Epigenetic regulation analyses of FOXP3 in olive pollen allergy

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Allergenicity and cross-reactivity of Senecio pollen: identification of novel allergens using the immunoproteomics approach.

BACKGROUND: The genus Senecio is the largest genus of the family Asteraceae (Compositae). The allergenicity of Senecio has not been assessed previously. OBJECTIVE: The aim of this study was to investigate the allergens of Senecio jacobea pollen and to determine their immunological characteristics and clinical relevance. METHODS: Fifty patients with rhinoconjunctivitis and a positive skin prick test (SPT) to Senecio were recruited. The clinical relevance of this pollen was assessed by means of a nasal provocation test (NPT). Allergens were characterized by one-dimensional electrophoresis (SDS-PAGE) and two-dimensional gel electrophoresis and immunoblotting. Furthermore, characterization and identification of the allergens were performed by mass spectrometry (MS). In vitro inhibition tests were performed to evaluate cross-reactivity with other pollen. RESULTS: Three predominant allergens, both in the intensity of reaction and the frequency of recognition by human-allergic sera, were 59 (60%), 42 (50%) and 31 kDa (50%). The two-dimensional analysis allowed the identification of several allergens. One spot around 42 kDa was identified as a protein homologous to pectate lyase and three other spots were homologous to malate dehydrogenase by MS. S. jacobea proteins showed cross-reactivity with other proteins of the Asteraceae family and also with Parietaria judaica. This was demonstrated by immunoblotting and ELISA inhibition studies. CONCLUSION: S. jacobea constitute a newly discovered allergenic source. It shows cross-reactivity with other members of the Asteraceae plant family as well as with P. judaica.

Components of the Mediterranean-type food pattern and serum inflammatory markers among patients at high risk for cardiovascular disease.

OBJECTIVE: To evaluate associations between components of the Mediterranean diet and circulating markers of inflammation in a large cohort of asymptomatic subjects at high risk for cardiovascular disease. SUBJECTS/METHODS: A total of 339 men and 433 women aged between 55 and 80 years at high cardiovascular risk because of presence of diabetes or at least three classical cardiovascular risk factors, food consumption was determined by a semi-quantitative food frequency questionnaire. Serum concentrations of high-sensitivity C-reactive protein (CRP) were measured by immunonephelometry and those of interleukin-6 (IL-6), intracellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) by enzyme-linked immunosorbent assay. RESULTS: After adjusting for age, gender, body mass index, diabetes, smoking, use of statins, non-steroidal antiinflammatory drugs and aspirin, a higher consumption of fruits and cereals was associated with lower concentrations of IL-6 (P for trend 0.005;both). Subjects with the highest consumption of nuts and virgin olive oil showed the lowest concentrations of VCAM-1, ICAM-1, IL-6 and CRP; albeit only for ICAM-1 was this difference statistically significant in the case of nuts (for trend 0.003) and for VCAM-1 in the case of virgin olive oil (P for trend 0.02). Participants with higher adherence to the Mediterranean-type diet did not show significantly lower concentrations of inflammatory markers (P<0.1 for VCAM-1 and ICAM-1). CONCLUSIONS: The consumption of some typical Mediterranean foods (fruits, cereals, virgin olive oil and nuts) was associated with lower serum concentrations of inflammatory markers especially those related to endothelial function, in subjects with high cardiovascular risk living in a Mediterranean country.

Modelling diseases: the allergens of Olea europaea pollen.

This study analyzes the influence of the IgE response to certain olive pollen allergens in the modulation of the different clinical phenotypes of allergic disease and their relationship with the level of exposure to pollen and genetic factors. Patients from high-exposure areas had a complex IgE antibody response to allergens of Olea euroapea, which included 3 or more allergens in 75% of cases. The majority allergens were Ole e 1, Ole e 2 (profilin), Ole e 7 (lipid transporting protein), Ole e 9 (glucanase), and Ole e 10. The existence of the antigen HLA-DR2 (15) led to a higher risk of sensitization to Ole e 10 and a greater trend towards the development of severe asthma, which increased in the presence of an anti-profilin IgE. Thirty percent of patients suffering from pollinosis simultaneously presented allergy to vegetable foods. Anti-Ole e 7 IgE was significantly associated with fruit anaphylaxis and anti-profilin IgE was detected in 90% of patients with oral syndrome. Finally, we analyzed the role of glucanase and Ole e 10 as causes of the pollen-latex-fruit syndrome.

Modulation of allergic response by gene-environment interaction: olive pollen allergy.

