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86 results on '"Benoît Polack"'

1. P1672: OCCUPATIONAL INTEGRATION OF ADULTS WITH SEVERE HAEMOPHILIA (INTHEMO): A STUDY BASED ON THE FRANCECOAG REGISTRY

Author: Ngoc Anh Thu Nguyen, Pascal Auquier, Any Beltran Anzola, Roseline D’oiron, Thierry Lambert, Céline Falaise, Christine Biron, Anne Lienhart, Jenny Goudemand, Antoine Rauch, Bénédicte Wibaut, Sabine Marie Castet, Dominique Desprez, Annie Harroche, Natalie Stieltjes, Annie Borel-Derlon, Marc Trossaërt, Amandine Celli, Benoît Guillet, Sophie Bayart, Marie-Anne Bertrand, Alexandra Fournel, Guillaume Mourey, Valérie Gay, Pierre Chamouni, Emmanuelle DE Raucourt, Birgit Frotscher, Michèle Martin, Mathieu Puyade, Brigitte Tardy, Stéphane Vanderbecken, Fabienne Genre-Volot, Philippe Nguyen, Benoît Polack, Caroline Oudot-Challard, Stéphane Girault, Annelise Voyer, Aurélien Lebreton, Abel Hassoun, Philippe Moreau, Pierre-Simon Rohrlich, Karine Baumstarck, Mohamed Boucekine, Vanessa Milien, Natacha Rosso, Clemence Tabele, Marie Viprey, Nicolas Giraud, Thomas Sannie, Hervé Chambost, and Noémie Resseguier
Subjects: Diseases of the blood and blood-forming organs, RC633-647.5
Published: 2023
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2. Fibrinography: A Multiwavelength Light-Scattering Assay of Fibrin Structure

Author: Carhel Dassi, Landry Seyve, Xabel García, Emmanuelle Bigo, Raphaël Marlu, François Caton, and Benoît Polack
Subjects: Diseases of the blood and blood-forming organs, RC633-647.5
Abstract: Abstract. We have previously developed a fibrin structural assay dedicated to purified fibrinogen-thrombin system. Here, we extend the pertinence of this test, called Fibrinography, to tissue factor-triggered plasma coagulation. We show that Fibrinography determines quantitatively the structure of fibrin fibers in plasma with an excellent reproducibility. We compare this assay with the commonly used single wavelength turbidity method, showing that the latter is not a proper structural assay, but determines essentially the fibrinogen content in plasma. In addition, we also show, in model plasmas, that Fibrinography is able to discriminate normal and hypocoagulant plasmas, and even between hypercoagulant plasmas. Therefore, Fibrinography, by measuring the final step of the coagulation cascade, may be used to evaluate patients’ plasma in hypo- or hypercoagulant diseases.
Published: 2019
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3. Catalytically inactive Gla-domainless factor Xa binds to TFPI and restores ex vivo coagulation in hemophilia plasma

Author: Atanur Ersayin, Aline Thomas, Landry Seyve, Nicole Thielens, Mathieu Castellan, Raphaël Marlu, Benoît Polack, and Marie-Claire Dagher
Subjects: Diseases of the blood and blood-forming organs, RC633-647.5
Published: 2017
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4. Combined sacB-based negative selection and cre-lox antibiotic marker recycling for efficient gene deletion in Pseudomonas aeruginosa

Author: Lauriane Quénée, Danièle Lamotte, and Benoît Polack
Subjects: Biology (General), QH301-705.5
Abstract: The complete genome of the bacterial pathogen Pseudomonas aeruginosa has now been sequenced, allowing gene deletion, one of the most frequently used methods in gene function study, to be fully exploited. In this study, we combine the sacB-based negative selection system with a cre-lox antibiotic marker recycling method. This methodology allows allelic exchange between a target gene and a gentamicin cassette flanked by the two lox sequences. A tetracycline plasmid expressing the cre recombinase is then introduced in the mutant strain to catalyze the excision of the lox-flanked resistance marker. We demonstrate here the efficiency of the combination of these two methods in P. aeruginosa by successively deleting ExoS and ExoT, which are two genetically independent toxins of the type-three secretion system (TTSS). This functional cre-lox recycling antibiotic marker system can create P. aeruginosa strains with multiple mutations without modifying the antibiotic resistance profile when compared to the parental strain.
Published: 2005
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5. Gla-domainless factor Xa: molecular bait to bypass a blocked tenase complex

