A latex agglutination assay to quantify the amount of hemagglutinin protein in adjuvanted low-dose influenza monovalent vaccines.

To formulate inactivated influenza vaccines, the concentration of hemagglutinin (HA) must be accurately determined. The standard test currently used to measure HA in influenza vaccines is the Single Radial Immunodiffusion (SRID) assay. We developed a very rapid, simple and sensitive alternative quantitative HA assay, namely the Latex Agglutination Assay (LAA). The LAA uses the Spherotest® technology, which is based on the agglutination of HA-specific immunoglobulin-coated latex beads. The amount of HA in a sample is calculated from the level of bead agglutination by a simple absorbance measurement at 405nm against a standard curve generated using a monovalent vaccine standard. In less than 2hours, tens of samples could be quantified using the LAA as opposed to 2days for the SRID assay. Ten steps are required to complete an SRID assay as compared to 6 steps for the LAA, from sample preparation through spectrophotometric analysis. Furthermore, the limit of detection of the LAA was found to be approximately 15ng HA/mL, similar to an ELISA, with the quantification of less than 1.8μg HA/mL. The quantification limit of the SRID is usually considered to be approximately 5μg HA/mL. The development of the assay and a comparison of the titers obtained by SRID and LAA for several monovalent vaccines corresponding to various strains were performed. For A/H5N1 and A/H1N1 monovalent vaccines, the LAA was found to be linear and accurate as compared to the SRID. The precision of the LAA was close to that of the standard test, and good reproducibility from one laboratory to another was observed. Moreover, the LAA enabled HA quantification in AlOOH-adjuvanted and in emulsion-adjuvanted low-dose vaccines as well as unadjuvanted vaccines. In conclusion, LAA may be useful to rapidly and accurately measure influenza HA protein in monovalent vaccines, especially in those containing less than 5μg/mL of HA in the presence of an adjuvant

Demonstration of the First Prototype of RUGBI, Design and Deployment of a Grid for Bioinformatics

présenté par N. Jacq, proceedings publiés par "Studies in health technology and informatics" seriesInternational audienceRUGBI is an industrial and academic project to design and deploy on top of existing technologies a computing grid offering a set of grid and bioinformatics services to analyse proteins. It aims to support life sciences SMEs for computing and storage, to deploy an interregional grid for bioinformatics and to create a biologists community in a grid environment. The proposed demonstration presents the first prototype of RUGBI architecture and bioinformatics services

Increase of the fecundity in Hermann’s Tortoise Testudo hermanni hermanni in insular conditions: an opposite case of the insular syndrome?

Increase of the fecundity in Hermann 's Tortoise Testudo hermanni hermanni in insular conditions: an opposite case of the insular syndrome? - The reproductive parameters of the Hermann's Tortoise Testudo hermanni were studied by radiography in Corsica (Porto-Vecchio -island population-) and in Provence (Maures Massif -mainland population-). The following parameters were measured: number of eggs per clutch, clutch frequency, annual fecundity (eggs produced per female per year) and size of the eggs. There was a significantly higher fecundity in the island area, i.e. more eggs per clutch (4 against 3), a higher clutch frequency (1.9 against 1.4) and a global fecundity of 7.7 eggs/female/year as compared to 4.2 eggs/female/year in the mainland area. This increase in fecundity did not change also after taking into account the size of the females, which were larger in the island area. Indeed, female size does not influence the size of the eggs. The observed increase in fecundity contradicts the theory of the insular syndrome which predicts a decrease in fecundity in insular conditions. This dissension may be the result of the originality of tortoises at the ecophysiological level (long lifetime herbivore species) or may partially depend on distinct local adaptations.The reproductive parameters of the Hermann’s Tortoise Testudo hermanni were studied by radiography in Corsica (Porto-Vecchio –island population–) and in Provence (Maures Massif –mainland population–). The following parameters were measured: number of eggs per clutch, clutch frequency, annual fecundity (eggs produced per female per year) and size of the eggs. There was a signifi cantly higher fecundity in the island area, i.e. more eggs per clutch (4 against 3), a higher clutch frequency (1.9 against 1.4) and a global fecundity of 7.7 eggs/female/year as compared to 4.2 eggs/female/year in the mainland area. This increase in fecundity did not change also after taking into account the size of the females, which were larger in the island area. Indeed, female size does not infl uence the size of the eggs. The observed increase in fecundity contradicts the theory of the insular syndrome which predicts a decrease in fecundity in insular conditions. This dissension may be the result of the originality of tortoises at the ecophysiological level (long lifetime herbivore species) or may partially depend on distinct local adaptation

