Center for Plant Biotechnology and Genomics

Pozuelo de Alarcón, Spain

Center for Plant Biotechnology and Genomics

Pozuelo de Alarcón, Spain

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Calderon A.,CSIC - Center of Edafology and Applied Biology of the Segura | Ortiz-Espin A.,CSIC - Center of Edafology and Applied Biology of the Segura | Iglesias-Fernandez R.,Center for Plant Biotechnology and Genomics | Carbonero P.,Center for Plant Biotechnology and Genomics | And 3 more authors.
Redox Biology | Year: 2017

Thioredoxins (Trxs), key components of cellular redox regulation, act by controlling the redox status of many target proteins, and have been shown to play an essential role in cell survival and growth. The presence of a Trx system in the nucleus has received little attention in plants, and the nuclear targets of plant Trxs have not been conclusively identified. Thus, very little is known about the function of Trxs in this cellular compartment. Previously, we studied the intracellular localization of PsTrxo1 and confirmed its presence in mitochondria and, interestingly, in the nucleus under standard growth conditions. In investigating the nuclear function of PsTrxo1 we identified proliferating cellular nuclear antigen (PCNA) as a PsTrxo1 target by means of affinity chromatography techniques using purified nuclei from pea leaves. Such protein–protein interaction was corroborated by dot-blot and bimolecular fluorescence complementation (BiFC) assays, which showed that both proteins interact in the nucleus. Moreover, PsTrxo1 showed disulfide reductase activity on previously oxidized recombinant PCNA protein. In parallel, we studied the effects of PsTrxo1 overexpression on Tobacco Bright Yellow-2 (TBY-2) cell cultures. Microscopy and flow-cytometry analysis showed that PsTrxo1 overexpression increases the rate of cell proliferation in the transformed lines, with a higher percentage of the S phase of the cell cycle at the beginning of the cell culture (days 1 and 3) and at the G2/M phase after longer times of culture (day 9), coinciding with an upregulation of PCNA protein. Furthermore, in PsTrxo1 overexpressed cells there is a decrease in the total cellular glutathione content but maintained nuclear GSH accumulation, especially at the end of the culture, which is accompanied by a higher mitotic index, unlike non-overexpressing cells. These results suggest that Trxo1 is involved in the cell cycle progression of TBY-2 cultures, possibly through its link with cellular PCNA and glutathione. © 2017 The Authors

Escribese M.M.,University of San Pablo - CEU | Gomez-Casado C.,Center for Plant Biotechnology and Genomics | Barber D.,University of San Pablo - CEU | Diaz-Perales A.,Center for Plant Biotechnology and Genomics
Journal of Investigational Allergology and Clinical Immunology | Year: 2015

Allergens come into contact with the immune system as components of a very diverse mixture. The most common sources are pollen grains, food, and waste. These sources contain a variety of immunomodulatory components that play a key role in the induction of allergic sensitization. The way allergen molecules bind to the cells of the immune system can determine the immune response. In order to better understand how allergic sensitization is triggered, we review the molecular mechanisms involved in the development of allergy and the role of immunomodulators in allergen recognition by innate cells. © 2015 Esmon Publicidad.

Barber D.,University of San Pablo - CEU | Diaz-Perales A.,Center for Plant Biotechnology and Genomics | Villalba M.,Complutense University of Madrid | Chivato T.,University of San Pablo - CEU
Current Allergy and Asthma Reports | Year: 2015

Over the past few decades, significant scientific progress has influenced clinical allergy practice. The biological standardization of extracts was followed by the massive identification and characterization of new allergens and their progressive use as diagnostic tools including allergen micro arrays that facilitate the simultaneous testing of more than 100 allergen components. Specific diagnosis is the basis of allergy practice and is always aiming to select the best therapeutic or avoidance intervention. As a consequence, redundant or irrelevant information might be adding unnecessary cost and complexity to daily clinical practice. A rational use of the different diagnostic alternatives would allow a significant improvement in the diagnosis and treatment of allergic patients, especially for those residing in complex pollen exposure areas. © 2014, Springer Science+Business Media New York.

