Ernesto Silva

Karolinska Institutet, Solna, Stockholm, Sweden

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Publications (3)13.04 Total impact

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    ABSTRACT: Alveolar macrophages (AMs) are important for granuloma formation, which is the histological hallmark in sarcoidosis. Their function as antigen presenting cell in sarcoidosis is also believed to be relevant to the outcome of disease, resulting either in remission or a prolonged chronic inflammation in the lungs.To study alterations in the expression levels of the soluble proteome of AMs in pulmonary sarcoidosis as compared to healthy controls, with the goal of identifying specific proteins and pathways important for the mechanisms of disease and/or disease phenotype.Quantitative proteomics approach using two-dimensional Difference Gel Electrophoresis coupled to mass spectrometry was applied. Data was evaluated using multivariate modelling and pathway analyses.Results: Sixty-nine protein spots were found to be significant altered between sarcoidosis and healthy controls. Among these, twenty-five unique proteins were identified. Several of the identified proteins were related to key AM functionality, including the Fcγ-mediated phagocytosis and Clathrin-mediated endocytosis pathways.Global proteomics analysis provided identification of alterations of a subset of proteins not previously reported in sarcoidosis. These alterations primarily affect biological pathways related to phagocytic macrophage functionality. These findings provide important insights into the role of macrophages in sarcoidosis pathogenesis.
    European Respiratory Journal 10/2012; 41(6):1331-9. · 6.36 Impact Factor
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    ABSTRACT: Historically, the use of two-dimensional electrophoresis (2-DE) in quantitative proteomics has been hampered by significant technical variance. Over the past decade, a range of technological leaps have reduced the overall variance of 2-DE, thus turning the technology into a robust platform for quantitative intact proteomics. However, as the confounding gel-to-gel variation improves, the variance arising from the subsequent image analysis becomes more prominent. Limitations in image alignment and spot detection of previous generations of 2-DE analysis software have demanded considerable user-intervention and manual editing, resulting in introduction of a large degree of subjectivity and software-induced variance. We evaluated the performance of SameSpots, representing a new generation of 2-DE image analysis software, using both DIGE and traditional single-stain 2-DE approaches. Evaluations of the software-induced variance in relation to other sources of variance, as well as the subjectivity through comparison of analyses performed by an expert user and a novice lab-user, were performed. In terms of statistical power, the less-experienced user achieved the better results, but no discernible difference was detected in multivariate comparisons between the users. In conclusion, we found that SameSpots represents improvements both in reproducibility and objectivity in relation to previous generations of 2-DE analysis software.
    Journal of Proteome Research 03/2010; 9(3):1522-32. · 5.06 Impact Factor
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    ABSTRACT: Sarcoidosis and chronic beryllium disease (CBD) are granulomatous disorders which can lead to development of chronic inflammation and fibrosis. These diseases have several similarities from their clinical aspects. The aim of this study was to compare the protein profile at the site of active inflammation i.e. the lungs of patients with sarcoidosis and CBD. Bronchoalveolar lavage (BAL) proteins from patients having sarcoidosis or CBD were studied using two dimensional gel based proteomics. In this study, we used Difference Gel Electrophoresis (DIGE) proteomics approach to analyse the protein expression profiles from sarcoidosis patients (n=4), CBD (n=4) and healthy controls (n = 5). Subsequently, differentially expressed proteins were identified by using mass spectrometry. We found 37 protein-spots with statistically different expression levels, and identified 14 of these proteins. The protein expression levels of peroxiredoxin 5, heat shock protein 70, complement C3, annexin A2 and transthyretin were significantly different in sarcoidosis versus control group. The proteins; hemopexin, beta2-microglobulin, alpha-1 antitrypsin, cystatin B, IgG kappa chain, apolipoprotein A1, and albumin were significantly different between CBD versus control group. When comparing CBD versus sarcoidosis, we found superoxide dismutase and hemoglobin upregulated in the CBD group. By using quantitative proteomics, we were able to find proteins with different expression levels in both diseases.
    Sarcoidosis, vasculitis, and diffuse lung diseases: official journal of WASOG / World Association of Sarcoidosis and Other Granulomatous Disorders 04/2007; 24(1):24-32. · 1.63 Impact Factor