This article summarizes the most important advances of recent years in the field of gene-environment interaction in allergic response. It specifically examines sensitization to olive pollen as an example of one of the main causes of allergic disease in the Mediterranean area. The presence of at least 20 proteins with allergic activity has been demonstrated in olive pollen, and 10 of these have been characterized (Ole e 1 to Ole e 10). Ole e 1, which is considered to be the majority allergen (causing sensitization in more than 70% of patients), has been the subject of many studies looking for risk factors and ways to protect against sensitization. Markers of the major histocompatibility complex and other genetic loci associated with the allergic response have been analyzed using population-based, family-based, and functional approaches, which have revealed the involvement of genetic regulation in this type of response. Furthermore, evaluation of environmental factors and their relationship with genetic factors is essential when attempting to understand this type of disease. In this review, we provide examples of how exposure to high doses of olive pollen allergen in a specific genetic context can trigger different allergic conditions (from asthma to nonresponse). We stress the importance of evaluating these factors in order to modulate this response correctly.

Biomethanization of mixtures of fruits and vegetables solid wastes and sludge from a municipal wastewater treatment plant.

The possible management of Fruit and Vegetable Solid Wastes (FVSWs) through their simultaneous digestion with the primary sludge of Municipal Wastewater Treatment plants is investigated. This alternative allows the recovery of energy and a solid product that can be used as an amendment for soils that generated the residue, while is not expensive. Results indicate that the ratio of FVSWs to sludge and the pH control are the main variables determining the methane production and concentration. NaHCO3 was selected to achieve the pH control. The results for a ratio of 50% sludge together with 10 g NaHCO3/kg of residue are among the best obtained, with a methane yield of about 90 L per kg of volatile solids, and a methane concentration of 40% (v/v) of the biogas. A 50% reduction of the total solids; 21% reduction of the volatile solids (in terms of total solids); and a pH value of the sludge, which is 6.9 indicate that the digested sludge can be a good material for soil amendment.

Allergy to cypress pollen.

Although Cupressus sempervirens has been spread over southern Europe since antiquity, cypress pollen allergy has not been reported until 1945. In France, the very first case reports were published in 1962. Since then, the prevalence of cypress pollinosis seems to demonstrate an upward trend, concomitantly with the increased use of cypress trees as ornamental plants, as wind breaks and as hedges. Hyposensitization, using improved pollen extracts, is increasingly prescribed. Besides, prevention measures begin to be implemented. Such measures include avoidance of planting new cypress trees, especially near human populations' centres, trimming of cypress hedges before the pollination season and agronomical research for hypoallergenic trees. Altogether, such new developments in cypress allergy deserve an update review.

Ole e 2 and Ole e 10: new clinical aspects and genetic restrictions in olive pollen allergy.

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 = 0.04) and Ole e 10 reactivities (OR 2.8, P = 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 = 0.002). Phenotypic frequency of DR7 (OR 5.4, Pc = 0.003) and DQ2 (OR 3.6, Pc = 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 = 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.

Cloning and expression of a major allergen from Cupressus arizonica pollen, Cup a 3, a PR-5 protein expressed under polluted environment.

BACKGROUND: This paper describes the cloning and expression of the Cupressus arizonica pollen protein Cup a 3. In addition, we present its modulation under polluted environmental conditions. Species of the Cupressaceae family are important because of their high sensitization prevalence. METHODS: Cup a 3 cloning is based on the sequence of the homologous protein Jun a 3. Cup a 3 was expressed with good yield in the methylotropic yeast Pichia pastoris. RESULTS: Recombinant Cup a 3 (rCup a 3) contains 199 amino acids, 10 potential phosphorylation sites and no glycosylation sites. By immunoblot 63% of cypress allergic patients had specific immunoglobulin E antibodies against rCup a 3 (n = 104). This major allergen is homologous to members of the pathogenesis-related proteins (PR-5 group) and contributes to the overall allergenicity of C. arizonica pollen. Our results show that the increased expression of Cup a 3 is dependent on the pollution in the area where the pollen has been collected, being higher under polluted conditions. CONCLUSIONS: Cup a 3 is a PR-5 protein derived from C. arizonica pollen. The expression of the protein under polluted conditions has a direct incidence on the pollen allergenicity, as has been demonstrated by skin tests and Radioallergosorbent test inhibition.

Immunochemical characterization of Russian thistle (Salsola kali) pollen extracts. Purification of the allergen Sal k 1.