Safety, Humoral and Cell Mediated Immune Responses to Two Formulations of an Inactivated, Split-Virion Influenza A/H5N1 Vaccine in Children

BACKGROUND:Highly pathogenic influenza A/H5N1 has caused outbreaks in wild birds and poultry in Asia, Africa and Europe. It has also infected people, especially children, causing severe illness and death. Although the virus shows limited ability to transmit between humans, A/H5N1 represents a potential source of the next influenza pandemic. This study assesses the safety and immunogenicity of aluminium hydroxide adjuvanted (Al) and non adjuvanted influenza A/Vietnam/1194/2004 NIBRG-14 (H5N1) vaccine in children. METHODS AND FINDINGS:In a Phase II, open, randomised, multicentre trial 180 children aged 6 months to 17 years received two injections, 21 days apart, of vaccine containing either: 30 microg haemagglutinin (HA) with adjuvant (30 microg+Al) or 7.5 microg HA without adjuvant. An additional 60 children aged 6-35 months received two "half dose" injections (ie 15 microg+Al or 3.8 microg). Safety was followed for 21 days after vaccination. Antibody responses were assessed 21 days after each injection and cellular immune responses were explored. Vaccination appeared well tolerated in all age groups. The 30 microg+Al formulation was more immunogenic than 7.5 microg in all age groups: in these two groups 79% and 46% had haemagglutinination inhibition antibody titres > or =32 (1/dil). Among 6-35 month-olds, the full doses were more immunogenic than their half dose equivalents. Vaccination induced a predominantly Th2 response against H5 HA. CONCLUSIONS:This influenza A(H5N1) vaccine was well tolerated and immunogenic in children and infants, with Al adjuvant providing a clear immunogenic advantage. These results demonstrate that an H5N1 Al-adjuvanted vaccine, previously shown to be immunogenic and safe in adults, can also be used in children, the group most at risk for pandemic influenza

Standardization of cytokine flow cytometry assays

BACKGROUND: Cytokine flow cytometry (CFC) or intracellular cytokine staining (ICS) can quantitate antigen-specific T cell responses in settings such as experimental vaccination. Standardization of ICS among laboratories performing vaccine studies would provide a common platform by which to compare the immunogenicity of different vaccine candidates across multiple international organizations conducting clinical trials. As such, a study was carried out among several laboratories involved in HIV clinical trials, to define the inter-lab precision of ICS using various sample types, and using a common protocol for each experiment (see additional files online). RESULTS: Three sample types (activated, fixed, and frozen whole blood; fresh whole blood; and cryopreserved PBMC) were shipped to various sites, where ICS assays using cytomegalovirus (CMV) pp65 peptide mix or control antigens were performed in parallel in 96-well plates. For one experiment, antigens and antibody cocktails were lyophilised into 96-well plates to simplify and standardize the assay setup. Results (CD4(+)cytokine(+ )cells and CD8(+)cytokine(+ )cells) were determined by each site. Raw data were also sent to a central site for batch analysis with a dynamic gating template. Mean inter-laboratory coefficient of variation (C.V.) ranged from 17–44% depending upon the sample type and analysis method. Cryopreserved peripheral blood mononuclear cells (PBMC) yielded lower inter-lab C.V.'s than whole blood. Centralized analysis (using a dynamic gating template) reduced the inter-lab C.V. by 5–20%, depending upon the experiment. The inter-lab C.V. was lowest (18–24%) for samples with a mean of >0.5% IFNγ + T cells, and highest (57–82%) for samples with a mean of <0.1% IFNγ + cells. CONCLUSION: ICS assays can be performed by multiple laboratories using a common protocol with good inter-laboratory precision, which improves as the frequency of responding cells increases. Cryopreserved PBMC may yield slightly more consistent results than shipped whole blood. Analysis, particularly gating, is a significant source of variability, and can be reduced by centralized analysis and/or use of a standardized dynamic gating template. Use of pre-aliquoted lyophilized reagents for stimulation and staining can provide further standardization to these assays

Processus historique de dissociation de l'agriculture et de la forêt

From the XIXth century to nowadays, a long process of dissociation between forest and agriculture has taken place. In order to allow the economic and industrial expansion, it was necessary to favor the development of a commercial agriculture and, at the same time, to create long term lignous resources. It was thus necessary to set up the first, then the second "agricultural revolution", to restrict the forest use, and to give other production goals to forestry. Within one and a half century, the agricultural and forest administrations, knowledge, regulations, activities and areas have been progressively separated. / Du XIXe siècle à nos jours, s'est opéré un long processus de dissociation de la forêt et de l'agriculture. Pour permettre l'essor économique et industriel, il fallait à la fois favoriser le développement d'une agriculture marchande intensive, et créer à long terme des ressources ligneuses. Il a donc été nécessaire de réaliser la première, puis la deuxième "révolution agricole", de limiter les droits d'usage en forêt, et de donner d'autres objectifs de production à la sylviculture. En un siècle et demi, les administrations, les savoirs, les législations, les activités, et les espaces agricoles et forestiers ont été progressivement séparés

