Background: Some recent familial studies have described a pattern of autosomal dominant inheritance for increased basal serum tryptase (BST), but no correlation with mRNA expression and gene dose have been reported. Objective: We analyzed TPSAB1 mRNA expression and gene dose in a four-member family with high BST and in two control subjects. Methods: Blood samples were collected from the family and control subjects. Complete morphologic, immunophenotypical, and molecular bone marrow mast cell (MC) studies were performed. mRNA gene expression and gene dose were performed in a LightCycler 480 instrument . Genotype and CNV were performed by quantitative real-time digital PCR (qdPCR). Results: CNV analysis revealed a hereditary copy number gain genotype (3β2α) present in all the family members studied. The elevated total BST in the family members correlated with a significant increase in tryptase gene expression and dose. Conclusions and Clinical Relevance: We present a family with hereditary α-tryptasemia and elevated BST which correlated with a high expression of tryptase genes and an increased gene dose. The family members presented with atypical MC-mediator release symptoms or were even asymptomatic. Clinicians should be aware that elevated BST does not always mean an MC disorder.

BACKGROUND:Predominantly antibody deficiencies (PADs) are the most prevalent primary immunodeficiencies, but their B-cell defects and underlying genetic alterations remain largely unknown.OBJECTIVE:We investigated patients with PADs for the distribution of 41 blood B-cell and plasma cell (PC) subsets, including subsets defined by expression of distinct immunoglobulin heavy chain subclasses.METHODS:Blood samples from 139 patients with PADs, 61 patients with common variable immunodeficiency (CVID), 68 patients with selective IgA deficiency (IgAdef), 10 patients with IgG subclass deficiency with IgA deficiency, and 223 age-matched control subjects were studied by using flow cytometry with EuroFlow immunoglobulin isotype staining. Patients were classified according to their B-cell and PC immune profile, and the obtained patient clusters were correlated with clinical manifestations of PADs.RESULTS:Decreased counts of blood PCs, memory B cells (MBCs), or both expressing distinct IgA and IgG subclasses were identified in all patients with PADs. In patients with IgAdef, B-cell defects were mainly restricted to surface membrane (sm)IgA+ PCs and MBCs, with 2 clear subgroups showing strongly decreased numbers of smIgA+ PCs with mild versus severe smIgA+ MBC defects and higher frequencies of nonrespiratory tract infections, autoimmunity, and affected family members. Patients with IgG subclass deficiency with IgA deficiency and those with CVID showed defects in both smIgA+ and smIgG+ MBCs and PCs. Reduced numbers of switched PCs were systematically found in patients with CVID (absent in 98%), with 6 different defective MBC (and clinical) profiles: (1) profound decrease in MBC numbers; (2) defective CD27+ MBCs with almost normal IgG3+ MBCs; (3) absence of switched MBCs; and (4) presence of both unswitched and switched MBCs without and; (5) with IgG2+ MBCs; and (6) with IgA1+ MBCs.CONCLUSION:Distinct PAD defective B-cell patterns were identified that are associated with unique clinical profiles.

The prevalence of adverse food reactions (AFR) has been increasing in the western world. Clinical manifestations are diversified and it may not be possible to clinically discriminate between IgE and non-IgE mediated AFR. In Portugal, the prevalence of AFR and food allergies in children is not known. Thus, the objectives of this study were to determine the prevalence of AFR in central Portugal.

AFFILIATIONS: 1. Department of Biomedics and Diagnostic Science, University of Salamanca. Spain 2. Biomedical Research Institute of Salamanca, IBSAL. Spain 3. Department of Clinical Biochemistry, Salamanca University Hospital. Spain 4. Department of Microbiology and Genetics, University of Salamanca. Spain 5. Universidade do Vale do Rio dos Sinos UNISINOS. Brasil 6. Department of Pediatrics, Salamanca University Hospital. Spain 7. Department of Medicine, University of Salamanca. Spain 8. Department of Immunoallergy, Salamanca University Hospital. Spain

Background: Several null-mutations in the FLG gene that produce a decrease or absence of filaggrin in the skin and predispose to atopic dermatitis and ichthyosis vulgaris have been described. The relationship with asthma is less clear and may be due to the influence of atopy in patients with associated asthma.Methods: Four hundred individuals were included, 300 patients diagnosed with asthma divided into two groups according to their phenotype (allergic and non-allergic asthma) and 100 strictly characterized controls. The coding region and flanking regions of the FLG gene were amplified by PCR. We proceeded to the characterization of potential gene variants in that region by RFLP and sequencing and analysed their association with lung function parameters, asthma control and severity, and quality of life.Results: We identified two null-mutations (R501X and 2282del4), seven SNPs previously described in databases and three SNPs that had not been previously described. One of the SNP identified in this study (1741A > T) was more frequently detected in patients with non-allergic asthma, worse FVC, FEV1 and PEF values and a higher treatment step. In addition, lowered spirometric values were observed in the non-allergic group carrying any of the nonsynonymous SNPs.Conclusions: In the association study of genetic variants of the FLG gene in our population the 1741A > T polymorphism seems to be associated with non-allergic asthma.