BACKGROUND: Salsola kali (Russian thistle) is a weed which belongs to the Chaenopodiacea family. It is widely distributed along the coasts of Europe, North Africa, USA and Australia. The objectives of this study were to study the allergenic composition of S. kali pollen and to purify an important allergen from the pollen extracts of this plant. METHODS: A population of 66 individuals with specific IgE-mediated allergic symptoms and positive skin tests to S. kali were included in the study. Specific IgE to S. kali was determined by direct enzyme-linked immunosorbent assay (ELISA). The antigenic and allergenic profile of S. kali was evaluated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focussing (IEF) and immunoblot. Allergen purification was conducted by preparative SDS-PAGE. The allergenicity of the protein was evaluated by skin testing, direct ELISA, ELISA inhibition and immunoblots. RESULTS: Specific IgE to S. kali was detected in 39 of the 66 individuals (59%). An allergen with a molecular weight of approximately 43 kDa was purified. This allergen was termed Sal k 1. A partial sequencing was obtained and no homology was found with other known proteins/allergens. The allergenicity of Sal k 1 was tested in vitro and in vivo. Of the 39 individuals with a positive specific IgE determination to S. kali, 26 (66.6%) had detectable specific IgE to Sal k 1. Twenty of these 39 individuals were skin-prick tested with the purified allergen (0.5 mg/ml) and all of them had a positive skin test to the purified allergen. Ten additional individuals, used as negative controls, had a negative response. CONCLUSIONS: Sal k 1, an important allergen of S. kali, is recognized, in vitro, by approximately 67% of the patients sensitized to S. kali. Twenty patients with a positive skin test to a standardized S. kali extract had a positive reaction to the purified allergen.

Molecular cloning of major allergen from Cupressus arizonica pollen: Cup a 1.

The family Cupressaceae is a relevant source of allergens that causes winter respiratory allergies. Cloning and sequencing the major antigen of Cupressus arizonica is important for a better diagnosis and treatment of sensitized patients. To obtain a full-length complementary DNA for Cup a 1, the major allergen of Cupressus arizonica pollen. It was cloned and sequenced and the recombinant protein was expressed. Messenger RNA from Cupressus arizonica pollen was obtained and the Cup a 1 sequence was established using a 3'-RACE system and primers based on the N-terminal amino acid sequence. Recombinant Cup a 1 was cloned in pBluescript and expressed in a glycosylated form in rabbit reticulocytes. The cDNA was subcloned in pGEX-5X-1 and expressed in Escherichia coli as a fusion protein with GST. Recombinant Cup a 1 is highly homologous with the major allergens of mountain cedar (Jun a 1), Japanese cypress (Cha o 1) and Japanese cedar (Cry j 1). Cup a 1 contains three potential N-glycosylation sites that are different from those found in Jun a 1 and Cry j 1. The cloned protein contains a pectate lyase active site identical to those of Cry j 1 and Jun a 1. The IgE from patients' sera recognizes recombinant Cup a 1, and this reactivity is higher with the glycosylated protein. Cup a 1 has been cloned and sequenced. As expected, the high degree of homology with Cha o 1, Jun a 1 and Cry j 1 explains the cross-reactivity of conifer pollens. Different IgE reactivity with the glycosylated and non-glycosylated protein suggests the importance of carbohydrate moieties in the IgE binding site.

Environment and sensitization to cypress in Israel.

Cypresses are part of the Israeli landscape, as well as they are over most of the Mediterranian area. Pollen dispersal starts in January and peaks in March-April. Assessment of hay-fever patients showed a prevalence of 24%-32% (depending on the region) sensitization to Cypress (Cupressus sempervirens) pollen extracts. 13% of all those who are sensitive to Cypress were monosensitized. They are symptomatic from February till April, when 70% of them have rhinitis, 30% also asthma and 18% conjunctivitis. In the monosensitized patients, anti-cypress immunotherapy was successful.

Molecular cloning and characterization of an IgE-reactive protein from Anisakis simplex: Ani s 1.

Ingestion of the parasitic nematode Anisakis simplex in undercooked fish can cause severe allergic reactions in some individuals. Using pooled human sera from sensitized patients we have probed an expression library for A. simplex antigens. One positive clone was found to encode a full length 21 kDa protein with strong homology to nematode troponins. The recombinant protein was expressed as a GST-fusion protein and found by immunoblot analysis to react with sera from 20% of allergic patients. The presence of functional EF-hand Ca(2+) binding motifs was demonstrated by gel-shift analysis.

DR2 antigens are associated with severity of disease in toxic oil syndrome (TOS).

