Analysis of gene-expression profiles by microarrays is useful for characterization of candidate g... more Analysis of gene-expression profiles by microarrays is useful for characterization of candidate genes, key regulatory networks, and to define phenotypes or molecular signatures which improve the diagnosis and/or classification of the allergic processes. We have used this approach in the study of olive pollen response in order to find differential molecular markers among responders and non-responders to this allergenic source. Five clinical groups, non-allergic, asymptomatic, allergic but not to olive pollen, untreated-olive-pollen allergic patients and olive-pollen allergic patients (under specific-immunotherapy), were assessed during and outside pollen seasons. Whole-genome gene expression analysis was performed in RNAs extracted from PBMCs. After assessment of data quality and principal components analysis (PCA), differential gene-expression, by multiple testing and, functional analyses by KEGG, for pathways and Gene-Ontology for biological processes were performed. Relevance was defined by fold change and corrected P values (less than 0.05). The most differential genes were validated by qRT-PCR in a larger set of individuals. Interestingly, gene-expression profiling obtained by PCA clearly showed five clusters of samples that correlated with the five clinical groups. Furthermore, differential gene expression and functional analyses revealed differential genes and pathways in the five clinical groups. The 93 most significant genes found were validated, and one set of 35 genes was able to discriminate profiles of olive pollen response. Our results, in addition to providing new information on allergic response, define a possible molecular signature for olive pollen allergy which could be useful for the diagnosis and treatment of this and other sensitizations.
Different molecular mechanisms may modulate sensitization and natural or induced tolerance to all... more Different molecular mechanisms may modulate sensitization and natural or induced tolerance to allergens. We have searched for differential mechanisms at humoral and cellular level in the olive pollen a~l ergi c response, checking the influence of exposure to allergens of subjects from an area with extremely high antigenic load during the pollen season. Sera and PBMCs were obtained during and outside the pollen season. Distinct Ig subtypes (total IgE and specific IgE, IgG4 and IgA), and Thl, Th2 and regulatory T cells (Treg) cytokines were analyzed in 5 groups of subjects: Group 1, non-allergic; Group 2, asymptomatic, sensitized to olive pollen; Group 3, allergic to pollen other than olive; Group 4, allergic to olive pollen (not treated); and Group 5, allergic to olive pollen, and getting specific immunotherapy. Asymptomatic subjects showed the highest total IgE levels. The major difference found between untreated and treated subjects was the high levels of non-inflammatory antibodies (lgG4) in treated patients. The main result of cytokine analyses was the statistically significant decrease in TGF-p levels in untreated olive pollen allergic subjects (pollen season) compared with treated. A slgnificnnt decrease In forkhead winged-helix transcription factor (FOXP3) mRNA expression (marker of regulatory response) and a lower presence of Treg cells in PBMCs of olive pollen allergic subjects was found. The results point to a decrease in the cellular regulatory mechanisms mediated by TGF-p and FOXP31n olive-pollen allergic patients that could be restored after specific-immunothernpy. The reason why exposure to common cnvironmcntal antigens induces allergic diseases in Some people and not in others remains undetermined. Allergen-specific CD4+ helper T-cell (Th) generation is the initial event leading to the development of allergic disease. Subtypes Th2 arc pivotal to the inflammatory cascade through production of IL-4 (essential for the development of Th2 cells), IL-S (for eosinophil recruitment), IL-13 (mediates isotype switching to IgE) and IL-9 (mucus hypersecretion). Th I cells (secreting mainly IFN-y) may contribute to the chronicity and effector phase
Journal of Investigational Allergy and Clinical Immunology
Introduction: Comorbidities can influence asthma control and promote asthma exacerbations (AEs). ... more Introduction: Comorbidities can influence asthma control and promote asthma exacerbations (AEs). However, the impact of multimorbidity in AEs, assessed based on long-term follow-up of patients with asthma of different degrees of severity, has received little attention in real-life conditions. Objective: To describe the epidemiological and clinical characteristics and predictors of AEs in patients who had presented at least 1 AE in the previous year in the MEchanism of Genesis and Evolution of Asthma (MEGA) cohort. Methods: The work-up included a detailed clinical examination, pulmonary function testing, fractional exhaled nitric oxide (FeNO), blood counts, induced sputum, skin prick-tests, asthma questionnaires, and assessment of multimorbidity. The number of moderate-severe AEs in the preceding year was registered for each patient. Results: The study population comprised 486 patients with asthma (23.7% mild, 35% moderate, 41.3% severe). Disease remained uncontrolled in 41.9%, and 4...