Allergic asthma is a multifactorial disorder in which activation and differentiation of B lymphocytes toward the production of IgE play an essential role. In these B cells, molecular mechanisms to generate IgE antibodies from IgM depend on a complex network of molecules that trigger class-switch recombination specifically at the Sε region of the IgH locus. Several transmembrane receptors and activating signaling cascades are known to physiologically participate in these processes, including the IL4R signaling pathway. In this regard, our group has studied in the past genetic polymorphisms in the IL4/IL4R pathway that might hold clinical correlations with atopy and asthma.1Isidoro-Garcia M. Davila I. Moreno E. Laffond E. Lorente F. Gonzalez-Sarmiento R. IL4RA gene polymorphism (Q576R) is associated with higher total IgE levels in Spanish patients with family history of atopy [in Spanish].Med Clin (Barc). 2005; 124: 211-212Crossref PubMed Scopus (14) Google Scholar, 2Isidoro-Garcia M. Davila I. Laffond E. Moreno E. Lorente F. Gonzalez-Sarmiento R. Interleukin-4 (IL4) and interleukin-4 receptor (IL4RA) polymorphisms in asthma: a case control study.Clin Mol Allergy. 2005; 3: 15Crossref PubMed Scopus (47) Google Scholar Because IgE antibodies play a key role as allergen recognizers, understanding the deregulation of IgE synthesis in B cells is crucial to elucidate the pathogenesis of allergic asthma and to identify B-cell–specific targets with therapeutic potential. In this sense, genome-wide association studies have been proven useful to identify some associated gene candidates and pathways involved in the pathogenesis of the disease.3Ober C. Yao T.C. The genetics of asthma and allergic disease: a 21st century perspective.Immunol Rev. 2011; 242: 10-30Crossref PubMed Scopus (475) Google Scholar Indeed, we have previously suggested that a more intricate interaction of genetic and epigenetic factors probably leads to a specific transcriptome, characteristic of the allergic B cell.4Pascual M. Suzuki M. Isidoro-Garcia M. Padron J. Turner T. Lorente F. et al.Epigenetic changes in B lymphocytes associated with house dust mite allergic asthma.Epigenetics. 2011; 6: 1131-1137Crossref PubMed Scopus (55) Google Scholar, 5Pascual M. Davila I. Isidoro-Garcia M. Lorente F. Epigenetic aspects of the allergic diseases.Front Biosci (Schol Ed). 2010; 2: 815-824Crossref PubMed Google Scholar For this reason, we decided to use “next-generation” massively parallel sequencing technologies for quantitative transcriptomics (RNA-seq) of circulating allergic human B cells. The aim of this study was to explore for the first time by using RNA-seq whether aberrant gene expression of B cells could reveal pathological hallmarks of allergy. To achieve this, we explored the transcriptome of CD19+ circulating lymphocytes from patients with house dust mite (HDM) allergic asthma. This group was selected because, unlike pollens, HDMs are permanently present in the environment, allowing us to minimize the effect of seasonal variations. A total of 41 samples corresponding to unrelated white individuals were analyzed in this study, with 24 samples from patients with HDM allergic asthma and 17 from control individuals. Each subject signed an informed written consent, and the local ethics committee approved this study. Controls had to fulfill the following criteria: (1) absence of symptoms or history of asthma or other pulmonary diseases, (2) no symptoms or history of allergy, (3) negative skin prick test results to a battery of common aeroallergens,1Isidoro-Garcia M. Davila I. Moreno E. Laffond E. Lorente F. Gonzalez-Sarmiento R. IL4RA gene polymorphism (Q576R) is associated with higher total IgE levels in Spanish patients with family history of atopy [in Spanish].Med Clin (Barc). 2005; 124: 211-212Crossref PubMed Scopus (14) Google Scholar and (4) absence of familial history of asthma or allergic diseases. Because both allergy and asthma were ruled out, they could be considered as control subjects. However, patients having a diagnosis of HDM allergic asthma had a clinical history compatible to HDM sensitization, a positive skin prick test result to HDM and specific IgE to HDM above class 2, and either a positive bronchodilator test result or a positive metacholine test result. Peripheral blood CD19+ B lymphocytes were isolated using immunomagnetic procedures, and fluorescence-activated cell sorting analysis was used to confirm a purity above 90% of the sorted cellular populations. Total RNA was isolated from purified CD19+ B lymphocytes, and bar-coded directional libraries were prepared using the dUTP/UNG system from 6 individuals (3 patients and 3 controls). The generated libraries were sequenced on the Illumina HiSeq 2000 platform (100 bp single-end reads), providing an average library size of approximately 30 million reads per sample (GEO accession no. GSE52742). Results were generated using the WASP system pipeline (http://wasp.einstein.yu.edu), which integrates the quart, gsnap, and htseq-count strategies for managing raw data, supervising quality metrics, and assigning the reads to known transcripts in the human ENSEMBL GTF annotation file. Differential expression analysis between control and allergic samples was performed by using DESeq, a Bioconductor package specifically designed for the analysis of replicated count-based expression data. DESeq calculates P values using a negative binomial distribution to test whether for a given transcript, the observed difference in read counts normalized to library sizes (base values) is greater than would be expected from random variation. We observed transcriptional differences between control and allergic groups (Fig 1, A; P < .05), which were most evident for a subgroup comprising the top 50 transcripts (P < .025). Indeed, when supervised hierarchical clustering analysis was applied to this set of transcripts, it succeeded in clustering allergic and nonallergic groups (Fig 1, B), suggesting that these genes are recurrently deregulated in B cells during the allergic response and might hold some translational value. The coding transcripts more differentially expressed between patients and controls are listed in Table I. Other noncoding transcripts and pseudogenes are listed in Table E1 in this article's Online Repository at www.jacionline.org. The interleukin 4 receptor (IL4R) attracted our attention because of its broadly studied role in B-cell activation, IgE production, and hypersensitivity reactions, as highlighted by Ingenuity Pathway Analysis (www.ingenuity.com) (Fig 1, C). Although other genes such as TCL1A or CD27 are also associated with hematologic and immunologic disease, we focused on IL4R as the best candidate allergic biomarker because it exhibited higher base mean reads by RNA-seq (Fig 1, D, right y-axis), which indeed supported a more robust detection when compared with the moderate fold increase in TCL1A or the subtle decrease in CD27 (Table I and Fig 1, D, left y-axis). To further explore whether the deregulation of IL4R could be validated in a larger cohort of independent allergic patients (full list in Fig 1, G), IL4R gene expression was analyzed by using qRT-PCR in 10 additional CD19+ samples. Consistent with the RNA-seq data, upregulation of the expression of IL4R was confirmed in purified B cells (unpaired t test, P = .024) (Fig 1, E). In this study, we extended the validation to 25 PBMC samples to explore IL4R as an easily detected marker by using a cheap, noninvasive, and