ViralORFeome: an integrated database to generate a versatile collection of viral ORFs

Large collections of protein-encoding open reading frames (ORFs) established in a versatile recombination-based cloning system have been instrumental to study protein functions in high-throughput assays. Such ‘ORFeome’ resources have been developed for several organisms but in virology, plasmid collections covering a significant fraction of the virosphere are still needed. In this perspective, we present ViralORFeome 1.0 (http://www.viralorfeome.com), an open-access database and management system that provides an integrated set of bioinformatic tools to clone viral ORFs in the Gateway® system. ViralORFeome provides a convenient interface to navigate through virus genome sequences, to design ORF-specific cloning primers, to validate the sequence of generated constructs and to browse established collections of virus ORFs. Most importantly, ViralORFeome has been designed to manage all possible variants or mutants of a given ORF so that the cloning procedure can be applied to any emerging virus strain. A subset of plasmid constructs generated with ViralORFeome platform has been tested with success for heterologous protein expression in different expression systems at proteome scale. ViralORFeome should provide our community with a framework to establish a large collection of virus ORF clones, an instrumental resource to determine functions, activities and binding partners of viral proteins

Simian virus 40 inhibits differentiation and maturation of rhesus macaque DC-SIGN+-dendritic cells

Dendritic cells (DC) are the initiators and modulators of the immune responses. Some species of pathogenic microorganisms have developed immune evasion strategies by controlling antigen presentation function of DC. Simian virus 40 (SV40) is a DNA tumor virus of rhesus monkey origin. It can induce cell transformation and tumorigenesis in many vertebrate species, but often causes no visible effects and persists as a latent infection in rhesus monkeys under natural conditions. To investigate the interaction between SV40 and rhesus monkey DC, rhesus monkey peripheral blood monocyte-derived DC were induced using recombinant human Interleukin-4 (rhIL-4) and infective SV40, the phenotype and function of DC-specific intracellular adhesion molecule-3 grabbing nonintegrin (DC-SIGN)+ DC were analyzed by flow cytometry (FCM) and mixed lymphocyte reaction (MLR). Results showed that SV40 can down-regulate the expression of CD83 and CD86 on DC and impair DC-induced activation of T cell proliferation. These findings suggest that SV40 might also cause immune suppression by influencing differentiation and maturation of DC

Th1 Disabled Function in Response to TLR4 Stimulation of Monocyte-Derived DC from Patients Chronically-Infected by Hepatitis C Virus

Background: Lack of protective antibodies and inefficient cytotoxic responses are characteristics of chronic hepatitis C infection. A defect in dendritic cell (DC) function has thus been suspected, but this remains a controversial issue. Methods and Findings: Here we show that monocyte-derived DC (MoDC) from chronically-infected patients can mature in response to TLR1/2, TLR2/6 or TLR3 ligands. In contrast, when stimulated with the TLR4 ligand LPS, MoDC from patients show a profound defect in inducing IFNc secretion by allogeneic T cells. This defect is not due to defective phenotypic maturation or to the presence of HCV-RNA in DC or monocytes but is correlated to reduced IL-12 secretion by DC. Restoration of DC ability to stimulate IFNc secretion can be obtained by blocking MEK activation in DC, indicating that MEK/ ERK pathway is involved in the Th1 defect of MoDC. Monocytes from HCV patients present increased spontaneous secretion of cytokines and chemokines, especially MIP-1b. Addition of MIP-1b on healthy monocytes during differentiation results in DC that have Th1 defect characteristic of MoDC from HCV patients, suggesting that MIP-1b secretion by HCV monocytes participates in the Th1 defect of DC. Conclusions: Our data indicate that monocytes from HCV patients are activated in vivo. This interferes with their differentiation into DC, leading to deficient TLR4 signaling in these cells that are enable to induce a Th1 response. Thi

T-cell recognition of chemicals, protein allergens and drugs: towards the development of in vitro assays

Chemicals can elicit T-cell-mediated diseases such as allergic contact dermatitis and adverse drug reactions. Therefore, testing of chemicals, drugs and protein allergens for hazard identification and risk assessment is essential in regulatory toxicology. The seventh amendment of the EU Cosmetics Directive now prohibits the testing of cosmetic ingredients in mice, guinea pigs and other animal species to assess their sensitizing potential. In addition, the EU Chemicals Directive REACh requires the retesting of more than 30,000 chemicals for different toxicological endpoints, including sensitization, requiring vast numbers of animals. Therefore, alternative methods are urgently needed to eventually replace animal testing. Here, we summarize the outcome of an expert meeting in Rome on 7 November 2009 on the development of T-cell-based in vitro assays as tools in immunotoxicology to identify hazardous chemicals and drugs. In addition, we provide an overview of the development of the field over the last two decades

Perovskite foams used in combination with dense ceramic membranes for oxygen transport membrane applications

International audienceThe future industrial implementation of membranes for oxygen transport requires new designs to increase oxygen semi-permeation, together with robust performance under operating conditions. This work describes the development of innovative membrane designs based on perovskite foams with a very large porosity (above 90%) supporting thin dense perovskite membranes (thickness of 50–100 µm). The preparation of such foam-supported membranes is described in this paper. The performances in terms of oxygen semi-permeation are also measured and compared with the best results reported in the literature up to date

The surface roughness effect on electrochemical properties of La0.5Sr0.5Fe0.7Ga0.3O3-δ perovskite for oxygen transport membranes

International audienc

Dilute Solution Structure of Bottlebrush Polymers

Bottlebrush polymers are a class of macromolecules that has recently found usein a wide variety of materials, ranging from lubricating brushes andnanostructured coatings to elastomeric gels that exhibit structural color. Thesepolymers are characterized by dense branches extending from a central backbone,and thus have properties distinct from linear polymers. It remains a challengeto specifically understand conformational properties of these molecules, due tothe wide range of architectural parameters that can be present in a system, andthus there is a need to accurately characterize and model these molecules. Inthis paper, we use a combination of viscometry, light scattering, and computersimulations to gain insight into the conformational properties of dilutebottlebrush polymers. We focus on a series of model bottlebrushes consisting ofa poly(norbornene) (PNB) backbone with poly(lactic acid) (PLA) side chains. Wedemonstrate that intrinsic viscosity and hydrodynamic radius are experimentalobservations \emph{sensitive} to molecular architecture, exhibiting distinctdifferences with different choices of branch and backbone lengths. Informed bythe atomistic structure of this PNB-PLA system, we rationalize a coarse-grainedsimulation model that we evaluate using a combination of Brownian Dynamics andMonte Carlo simulations. We show that this exhibits quantitative matching toexperimental results, enabling us to characterize the overall shape of thebottlebrush via a number of metrics that can be extended to more generalbottlebrush architectures.</div