Asthma is a complex condition resulting from the interaction of genes and environment. Obesity is... more Asthma is a complex condition resulting from the interaction of genes and environment. Obesity is a risk factor to develop asthma and contributes to poor response to asthma therapy and severity. The aim of the study was to evaluate the effect of obesity on the expression levels of genes previously associated with severe asthma. Three groups of subjects were studied: non-obese asthmatics (NOA), obese asthma patients (OA), and non-asthmatic obese subjects (O). Previously reported overexpressed (IL-10, MSR1, PHLDA1, SERPINB2, and CD86) and underexpressed genes (CHI3L1, CPA3, IL-8, and PI3) in severe asthma were analyzed by RT-qPCR in peripheral blood mononuclear cells (PBMCs). In the overexpressed genes, obesity significantly decreased the expression of MSR1 and PHLDA1 and had no effects on CD86, IL-10, and SERPINB2. In underexpressed genes, obesity did not affect PI3, CHI3L1, and IL-8 and significantly reduced CPA3 expression. The results of this study show that obesity should be incl...
Journal of investigational allergology & clinical immunology: official organ of the International Association of Asthmology (INTERASMA) and Sociedad Latinoamericana de Alergia e Inmunología
We studied the early response to ovalbumin challenge in sensitized Brown-Norway rats through its ... more We studied the early response to ovalbumin challenge in sensitized Brown-Norway rats through its effect on N 2 , He, and SF 6 phase III slopes of the single-breath washout and on indexes of lung function. Sensitized rats showed varying degrees of response in terms of pulmonary pressure (PL), with increases ranging between 125 and 225% of baseline. The sensitized rats presented decreased quasistatic compliance, forced vital capacity, and end-expiratory flow, with all three lung function indexes showing a significant negative correlation with corresponding PL values. They also showed significant positive correlations of PL with the N 2 , He, and SF 6 phase III slopes, reflecting diffusion-convection-dependent inhomogeneities generated by conformation changes throughout the entire rat lung. In addition, the rats showing the most marked PL increases (Ͼ150% baseline PL) also revealed a reversal of the SF 6-He slope difference because of a more marked SF 6 than He slope increase. This latter finding suggests that the degree of structural heterogeneity during early response is even more marked in the most peripheral rat lung generations. Brown-Norway rats; early response; diffusion-convectiondependent inhomogeneity
Olive pollen is one of the most important causes of respiratory allergy in the Mediterranean area... more Olive pollen is one of the most important causes of respiratory allergy in the Mediterranean area (1, 2). Although several reports have described the high prevalence of Olea europaea-induced nasal and conjunctival symptoms (3), this pollen may also induce epidemics of pollen-induced asthma exacerbations between late April and early June. However, the threshold level of O. europaea pollen required to elicit symptoms of seasonal allergic rhinitis is extremely high (around 400 grains/m 3) compared with the 50 grains/m 3 needed by patients clinically sensitive to grasses (4). In Jae´n (southern Spain), the atmospheric pollen concentration varies between 500 and 1000 grains/m 3 during at least half of the pollination season, with peaks of more than 5000 grains/m 3 by mid-May. For this reason, the patients have rhinitis and/or bronchial asthma of an increased severity when they are exposed to these high levels of pollen counts (5). According to this fact, there is some recent evidence that suggest a more complex IgE response against the olive pollen allergens in allergic patients from Jae´n (with extremely high levels of pollen exposure) than that observed in pollinic patients from other places with a lower level of exposure (6). Using different standard laboratory methods, olive pollen extract has showed the presence of at least 20 protein bands with allergenic activity (7). Amongst them, Ole e 1 is the most frequent sensitizing allergen and it is recognized for more than 70% of the patients (8, 9). Besides Ole e 1, nine additional olive pollen allergens have been also isolated and purified from O. europaea