Author: Raphaël Marlu and Benoît Polack
Subjects: Diseases of the blood and blood-forming organs, RC633-647.5
Abstract: Background Hemophilia is caused by deficiencies in coagulation factor VIII or IX, resulting in direct blockade of the intrinsic tenase complex and indirect blockade of the extrinsic tenase complex which is rapidly inhibited upon binding of factor Xa to tissue factor pathway inhibitor. We evaluated the ability of Gla-domainless factor Xa, a truncated form of factor Xa devoid of procoagulant properties, to bind to tissue factor pathway inhibitor and to alleviate the physiological inhibition of the extrinsic tenase.Design and Methods Using a thrombin generation assay triggered by a low concentration of tissue factor, we evaluated the ability of Gla-domainless factor Xa to restore blood coagulation in plasma from hemophilia A and B patients without and with inhibitors. We then compared its efficacy to generate thrombin to depletion of antithrombin or tissue factor pathway inhibitor by specific antibodies. Finally, we compared the kinetics of neutralization of factor Xa and Gla-domainless factor Xa by antithrombin and tissue factor pathway inhibitor.Results Gla-domainless factor Xa was able to restore thrombin generation in plasma samples from hemophiliacs. This effect was observed for plasma from hemophilia A patients without or with inhibitors and for plasma from hemophilia B patients. Gla-domainless factor Xa had a lower affinity than factor Xa for tissue factor pathway inhibitor whereas the affinities of both proteins for antithrombin were similar. Finally, despite a short half-life in plasma, the effect of Gla-domainless factor Xa on thrombin generation was sustained for at least 1 hour.Conclusions As Gla-domainless factor Xa was able to restore thrombin generation in plasma from hemophilia patients, our results suggest that it may be an effective alternative to current treatments for hemophilia with or without an inhibitor.
Published: 2012
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6. Molecular interface of S100A8 with cytochrome b558 and NADPH oxidase activation.

Author: Sylvie Berthier, Minh Vu Chuong Nguyen, Athan Baillet, Marc-André Hograindleur, Marie-Hélène Paclet, Benoît Polack, and Françoise Morel
Subjects: Medicine, Science
Abstract: S100A8 and S100A9 are two calcium binding Myeloid Related Proteins, and important mediators of inflammatory diseases. They were recently introduced as partners for phagocyte NADPH oxidase regulation. However, the precise mechanism of their interaction remains elusive. We had for aim (i) to evaluate the impact of S100 proteins on NADPH oxidase activity; (ii) to characterize molecular interaction of either S100A8, S100A9, or S100A8/S100A9 heterocomplex with cytochrome b(558); and (iii) to determine the S100A8 consensus site involved in cytochrome b(558)/S100 interface. Recombinant full length or S100A9-A8 truncated chimera proteins and ExoS-S100 fusion proteins were expressed in E. coli and in P. aeruginosa respectively. Our results showed that S100A8 is the functional partner for NADPH oxidase activation contrary to S100A9, however, the loading with calcium and a combination with phosphorylated S100A9 are essential in vivo. Endogenous S100A9 and S100A8 colocalize in differentiated and PMA stimulated PLB985 cells, with Nox2/gp91(phox) and p22(phox). Recombinant S100A8, loaded with calcium and fused with the first 129 or 54 N-terminal amino acid residues of the P. aeruginosa ExoS toxin, induced a similar oxidase activation in vitro, to the one observed with S100A8 in the presence of S100A9 in vivo. This suggests that S100A8 is the essential component of the S100A9/S100A8 heterocomplex for oxidase activation. In this context, recombinant full-length rS100A9-A8 and rS100A9-A8 truncated 90 chimera proteins as opposed to rS100A9-A8 truncated 86 and rS100A9-A8 truncated 57 chimeras, activate the NADPH oxidase function of purified cytochrome b(558) suggesting that the C-terminal region of S100A8 is directly involved in the molecular interface with the hemoprotein. The data point to four strategic (87)HEES(90) amino acid residues of the S100A8 C-terminal sequence that are involved directly in the molecular interaction with cytochrome b(558) and then in the phagocyte NADPH oxidase activation.
Published: 2012
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7. Rationally designed Gla-domainless FXa as TFPI bait in hemophilia

Author: Marie-Claire Dagher, Atanur Ersayin, Landry Seyve, Mathieu Castellan, Cyril Moreau, Luc Choisnard, Nicole Thielens, Raphaël Marlu, Benoît Polack, Aline Thomas, Institut de biologie structurale (IBS - UMR 5075), Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA), Translational Innovation in Medicine and Complexity / Recherche Translationnelle et Innovation en Médecine et Complexité - UMR 5525 (TIMC ), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP ), Université Grenoble Alpes (UGA), Département de pharmacochimie moléculaire (DPM), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), ISBG, ANR-13-RPIB-0011,MINITEN,Le GD-Xa comme nouveau traitement anti-hémorragique(2013), and European Project: IRS-ARCANE
Subjects: [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM]
Abstract: Gla-domainless factor Xa (GD-FXa) was proposed as a trap to the endogenous anticoagulant Tissue Factor Pathway Inhibitor (TFPI) to restore thrombin generation in hemophilia. Using computational chemistry and experimental approaches, we previously showed that S195A GD-FXa also binds TFPI and restores ex vivo coagulation in hemophilia plasmas.To design a GD-FXa variant with improved anti-TFPI activity and identify suitable sites for mutagenesis, we performed molecular dynamics simulations. The calculations identified residues R150FXa and K96FXa as cold-spots of interaction between GD-FXa and the K2 domain of TFPI. In the three-dimensional model, both residues are facing TFPI hydrophobic residues and are thus potential candidates for mutagenesis into hydrophobic residues to favor an improved protein-protein interaction.Catalytically inactive GD-FXa variants containing the S195A mutation and additional mutations as K96Y, R150I, R150G and R150F were produced to experimentally confirm these computational hypotheses. Among these mutants, the R150FFXA showed increased affinity for TFPI as theoretically predicted, and was also more effective than S195A GD-FXa in restoring coagulation in FVIII deficient plasmas. Moreover, the R150 mutants lost interaction with antithrombin, which is favorable to extend their half-life.
Published: 2022
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8. Are coagulation profiles in Andean highlanders with excessive erythrocytosis favouring hypercoagulability?