widely applicable approach. The results were also confirmed in bulk PBMCs (unpaired t test, P = .043) (Fig 1, F), validating the discovery potential of our genome-wide approach in patients with allergic asthma. Interestingly, the relative IL4R expression levels were significantly higher in the sorted B cells than in peripheral blood (33.3 ± 23 vs 1.9 ± 1.6; P = .038). Although both cellular populations are not directly comparable because cells were obtained from different subjects, this observation could also be due to a masking effect from other cell types in the PBMCs. This could also explain why differences observed between patients and controls were statistically more significant in CD19+ than in PBMCs.Table IProtein-coding transcripts most differentially expressed between the group of patients with allergic asthma and the controls, listed by their base mean value in the control groupEnsemble IDExternal gene IDBase meanFold changeP valueControlsPatientsENSG00000077238IL4R454.8798.51.8.024ENSG00000100721TCL1A189.7457.42.4.001ENSG00000187231SESTD1127.9311.62.4.002ENSG00000139193CD2737.796.92.6.025ENSG00000196374HIST1H2BM15.345.73.0.002ENSG00000150625GPM6A85.741.20.5.003ENSG00000134909ARHGAP321.538.826.7.011ENSG00000064886CHI3L270.822.30.3.012ENSG00000107104KANK13.721.15.7.023ENSG00000185015CA130.614.626.6.014ENSG00000042317SPATA743.914.60.3.012ENSG00000154380ENAH2.513.45.5.003ENSG00000122970IFT810.310.434.5.024ENSG00000173210ABLIM331.510.10.3.010ENSG00000185666SYN333.810.10.3.006ENSG00000105507CABP50.69.215.2.012ENSG00000151838CCDC1750.05.2Inf.021ENSG00000174599TRAM1L112.11.20.1.023 Open table in a new tab In summary, our study revealed an increased expression of IL4R in B cells of patients with HDM allergic asthma. It is well known that human B cells undergo isotype class switching and secrete IgE when stimulated with IL-4 and anti-CD40.6Jabara H.H. Fu S.M. Geha R.S. Vercelli D. CD40 and IgE: synergism between anti-CD40 monoclonal antibody and interleukin 4 in the induction of IgE synthesis by highly purified human B cells.J Exp Med. 1990; 172: 1861-1864Crossref PubMed Scopus (454) Google Scholar Consequently, it had been speculated that upregulation of IL4R would facilitate signaling through IL-4 and favor class-switch recombination to increase synthesis of IgE,7Chan M.A. Gigliotti N.M. Meng J. Rosenwasser L.J. Asthma-related SNP in FCER2 is associated with increased expression of IL-4R on human B cells.Int J Immunogenet. 2011; 38: 533-538Crossref PubMed Scopus (6) Google Scholar which indeed is in accordance with our observation that allergic patients with higher IL4R levels tend to also exhibit high levels of serum IgE (>100 KU/L) (Fig 1, H). Previous genome-wide studies in allergic asthma were developed using primarily microarrays in peripheral blood, highlighting that additional important information would be obtained from specifically studying the lymphoid tissue where IgE is produced. Here, we strictly selected control subjects to identify 50 transcripts differentially expressed in peripheral CD19+ cells from allergic patients. Although only a few of those (n = 7) appeared downregulated in the allergic group, the majority of the identified transcripts appeared upregulated when compared with control subjects (n = 43). Many of these deregulated genes could be ascribed to biological pathways involved in hematologic and immunologic functions, suggesting a potential implication of those genes in the pathophysiology of allergic diseases. Our results revealed IL4R as a robust protein-coding biomarker of allergic B cells, which is consistent with previous work suggesting a potential role of genetic variations of the IL4/IL4R pathway in allergy.1Isidoro-Garcia M. Davila I. Moreno E. Laffond E. Lorente F. Gonzalez-Sarmiento R. IL4RA gene polymorphism (Q576R) is associated with higher total IgE levels in Spanish patients with family history of atopy [in Spanish].Med Clin (Barc). 2005; 124: 211-212Crossref PubMed Scopus (14) Google Scholar, 2Isidoro-Garcia M. Davila I. Laffond E. Moreno E. Lorente F. Gonzalez-Sarmiento R. Interleukin-4 (IL4) and interleukin-4 receptor (IL4RA) polymorphisms in asthma: a case control study.Clin Mol Allergy. 2005; 3: 15Crossref PubMed Scopus (47) Google Scholar, 8Loza M.J. Chang B.L. Association between Q551R IL4R genetic variants and atopic asthma risk demonstrated by meta-analysis.J Allergy Clin Immunol. 2007; 120: 578-585Abstract Full Text Full Text PDF PubMed Scopus (46) Google Scholar In our population, however, the exact effect of IL4R polymorphisms on the allergic phenotype has been difficult to elucidate because of complex significance in meta-analysis.3Ober C. Yao T.C. The genetics of asthma and allergic disease: a 21st century perspective.Immunol Rev. 2011; 242: 10-30Crossref PubMed Scopus (475) Google Scholar Importantly, the fact that the increase in IL4R can also be detected in peripheral blood, although with an attenuated effect, could provide a more easily detected, noninvasive, and widely applicable diagnostic approach. The translational significance of our results is further strengthened by the fact that several therapeutic strategies to target the IL4R pathway have recently being proposed. For example, a soluble form of the IL4 receptor (shIL-4Ralpha) is able to block B-cell binding of IL-4 or other IL-4R antagonists9Andrews A.L. Holloway J.W. Holgate S.T. Davies D.E. IL-4 receptor alpha is an important modulator of IL-4 and IL-13 receptor binding: implications for the development of therapeutic targets.J Immunol. 2006; 176: 7456-7461Crossref PubMed Scopus (138) Google Scholar and it has been recently reported that omalizumab may have an effect over the in vitro expression of IL4R in B cells.10Chan M.A. Gigliotti N.M. Dotson A.L. Rosenwasser L.J. Omalizumab may decrease IgE synthesis by targeting membrane IgE+ human B cells.Clin Transl Allergy. 2013; 3: 29Crossref PubMed Scopus (52) Google Scholar Although genetic variations and aberrant epigenetic mechanisms might play a role in IL4R overexpression in allergic B cells, we could not prove a clear correlation between missense mutations, such as the Q576R,1Isidoro-Garcia M. Davila I. Moreno E. Laffond E. Lorente F. Gonzalez-Sarmiento R. IL4RA gene polymorphism (Q576R) is associated with higher total IgE levels in Spanish patients with family history of atopy [in Spanish].Med Clin (Barc). 2005; 124: 211-212Crossref PubMed Scopus (14) Google Scholar, 2Isidoro-Garcia M. Davila I. Laffond E. Moreno E. Lorente F. Gonzalez-Sarmiento R. Interleukin-4 (IL4) and interleukin-4 receptor (IL4RA) polymorphisms in asthma: a case control study.Clin Mol Allergy. 2005; 3: 15Crossref PubMed Scopus (47) Google Scholar and final IL4R expression levels (Fig 1, I). Reanalysis of analogous epigenetic studies on the B-cell methylome of HDM allergic patients4Pascual M. Suzuki M. Isidoro-Garcia M. Padron J. Turner T. Lorente F. et al.Epigenetic changes in B lymphocytes associated with house dust mite allergic asthma.Epigenetics. 2011; 6: 1131-1137Crossref PubMed Scopus (55) Google Scholar also failed to identify a distinct DNA methylation signature at the IL4R locus of allergic samples (Fig 1, J). To conclude, although further studies are required to explore the causes for its transcriptional deregulation, it is plausible to suggest that the IL4R might hold translational potential as both a biomarker and a therapeutic target in patients with allergic asthma to HDM. Download .pdf (.05 MB) Help with pdf files Table E1

