziców.

UNLABELLED: Hymenoptera venom allergy, although rare in children, by its potential fatalities, leads to many psychosocial consequences, influencing quality of life of children and their parents. Aim of this paper is the estimation of health-related quality of life of venom allergic children treated with specific immunotherapy, and their parents. Assessment of factors influencing health-related quality of life levels was also performed. MATERIAL AND METHODS: Sample under study consisted of 73 children: mean age 10.6, SD 2, 4, treated because of Hymenoptera venom allergy in 5 clinical allergy centers in Poland. Data was collected using VQLQ questionnaire adapted for children and their parents. Determinants of quality of life were assessed with multivariate linear and logistic regression models. Analysis were done with SPSS 15 for Windows package. RESULTS: Girls reported higher level of anxiety than boys (B = 0.47; 95% CI = (0.01; 0.94)). Level of caution in children increased along with increase of their anxiety against re-sting (B = 0.49; 95% CI = (0.27; 0.71)). Level of anxiety of children who were under treatment from 6 months to 2 years was lower than level of anxiety of parents of children treated shorter than 6 months (B = -1.21; 95% CI = (-2.16; -0.25)). The lowest level of caution was reported by parents of children aged 10 year or less (B = -0.86; 95% CI = (-1.67; -0.05)), while the highest was reported by parents of children aged 11 years (B = 0.86; 95% CI = (0.20; 1.53)) in comparison to parents of children aged 12 years or more. Parents' caution increased along with increase of their anxiety (B = 0.61; 95% CI = (0.40; 0.83)). Higher level of limitations was imposed by parents of children treated with rush or ultra rush method, in comparison to parents of children treated with conventional method (B = 1.27; 95% CI = (0.21; 2.33)). Levels of quality of life in children and their parents were strongly dependent in the same dimensions. CONCLUSIONS: 1. Levels of quality of life in particular dimension in children is related to level of the same dimension in parents. 2. Age of children influenced level of caution of their parents. 3. Treatment duration influenced level of anxiety of parents. 4. Safety feeling acquired by parents at the beginning of treatment improves their quality of life in all dimensions.