Author: Benoit Champigneulle, François Caton, Landry Seyve, Émeric Stauffer, Aurélien Pichon, Julien V. Brugniaux, Michael Furian, Ivan Hancco, Blandine Deschamps, Lars Kaestner, Paul Robach, Philippe Connes, Pierre Bouzat, Benoit Polack, Raphael Marlu, and Samuel Verges
Subjects: blood coagulation, chronic mountain sickness, excessive erythrocytosis, hypoxia, thromboelastometry, Physiology, QP1-981
Abstract: Abstract Chronic mountain sickness is a maladaptive syndrome that affects individuals living permanently at high altitude and is characterized primarily by excessive erythrocytosis (EE). Recent results concerning the impact of EE in Andean highlanders on clotting and the possible promotion of hypercoagulability, which can lead to thrombosis, were contradictory. We assessed the coagulation profiles of Andeans highlanders with and without excessive erythrocytosis (EE+ and EE−). Blood samples were collected from 30 EE+ and 15 EE− in La Rinconada (Peru, 5100–5300 m a.s.l.), with special attention given to the sampling pre‐analytical variables. Rotational thromboelastometry tests were performed at both native and normalized (40%) haematocrit using autologous platelet‐poor plasma. Thrombin generation, dosages of clotting factors and inhibitors were measured in plasma samples. Data were compared between groups and with measurements performed at native haematocrit in 10 lowlanders (LL) at sea level. At native haematocrit, in all rotational thromboelastometry assays, EE+ exhibited hypocoagulable profiles (prolonged clotting time and weaker clot strength) compared with EE− and LL (all P
Published: 2024
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9. Cost‐effectiveness of emicizumab vs bypassing agents in the prevention of bleeding episodes in haemophilia A patients with anti‐FVIII inhibitors in France

Author: Mathias Cousin, Alexandra Pruvot, Benoît Polack, Sandrine Baffert, Chloé Godard, and Marc Trossaert
Subjects: medicine.medical_specialty, Cost effectiveness, Cost-Benefit Analysis, Haemophilia A, 030204 cardiovascular system & hematology, Antibodies, Monoclonal, Humanized, Hemophilia A, 03 medical and health sciences, 0302 clinical medicine, Quality of life, Antibodies, Bispecific, Humans, Medicine, Adverse effect, Intensive care medicine, Genetics (clinical), Emicizumab, Cost–benefit analysis, business.industry, Hematology, General Medicine, medicine.disease, Quality of Life, France, business, Complication, 030215 immunology, Rare disease
Abstract: Introduction The development of an anti-FVIII inhibitor is the most serious complication of haemophilia A occurring in up to 30% of severe haemophilic patients. The current management of haemophilia A with inhibitor uses bypassing agents (BPA) and represents a significant therapeutic burden together with a limited adherence to prophylactic treatment. Emicizumab is the first monoclonal antibody developed in haemophilia A approved for the prevention of bleeding episodes in patients with anti-FVIII inhibitor. Aim The purpose of this study is to evaluate the incremental cost-effectiveness ratio (ICER) of emicizumab versus BPAs. Methods A Markov model was developed over a five-year time horizon to estimate the comparative costs and benefits of the different therapeutic approaches in this rare disease. Model inputs were clinical, including annual bleeding rate and quality of life, and economical including mainly costs of prophylaxis, bleeds and adverse events. Results Emicizumab treatment is dominant, ie lest costly and more effective, in the base-case analysis saving 234 191 € for a gain of 0.88 QALY. This is confirmed by both the deterministic and probabilistic sensitivity analyses. The main limit of the study remains the absence of long-term clinical data allowing to relate treatment consumption to clinical benefit, especially in the progression of haemophilic arthropathy. Conclusion Our results show that emicizumab is a cost-effective treatment allowing to consider an easy to implement prophylactic treatment for haemophilia A patients with anti-FVIII inhibitors.
Published: 2020
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10. Extracellular vesicles from myelodysplastic mesenchymal stromal cells induce DNA damage and mutagenesis of hematopoietic stem cells through miRNA transfer