Various nonpharmacological strategies to relieve hospitalized children's pain propose play as a central element. Play is considered an essential resource to improve the negative psychosocial effects of the disease and the hospitalization itself. However, the empirical research of play in health settings has not received much attention. The goal of this study was to determine the effect of a program to promote play in the hospital on postsurgical pain in pediatric patients. The research hypothesis was that children will manifest less pain if they are distracted through play during the postsurgical period. We carried out a randomized parallel trial with two groups, an experimental group and a control group. The control group did not receive any specific treatment, only the standard attention contemplated in the hospital. The parents of the children from the experimental group received instructions to play with their children in the postsurgical period and specific play material with which to play. The results obtained support the research hypothesis. On average, the children from the experimental group scored lower on a pain scale than the children from the control group. This occurred in the three postsurgical measurements of pain. It is concluded that the program to promote play can decrease children's perception of pain.

β-Lactam antibiotics are the most frequent cause of drug allergic reactions. During the last few decades, changes in prescribing habits have caused amoxicillin and amoxicillin-clavulanic to replace benzylpenicillin as the most frequently involved agents. 1 Blanca M. Romano A. Torres M.J. et al. Update on the evaluation of hypersensitivity reactions to betalactams. Allergy. 2009; 64: 183-193 Crossref PubMed Scopus (379) Google Scholar Skin tests with major (penicilloyl-polylysine [PPL]) and minor (minor determinant mixture [MDM]) penicillin determinants are considered the first diagnostic procedure in the assessment of immediate allergic reactions. 2 Joint Task Force on Practice ParametersAmerican Academy of Allergy, Asthma and ImmunologyAmerican Academy of Allergy, Asthma and ImmunologyJoint Council of Allergy, Asthma and ImmunologyExecutive summary of disease management of drug hypersensitivity: a practice parameter. Ann Allergy Asthma Immunol. 1999; 83: 665-700 Abstract Full Text PDF PubMed Google Scholar , 3 Torres M.J. Blanca M. Fernandez J. et al. Diagnosis of immediate allergic reactions to beta-lactam antibiotics. Allergy. 2003; 58: 961-972 Crossref PubMed Scopus (502) Google Scholar However, in the last few years several authors have described a decrease in the diagnostic sensitivity of these determinants. 1 Blanca M. Romano A. Torres M.J. et al. Update on the evaluation of hypersensitivity reactions to betalactams. Allergy. 2009; 64: 183-193 Crossref PubMed Scopus (379) Google Scholar This could be because a significant proportion of patients produce specific and selective antibodies against the side chain of amoxicillin or other β-lactam antibiotics. 1 Blanca M. Romano A. Torres M.J. et al. Update on the evaluation of hypersensitivity reactions to betalactams. Allergy. 2009; 64: 183-193 Crossref PubMed Scopus (379) Google Scholar There is some controversy about what determinants should be included in the evaluation of an allergic reaction to β-lactam. 3 Torres M.J. Blanca M. Fernandez J. et al. Diagnosis of immediate allergic reactions to beta-lactam antibiotics. Allergy. 2003; 58: 961-972 Crossref PubMed Scopus (502) Google Scholar , 4 Fox S. Park M.A. Penicillin skin testing in the evaluation and management of penicillin allergy. Ann Allergy Asthma Immunol. 2011; 106: 1-7 Abstract Full Text Full Text PDF PubMed Scopus (57) Google Scholar , 5 Lin E. Saxon A. Riedl M. Penicillin allergy: value of including amoxicillin as a determinant in penicillin skin testing. Int Arch Allergy Immunol. 2010; 152: 313-318 Crossref PubMed Scopus (40) Google Scholar