pollen extract (10). Two of these allergenic proteins, Ole e 4 (11) and Ole e 6 (12) fail to show any homology to known protein sequences and therefore the biochemical function of these gene products remains unknown. Many other allergens correspond to protein showing a high degree of sequence homology to proteins cloned from different vegetable tissues, such as superoxide dismutase (Ole e 5) (11), calcium-binding proteins (Ole e 3 and Ole e 8) (13, 14), lipid transfer proteins (Ole e 7) (15, 16) and b1-3 glucanases (Ole e 9) (17). The present work focuses on two olive allergens Ole e 2 and Ole e 10. Olive profilin (Ole e 2) has been purified from olive pollen (18), as well as cloned and sequenced (19), with a recognition frequency estimated at 24% of olive-sensitized patients. A strong cross-reactivity was detected with Poaceae and other Oleaceae profilins (18). Background: The clinical characteristics in olive pollen allergy are dependent on the antigenic load, the allergens profile, and the genetic restrictions. Our objective was to determine specific response pattern in Ole e 2 and Ole e 10 sensitization at those levels. Methods: We studied 146 patients with seasonal rhinitis and/or asthma and positive prick test to Olea europaea pollen. IgE against Ole e 2 and Ole e 10 were detected by skin prick test and ELISA. HLA-DRB1 and HLA-DQB1 loci were typed by polymerase chain reaction sequence-specific primers method. Results: A total of 102 (69.9%) and 79 (54.0%) patients showed significant IgE antibody response against Ole e 2 and Ole e 10, respectively. There was a significant association between Ole e 2 (OR 2.2, P ¼ 0.04) and Ole e 10 reactivities (OR 2.8, P ¼ 0.007) with asthma. In addition, total and specific IgE antibody levels significantly correlated with asthma (P < 0.05). Patients who reacted to both allergens reached the highest asthma risk factor (OR 4.3, P ¼ 0.002). Phenotypic frequency of DR7 (OR 5.4, Pc ¼ 0.003) and DQ2 (OR 3.6, Pc ¼ 0.02) were increased in positive Ole e 2 patients compared with control subjects. DR2(15) phenotypic frequency was significantly increased (OR 5.6, Pc ¼ 0.02) in positive Ole e 10 patients compared with control subjects. Conclusions: Our data suggest an association of Ole e 2 and Ole e 10 with bronchial asthma. Also, we found a genetic control of Ole e 2 and Ole e 10 IgE-specific responses that could be relevant to clinical disease in olive pollen allergy.
Background and Aims: Asthma is a heterogeneous respiratory disease that encompasses different inf... more Background and Aims: Asthma is a heterogeneous respiratory disease that encompasses different inflammatory and functional endophenotypes. Many non-invasive biomarkers has been investigated to its pathobiology. Heany et al proposed a clinical algorithm that classifies severe asthmatic patients into likely-eosinophilic phenotypes, based on accessible biomarkers: PBE, current treatment, FeNO, presence of nasal polyps (NP) and age of onset. Materials and Methods: We assessed the concordance between the algorithm proposed by Heany et al. with sputum examination, the gold standard, in 145 asthmatic patients of the MEGA cohort with varying grades of severity. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
ADAM10 is the main α-secretase acting in the non-amyloidogenic processing of APP. We hypothesized... more ADAM10 is the main α-secretase acting in the non-amyloidogenic processing of APP. We hypothesized that certain rare ADAM10 variants could increase the risk for AD by conferring the age-related downregulation of α-secretase. The ADAM10 gene was sequenced in 103 AD cases (82% familial) and 96 cognitively preserved nonagenarians. We examined rare variants (MAF < 0.01) and determined their potential association in the AD group with lower CSF protein levels, as analyzed by means of ELISA, and Western blot (species of 50 kDa, 55 kDa, and 80 kDa). Rare variants were found in 15.5% of AD cases (23% early-onset, 8% late-onset) and in 12.5% of nonagenarians, and some were group-specific. All were intronic variants except Q170H, found in three AD cases and one nonagenarian. The 3′UTR rs74016945 (MAF = 0.01) was found in 6% of the nonagenarians (OR 0.146, p = 0.057). Altogether, ADAM10 total levels or specific species were not significantly different when comparing AD with controls or carrie...