Bases Moléculaires de l’activité oncogénique de BCL2L10/Nrh dans le contexte du cancer du sein

Apoptosis, also called “Programmed Cell Death”, plays a key role in many biological processes and pathologies. The B-cell lymphoma 2 (Bcl-2) proteins, whose expression is often altered in tumor cells, are the main regulators of apoptosis.Among this family, the actual physiological function of the human apoptosis inhibitor Nrh, also referred to as BCL2L10 or Bcl-B, remains elusive. Although in most healthy tissues the Nrh protein is nearly undetectable, clinical studies have shown that Nrh expression is correlated with poor prognosis in breast and prostate carcinomas. We have shed light on a novel mechanism by which Nrz, the zebrafish ortholog of Nrh, was found to interact with the Ligand Binding Domain (LBD) of the Inositol-1,4,5-triphosphate receptor (IP3R) type-I Ca2+ channel. Indeed, the regulation of IP3Rs-mediated Ca2+ signaling by Nrz was shown to be critical during zebrafish embryogenesis. We used the knowledge gained with the zebrafish model to investigate Nrh function in cancer. We showed that Nrh interacts with the LBD of IP3Rs via its BH4 (Bcl-2 Homology 4) domain, which is critical to regulate intracellular Ca2+ trafficking and cell death. Actually, this interaction seems to be unique among the Bcl-2 family, and sets Nrh as the only Bcl-2 homolog to negatively regulate apoptosis by acting exclusively at the Endoplasmic Reticulum. Furthermore, we showed that disruption of the Nrh/IP3Rs complex primes Nrh-dependent cells to apoptotic cell death and enhances chemotherapy efficiency in breast cancer cell lines.Lastly our results bring a new insight to the role of Nrh regarding chemotherapy resistanceL'apoptose, ou « mort cellulaire programmée », joue un rôle clé dans de nombreux processus biologiques. Les protéines de la famille Bcl-2, dont l'expression est souvent altérée dans les cellules tumorales, sont les principaux régulateurs de l'apoptose. Parmi cette famille, la fonction exacte du répresseur apoptotique Nrh, aussi appelé BCL2L10 ou Bcl-B, reste à ce jour mal comprise. Bien que son expression ne soit pas détectable dans la plupart des tissus sains, on retrouve des niveaux élevés de Nrh corrélés à un mauvais pronostique dans les cancers du sein et de la prostate. Nous avons mis au jour un nouveau mécanisme selon lequel Nrz, l'orthologue de Nrh chez le poisson zèbre, interagit avec le domaine de liaison du ligand IP3 du canal calcique IP3R1. Il s'est avéré que la régulation négative des flux calciques par Nrz est critique lors du développement embryonnaire du poisson zèbre. Grâce à ces nouvelles données, nous avons cherché à comprendre la fonction de Nrh chez l'Homme, dans un contexte pathologique. Nous avons montré que Nrh interagit via son domaine BH4 avec le domaine de liaison du ligand du récepteur IP3R1 humain pour réguler l'homéostasie calcique et la mort cellulaire. Cette interaction définit Nrh comme la seule protéine de la famille Bcl-2 à réguler négativement la mort cellulaire exclusivement au niveau du réticulum endoplasmique. Pour aller plus loin, nous avons montré que la dissociation du complexe Nrh/IP3Rs sensibilise des cellules tumorales mammaires à l'action d'agents chimiothérapeutiques. Pour finir, nos résultats apportent une explication moléculaire sur la contribution de Nrh dans la résistance aux thérapies anti-tumorale

Le Christ, une figure subversive de l’autorité ? Quelques propos sur Le Gibet de Victor Hugo

Lorsque Hugo entreprend La Fin de Satan, il vient d’achever les Châtiments, le livre de l’immense colère. Il lui faut revenir à l’essentiel et renouer avec une écriture qui ne se contente pas de réagir, de fustiger, de dénoncer, si violemment que ce soit, mais qui se projette vers l’avenir. C’est ce qu’il va s’efforcer de faire en outrepassant sa propre colère pour atteindre l’essence même de ce qui a permis 1851, en l’inscrivant dans ce qui est de l’ordre de l’universel, dominant ainsi ce qu..