Although there is no doubt about the influence of the genetic background in the onset of the allergic diseases, Epigenome-Wide Association Studies are needed to elucidate the possible relationship between allergic diseases and epigenomic dysregulation. In this study we aimed to analyze the epigenetic patterns, in terms of DNA methylation, of three well-characterized populations of house dust mite allergic subjects, aspirin-intolerant asthmatics and controls. As a first, genome-wide phase, we used the HELP assay to study the methylation patterns in CD19(+) B lymphocytes in these populations, and found that there are reproducible epigenetic differences at limited numbers of loci distinguishing the groups, corroborated by bisulphite MassArray in a second validation phase of an expanded 40 subject group. These validated epigenetic changes occur at loci characterized as important for the immune response. One such locus is a new candidate gene, CYP26A1, which shows differential methylation patterns and expression levels between groups. Our results suggest that epigenomic dysregulation may contribute to the susceptibility to allergic diseases, showing for the first time differences in DNA methylation between allergic and non-allergic healthy subjects, both globally and at specific loci. These observations indicate that the epigenome may offer new pathophysiological insights and therapeutic targets in atopic diseases.

The data of the ISAAC project in Spain show a prevalence of childhood asthma ranging from 7.1% to 15.3%, with regional differences; a higher prevalence, 22.6% to 35.8%, is described for rhinitis, and atopic dermatitis is found in 4.1% to 7.6% of children. The prevalence of food allergy is 3%. All children in Spain have the right to be visited in the National Health System. The medical care at the primary level is provided by pediatricians, who have obtained their titles through a 4-yr medical residency training program. The education on pediatric allergy during that period is not compulsory and thus very variable. There are currently 112 certified European pediatric allergists in Spain, who have obtained the accreditation of the European Union of Medical Specialist for proven skills and experience in pediatric allergy. Future specialists in pediatric allergy should obtain their titles through a specific education program to be developed in one of the four accredited training units on pediatric allergy, after obtaining the title on pediatrics. The Spanish Society of Pediatric Allergy and Clinical Immunology (SEICAP) gathers over 350 pediatric allergists and pediatricians working in this field. SEICAP has a growing activity including yearly congresses, continued education courses, elaboration of technical clinical documents and protocols, education of patients, and collaboration with other scientific societies and associations of patients. The official journal of SEICAP is Allergologia et Immunophatologia, published every 2 months since 1972. The web site of SEICAP, http://www.seicap.es, open since 2004, offers information for professionals and extensive information on pediatric allergic and immunologic disorders for the lay public; the web site is receiving 750 daily visits during 2011. The pediatric allergy units are very active in clinical work, procedures as immunotherapy or induction of oral tolerance in food allergy, contribution to scientific literature, and collaboration in international projects.

Several studies have proven the important influence that environmental exposures have in the individual's susceptibility to suffer allergy and other related diseases, mostly during embryonic or early life. Although the relationship between the environment and allergic diseases had been previously reported, one interesting attempt to describe this relationship was Strachan's hygiene hypothesis, proposed almost two decades ago. Since then, several studies have identified new environmental factors related to an increased risk of allergic disease. In this context, epigenetic modifications appear as a possible link between the environment and genome, providing a plausible mechanism for explaining how the recent changes in lifestyle can modify gene expression, and thus, lead to a disease state. Here, we will focus on the environmental modifiers that have been described and the possible role of epigenetic modifications.

Understanding how class switch recombination (CSR) is regulated to produce immunoglobulin E (IgE) has become fundamental because of the dramatic increase in the prevalence of IgE-mediated hypersensitivity reactions. CSR requires the induction of the enzyme AICDA in B cells. Mutations in AICDA have been linked to Hyper-IgM syndrome (HIGM2), which shows absence of switching to IgE as well as to IgG and IgA. Although isolated IgE deficiency is a rare entity, here we show some individuals with normal serum IgM, IgG, and IgA levels that had undetectable total serum IgE levels. We have analyzed the AICDA gene in these individuals to determine if there are mutations in AICDA that could lead to selective IgE deficiency. Conformational sensitive gel electrophoresis (CSGE) and sequencing analysis of AICDA coding sequences demonstrated sequence heterogeneity due to 5923A/G and 7888C/T polymorphisms, but did not reveal any novel mutation that might explain the selective IgE deficit.