RATIONALE: Our previous studies demonstrated that galectin-3, a beta galactoside binding lectin, ... more RATIONALE: Our previous studies demonstrated that galectin-3, a beta galactoside binding lectin, induces selective down-regulation of IL-5 gene expression on different cells. Also, intra-tracheal instillation of plasmid DNA encoding galectin-3 in asthmatic rats, inhibits inflammatory cells in BAL fluid with recovery of pulmonary function (Am J Resp Crit Care Med, accepted for publication July 2002). Now we investigate this treatment in a time dependent manner. METHODS: Rats were previously sensitized to OA in alum and galectin-3 gene was introduced by orotracheal instillation, with 0.5 ml of plasmid with: a) galectin-3, b) antisense galectin-3 and c) without insert, or with saline instead of plasmid instillation as positive control. After that, rats were exposed to aerosolized OVA solution for fifteen minutes per day. Functional tests were performed at different time. Cells were obtained by BAL and used for RNA extraction and for cytometric studies. lmmunoblotting and PCR were used for galectin-3 expression in lungs. RESULTS: Our results demonstrate that intra-tracheal instillation of plasmid-galectin-3 in sensitized and antigen challenged rats leads to an improvement on the cellular inflammatory infiltrate and in pulmonary function with normalization in both eosinophil and T cell numbers in BALE Time-dependent studies showed that the effects of the gene therapy are demonstrated for more than one month after instillation. Now we are analyzing the expression of galectin-3 in different sites by confocal microscopy. CONCLUSIONS: New treatment with plasmid encoding galectin-3, obtaining a marked inhibition of eosinophil airway accumulation with improved lung function. This open a new approach for future therapies.
TRIM25 mutation (p.C168*), coding for an E3 ubiquitin ligase, is a cause of early-onset autosomal... more TRIM25 mutation (p.C168*), coding for an E3 ubiquitin ligase, is a cause of early-onset autosomal dominant dementia with amyloid load and parkinsonism
Background: Macrophage scavenger receptor 1 (MSR1) has mostly been described in macrophages, but ... more Background: Macrophage scavenger receptor 1 (MSR1) has mostly been described in macrophages, but we previously found a significant gene expression increase in peripheral blood mononuclear cells (PBMCs) of asthmatic patients. Objective: To confirm those results and to define its cellular origin in PBMCs. Methods: Four groups of subjects were studied: healthy controls (C), nonallergic asthmatic (NA), allergic asthmatic (AA), and chronic obstructive pulmonary disease (COPD) patients. RNA was extracted from PBMCs. MSR1 gene expression was analyzed by RT-qPCR. The presence of MSR1 on the cellular surface of PBMC cellular subtypes was analyzed by confocal microscopy and flow cytometry. Results: MSR1 gene expression was significantly increased in the three clinical conditions compared to the healthy control group, with substantial variations according to disease type and severity. MSR1 expression on T cells (CD4+ and CD8+), B cells, and monocytes was confirmed by confocal microscopy and fl...
The immune system regulates itself to establish an appropriate immune response to potentially har... more The immune system regulates itself to establish an appropriate immune response to potentially harmful pathogens while tolerating harmless environmental antigens and self-antigens. A central role in this balance is played by regulatory T cells (Tregs) through various ways of actions. By means of molecule secretion and cell-cell contact mechanisms, Tregs may have the capacity to modulate effector T cells and suppress the action of proinflammatory cytokines across a broad range of cell types. As a result, abnormal regulatory T cell function has been pointed as a main cause in the development of allergic diseases, a major public health problem in industrialized countries, with a high socioeconomic impact. This prevalence and impact have created an international interest in improving the allergy diagnosis and therapy. Additionally, research has sought to gain a better understanding of the molecular mechanisms underlining this kind of disease, in order to a better management. At this resp...
Introduction. Asthma and respiratory allergy are chronic inflammatory diseases, with high prevale... more Introduction. Asthma and respiratory allergy are chronic inflammatory diseases, with high prevalence and a wide clinical spectrum. Due to their heterogeneity, it is difficult to diagnose some patients and predict their response to treatments. Moreover, although allergic mechanisms have been implicated in most asthma diagnostics, there are still a 10-33% of patients with nonallergic asthma, less studied and understood. Thus, there is a need to define new biomarkers capable of classify patients correctly. At this respect, we defined a group of 94 potential biomarkers with the ability to differentiate clinical phenotypes and disease severity. Here, the objective was to theoretically prioritize those biomarkers using systems biology, based on their association with the studied diseases. Methods. Anaxomics’ TPMS technology (Therapeutic Performance Mapping System) was used to create one mathematical model, according to molecular motifs, per disease: respiratory allergy (RA), allergic asth...