Author: Rémy Sadoul, Mathieu Meunier, Karine Laulagnier, Jean-Paul Issartel, Sylvie Tondeur, Sophie Park, Claire Jouzier, Quentin Testard, Audrey Guttin, Christine Lefebvre, Sarah Ancelet, Christine Chatellard, Julien Thevenon, Pierre Hainaut, Virginie Persoons, Benoît Polack, Johanna Zannoni, Jean-François Deleuze, Jean-Yves Cahn, Florent Chuffart, Mylène Pezet, Sophie Rousseaux, David Laurin, Karin Pernet-Gallay, University Clinic of Hematology, Centre Hospitalier Universitaire [Grenoble] (CHU), Institute for Advanced Biosciences / Institut pour l'Avancée des Biosciences (Grenoble) (IAB), Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Etablissement français du sang - Auvergne-Rhône-Alpes (EFS), Institut de Biologie et Pathologie [CHU Grenoble] (IBP), CHU Grenoble-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), [GIN] Grenoble Institut des Neurosciences (GIN), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes (UGA), Equipe GAD (LNC - U1231), Lipides - Nutrition - Cancer [Dijon - U1231] (LNC), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement, FHU TRANSLAD (CHU de Dijon), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon), Université Grenoble Alpes - UFR Médecine (UGA UFRM), Université Grenoble Alpes (UGA), Centre National de Recherche en Génomique Humaine (CNRGH), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), PDC*line Pharma SAS, Groupe d'imagerie neurofonctionnelle (GIN), Institut des Maladies Neurodégénératives [Bordeaux] (IMN), Université de Bordeaux (UB)-Centre National de la Recherche Scientifique (CNRS)-Université de Bordeaux (UB)-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Thérapeutique Recombinante Expérimentale (TIMC-IMAG-TheREx ), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP ), Université Grenoble Alpes (UGA)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP ), and CHU Grenoble
Subjects: Ribonuclease III, Cancer Research, DNA damage, [SDV]Life Sciences [q-bio], Extracellular vesicles, DEAD-box RNA Helicases, Extracellular Vesicles, microRNA, Humans, Chemistry, Mesenchymal stem cell, Mutagenesis, Mesenchymal Stem Cells, Hematology, Hematopoietic Stem Cells, Microenvironement tumoral, Cell biology, MicroRNAs, Haematopoiesis, Mesenchymal Stem Cell, Oncology, Myelodysplastic Syndromes, Stem cell, Myelodysplastic syndrome, DNA Damage
Abstract: International audience; Physiopathology of myelodysplastic syndrome (MDS) remains poorly understood and the role of the microenvironment is increasingly highlighted. Recent studies in mouse models demonstrate that abnormalities of mesenchymal stromal cells (MSC) contribute to the physiopathology of MDS. In particular, genetic deletion of dicer1, a gene encoding a type III RNase essential for the genesis of miRNA, in murine MSC-derived osteoprogenitors led to a pathological microenvironment generating myelodysplastic features in hematopoietic progenitors and ultimately leading to acute myeloid leukemia [1]. In human, there is an increased susceptibility to senescence of the MDS mesenchymal stem cells and defects in the support properties of the growth of hematopoietic stem cells (HSC) [2]. These observations establish a causal relationship between deregulation of the hematopoietic niche and MDS pathogenesis. However, so far only few studies have addressed the mechanisms by microenvironmental MSC and HSC exchange signals that may interfere with miRNA processing, specifically in the human MDS microenvironment.
Published: 2020
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11. The influence of coagulation on the drying dynamics of blood pools

Author: Nick Laan, Charles Compain, Benoît Polack, François Caton, Landry Seyve, Celine Nicloux, Institut de Recherche Criminelle de la Gendarmerie Nationale (IRCGN), Ministère de l'intérieur, de l'outre-mer et des collectivités territoriales, Thérapeutique Recombinante Expérimentale (TIMC-IMAG-TheREx), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Laboratoire Rhéologie et Procédés (LRP), and Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])
Subjects: Chromatography, Chemistry, 010401 analytical chemistry, [SPI.MECA.BIOM]Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], Forensic Medicine, 01 natural sciences, Specimen Handling, 0104 chemical sciences, Pathology and Forensic Medicine, [SPI.MAT]Engineering Sciences [physics]/Materials, 03 medical and health sciences, 0302 clinical medicine, Blood Stains, Hemorheology, Photography, Humans, Coagulation (water treatment), 030216 legal & forensic medicine, Desiccation, Blood Coagulation, Law, ComputingMilieux_MISCELLANEOUS, circulatory and respiratory physiology
Abstract: Little is currently known about the importance of clotting during the drying of blood pools. While this is of little moment for droplets where drying occurs faster than contact-phase-induced clotting, clotting may significantly influence blood pools drying as it transform a liquid into a gel. To investigate this influence, we compare the drying of citrated and unmodified blood pools at constant haematocrit, showing large morphological differences during drying, both in the surface appearance, in the colour lightness, as well as in the generation and location of cracks. Further, we designed a clotting-reactivation protocol which allowed recovering the morphological evolution of pure blood drying while using citrate-sampled blood. This result opens the way to the use of citrated blood in blood pools investigations.
Published: 2019
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12. Fibrinography: A Multiwavelength Light-Scattering Assay of Fibrin Structure

