Clinical significance of fluoroscopic patterns specific for the mitotic spindle in patients with reumatic diseases

Objective: we proposed to determine the clinical significance of anti-NuMA and anti-HsEg5 antibodies in a group of patients affected with rheumatic diseases. Materials and methods: indirect immunofluorescence on HEp-2000 cells at serum dilution of 1:40 was used to examin 26 sera which had previously showed a "mitotic spindle" fluoroscopic pattern type during laboratory routine. Results: 21 sera (80,7%) were identified with NuMA and 5 (19,3%) with HsEg5 patterns alone or associated with other ANA patterns. However only patients with isolated positiveness and that is 15 with NuMA and 4 with HsEg5 stainings were included in this study. Of the NuMA positive patients 5 were affected with arthropathies associated to different forms of thyroiditis, 2 with seronegative arthritis, 2 with antiphospholipid syndrome, 1 with systemic lupus erythematosus (SLE), 1 with rheumatoid arthritis, 1 with sicca syndrome, 1 with undifferentiated connective tissue disease, 1 with Mycoplasma pneumaniae infection and 1 with retinal thrombosis. Of the HsEg5 positive patients 3 were affected with SLE and 1 with seronegative arthritis. Conclusions: NuMA does not prevail in any defined rheumatic disease, while HsEg5 staining were more frequent (75%) in patients affected with SLE all of whom showing high antibody titres

The Genopolis Microarray Database

<p>Abstract</p> <p>Background</p> <p>Gene expression databases are key resources for microarray data management and analysis and the importance of a proper annotation of their content is well understood.</p> <p>Public repositories as well as microarray database systems that can be implemented by single laboratories exist. However, there is not yet a tool that can easily support a collaborative environment where different users with different rights of access to data can interact to define a common highly coherent content. The scope of the Genopolis database is to provide a resource that allows different groups performing microarray experiments related to a common subject to create a common coherent knowledge base and to analyse it. The Genopolis database has been implemented as a dedicated system for the scientific community studying dendritic and macrophage cells functions and host-parasite interactions.</p> <p>Results</p> <p>The Genopolis Database system allows the community to build an object based MIAME compliant annotation of their experiments and to store images, raw and processed data from the Affymetrix GeneChip^®platform. It supports dynamical definition of controlled vocabularies and provides automated and supervised steps to control the coherence of data and annotations. It allows a precise control of the visibility of the database content to different sub groups in the community and facilitates exports of its content to public repositories. It provides an interactive users interface for data analysis: this allows users to visualize data matrices based on functional lists and sample characterization, and to navigate to other data matrices defined by similarity of expression values as well as functional characterizations of genes involved. A collaborative environment is also provided for the definition and sharing of functional annotation by users.</p> <p>Conclusion</p> <p>The Genopolis Database supports a community in building a common coherent knowledge base and analyse it. This fills a gap between a local database and a public repository, where the development of a common coherent annotation is important. In its current implementation, it provides a uniform coherently annotated dataset on dendritic cells and macrophage differentiation.</p

Perfluorocarbon Particle Size Influences Magnetic Resonance Signal and Immunological Properties of Dendritic Cells

The development of cellular tracking by fluorine (19F) magnetic resonance imaging (MRI) has introduced a number of advantages for following immune cell therapies in vivo. These include improved signal selectivity and a possibility to correlate cells labeled with fluorine-rich particles with conventional anatomic proton (1H) imaging. While the optimization of the cellular labeling method is clearly important, the impact of labeling on cellular dynamics should be kept in mind. We show by 19F MR spectroscopy (MRS) that the efficiency in labeling cells of the murine immune system (dendritic cells) by perfluoro-15-crown-5-ether (PFCE) particles increases with increasing particle size (560>365>245>130 nm). Dendritic cells (DC) are professional antigen presenting cells and with respect to impact of PFCE particles on DC function, we observed that markers of maturation for these cells (CD80, CD86) were also significantly elevated following labeling with larger PFCE particles (560 nm). When labeled with these larger particles that also gave an optimal signal in MRS, DC presented whole antigen more robustly to CD8+ T cells than control cells. Our data suggest that increasing particle size is one important feature for optimizing cell labeling by PFCE particles, but may also present possible pitfalls such as alteration of the immunological status of these cells. Therefore depending on the clinical scenario in which the 19F-labeled cellular vaccines will be applied (cancer, autoimmune disease, transplantation), it will be interesting to monitor the fate of these cells in vivo in the relevant preclinical mouse models

The role of dendritic cells in the immunopathogenesis of psoriasis

Psoriasis vulgaris is a chronic inflammatory skin disease that is marked by a complex interplay of dendritic cells (DCs), T-cells, cytokines, and downstream transcription factors as part of a self-sustaining type 1 cytokine network. As integral players of the immune system, DCs represent antigen-presenting cells that are crucial for efficient activation of T-cells and B-cells. DCs have also been linked to distinct chronic inflammatory conditions, including psoriasis. In the setting of psoriasis therapy, DC/T cell interactions serve as a potential target for biologic response modifiers. Here we describe the major DC subsets as well as the immunologic involvement of DCs within the context of psoriatic lesions

Regulatory T-cells are more resistant to genotoxic agents compared to effector T-cells

Background: Both host and graft regulatory T cells (Treg) may provide protection from graft-versus-host disease (GVHD) in the mouse system. Findings from a murine model of syngeneic GVHD suggest that host Treg preferentially survive a lethal dose of irradiation and confer protection from disease. In humans, various combinations of genotoxic agents are actually used for myeloablative conditioning. The sensitivity of human Treg to DNA damage induced apoptosis has so far not been investigated. Objective:To evaluate the resistance of Treg to apoptosis induced by genotoxic agents compared to effector T cells (Teff) in vitro. Materials and methods: PBMC of healthy donors were exposed to either ionizing irradiation or etoposide. After incubation, samples were labelled with annexin V FITC or anti-activated caspase 3 FITC combined with monoclonal antibody conjugates to CD4, CD25 and CD127 or to surface antigen CD4 and transcription factor FoxP3. Propidium iodide or fixable infrared viability dye were included in the staining protocol as dead cell markers. The frequency of apoptotic cells among Treg and effector CD4+ T lymphocytes was determined by flow cytometric analysis at different time points (up to 72h). CD45RA staining was used to analyse the role of differentiation for resistance to apoptosis. Expression of anti-apoptotic proteins bcl-2 and Bcl-xL by Treg and Teff was compared. Results: Both activation of caspase 3 and exposure of phosphatidylserine following DNA damage in vitro was observed with significantly higher frequency in Treg compared to effector CD4 lymphocytes. This effect was observed in both etoposide treated cultures and those exposed to ionizing irradiation. Similar results were obtained by identifying Treg as CD25highCD127- or as FoxP3+ events. The resistance to DNA damage by Treg could not be explained by their differentiation to a CD45RA negative memory phenotype. When measured as mean fluorescence intensity, anti-apoptotic proteins Bcl-2 and bcl-xL were not more expressed by Treg compared to Teff. Conclusion: Our observations suggest a higher resistance to DNA damage of regulatory T cells compared to effector T cells in vitro. The higher resistance cannot be explained by differentiation to a memory phenotype or differential expression of anti-apopotic proteins bcl-2 and bcl-xL. The higher resistance to genotoxic agents of Treg could affect the relative size ratio of Treg to effector T cells following conditioning