Toxic oil syndrome (TOS) was an epidemic which broke out in Spain in 1981, caused by the ingestion of rapeseed oil denatured with 2% aniline and sold illegally as edible oil. More than 20,000 people were affected and mortality rate was 8.4%. Genetic susceptibility appears to be involved in the pathology of this disease. Several reports have described association between the chronic stage of the disease and DR-DQ antigens (DR3, DR4, DR2 and DQ8). In the present work, we have reassessed the HLA class II antigens in a well-designed case-control study. Triplets of subjects (n=265) composed by chronic patients (n=117), non-affected family members (n=71) and non-related controls (n=77) were studied. Also, HLA class II antigens were analyzed in patients who had died from TOS (n= 34) and in TOS control patients who died from other non-TOS related causes (n=13). Regarding surviving patients no significant association was found between HLA and disease. In contrast, an increase in phenotypic frequency of DR2 antigen, was found in patients who had died from TOS (73.5%) compared with the whole study group: TOS-affected alive patients (25.6%, corrected P<0.001), non-affected family members (28.5%, corrected P<0.001), non-related controls (23.9%, corrected P<0.001) and dead controls (38.4%, P=0.03).

Genetic restrictions in olive pollen allergy.

BACKGROUND: The major antigen of olive tree pollen, Ole e 1, produces an IgE response restricted by DQ2. OBJECTIVE: Our purpose was to further analyze the genetic restrictions associated with IgE and IgG antibodies against Ole e 1 and IgE against the recently described antigen Ole e 3. METHODS: Twenty-two nuclear olive pollen-allergic families (n = 88) were selected. DRB1 and DQB1, TCR-Valpha 8.1, the high-affinity receptor of IgE (FcepsilonRI-beta) Rsa I exon 7 and intron 2 and TNF-beta (LTalpha-Nco I) polymorphisms were determined by PCR and analyzed for association with allergic traits by the multiallelic transmission disequilibrium test. RESULTS: Significant associations were found among HLA-DQB1*0201 (n = 29) and high levels of IgG (P =.023) and IgE (P =.0136) antibodies to Ole e 1 and with IgE specific to Ole e 3 (P =.0368). DRB1*0701 was associated with high levels of total serum IgE (P =.04) and IgG against Ole e 1 (P =.025). The FcepsilonRI-beta Rsa I exon 7, allele 1 (n = 39), was associated with high levels of total serum IgE (P =. 01), IgE antibodies against Olea europaea extract (P =.004), and specific antibodies to Ole e 1, IgG (P =.04), and IgE (P =.006). The FcepsilonRI-beta Rsa I intron 2, allele 2 (n = 33), was associated with IgE antibodies to O europaea extract (P =.003) and specific antibodies to Ole e 1, IgG (P =.025), and IgE (P =.05). CONCLUSIONS: We found a new association between IgE antibody response to Ole e 3 and DQB1*0201 and verified the previously reported association between Ole e 1-specific response and DQB1*0201. Also, the association between FcepsilonRI-beta and IgE antibodies against Ole e 1 was demonstrated.

[Diets enriched with monounsaturated fatty acids and omega-3 polyunsaturated fatty acids decrease blood pressure without changing the plasma insulin concentration in healthy subjects]. / Las dietas enriquecidas en ácidos grasos monoinsaturados y ácidos grasos poliinsaturados omega 3 disminuyen la presión arterial, sin modificar la concentración de insulina plasmática en sujetos sanos.

BACKGROUND: Plasma glucose, insulin and blood pressure are wellknown cardiovascular risk factors, which may be influence by dietary factors. The aim of the study was to investigate whether changes in dietary fatty acids could modify plasma concentration of glucose, insulin and mean blood pressure (MBP). SUBJECTS AND METHODS: Forty two subjects (18 women and 24 men) were placed in four consecutive five week diet periods. Energy intake from proteins, carbohydrates and fats was constant during the study and there was only changes on fatty acids composition. First period was enriched on saturated fatty acids (SFA), second period on monounsaturated fatty acids (MUFA) and third and fourth periods were enriched on polyunsaturated fatty acids (PUFA). Fourth period was also enriched on PUFA n-3. RESULTS: No significant changes were found on glucose and insulin plasma concentration. However, a significant effect was detected on MBP on total population (p < 0.0001) and by gender. MUFA and PUFA n-3 enriched diet decreased significantly MBP compared to SFA enriched diet AGS (85.7, SD 9.1, 87.3, SD 8.7 y 90.3, SD 8.8 mmHg, respectively). In addition, a weak (r = 0.28) but significant (p = 0.002) correlation was found between MBP and plasma insulin. CONCLUSIONS: Diets enriched on MUFA fatty acids and n-3 fatty acids decrease significantly MBP without modifying glucose and insulin plasma concentration.