Recently, functional genetic variants of the PTGDR gene have been associated with asthma. The objective of this work was to study polymorphisms of the promoter region of PTGDR and their haplotype and diplotype combinations in a Spanish population of children with asthma. In this study, 200 Caucasian individuals were included. Asthma was specialist-physician diagnosed according to the ATS criteria. The polymorphisms were analyzed by direct sequencing. In the study, the new polymorphism (-613C > T) in the promoter region of PTGDR was analyzed. The CT genotype was more common in controls (17%) than in patients with asthma (1%) (p-value = 0.0003; OR, 0.057; 95% CI, 0.007-0.441). The CCCT CCCC diplotype (promoter positions -613, -549, -441, and -197) was more frequent in the group of patients with asthma [Fisher's p-value = 0.012; OR, 10.24; 95% CI (1.25-83.68)]; this diplotype is unambiguous. To our knowledge, this is the first study of -613C > T PTGDR polymorphism in patients. This analysis provides more complete information on influence of diplotype combinations of PTGDR polymorphisms in asthma.

At present, cephalosporins represent one of the most prescribed classes of antibiotics. Although allergic reactions have been estimated to be infrequent, the number of reactions to cephalosporins is increasing due to their wide use. Cross-reactivity with penicillins has mainly been evaluated in patients with penicillin allergy. It is higher between first- and second-generation cephalosporins with the same or similar side chain than between cephalosporins with different side chains. Unlike penicillins, cephalosporin haptens or determinants have not been defined, and therefore the diagnosis is complicated. Nevertheless, skin tests with cephalosporins are useful in the evaluation of several allergic reactions. Although more studies are necessary, a negative result in skin testing to penicillin and cephalosporins with different side chains seems to be a good predictor of tolerance, and could be used in select cases.

Background: Allergic diseases are (IgE)-mediated hypersensitivity reactions affecting more than 25% of the world's population. Proteomic technologies have been increasingly used in the field of allergy and include the use of protein microarrays and two-dimensional gel electrophoresis coupled with immunoblotting.Methods: The literature relevant to proteomic approaches to allergic diseases was searched using MEDLINE database. We reviewed proteomics approaches and applications, focusing specifically on two-dimensional immunoblotting techniques and allergen microarrays.Results: The results obtained show that proteomic approaches using two-dimensional immunoblotting appear to be a powerful strategy for the identification of allergenic proteins. Likewise, the use of allergen microarrays allows a large number of IgE antibodies to be simultaneously identified.Conclusions: Proteomic approaches are only beginning to be applied to the study of allergy. In the field of in vitro diagnosis, allergen microarrays provide a promising tool not routinely used in the allergy laboratory. In the near future this powerful technique will be used as a standard technique for in vitro diagnosis of allergy. (c) 2007 Elsevier B.V. All rights reserved.

The prevalence of latex allergy has been increasing not only in risk groups but also in the general population, where it is accepted to average 1%. In children, latex sensitization prevalence studies are scarce and involve different population sampling and allergy testing methods, which makes it difficult to compare across studies. Nevertheless, existing studies point towards a low prevalence of latex allergy in children, which still needs to be confirmed in the Portuguese population. Aiming at studying the prevalence of latex sensitization and allergy in a sample of Portuguese children, we studied 182 children from two different hospital outpatient clinics. A standardized questionnaire focusing on atopic background, previous history and allergic signs or symptoms on exposure to latex or fruits was given to all children and parents. Skin prick testing was performed with a battery of common aeroallergens as well as latex. Serum total IgE, Phadiatop, F x 5E and latex-specific IgE were determined in all children. Specific IgE to latex-crossreacting fruits was determined in latex-sensitized children. Based upon the questionnaire, the prevalence of latex allergy would be 0.5%. The prevalence of latex sensitization would be 3.8%, when based solely upon skin prick testing, and 12.1% (>/=0.35 IU/ml) or 6.6% (>/=0.70 IU/ml) when based singly upon determination of latex-specific IgE. When positive results for either test were considered, the prevalence of latex sensitization was 14.3%. All latex-sensitized children were atopic. Sensitivity to latex-crossreacting foodstuffs was demonstrated in 61.5% of latex-sensitized children (16/26). This study shows that the prevalence of latex allergy and sensitization in Portuguese atopic and non-atopic children, as analysed using various diagnostic methods, is similar to that observed in other countries. In addition, the assessment of latex allergy and sensitization should always include skin prick testing and determination of serum IgE.

Asthma and atopic dermatitis share several common features and Cysteinyl-leukotrienes are mediators that participate in the pathogenesis of both diseases. Recently, a new polymorphism (927T>C) has been identified in cysteinyl-leukotriene type-1 receptor (CYSLTR1) gene. This gene is found on the X chromosome. The aim of this study was to analyze this SNP in a population of children with asthma and atopic dermatitis. In this study, 166 individuals, 79 adult controls (CTR) and 87 children with asthma (AA) were included. Forty-one patients with asthma presented atopic dermatitis (AA-AD). Adults were chosen as controls to confirm lack of development of asthma and allergy during childhood. Standardized history, physical examination, skin prick tests, and lung function measurements were performed in all patients. The 927T>C CYSLTR1 SNP was analyzed by direct sequencing after PCR amplification. In males (53 individuals), the C allele was significantly more common among AA-AD patients (47%) than in CTR (8%) (Fisher's p < 0.005; Monte Carlo p < 0.008; OR:9.78; 95%CI: 1.73-55.30). When comparing AA-AD vs. AA-NAD (patients with asthma but not atopic dermatitis), significant differences were observed, (47% vs. 15%, Fisher's p = 0.014; Monte Carlo p = 0.022; OR: 4.97; 95%CI: 1.29-19.13). No differences in allele distribution were observed between these disease sub-groups in females. The 927T>C is a silent SNP; however, it could affect transcription or translation or may be linked to an unidentified, functional polymorphism and thus may pre-dispose male children to asthma and atopic dermatitis in our population. Further studies are needed to confirm these findings.