Author: Raphaël Marlu, Benoît Polack, Francois Caton, Landry Seyve, Carhel Dassi, Emmanuelle Bigo, Xabel García, Thérapeutique Recombinante Expérimentale (TIMC-IMAG-TheREx), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Laboratoire Rhéologie et Procédés (LRP), and Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])
Subjects: Materials science, 030204 cardiovascular system & hematology, Fibrinogen, Article, Light scattering, Fibrin, 03 medical and health sciences, 0302 clinical medicine, Coagulation cascade, medicine, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Reproducibility, biology, lcsh:RC633-647.5, [SDV.MHEP.HEM]Life Sciences [q-bio]/Human health and pathology/Hematology, lcsh:Diseases of the blood and blood-forming organs, Hematology, Plasma, Wavelength, Coagulation, Biophysics, biology.protein, [SDV.IB]Life Sciences [q-bio]/Bioengineering, medicine.drug
Abstract: We have previously developed a fibrin structural assay dedicated to purified fibrinogen-thrombin system. Here, we extend the pertinence of this test, called Fibrinography, to tissue factor-triggered plasma coagulation. We show that Fibrinography determines quantitatively the structure of fibrin fibers in plasma with an excellent reproducibility. We compare this assay with the commonly used single wavelength turbidity method, showing that the latter is not a proper structural assay, but determines essentially the fibrinogen content in plasma. In addition, we also show, in model plasmas, that Fibrinography is able to discriminate normal and hypocoagulant plasmas, and even between hypercoagulant plasmas. Therefore, Fibrinography, by measuring the final step of the coagulation cascade, may be used to evaluate patients’ plasma in hypo- or hypercoagulant diseases.
Published: 2019
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13. Tumor microenvironment and clonal monocytes from chronic myelomonocytic leukemia induce a procoagulant climate

Author: Johanna Zannoni, Claire Jouzier, Martine Pernollet, Julie Brault, Fabrice Cognasse, Jean-Yves Cahn, Landry Seyve, Benoît Polack, Mathieu Meunier, David Laurin, Sophie Park, Karin Pernet-Gallay, Natacha Mauz, and Mylène Pezet
Subjects: 0301 basic medicine, Myeloid, Chronic myelomonocytic leukemia, Monocytes, Thromboplastin, Thrombosis and Hemostasis, 03 medical and health sciences, Tissue factor, Extracellular Vesicles, 0302 clinical medicine, hemic and lymphatic diseases, medicine, Tumor Microenvironment, Homeostasis, Humans, Cells, Cultured, Tumor microenvironment, business.industry, Myelodysplastic syndromes, Monocyte, Mesenchymal stem cell, Hematopoietic stem cell, Leukemia, Myelomonocytic, Chronic, Mesenchymal Stem Cells, Hematology, medicine.disease, Hematopoietic Stem Cells, Blood Coagulation Factors, 030104 developmental biology, medicine.anatomical_structure, 030220 oncology & carcinogenesis, Cancer research, Nanoparticles, business
Abstract: Chronic myelomonocytic leukemia (CMML) is a myeloid hematological malignancy with overlapping features of myelodysplastic syndromes (MDSs) and myeloproliferative neoplasms (MPNs). The knowledge of the role of the tumor microenvironment (TME), particularly mesenchymal stromal cells (MSCs), in MDS pathogenesis is increasing. Generally, cancer is associated with a procoagulant state participating in tumor development. Monocytes release procoagulant, tissue factor (TF)–bearing microparticles. We hypothesized that MSCs and clonal monocytes release procoagulant extracellular vesicles (EVs) within the CMML TME, inducing a procoagulant state that could modify hematopoietic stem cell (HSC) homeostasis. We isolated and cultured MSCs and monocytes from CMML patients and MSCs from healthy donors (HDs). Their medium EVs and small EVs (sEVs) were collected after iterative ultracentrifugations and characterized by nanoparticle tracking analysis. Their impact on hemostasis was studied with a thrombin generation assay and fibrinography. CMML or HD HSCs were exposed to sEVs from either CMML or HD MSCs. CMML MSC sEVs increased HD HSC procoagulant activity, suggesting a transfer of TF from the CMML TME to HD HSCs. The presence of TF on sEVs was shown by electron microscopy and western blot. Moreover, CMML monocyte EVs conferred a procoagulant activity to HD MSCs, which was reversed by an anti-TF antibody, suggesting the presence of TF on the EVs. Our findings revealed a procoagulant “climate” within the CMML environment related to TF-bearing sEVs secreted by CMML MSCs and monocytes.
Published: 2018

14. Determinants of adherence and consequences of the transition from adolescence to adulthood among young people with severe haemophilia (TRANSHEMO): study protocol for a multicentric French national observational cross-sectional study