Gomez F.,University of Malaga | Aranda A.,University of Malaga | Campo P.,University of Malaga | Diaz-Perales A.,Center for Plant Biotechnology and Genomics | And 6 more authors.
PloS one | Year: 2014

BACKGROUND: Apple allergy manifests as two main groups of clinical entities reflecting different patterns of allergen sensitization: oral allergy syndrome (OAS) and generalized symptoms (GS).OBJECTIVE: We analysed the sensitization profile to a wide panel of different components of food allergens (rMal d 1, Mal d 2, rMal d 3, rMal d 4, rPru p 3, rBet v 1 and Pho d 2) for a population of Mediterranean patients with OAS and GS to apple.METHODS: Patients (N = 81) with a history of apple allergy that could be confirmed by positive prick-prick test and/or double-blind-placebo-controlled food challenge (DBPCFC), were included. Skin prick test (SPT) and ELISA were performed using a panel of inhalant, fruit and nut allergens. ELISA and ELISA inhibition studies were performed in order to analyse the sensitization patterns.RESULTS: Thirty-five cases (43.2%) had OAS and 46 (56.8%) GS. SPT showed a significantly higher number of positive results with peach, cherry and hazelnut in those with GS. ELISA showed a significantly high percentage of positive cases to rMal d 3, rMal d 4, rPru p 3 and Pho d 2 in patients with OAS and GS compared to controls, and to rBet v 1 in patients with OAS vs controls and between OAS and GS patients. Three different patterns of recognition were detected: positive to LTP (rMal d 3 or rPru p 3), positive to profilin (rMal d 4 and Pho d 2), or positive to both. There were also patients with rMal d 1 recognition who showed cross-reactivity to rBet v 1.CONCLUSION: In an apple allergy population with a high incidence of pollinosis different patterns of sensitization may occur. LTP is most often involved in those with GS. Profilin, though more prevalent in patients with OAS, has been shown to sensitise patients with both types of symptoms.

Aranda A.,Carlos Haya Hospital | Campo P.,Carlos Haya Hospital | Palacin A.,Center for Plant Biotechnology and Genomics | Dona I.,Carlos Haya Hospital | And 4 more authors.
PLoS ONE | Year: 2013

Background: Obeche wood dust is a known cause of occupational asthma where an IgE-mediated mechanism has been demonstrated. Objective: To characterize the allergenic profile of obeche wood dust and evaluate the reactivity of the proteins by in vitro, ex vivo and in vivo assays in carpenters with confirmed rhinitis and/or asthma Materials and methods: An in-house obeche extract was obtained, and two IgE binding bands were purified (24 and 12 kDa) and sequenced by N-terminal identity. Specific IgE and IgG, basophil activation tests and skin prick tests (SPTs) were performed with whole extract and purified proteins. CCD binding was analyzed by ELISA inhibition studies. Results: Sixty-two subjects participated: 12 with confirmed occupational asthma/rhinitis (ORA+), 40 asymptomatic exposed (ORA-), and 10 controls. Of the confirmed subjects, 83% had a positive SPT to obeche. There was a 100% recognition by ELISA in symptomatic subjects vs. 30% and 10% in asymptomatic exposed subjects and controls respectively (p<0.05). Two new proteins were purified, a 24 kDa protein identified as a putative thaumatin-like protein and a 12 kDa gamma-expansin. Both showed allergenic activity in vitro, with the putative thaumatin being the most active, with 92% recognition by ELISA and 100% by basophil activation test in ORA+ subjects. Cross-reactivity due to CCD was ruled out in 82% of cases. Conclusions: Two proteins of obeche wood were identified and were recognized by a high percentage of symptomatic subjects and by a small proportion of asymptomatic exposed subjects. Further studies are required to evaluate cross reactivity with other plant allergens. © 2013 Aranda et al.

Lavilla M.,Food Research Division | Orcajo J.,Food Research Division | Diaz-Perales A.,Center for Plant Biotechnology and Genomics | Gamboa P.,Basurto Hospital
Innovative Food Science and Emerging Technologies | Year: 2016

The aim of this work is to study the effect of High Pressure Processing (HPP) on the allergenicity of the main protein involved in peach allergy (Pru p 3). Results obtained showed that most pressure/time combinations slightly enhanced in vitro IgE-binding to Pru p 3 and peach extract. Moreover, additional tests were carried out by means of skin prick tests on peach allergic patients. Different from in vitro results, in vivo effects evidenced that HPP (600. MPa/5. min) can either reduce or increase Pru p 3 allergenicity, depending on the particular sensitisation of each patient. Notwithstanding this variability, it is highly remarkable that the skin response to pressurized peach extract was stronger in more than half of individuals. These results would suggest a higher risk of HPP-treated peach products to elicit an allergenic reaction. However, it has been also proved that matrix plays an important role in peach allergenicity modification. Consequently, further investigations are needed before extrapolating results to more complex products. Industrial relevance: Peach allergy is one of the most frequent allergies in Mediterranean countries among adult patients. However, up to date, there is no information about the effect of HPP on peach allergenicity. This study increases understanding about the impact of processing on an important fruit allergen in order to advance knowledge-based ways to managing allergens risks in industry, as well as it opens new opportunities of research in other technologies or strategies in the attempt to reduce peach allergenicity. © 2016 Elsevier Ltd.

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