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Olive (Olea europaea) pollen constitutes one of the most important allergen sources in the Mediterranean countries and some areas of the United States, South Africa, and Australia. Recently, we provided evidence that olive pollen releases nanovesicles of respirable size, named generically pollensomes, during in vitro germination. Olive pollensomes contain allergens, such as Ole e 1, Ole e 11, and Ole e 12, suggesting a possible role in allergy. The aim of this study was to assess the contribution of pollensomes to the allergic reaction. We show that pollensomes exhibit allergenic activity in terms of patients’ IgE-binding capacity, human basophil activation, and positive skin reaction in sensitized patients. Furthermore, allergen-containing pollensomes have been isolated from three clinically relevant nonphylogenetically related species: birch (Betula verrucosa), pine (Pinus sylvestris), and ryegrass (Lolium perenne). Most interesting, pollensomes were isolated from aerobiological samples collected with an eight-stage cascade impactor collector, indicating that pollensomes secretion is a naturally occurring phenomenon. Our findings indicate that pollensomes may represent widespread vehicles for pollen allergens, with potential implications in the allergic reaction.

The basophil activation test (BAT) has become a pervasive test for allergic response through the development of flow cytometry, discovery of activation markers such as CD63 and unique markers identifying basophil granulocytes. Basophil activation test measures basophil response to allergen cross-linking IgE on between 150 and 2000 basophil granulocytes in

Background The ImmunoCAP ISAC 112 is a fluoro-immunoassay that allows detection of specific IgE to 112 molecular components from 51 allergenic sources. We studied the reliability of this technique intra- and inter- assay, as well as inter-batch- and inter-laboratory-assay. Methods Twenty samples were studied, nineteen sera from polysensitized allergic patients, and the technique calibrator provided by the manufacturer (CTR02). We measured the sIgE from CTR02 and three patients' sera ten times in the same and in different assays. Furthermore, all samples were tested in two laboratories and with two batches of ISAC kit. To evaluate the accuracy of ISAC 112, we contrasted the determinations of CTR02 calibrator with their expected values by T Student test. To analyse the precision, we calculated the coefficient of variation (CV) of the 15 allergens that generate the calibration curve, and to analyse the repeatability and the reproducibility, we calculated the intraclass coefficient correlation (ICC) to each allergen. Results The results obtained for CTR02 were similar to those expected in 7 of 15 allergens that generate the calibration curve, whereas in 8 allergens the results showed significant differences. The mean CV obtained in the CTR02 determinations was of 9.4%, and the variability of sera from patients was of 22.9%. The agreement in the intra- and inter-assay analysis was very good to 94 allergens and good to one. In the inter-batch analyse, we obtained a very good agreement to 82 allergens, good to 14, moderate to 5 allergens, poor to one, and bad to 1 allergen. In the inter-laboratory analyse, we obtained a very good agreement to 73 allergens, good to 22, moderate to 6 and poor to two allergens. Conclusion The allergen microarray immunoassay, ISAC 112, is a repeatable and reproducible in vitro diagnostic tool for determination of sIgE beyond the own laboratory.

This is an open-access article distributed under the terms of the Creative Commons Attribution License.-- et al., Here we perform a large-scale study of the structural properties and the expression of proteins that constitute the human Centrosome. Centrosomal proteins tend to be larger than generic human proteins (control set), since their genes contain in average more exons (20.3 versus 14.6). They are rich in predicted disordered regions, which cover 57% of their length, compared to 39% in the general human proteome. They also contain several regions that are dually predicted to be disordered and coiled-coil at the same time: 55 proteins (15%) contain disordered and coiled-coil fragments that cover more than 20% of their length. Helices prevail over strands in regions homologous to known structures (47% predicted helical residues against 17% predicted as strands), and even more in the whole centrosomal proteome (52% against 7%), while for control human proteins 34.5% of the residues are predicted as helical and 12.8% are predicted as strands. This difference is mainly due to residues predicted as disordered and helical (30% in centrosomal and 9.4% in control proteins), which may correspond to alpha-helix forming molecular recognition features (α-MoRFs). We performed expression assays for 120 full-length centrosomal proteins and 72 domain constructs that we have predicted to be globular. These full-length proteins are often insoluble: Only 39 out of 120 expressed proteins (32%) and 19 out of 72 domains (26%) were soluble. We built or retrieved structural models for 277 out of 361 human proteins whose centrosomal localization has been experimentally verified. We could not find any suitable structural template with more than 20% sequence identity for 84 centrosomal proteins (23%), for which around 74% of the residues are predicted to be disordered or coiled-coils. The three-dimensional models that we built are available at http://ub.cbm.uam.es/centrosome/models/i​ndex.php, The authors gratefully acknowledge financial support from the Spanish Ministry of Science, grant CSD2006-00023, and from the Madrid Community, grant S2010/BMD-2305.

Pru p 3 has been suggested to be the primary sensitizing allergen in patients with peanut allergy in the Mediterranean area. We aimed to confirm this hypothesis, studying 79 subjects.