Author: Noémie Resseguier, Natacha Rosso-Delsemme, Any Beltran Anzola, Karine Baumstarck, Vanessa Milien, Laurent Ardillon, Sophie Bayart, Claire Berger, Marie-Anne Bertrand, Christine Biron-Andreani, Annie Borel-Derlon, Sabine Castet, Pierre Chamouni, Ségolène Claeyssens Donadel, Emmanuelle De Raucourt, Dominique Desprez, Céline Falaise, Birgit Frotscher, Valérie Gay, Jenny Goudemand, Yves Gruel, Benoît Guillet, Annie Harroche, Abel Hassoun, Yoann Huguenin, Thierry Lambert, Aurélien Lebreton, Anne Lienhart, Michèle Martin, Sandrine Meunier, Fabrice Monpoux, Guillaume Mourey, Claude Negrier, Philippe Nguyen, Placide Nyombe, Caroline Oudot, Brigitte Pan-Petesch, Benoît Polack, Anne Rafowicz, Antoine Rauch, Delphine Rivaud, Pascale Schneider, Alexandra Spiegel, Cecile Stoven, Brigitte Tardy, Marc Trossaërt, Jean-Baptiste Valentin, Stéphane Vanderbecken, Fabienne Volot, Annelise Voyer-Ebrard, Bénédicte Wibaut, Tanguy Leroy, Thomas Sannie, Hervé Chambost, Pascal Auquier, Centre d'études et de recherche sur les services de santé et la qualité de vie (CEReSS), Aix Marseille Université (AMU), Hématogoie pédiatrique, hôpital Sud, Département d'hématologie biologique[Montpellier], Université Montpellier 1 (UM1)-Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)-Hôpital Saint Eloi (CHRU Montpellier), Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier), Laboratoire d'Hématologie Biologique [CHU Caen], Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), Laboratoire d'hématologie [Rouen], CHU Rouen, Normandie Université (NU)-Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU), Centre Hospitalier de Versailles André Mignot (CHV), Service de pédiatrie, d'hématologie et d'oncologie [Hôpital de La Timone - APHM], Assistance Publique - Hôpitaux de Marseille (APHM)- Hôpital de la Timone [CHU - APHM] (TIMONE), Service d’Hématologie Clinique [CHRU Nancy], Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy), Service d'Hématologie, Centre Hospitalier Chambéry, Hôpital cardiologique, Université de Lille, Droit et Santé-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), Groupe innovation et ciblage cellulaire (GICC), EA 7501 [2018-...] (GICC EA 7501), Université de Tours (UT), Institut de recherche en santé, environnement et travail (Irset), Université d'Angers (UA)-Université de Rennes (UR)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Unité de Nutrition Humaine (UNH), Institut National de la Recherche Agronomique (INRA)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020]), Centre Hospitalier Universitaire de Nancy (CHU Nancy), Hôpital Louis Pradel [CHU - HCL], Hospices Civils de Lyon (HCL), Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC (UMR INSERM 1098) (RIGHT), Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS BFC)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Centre Hospitalier Universitaire de Lyon (CHU Lyon), Service de Pédiatrie médicale - Spécialités médicales [CHU Limoges], CHU Limoges, Hôpital Morvan [Brest], Groupe d'Etude de la Thrombose de Bretagne Occidentale (GETBO), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO)-Université de Brest (UBO), CIC Brest, Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Hôpital de la Cavale Blanche, Thérapeutique Recombinante Expérimentale (TIMC-IMAG-TheREx), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Service d'hémato-immuno-oncologie pédiatrique [Rouen], Groupe de recherche sur la thrombose, pharmacologie des antithrombotiques et situations à risque (GRT), Université Jean Monnet - Saint-Étienne (UJM), Centre hospitalier universitaire de Nantes (CHU Nantes), Groupe de Recherche en Psychologie Sociale (GRePS), Université Lumière - Lyon 2 (UL2), Service d'hématologie pédiatrique, Université de la Méditerranée - Aix-Marseille 2-Assistance Publique - Hôpitaux de Marseille (APHM)- Hôpital de la Timone [CHU - APHM] (TIMONE), Laboratoire de Santé Publique, Université de la Méditerranée - Aix-Marseille 2, Ministère des Affaires Sociales et de la Santé, Filière MHEMO, Université Aix-Marseille, Université Montpellier 1 (UM1)-Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)-CHU Saint-Eloi, Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de la Santé et de la Recherche Médicale (INSERM)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Université d'Angers (UA), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté]), Université de Brest (UBO)-Institut Brestois Santé Agro Matière (IBSAM), Groupe de recherche sur la thrombose (GRT (EA 3065)), Université Jean Monnet [Saint-Étienne] (UJM), Hôpital Sud, Centre hospitalier de Versailles André-Mignot, 177, rue de Versailles, 78150 Le Chesnay, France, parent, Université de Tours, Université d'Angers (UA)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Unité de Nutrition Humaine - Clermont Auvergne (UNH), Institut National de la Recherche Agronomique (INRA)-Université Clermont Auvergne (UCA), CHU de Nancy - Hôpitaux de Brabois, Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC (UMR INSERM 1098) (HOTE GREFFON), Université de Franche-Comté (UFC)-Etablissement français du sang [Bourgogne-France-Comté] (EFS [Bourgogne-France-Comté])-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Brest (UBO), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP)-IMAG-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP)-IMAG-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Université de Franche-Comté (UFC), and Jonchère, Laurent
Subjects: [SDV.MHEP.HEM] Life Sciences [q-bio]/Human health and pathology/Hematology, Male, Activities of daily living, Cross-sectional study, [SDV]Life Sciences [q-bio], 030204 cardiovascular system & hematology, 0302 clinical medicine, Risk Factors, Academic Performance, Protocol, Early childhood, adherence, adolescents, Young adult, ComputingMilieux_MISCELLANEOUS, Qualitative Research, [SDV.MHEP] Life Sciences [q-bio]/Human health and pathology, transition, [SDV.MHEP.HEM]Life Sciences [q-bio]/Human health and pathology/Hematology, General Medicine, 3. Good health, [SDV] Life Sciences [q-bio], Patient Satisfaction, Female, Family Relations, France, Psychosocial, Attitude to Health, Haematology (Incl Blood Transfusion), Adult, young adults, medicine.medical_specialty, Transition to Adult Care, Adolescent, haemophilia, Haemophilia, Hemophilia A, 03 medical and health sciences, Young Adult, Quality of life (healthcare), 030225 pediatrics, medicine, Humans, business.industry, Protective Factors, medicine.disease, Treatment Adherence and Compliance, Cross-Sectional Studies, Social Class, Family medicine, Quality of Life, Observational study, business, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract: IntroductionSevere haemophilia is a rare disease characterised by spontaneous bleeding from early childhood, which may lead to various complications, especially in joints. It is nowadays possible to avoid these complications thanks to substitutive therapies for which the issue of adherence is major. The transition from adolescence to adulthood in young people with severe haemophilia is a critical period as it is associated with a high risk of lack of adherence to healthcare, which might have serious consequences on daily activities and on quality of life.Methods and analysisWe present the protocol for a cross-sectional, observational, multicentric study to assess the differences between adolescents and young adults with severe haemophilia in France through the transition process, especially on adherence to healthcare. This study is based on a mixed methods design, with two complementary and consecutive phases, comparing data from a group of adolescents (aged 14–17 years) with those from a group of young adults (aged 20–29 years). The quantitative phase focuses on the determinants (medical, organisational, sociodemographic and social and psychosocial and behavioural factors) of adherence to healthcare (considered as a marker of the success of transition). The qualitative phase explores participants’ views in more depth to explain and refine the results from the quantitative phase. Eligible patients are contacted by the various Haemophilia Treatment Centres participating in the French national registry FranceCoag.Ethics and disseminationThe study was approved by the French Ethics Committee and by the French National Agency for Medicines and Health Products Safety (number: 2016-A01034-47). Study findings will be disseminated to the scientific and medical community in peer-reviewed journals and presented at scientific meetings. Results will be popularised to be communicated via the French association for people with haemophilia to participants and to the general public.Trial registration numberNCT02866526; Pre-results.
Published: 2018
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15. Corrigendum to 'Effect of double-filtration plasmapheresis for antibody-mediated rejection on hemostasis parameters and thrombin generation' [Thromb. Res. 166 (2018) 113-121]