To the Editor:In a recent study, Kedda et al1Kedda M.A. Shi J. Duffy D. Phelps S. Yang I. O' Hara K. et al.Characterization of two polymorphisms in the leukotriene C4 synthase gene in an Australian population of subject with mild, moderate and severe asthma.J Allergy Clin Immunol. 2004; 113: 889-895Abstract Full Text Full Text PDF PubMed Scopus (60) Google Scholar analyzed the A−444C polymorphism in the leukotriene C4 synthase (LTC4S) gene in an Australian population. They concluded that this polymorphism was weakly associated with the asthma phenotype but was not related to aspirin intolerance or disease severity. We have analyzed this polymorphism in a Spanish population of 226 individuals: 123 patients with asthma and 103 controls without asthma from the Allergy Department of the University Hospital of Salamanca. The allergic status was based on a positive skin test to at least 1 allergen of a battery of common aeroallergens. Lung function was measured by spirometry, and asthma was diagnosed according to the criteria of the American Thoracic Society and classified following the Global Initiative for Asthma guidelines. The A−444C polymorphism, firstly described by Sanak et al,2Sanak M. Simon H.U. Szczeklik A. Leukotriene C(4) synthase promoter polymorphism and risk of aspirin-induced asthma.Lancet. 1997; 350: 1599-1600Abstract Full Text Full Text PDF PubMed Scopus (330) Google Scholar was analyzed by PCR-RFLP by using MspI restriction endonuclease. Allelic and genetic frequencies are shown in Table I. Although we have not found a statistically significant association between the C−444 allele and the asthma phenotype, we have detected a discrete increase of C−444 allele in patients with asthma. This trend was clearer observed in patients with severe asthma, but these data were not statistically significant. Association of this allele with severe asthma has been previously reported by Sampson et al.3Sampson A.P. Siddiqui S. Buchanan D. Howarth P.H. Holgate S.T. Holloway J.W. et al.Variant LTC(4) synthase allele modifies cysteinyl leukotriene synthesis in eosinophils and predicts clinical response to zafirlukast.Thorax. 2000; 55: S28-S31Crossref PubMed Google Scholar In aspirin-intolerant asthma (AIA), Sanak et al2Sanak M. Simon H.U. Szczeklik A. Leukotriene C(4) synthase promoter polymorphism and risk of aspirin-induced asthma.Lancet. 1997; 350: 1599-1600Abstract Full Text Full Text PDF PubMed Scopus (330) Google Scholar have described an association between C−444 allele and AIA in a Polish population. We considered positive those patients who had undergone a formal aspirin challenge or who reported more than 3 episodes with characteristic symptoms. We have not found any association between AIA and C−444 allele (Table I). These results are in agreement with those of Kedda et al1Kedda M.A. Shi J. Duffy D. Phelps S. Yang I. O' Hara K. et al.Characterization of two polymorphisms in the leukotriene C4 synthase gene in an Australian population of subject with mild, moderate and severe asthma.J Allergy Clin Immunol. 2004; 113: 889-895Abstract Full Text Full Text PDF PubMed Scopus (60) Google Scholar and other authors. In conclusion, we have not detected any association between the C−444 allele and asthma phenotype, degree of severity, or AIA in our population. Nevertheless, we have observed a slight increase of carriers of C−444 allele in the group of patients with asthma compared with controls, particularly in patients with severe asthma. This is the first study of the A−444C polymorphism of the LTC4S gene performed in a population of southern Europe with asthma and confirms the results obtained in American, Dutch, Japanese, and Australian populations.Table IGenotype and allele frequencies of leukotriene C4 synthase A−444C promoter polymorphismGenotypeAlleleHardy-Weinberg equilibrium P valuePhenotypeNAAACCCACNonasthma controls1030.530.400.070.730.27.86All asthma1230.470.440.090.690.31.75Intermittent asthma570.530.420.050.740.26.52Mild asthma270.480.370.150.670.33.39Moderate asthma260.420.500.080.670.33.49Severe asthma120.330.500.170.580.42.92Aspirin-intolerant asthma430.510.470.020.740.26.15Aspirin-tolerant asthma760.450.450.100.670.33.91 Open table in a new tab To the Editor: In a recent study, Kedda et al1Kedda M.A. Shi J. Duffy D. Phelps S. Yang I. O' Hara K. et al.Characterization of two polymorphisms in the leukotriene C4 synthase gene in an Australian population of subject with mild, moderate and severe asthma.J Allergy Clin Immunol. 2004; 113: 889-895Abstract Full Text Full Text PDF PubMed Scopus (60) Google Scholar analyzed the A−444C polymorphism in the leukotriene C4 synthase (LTC4S) gene in an Australian population. They concluded that this polymorphism was weakly associated with the asthma phenotype but was not related to aspirin intolerance or disease severity. We have analyzed this polymorphism in a Spanish population of 226 individuals: 123 patients with asthma and 103 controls without asthma from the Allergy Department of the University Hospital of Salamanca. The allergic status was based on a positive skin test to at least 1 allergen of a battery of common aeroallergens. Lung function was measured by spirometry, and asthma was diagnosed according to the criteria of the American Thoracic Society and classified following the Global Initiative for Asthma guidelines. The A−444C polymorphism, firstly described by Sanak et al,2Sanak M. Simon H.U. Szczeklik A. Leukotriene C(4) synthase promoter polymorphism and risk of aspirin-induced asthma.Lancet. 1997; 350: 1599-1600Abstract Full Text Full Text PDF PubMed Scopus (330) Google Scholar was analyzed by PCR-RFLP by using MspI restriction endonuclease. Allelic and genetic frequencies are shown in Table I. Although we have not found a statistically significant association between the C−444 allele and the asthma phenotype, we have detected a discrete increase of C−444 allele in patients with asthma. This trend was clearer observed in patients with severe asthma, but these data were not statistically significant. Association of this allele with severe asthma has been previously reported by Sampson et al.3Sampson A.P. Siddiqui S. Buchanan D. Howarth P.H. Holgate S.T. Holloway J.W. et al.Variant LTC(4) synthase allele modifies cysteinyl leukotriene synthesis in eosinophils and predicts clinical response to zafirlukast.Thorax. 2000; 55: S28-S31Crossref PubMed Google Scholar In aspirin-intolerant asthma (AIA), Sanak et al2Sanak M. Simon H.U. Szczeklik A. Leukotriene C(4) synthase promoter polymorphism and risk of aspirin-induced asthma.Lancet. 1997; 350: 1599-1600Abstract Full Text Full Text PDF PubMed Scopus (330) Google Scholar have described an association between C−444 allele and AIA in a Polish population. We considered positive those patients who had undergone a formal aspirin challenge or who reported more than 3 episodes with characteristic symptoms. We have not found any association between AIA and C−444 allele (Table I). These results are in agreement with those of Kedda et al1Kedda M.A. Shi J. Duffy D. Phelps S. Yang I. O' Hara K. et al.Characterization of two polymorphisms in the leukotriene C4 synthase gene in an Australian population of subject with mild, moderate and severe asthma.J Allergy Clin Immunol. 2004; 113: 889-895Abstract Full Text Full Text PDF PubMed Scopus (60) Google Scholar and other authors. In conclusion, we have not detected any association between the C−444 allele and asthma phenotype, degree of severity, or AIA in our population. Nevertheless, we have observed a slight increase of carriers of C−444 allele in the group of patients with asthma compared with controls, particularly in patients with severe asthma. This is the first study of the A−444C polymorphism of the LTC4S gene performed in a population of southern Europe with asthma and confirms the results obtained in American, Dutch, Japanese, and Australian populations.

