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Publications (12)

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    Ulrika Rolén · Elio Freda · Jianjun Xie · [...] · Maria G Masucci
    [Show abstract] [Hide abstract] ABSTRACT: The ubiquitin C-terminal hydrolase-L1 (UCH-L1) is a deubiquitinating enzyme that catalyses the hydrolysis of polyubiquitin precursors and small ubiquitin adducts. UCH-L1 has been detected in a variety of malignant and metastatic tumours but its biological function in these cells is unknown. We have previously shown that UCH-L1 is highly expressed in Burkitt's lymphoma (BL) and is up-regulated upon infection of B lymphocytes with Epstein-Barr virus (EBV). Here we show that knockdown of UCH-L1 by RNAi inhibits the proliferation of BL cells in suspension and semisolid agar and activates strong LFA-1-dependent homotypic adhesion. Induction of cell adhesion correlated with cation-induced binding to ICAM-1, clustering of LFA-1 into lipid rafts and constitutive activation of the Rap1 and Rac1 GTPases. Expression of a catalytically active UCH-L1 promoted the proliferation of a UCH-L1-negative EBV transformed lymphoblastoid cell line (LCL) and inhibited cell adhesion, whereas a catalytic mutant had no effect, confirming the requirement of UCH-L1 enzymatic activity for the regulation of these phenotypes. Our results identify UCH-L1 as a new player in the signalling pathways that promote the proliferation and invasive capacity of malignant B cells.
    Full-text Article · Aug 2009 · Journal of Cellular and Molecular Medicine
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    Dataset: Table S4
    [Show abstract] [Hide abstract] ABSTRACT: (0.03 MB DOC)
    Full-text Dataset · Nov 2007
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    Dataset: Table S3
    [Show abstract] [Hide abstract] ABSTRACT: (3.18 MB DOC)
    Full-text Dataset · Nov 2007
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    Dataset: Figure S1
    [Show abstract] [Hide abstract] ABSTRACT: Characteristics of NPC primary culture cells. Morphology of NPC primary culture cells under light microscope (A and B). Keratin staining (C). EBERs in situ hybridization in NPC primary cells (C). Negative control for EBERs staining in normal nasopharyngeal epithelia (D) (3.45 MB DOC)
    Full-text Dataset · Nov 2007
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    Dataset: Figure S2
    [Show abstract] [Hide abstract] ABSTRACT: The frequency of T cells specific for HLA-A2 restriction epitopes in EBV LMP1 and LMP2 in HLA-A2 positive NPC patient and healthy donor. The frequencies of tetramer positive cells in CD3+CD8high PBMCs and TILs from NPC patient P24, and in PBMCs and auto-LCL stimulated PBMCs from healthy donor N16 are shown in this figure. (3.27 MB TIF)
    Full-text Dataset · Nov 2007
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    Dataset: Table S1
    [Show abstract] [Hide abstract] ABSTRACT: (0.06 MB DOC)
    Full-text Dataset · Nov 2007
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    Dataset: Table S2
    [Show abstract] [Hide abstract] ABSTRACT: (0.08 MB DOC)
    Full-text Dataset · Nov 2007
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    Dataset: Figure S3
    [Show abstract] [Hide abstract] ABSTRACT: Cytotoxicity analysis of auto-LCL stimulated PBMCs from NPC patient 1. Polyclonal CTL cultures were tested for cytotoxic activity against autologous PHA blasts loaded with LMP1 or LMP2 peptides in 4 hrs 51Cr release assays(A); or stimulated with LMP1 or LMP2 peptides for 4 hrs in round 96-well tissue culture plate followed by intracellular staining for CD107a and IFN-γ(B). (3.18 MB TIF)
    Full-text Dataset · Nov 2007
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    Dataset: Figure S4
    [Show abstract] [Hide abstract] ABSTRACT: Cytotoxicity analysis of TILs from NPC patient 31. Tumor infiltrating lymphocyte (P31) expanded for 1 to 4 weeks in IL-2 medium without antigen stimulation, were tested for cytotoxic activity against autologous PHA blasts loaded with LMP1 or LMP2 peptides in 4 hrs 51Cr release assays (A); or co-cultured with different targets cells (E∶T = 10∶1) for 4 hrs in round 96-well tissue culture plate followed by intracellular staining for CD107a and IFN-γ(B). (3.22 MB TIF)