Analysis of gene-expression profiles by microarrays is useful for characterization of candidate g... more Analysis of gene-expression profiles by microarrays is useful for characterization of candidate genes, key regulatory networks, and to define phenotypes or molecular signatures which improve the diagnosis and/or classification of the allergic processes. We have used this approach in the study of olive pollen response in order to find differential molecular markers among responders and non-responders to this allergenic source. Five clinical groups, non-allergic, asymptomatic, allergic but not to olive pollen, untreated-olive-pollen allergic patients and olive-pollen allergic patients (under specific-immunotherapy), were assessed during and outside pollen seasons. Whole-genome gene expression analysis was performed in RNAs extracted from PBMCs. After assessment of data quality and principal components analysis (PCA), differential gene-expression, by multiple testing and, functional analyses by KEGG, for pathways and Gene-Ontology for biological processes were performed. Relevance was defined by fold change and corrected P values (less than 0.05). The most differential genes were validated by qRT-PCR in a larger set of individuals. Interestingly, gene-expression profiling obtained by PCA clearly showed five clusters of samples that correlated with the five clinical groups. Furthermore, differential gene expression and functional analyses revealed differential genes and pathways in the five clinical groups. The 93 most significant genes found were validated, and one set of 35 genes was able to discriminate profiles of olive pollen response. Our results, in addition to providing new information on allergic response, define a possible molecular signature for olive pollen allergy which could be useful for the diagnosis and treatment of this and other sensitizations.
Different molecular mechanisms may modulate sensitization and natural or induced tolerance to all... more Different molecular mechanisms may modulate sensitization and natural or induced tolerance to allergens. We have searched for differential mechanisms at humoral and cellular level in the olive pollen a~l ergi c response, checking the influence of exposure to allergens of subjects from an area with extremely high antigenic load during the pollen season. Sera and PBMCs were obtained during and outside the pollen season. Distinct Ig subtypes (total IgE and specific IgE, IgG4 and IgA), and Thl, Th2 and regulatory T cells (Treg) cytokines were analyzed in 5 groups of subjects: Group 1, non-allergic; Group 2, asymptomatic, sensitized to olive pollen; Group 3, allergic to pollen other than olive; Group 4, allergic to olive pollen (not treated); and Group 5, allergic to olive pollen, and getting specific immunotherapy. Asymptomatic subjects showed the highest total IgE levels. The major difference found between untreated and treated subjects was the high levels of non-inflammatory antibodies (lgG4) in treated patients. The main result of cytokine analyses was the statistically significant decrease in TGF-p levels in untreated olive pollen allergic subjects (pollen season) compared with treated. A slgnificnnt decrease In forkhead winged-helix transcription factor (FOXP3) mRNA expression (marker of regulatory response) and a lower presence of Treg cells in PBMCs of olive pollen allergic subjects was found. The results point to a decrease in the cellular regulatory mechanisms mediated by TGF-p and FOXP31n olive-pollen allergic patients that could be restored after specific-immunothernpy. The reason why exposure to common cnvironmcntal antigens induces allergic diseases in Some people and not in others remains undetermined. Allergen-specific CD4+ helper T-cell (Th) generation is the initial event leading to the development of allergic disease. Subtypes Th2 arc pivotal to the inflammatory cascade through production of IL-4 (essential for the development of Th2 cells), IL-S (for eosinophil recruitment), IL-13 (mediates isotype switching to IgE) and IL-9 (mucus hypersecretion). Th I cells (secreting mainly IFN-y) may contribute to the chronicity and effector phase
Journal of Investigational Allergy and Clinical Immunology
Introduction: Comorbidities can influence asthma control and promote asthma exacerbations (AEs). ... more Introduction: Comorbidities can influence asthma control and promote asthma exacerbations (AEs). However, the impact of multimorbidity in AEs, assessed based on long-term follow-up of patients with asthma of different degrees of severity, has received little attention in real-life conditions. Objective: To describe the epidemiological and clinical characteristics and predictors of AEs in patients who had presented at least 1 AE in the previous year in the MEchanism of Genesis and Evolution of Asthma (MEGA) cohort. Methods: The work-up included a detailed clinical examination, pulmonary function testing, fractional exhaled nitric oxide (FeNO), blood counts, induced sputum, skin prick-tests, asthma questionnaires, and assessment of multimorbidity. The number of moderate-severe AEs in the preceding year was registered for each patient. Results: The study population comprised 486 patients with asthma (23.7% mild, 35% moderate, 41.3% severe). Disease remained uncontrolled in 41.9%, and 4...