Author: Paolo Malvezzi, Raphaël Marlu, Lionel Rostaing, Benoît Polack, Jocelyne Maurizi, F. Defendi, Maryvonne Christophe, A. Le Gouellec, Thomas Jouve, Pierre-Louis Carron, and L. Seyve
Subjects: Chemistry, Hemostasis, Antibody mediated rejection, Hematology, Pharmacology, Thrombin generation, Double filtration plasmapheresis
Published: 2018

16. Effect of double-filtration plasmapheresis for antibody-mediated rejection on hemostasis parameters and thrombin generation

Author: Lionel Rostaing, Pierre-Louis Carron, Jocelyne Maurizi, Maryvonne Christophe, A. Le Gouellec, L. Seyve, Benoît Polack, F. Defendi, Thomas Jouve, Raphaël Marlu, Paolo Malvezzi, Université Grenoble Alpes - UFR Médecine (UGA UFRM), Université Grenoble Alpes (UGA), Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Thérapeutique Recombinante Expérimentale (TIMC-IMAG-TheREx), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Biostatistiques santé, Département biostatistiques et modélisation pour la santé et l'environnement [LBBE], Laboratoire de Biométrie et Biologie Evolutive - UMR 5558 (LBBE), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de Recherche en Informatique et en Automatique (Inria)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de Recherche en Informatique et en Automatique (Inria)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Laboratoire de Biométrie et Biologie Evolutive - UMR 5558 (LBBE), Université de Lyon-Université de Lyon-Institut National de Recherche en Informatique et en Automatique (Inria)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS), Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Department of Multiorgan Transplantation, CHU Toulouse [Toulouse], Département de Néphrologie et Transplantation d'organes [CHU Toulouse], Pôle Urologie - Néphrologie - Dialyse - Transplantations - Brûlés - Chirurgie plastique - Explorations fonctionnelles et physiologiques [CHU Toulouse], and Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)
Subjects: Adult, Male, [SDV]Life Sciences [q-bio], 030204 cardiovascular system & hematology, 030230 surgery, Pharmacology, Fibrinogen, Thrombomodulin, [SDV.MHEP.UN]Life Sciences [q-bio]/Human health and pathology/Urology and Nephrology, 03 medical and health sciences, Tissue factor, 0302 clinical medicine, Thrombin, Von Willebrand factor, medicine, Humans, Platelet, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, ComputingMilieux_MISCELLANEOUS, Aged, Clotting factor, Hemostasis, biology, business.industry, [SDV.MHEP.HEM]Life Sciences [q-bio]/Human health and pathology/Hematology, Plasmapheresis, Hematology, Middle Aged, 3. Good health, biology.protein, Female, business, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology, medicine.drug
Abstract: Donor-specific alloantibodies (DSAs) cause kidney-allograft loss in chronic antibody-mediated rejection (CAMR). Treatment relies on blocking antibody-producing cells and removing DSAs by apheresis: e.g., double-filtration plasmapheresis (DFPP).To determine the impact of DFPP (6 or 8 sessions/patient) on clotting factors and natural anticoagulants, and on thrombin generation, we performed a prospective and observational study in five CAMR kidney-transplant patients who received DFPP plus rituximab therapy. Thrombin generation was performed in poor platelet plasma (PPP) with 5 pM tissue factor without and with 2 nM recombinant human thrombomodulin.After the first DFPP session, median levels of high molecular-weight proteins (fibrinogen, FV, FVIII, FXI, FXIII, von Willebrand factors and α2-MG) decreased significantly to50% of baseline values, whereas levels of low molecular-weight factors (100 kDa) were not significantly modified, except for protein S and TFPI. Of note, binding-protein (BP) S, i.e., C4BP, was significantly decreased. Over the course of successive DFPP sessions, both high and lower molecular-weight proteins (100 kDa) with longer half-lives (2 days, prothrombin and factor XII) were significantly decreased. DFPP also highly affected thrombin generation in the absence of thrombomodulin but not significantly in the presence of thrombomodulin. After the first DFPP session, mean endogenous thrombin potential (ETP) and peak thrombin (PH) significantly decreased when the thrombin generation assay was performed without thrombomodulin (respectively, 1084 nM·min for ETP and 210 nM for PH after the first DFPP session compared to 1616 nM·min and 264 nM at baseline). In the presence of thrombomodulin, there was only a slight decrease in ETP and PH (respectively 748 nM·min, and 172 nM after the first DFPP session compared to 822 nM·min and 179 nM at baseline). After the last session, median ETP and PH decreased respectively to 646 nM·min and 143 nM without thrombomodulin, and, to 490 nM·min and 117 nM with thrombomodulin.DFPP significantly removed high molecular-weight proteins from the haemostatic system and profoundly decreased levels of protein S and TFPI. Overall thrombin-generation balance was only moderately affected in the presence of thrombomodulin. Nevertheless, high depletion of fibrinogen, FXIII and Von Willebrand Factor may expose patients to an increased risk of bleeding.
Published: 2018
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17. Regulation of the Expression of Type III Secretion Systems: an Example from Pseudomonas aeruginosa