In the last decades, significant progress in research and clinics has been made to offer possible innovative therapeutics for the management of allergic diseases. However, current allergen immunotherapy shows limitations concerning the long-term efficacy and safety due to local side effects and risk of anaphylaxis. Thus, effective and safe vaccines with reduced dose of allergen have been developed using adjuvants. Nevertheless, the use of adjuvants still has several disadvantages, which limits its use in human vaccines. In this context, several novel adjuvants for allergen immunotherapy are currently being investigated and developed. Currently, nanoparticles-based allergen-delivery systems have received much interest as potential adjuvants for allergen immunotherapy. It has been demonstrated that the incorporation of allergens into a delivery system plays an important role in the efficacy of allergy vaccines. Several nanoparticles-based delivery systems have been described, including biodegradable and nondegradable polymeric carriers. Therefore, this paper provides an overview of the current adjuvants used for allergen immunotherapy. Furthermore, nanoparticles-based allergen-delivery systems are focused as a novel and promising strategy for allergy vaccines.

To my opinion, some patients may have reversible tolerance to food allergens. This tolerance that might appear or disappear can be related to the presence of triggers that activate the mast cell, and include exercise, NSAIDs, alcohol or stress. Different authors have already stated that food allergy can be activated by NSAIDs, exercise, alcohol or stress. These authors demonstrated increase of food absortion as an explanation. It can also suggest that these triggers lowered threshold for the release of cytoplasmatic mediators from mast cell As we can not work with mast cell we did it with basophils and we evaluate the effect of NSAID on basophile response to food allergens using BAT.

Background: Specific immunotherapy implies certain drawbacks which could be minimized by the use of good adjuvants, capable of amplifying the appropriate immune response with minimal adverse effects. Objective: To evaluate the adjuvant capacity of the association of the rough lipopolysaccharide of Brucella ovis with Gantrez® AN nanoparticles. Methods: Ovalbumin (allergen model)-containing Gantrez® AN nanoparticles with either encapsulated or coated lipopolysaccharide were prepared and administered intradermally to BALB/c mice in order to evaluate the immune response. Pre-sensitized BALB/c mice were also administered with the formulations and they were challenged with an intraperitoneal injection of ovalbumin. Anaphylactic symptoms, including mortality rates, were evaluated after the challenge. Results: The intradermal administration of mice with ovalbumin-containing nanoparticles elicited high and sustained specific IgG1 and IgG2a responses. However, the only treatment that totally protected the mice from death after the challenge of induced allergic mice to ovalbumin, required the co-administration of lipopolysaccharide entrapped inside the nanoparticles. Conclusion: Gantrez® AN nanoparticles with entrapped rough lipopolysaccharide of Brucella ovis protected ovalbumin pre-sensitized BALB/c mice from anaphylactic shock. Clinical Implications: These results are highly suggestive for the valuable use of Gantrez® nanoparticles combined with lipopolysaccharide of Brucella ovis in immunotherapy with allergens.

Performance of commercially available component resolved microarray has not been widely evaluated in food allergy. Nut allergy is a prevalent and increasing food allergy in which component resolved diagnosis is crucial. In the context of a multicentric study on pollen and vegetable food allergy in Spain, we enrolled patients (n=100) who had suffered at least two episodes of immediate symptoms after ingestion of nuts (peanut, walnut or hazelnut). Skin prick test was positive for the corresponding nut. ImmunoCAP ISAC 112 was analyzed in all patients and ImmunoCAP to the available components from the studied sources were analyzed (Ara h 1, Ara h 2, Ara h 3 and Ara h 8 and Ara h 9, Jug r 1, Jug r 3, Cor a 1, Cor a 8, Cor a 9, Cor a 14) in those patients showing negative result to ISAC. Among the 67 patients allergic to peanut 3 were sensitized to Ara h 1, 3 to Ara h 2, 1 to Ara h 3, 9 to Ara h 6, 1 to Ara h 8 and 36 to Ara h 9. Twenty-seven peanut allergic patients showed no sensitization to any components in the microarray. Among these 27 ISAC negative peanut allergic patients, 3 were sensitized to Ara h 1, 1 to Arah 2, 1 to Ara h 3, 1 to Ara h 8 and 23 to Ara h 9 by CAP. Among the 69 patients allergic to walnut 7 were sensitized to Jug r 1, 6 to Jug r 2 and 45 to Jug r 3. Eighteen walnut allergic patient showed no sensitization to any component in the microarray. Among these 18 ISAC negative walnut allergic patients, 10 were sensitized to Jug r 3 by CAP (none to Jug r 1). Among the 56 patients allergic to hazelnut one was sensitized to Cor a 1, 33 to Cor a 8 and 2 to Cor a 9. Twenty hazelnut allergic patient showed no sensitization to any components in the microarray. Among these 20 ISAC negative hazelnut allergic patients, 10 were sensitized to Cor a 8 by CAP (none to Cor a 1, Cor a 9 or Cor a 14). Among the 100 patients allergic to peanut, walnut or hazelnut, 40 patients did not show specific IgE against the studied nuts/ components in ISAC. Only a sixty per cent of the patients allergic to nuts (peanut, walnut or hazelnut) are diagnosed by ISAC. Sensitization to the lipid transfer proteins Ara h9, Jug r 3 and Cor a 8 was not detected by ISAC but it was recovered by CAP analysis.