Asthma is a complex condition resulting from the interaction of genes and environment. Obesity is... more Asthma is a complex condition resulting from the interaction of genes and environment. Obesity is a risk factor to develop asthma and contributes to poor response to asthma therapy and severity. The aim of the study was to evaluate the effect of obesity on the expression levels of genes previously associated with severe asthma. Three groups of subjects were studied: non-obese asthmatics (NOA), obese asthma patients (OA), and non-asthmatic obese subjects (O). Previously reported overexpressed (IL-10, MSR1, PHLDA1, SERPINB2, and CD86) and underexpressed genes (CHI3L1, CPA3, IL-8, and PI3) in severe asthma were analyzed by RT-qPCR in peripheral blood mononuclear cells (PBMCs). In the overexpressed genes, obesity significantly decreased the expression of MSR1 and PHLDA1 and had no effects on CD86, IL-10, and SERPINB2. In underexpressed genes, obesity did not affect PI3, CHI3L1, and IL-8 and significantly reduced CPA3 expression. The results of this study show that obesity should be incl...
Journal of investigational allergology & clinical immunology: official organ of the International Association of Asthmology (INTERASMA) and Sociedad Latinoamericana de Alergia e Inmunología
We studied the early response to ovalbumin challenge in sensitized Brown-Norway rats through its ... more We studied the early response to ovalbumin challenge in sensitized Brown-Norway rats through its effect on N 2 , He, and SF 6 phase III slopes of the single-breath washout and on indexes of lung function. Sensitized rats showed varying degrees of response in terms of pulmonary pressure (PL), with increases ranging between 125 and 225% of baseline. The sensitized rats presented decreased quasistatic compliance, forced vital capacity, and end-expiratory flow, with all three lung function indexes showing a significant negative correlation with corresponding PL values. They also showed significant positive correlations of PL with the N 2 , He, and SF 6 phase III slopes, reflecting diffusion-convection-dependent inhomogeneities generated by conformation changes throughout the entire rat lung. In addition, the rats showing the most marked PL increases (Ͼ150% baseline PL) also revealed a reversal of the SF 6-He slope difference because of a more marked SF 6 than He slope increase. This latter finding suggests that the degree of structural heterogeneity during early response is even more marked in the most peripheral rat lung generations. Brown-Norway rats; early response; diffusion-convectiondependent inhomogeneity
Olive pollen is one of the most important causes of respiratory allergy in the Mediterranean area... more Olive pollen is one of the most important causes of respiratory allergy in the Mediterranean area (1, 2). Although several reports have described the high prevalence of Olea europaea-induced nasal and conjunctival symptoms (3), this pollen may also induce epidemics of pollen-induced asthma exacerbations between late April and early June. However, the threshold level of O. europaea pollen required to elicit symptoms of seasonal allergic rhinitis is extremely high (around 400 grains/m 3) compared with the 50 grains/m 3 needed by patients clinically sensitive to grasses (4). In Jae´n (southern Spain), the atmospheric pollen concentration varies between 500 and 1000 grains/m 3 during at least half of the pollination season, with peaks of more than 5000 grains/m 3 by mid-May. For this reason, the patients have rhinitis and/or bronchial asthma of an increased severity when they are exposed to these high levels of pollen counts (5). According to this fact, there is some recent evidence that suggest a more complex IgE response against the olive pollen allergens in allergic patients from Jae´n (with extremely high levels of pollen exposure) than that observed in pollinic patients from other places with a lower level of exposure (6). Using different standard laboratory methods, olive pollen extract has showed the presence of at least 20 protein bands with allergenic activity (7). Amongst them, Ole e 1 is the most frequent sensitizing allergen and it is recognized for more than 70% of the patients (8, 9). Besides Ole e 1, nine additional olive pollen allergens have been also isolated and purified from O. europaea