BACKGROUND AND OBJECTIVE:Atopy is a common immunological disorder underlying allergic rhinitis, atopic dermatitis and allergic asthma. There is an association between atopy and the polymorphism Q576R in the IL4RA gene. The aim of this study is to analyze the allelic distribution of the Q576R polymorphism in an atopic and non atopic population and the relationship with total IgE levels and the family history of atopy.PATIENTS AND METHOD:Q576R polymorphism of IL4RA gene was analyzed by PCR-restriction fragment length polymorphism (RFLP) using MspI restriction enzyme in 154 patients from the Allergy Department of the University Hospital of Salamanca.RESULTS:We have not found an association between the R576 allele and higher serum IgE levels nor atopy in this population. Nevertheless, there is an association between this allele and IgE levels in patients with positive skin prick test and family history of atopy.CONCLUSIONS:Our results suggest that the R576 allele could characterize a specific group of patients with a familial history of atopy in whom the presence of this allele may be related to higher levels of serum IgE.

Abstract Background IL4/IL4RA pathway plays an important role in atopy and asthma. Different polymorphisms in IL4 and IL4RA genes have been described. Particularly, -33C>TIL4 and 576Q>RIL4RA SNPs have been independently associated to atopy and asthma. The purpose of this study was to analyse these polymorphisms in a population of patients with a well-characterized asthma phenotype. Methods A total of 212 unrelated Caucasian individuals, 133 patients with asthma and 79 healthy subjects without symptoms or history of asthma or atopy and with negative skin prick tests were recruited. Lung function was measured by spirometry and asthma was specialist physician-diagnosed according to the ATS (American Thoracic Society) criteria and classified following the GINA (Global Initiative for Asthma) guidelines. Skin prick tests were performed according to EAACI recommendations. -33C>TIL4 was studied with TaqMan assay and 576Q>RIL4RA by PCR-RFLP technique. Hardy-Weinberg equilibrium was analysed in all groups. Dichotomous variables were analysed using χ2, Fisher exact test, Monte Carlo simulation test and odds ratio test. To model the effects of multiple covariates logistic regression was used. Results No statistically significant differences between the group of patients with asthma and the controls were found when the allele and genotype distribution of -33C>TIL4 and 576Q>RIL4RA polymorphisms were compared. However, the T allele of the -33C>TIL4 SNP was more frequent in patients with persistent asthma. Multivariate analysis adjusted for age and sex confirmed that carriers of allele T had an increased risk of persistent asthma (OR:2.77, 95%CI:1.18–6.49; p = 0.019). Analysis of combination of polymorphisms showed that patients carrying both the T allele of -33C>TIL4 and the A allele of 576Q>RIL4RA had an increased risk of asthma. This association was particularly observed in persistent asthma [Fisher's p value = 0.0021, Monte Carlo p value (after 104 simulations) = 0.0016, OR:3.39; 95% CI:1.50–7.66]. Conclusion Our results show a trend of association between the genetic combination of the T allele of -33C>TIL4 and the A allele of 576Q>RIL4RA with asthma. This genetic variant was more frequently observed in patients with persistent asthma. As long as this study was performed in a small population, further studies in other populations are needed to confirm these results.