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Cabal A.,Hopsital Universitario Miguel Servet | Strunk M.,CIBER ISCIII | Dominguez J.,Institute dInvestigacio Germans Trias i Pujol | Dominguez J.,Autonomous University of Barcelona | And 14 more authors.
BMC Microbiology | Year: 2014

Background: Different polymorphisms have been described as markers to classify the lineages of the Mycobacterium tuberculosis complex. The analysis of nine single nucleotide polymorphisms (SNPs) was used to describe seven SNPs cluster groups (SCGs). We attempted to classify those strains that could not been categorized into lineages by the genotyping methods used in the routine testing. Results: The M. tuberculosis complex isolates collected in 2010 in our region were analysed. A new method based on multiplex-PCRs and pyrosequencing to analyse these SNPs was designed. For the pyrosequencing assay nine SNPs that defined the seven SCGs were selected from the literature: 1977, 74092, 105139, 232574, 311613, 913274, 2460626, 3352929 and gyrA95. In addition, SNPs in katG463,mgtC182,Ag85C103 and RDRio deletion were detected. Conclusions: This work has permitted to achieve a better classification of Aragonian strains into SCGs and in some cases, to assign strains to its certain lineage. Besides, the description of a new pattern shared by two isolates "SCG-6c" reinforces the interest of SNPs to follow the evolution of M. tuberculosis complex. © 2014 Cabal et al.; licensee BioMed Central Ltd. Source


Karachaliou N.,Quiron Dexeus Hospital Universitario | Rosell R.,Catalan Institute of Nanoscience and Nanotechnology | Rosell R.,Molecular Oncology Research MORe Foundation | Morales-Espinosa D.,Institute dInvestigacio Germans Trias i Pujol | Viteri S.,Quiron Dexeus Hospital Universitario
Expert Review of Anticancer Therapy | Year: 2014

Over a short period of time, translational research has described a new clinically relevant molecular subset of non-small-cell lung cancer (NSCLC) that is defined by EGFR mutations or EML4-ALK fusions. Today, patients with metastatic disease can achieve survival rates at least double that of patients with wild-type tumors. Through the rational dissection of the mechanisms of drug sensitivity and resistance, promising strategies have been defined to further improve the outcomes of patients with NCSLC. This review adds to a growing body of knowledge into mechanisms of resistance that can be interrogated in NSCLC patients with EGFR mutations or EML4-ALK fusions, as well as strategies to overcome resistance to TKIs. © 2014 Informa UK, Ltd. Source


Perez-Cabezas B.,Autonomous University of Barcelona | Naranjo-Gomez M.,Autonomous University of Barcelona | Ruiz-Riol M.,Autonomous University of Barcelona | Bastos-Amador P.,Autonomous University of Barcelona | And 5 more authors.
Journal of Leukocyte Biology | Year: 2012

Cooperative events between DC subsets involve cell contact and soluble factors. Upon viral challenge, murine pDCs induce cDC cooperation through CD40-CD40L interactions and IL-15 secretion, whereas in humans, the same effect is mediated by IFN-α. Conversely, during bacterial infections, pDC maturation may be induced by activated cDCs, although no mechanisms had been described so far. Here, we investigate how human pDCs are "conditioned" by cDCs. Blood-borne DC subsets (cDCs and pDCs) were sorted from healthy donors. IL-3-maintained pDCs were cocul-tured with LPS-activated, poly (I:C)-activated, or control cDCs [cDCLPS, cDCP(I:C), cDCCTRL]. Coculture experiments showed that cDCLPS-conditioned pDCs up-regulated maturation markers, such as CD25 and CD86, whereas SNs contained higher amounts of IL-6 and CCL19 compared with control conditions. Gene-expression analyses on sorted cDCLPS or cDCP(I:C) conditioned pDCs confirmed the induction of several genes, including IL-6 and CCL19 and remarkably, several Notch target genes. Further studies using the γ-secretase/Notch inhibitor DAPT and soluble Notch ligands resulted in a significantly reduced expression of canonical Notch target genes in conditioned pDCs. DAPT treatment also hampered the secretion of CCL19 (but not of IL-6) by cDCLPS conditioned pDCs. These results reveal the involvement of γ-secretase-mediated mechanisms, including the Notch pathway, in the cell contact-dependent communication between human DC subsets. The resulting partial activation of pDCs after encountering with mature cDCs endows pDCs with an accessory function that may contribute to T cell recruitment and activation. © Society for Leukocyte Biology. Source


Nogales-Gadea G.,Maastricht University | Nogales-Gadea G.,Institute dInvestigacio Germans Trias i Pujol | Saxena A.,Maastricht University | Hoffmann C.,Maastricht University | And 5 more authors.
Journal of Visualized Experiments | Year: 2015

Finding new methods for generating human monoclonal antibodies is an active research field that is important for both basic and applied sciences, including the development of immunotherapeutics. However, the techniques to identify and produce such antibodies tend to be arduous and sometimes the heavy and light chain pair of the antibodies are dissociated. Here, we describe a relatively simple, straightforward protocol to produce human recombinant monoclonal antibodies from human peripheral blood mononuclear cells using immortalization with Epstein-Barr Virus (EBV) and Toll-like receptor 9 activation. With an adequate staining, B cells producing antibodies can be isolated for subsequent immortalization and clonal expansion. The antibody transcripts produced by the immortalized B cell clones can be amplified by PCR, sequenced as corresponding heavy and light chain pairs and cloned into immunoglobulin expression vectors. The antibodies obtained with this technique can be powerful tools to study relevant human immune responses, including autoimmunity, and create the basis for new therapeutics. © 2015 Journal of Visualized Experiments. Source


De Souza-Galvao M.L.,Hospital Universitari Vall dHebron | De Souza-Galvao M.L.,IDIAP Jordi Gol Research Foundation | De Souza-Galvao M.L.,University of Barcelona | Latorre I.,University of Barcelona | And 15 more authors.
BMC Infectious Diseases | Year: 2014

Background: The aim of the study was to assess the correlation between the tuberculin skin test (TST) and in vitro interferon-gamma released assays (IGRAs) with risk factors for the spread of infection in smear positive pulmonary tuberculosis (TB) contacts.Methods: We recruited prospective contacts with smear positive pulmonary TB cases. We looked at human immunodeficiency virus (HIV) infection and other conditions of immunosuppression, presence of BCG vaccination and the degree of exposure to the index case. Patients underwent the TST, chest radiography, sputum analysis when necessary, and IGRA assays (QFN-G-IT and T-SPOT.TB). Presence of cough, diagnostic delay (days between first symptoms and TB diagnostic), contact conditions: room size (square meters) and index of overcrowding (square meters per person) were investigated in the index case.Results: 156 contacts (119 adults, 37 children) of 66 TB patients were enrolled, 2.4 (1-14) contacts per TB case. The positivity of the TST did not correlate with the risk factors studied: presence of cough (p = 0.929); delayed diagnosis (p = 0.244); room size (p = 0.462); overcrowding (p = 0.800). Both QFN-G-IT and T-SPOT.TB, showed significant association with cough (p = 0.001, and p = 0.007) and room size (p = 0.020, and p = 0.023), respectively.Conclusions: Both IGRA associated better than TST with certain host-related risk factors involved in the transmission of disease, such as the presence of cough. © 2014 de Souza-Galvão et al.; licensee BioMed Central Ltd. Source

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