pollen extract (10). Two of these allergenic proteins, Ole e 4 (11) and Ole e 6 (12) fail to show any homology to known protein sequences and therefore the biochemical function of these gene products remains unknown. Many other allergens correspond to protein showing a high degree of sequence homology to proteins cloned from different vegetable tissues, such as superoxide dismutase (Ole e 5) (11), calcium-binding proteins (Ole e 3 and Ole e 8) (13, 14), lipid transfer proteins (Ole e 7) (15, 16) and b1-3 glucanases (Ole e 9) (17). The present work focuses on two olive allergens Ole e 2 and Ole e 10. Olive profilin (Ole e 2) has been purified from olive pollen (18), as well as cloned and sequenced (19), with a recognition frequency estimated at 24% of olive-sensitized patients. A strong cross-reactivity was detected with Poaceae and other Oleaceae profilins (18). Background: The clinical characteristics in olive pollen allergy are dependent on the antigenic load, the allergens profile, and the genetic restrictions. Our objective was to determine specific response pattern in Ole e 2 and Ole e 10 sensitization at those levels. Methods: We studied 146 patients with seasonal rhinitis and/or asthma and positive prick test to Olea europaea pollen. IgE against Ole e 2 and Ole e 10 were detected by skin prick test and ELISA. HLA-DRB1 and HLA-DQB1 loci were typed by polymerase chain reaction sequence-specific primers method. Results: A total of 102 (69.9%) and 79 (54.0%) patients showed significant IgE antibody response against Ole e 2 and Ole e 10, respectively. There was a significant association between Ole e 2 (OR 2.2, P ¼ 0.04) and Ole e 10 reactivities (OR 2.8, P ¼ 0.007) with asthma. In addition, total and specific IgE antibody levels significantly correlated with asthma (P < 0.05). Patients who reacted to both allergens reached the highest asthma risk factor (OR 4.3, P ¼ 0.002). Phenotypic frequency of DR7 (OR 5.4, Pc ¼ 0.003) and DQ2 (OR 3.6, Pc ¼ 0.02) were increased in positive Ole e 2 patients compared with control subjects. DR2(15) phenotypic frequency was significantly increased (OR 5.6, Pc ¼ 0.02) in positive Ole e 10 patients compared with control subjects. Conclusions: Our data suggest an association of Ole e 2 and Ole e 10 with bronchial asthma. Also, we found a genetic control of Ole e 2 and Ole e 10 IgE-specific responses that could be relevant to clinical disease in olive pollen allergy.
Background and Aims: Asthma is a heterogeneous respiratory disease that encompasses different inf... more Background and Aims: Asthma is a heterogeneous respiratory disease that encompasses different inflammatory and functional endophenotypes. Many non-invasive biomarkers has been investigated to its pathobiology. Heany et al proposed a clinical algorithm that classifies severe asthmatic patients into likely-eosinophilic phenotypes, based on accessible biomarkers: PBE, current treatment, FeNO, presence of nasal polyps (NP) and age of onset. Materials and Methods: We assessed the concordance between the algorithm proposed by Heany et al. with sputum examination, the gold standard, in 145 asthmatic patients of the MEGA cohort with varying grades of severity. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
ADAM10 is the main α-secretase acting in the non-amyloidogenic processing of APP. We hypothesized... more ADAM10 is the main α-secretase acting in the non-amyloidogenic processing of APP. We hypothesized that certain rare ADAM10 variants could increase the risk for AD by conferring the age-related downregulation of α-secretase. The ADAM10 gene was sequenced in 103 AD cases (82% familial) and 96 cognitively preserved nonagenarians. We examined rare variants (MAF < 0.01) and determined their potential association in the AD group with lower CSF protein levels, as analyzed by means of ELISA, and Western blot (species of 50 kDa, 55 kDa, and 80 kDa). Rare variants were found in 15.5% of AD cases (23% early-onset, 8% late-onset) and in 12.5% of nonagenarians, and some were group-specific. All were intronic variants except Q170H, found in three AD cases and one nonagenarian. The 3′UTR rs74016945 (MAF = 0.01) was found in 6% of the nonagenarians (OR 0.146, p = 0.057). Altogether, ADAM10 total levels or specific species were not significantly different when comparing AD with controls or carrie...