    Full-text Dataset · Nov 2007
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    [Show abstract] [Hide abstract] ABSTRACT: Nasopharyngeal carcinoma (NPC) is an Epstein-Barr virus (EBV) associated malignancy with high prevalence in Southern Chinese. In order to assess whether defects of EBV-specific immunity may contribute to the tumor, the phenotype and function of circulating T-cells and tumor infiltrating lymphocytes (TILs) were investigated in untreated NPC patients. Circulating naïve CD3+CD45RA+ and CD4+CD25- cells were decreased, while activated CD4+CD25+ T-cells and CD3-CD16+ NK-cells were increased in patients compared to healthy donors. The frequency of T-cells recognizing seven HLA-A2 restricted epitopes in LMP1 and LMP2 was lower in the patients and remained low after stimulation with autologous EBV-carrying cells. TILs expanded in low doses of IL-2 exhibited an increase of CD3+CD4+, CD3+CD45RO+ and CD4+CD25+ cells and 2 to 5 fold higher frequency of LMP1 and LMP2 tetramer positive cells compared to peripheral blood. EBV-specific cytotoxicity could be reactivated from the blood of most patients, whereas the TILs lacked cytotoxic activity and failed to produce IFNgamma upon specific stimulation. Thus, EBV-specific rejection responses appear to be functionally inactivated at the tumor site in NPC.
    Full-text Article · Feb 2007 · PLoS ONE
  • Ulrika Rolén · Vera Kobzeva · Natalja Gasparjan · [...] · Maria G Masucci
    [Show abstract] [Hide abstract] ABSTRACT: Ubiquitin specific proteases (USPs) regulate the production and recycling of ubiquitin and are thereby critically involved in the control of cell growth, differentiation, and apoptosis. Increasing evidence implicates deregulation of USPs in malignant transformation but there is very little information on the overall and specific activity of USPs in normal and tumor tissues. We have used a chemistry-based functional proteomics approach to profile the activities of individual USPs in biopsies of human papillomavirus (HPV) carrying cervical carcinoma and adjacent normal tissue. To assess the contribution of HPV proteins, USP activity was also compared in HPV positive and negative cervical carcinoma cell lines and HPV E6/E7 immortalized human keratinocytes. The activity of the C-terminal hydrolases UCH-L3 and UCH37 was upregulated in the majority of tumor tissues compared to the adjacent normal tissues. UCH-L1 activity was lower in a significant proportion of the tumors but to a less extent in advanced tumors. In accordance with the relatively low UCH-L1 activity in tumor biopsies, UCH-L1 was detected only in one out of eight cervical carcinoma lines. UCH-L1, UCH-L3, USP7, and USP9X activity was upregulated following HPV E6/E7 immortalization of keratinocytes, suggesting a role of these enzymes in growth transformation.
    Article · Apr 2006 · Molecular Carcinogenesis
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    Huib Ovaa · Benedikt M Kessler · Ulrika Rolén · [...] · Maria G Masucci
    [Show abstract] [Hide abstract] ABSTRACT: The family of ubiquitin (Ub)-specific proteases (USP) removes Ub from Ub conjugates and regulates a variety of cellular processes. The human genome contains many putative USP-encoding genes, but little is known about USP tissue distribution, pattern of expression, activity, and substrate specificity. We have used a chemistry-based functional proteomics approach to identify active USPs in normal, virus-infected, and tumor-derived human cells. Depending on tissue origin and stage of activation/differentiation, different USP activity profiles were revealed. The activity of specific USPs, including USP5, -7, -9, -13, -15, and -22, was up-regulated by mitogen activation or virus infection in normal T and B lymphocytes. UCH-L1 was highly expressed in tumor cell lines of epithelial and hematopoietic cell origin but was not detected in freshly isolated and mitogen-activated cells. Up-regulation of this USP was a late event in the establishment of Epstein-Barr virus-immortalized lymphoblastoid cell lines and correlated with enhanced proliferation, suggesting a possible role in growth transformation.
    Full-text Article · Mar 2004 · Proceedings of the National Academy of Sciences