RATIONALE: Our previous studies demonstrated that galectin-3, a beta galactoside binding lectin, ... more RATIONALE: Our previous studies demonstrated that galectin-3, a beta galactoside binding lectin, induces selective down-regulation of IL-5 gene expression on different cells. Also, intra-tracheal instillation of plasmid DNA encoding galectin-3 in asthmatic rats, inhibits inflammatory cells in BAL fluid with recovery of pulmonary function (Am J Resp Crit Care Med, accepted for publication July 2002). Now we investigate this treatment in a time dependent manner. METHODS: Rats were previously sensitized to OA in alum and galectin-3 gene was introduced by orotracheal instillation, with 0.5 ml of plasmid with: a) galectin-3, b) antisense galectin-3 and c) without insert, or with saline instead of plasmid instillation as positive control. After that, rats were exposed to aerosolized OVA solution for fifteen minutes per day. Functional tests were performed at different time. Cells were obtained by BAL and used for RNA extraction and for cytometric studies. lmmunoblotting and PCR were used for galectin-3 expression in lungs. RESULTS: Our results demonstrate that intra-tracheal instillation of plasmid-galectin-3 in sensitized and antigen challenged rats leads to an improvement on the cellular inflammatory infiltrate and in pulmonary function with normalization in both eosinophil and T cell numbers in BALE Time-dependent studies showed that the effects of the gene therapy are demonstrated for more than one month after instillation. Now we are analyzing the expression of galectin-3 in different sites by confocal microscopy. CONCLUSIONS: New treatment with plasmid encoding galectin-3, obtaining a marked inhibition of eosinophil airway accumulation with improved lung function. This open a new approach for future therapies.
TRIM25 mutation (p.C168*), coding for an E3 ubiquitin ligase, is a cause of early-onset autosomal... more TRIM25 mutation (p.C168*), coding for an E3 ubiquitin ligase, is a cause of early-onset autosomal dominant dementia with amyloid load and parkinsonism
Background: Macrophage scavenger receptor 1 (MSR1) has mostly been described in macrophages, but ... more Background: Macrophage scavenger receptor 1 (MSR1) has mostly been described in macrophages, but we previously found a significant gene expression increase in peripheral blood mononuclear cells (PBMCs) of asthmatic patients. Objective: To confirm those results and to define its cellular origin in PBMCs. Methods: Four groups of subjects were studied: healthy controls (C), nonallergic asthmatic (NA), allergic asthmatic (AA), and chronic obstructive pulmonary disease (COPD) patients. RNA was extracted from PBMCs. MSR1 gene expression was analyzed by RT-qPCR. The presence of MSR1 on the cellular surface of PBMC cellular subtypes was analyzed by confocal microscopy and flow cytometry. Results: MSR1 gene expression was significantly increased in the three clinical conditions compared to the healthy control group, with substantial variations according to disease type and severity. MSR1 expression on T cells (CD4+ and CD8+), B cells, and monocytes was confirmed by confocal microscopy and fl...
The immune system regulates itself to establish an appropriate immune response to potentially har... more The immune system regulates itself to establish an appropriate immune response to potentially harmful pathogens while tolerating harmless environmental antigens and self-antigens. A central role in this balance is played by regulatory T cells (Tregs) through various ways of actions. By means of molecule secretion and cell-cell contact mechanisms, Tregs may have the capacity to modulate effector T cells and suppress the action of proinflammatory cytokines across a broad range of cell types. As a result, abnormal regulatory T cell function has been pointed as a main cause in the development of allergic diseases, a major public health problem in industrialized countries, with a high socioeconomic impact. This prevalence and impact have created an international interest in improving the allergy diagnosis and therapy. Additionally, research has sought to gain a better understanding of the molecular mechanisms underlining this kind of disease, in order to a better management. At this resp...
Introduction. Asthma and respiratory allergy are chronic inflammatory diseases, with high prevale... more Introduction. Asthma and respiratory allergy are chronic inflammatory diseases, with high prevalence and a wide clinical spectrum. Due to their heterogeneity, it is difficult to diagnose some patients and predict their response to treatments. Moreover, although allergic mechanisms have been implicated in most asthma diagnostics, there are still a 10-33% of patients with nonallergic asthma, less studied and understood. Thus, there is a need to define new biomarkers capable of classify patients correctly. At this respect, we defined a group of 94 potential biomarkers with the ability to differentiate clinical phenotypes and disease severity. Here, the objective was to theoretically prioritize those biomarkers using systems biology, based on their association with the studied diseases. Methods. Anaxomics’ TPMS technology (Therapeutic Performance Mapping System) was used to create one mathematical model, according to molecular motifs, per disease: respiratory allergy (RA), allergic asth...
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Papers by Blanca Cárdaba