Author: Audrey Le Gouellec, Benoît Polack, Bertrand Toussaint, Dakang, Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), and Centre Hospitalier Universitaire [Grenoble] (CHU)
Subjects: 0303 health sciences, biology, 030306 microbiology, Operon, Pseudomonas aeruginosa, Virulence, Pathogenic bacteria, Yersinia, biology.organism_classification, medicine.disease_cause, Type three secretion system, Microbiology, 03 medical and health sciences, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Gene expression, medicine, bacteria, ComputingMilieux_MISCELLANEOUS, Bacteria, 030304 developmental biology
Abstract: This chapter presents a general overview of type III secretion system (T3SS) regulation in the main pathogenic bacteria, and focuses on the different aspects of the regulation of T3SS gene expression in Pseudomonas aeruginosa. Pathogenic bacteria occupy very different infection foci; for instance, the animal pathogens Shigella spp. and Salmonella spp. live intracellularly after successful invasion, whereas Yersinia spp., P. aeruginosa, and enteropathogenic Escherichia coli (EPEC) predominantly remain extracellular. Therefore, different stimuli could be used to up or downregulate the expression of T3SS genes: temperature, divalent cations, host cell contact, serum, or other factors. Although the mechanism by which metabolic pathways influence virulence gene expression in bacteria is unclear, the example of P. aeruginosa detailed could facilitate progress in that field. It has been shown that T3SS expression is dependent on cell density and that exsCEBA operon expression decreases rapidly in the second part of stationary phase, and also that indole-3-acetic acid (IAA), naphthalenacetic, and 3-hydroxykynurenine inhibit exsCEBA operon expression at millimolar concentrations. In 2005, Wu and coworkers proposed that the opportunistic pathogen P. aeruginosa could adapt to the host by sensing alterations in the host immune function and respond by enhancing the virulence phenotype. Despite the large number of genes that influence the regulation of the expression of T3SS genes in P. aeruginosa, only a minority have been shown to have a precise role. Both fundamental and applied research can make the inhibition of T3SS expression as one of the first new antibacterial therapeutics.
Published: 2016
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18. Bacterial vectors for active immunotherapy reach clinical and industrial stages

Author: Bertrand Toussaint, Audrey Le Gouellec, Laurent Buffat, Benoît Polack, Xavier Chauchet, Laboratoire d'Ingénierie des Macromolécules (LIM), Institut de biologie structurale (IBS - UMR 5075 ), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), TheREx, Université Grenoble Alpes - UFR Pharmacie (UGA UFRP), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Valigen, Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), and Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)
Subjects: [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Listeria, medicine.medical_treatment, Immunology, Active immunotherapy, Biology, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, 03 medical and health sciences, 0302 clinical medicine, Immune system, Antigen, Pseudomonas, medicine, Humans, Immunology and Allergy, Cytotoxic T cell, Engineering tool, [INFO.INFO-BT]Computer Science [cs]/Biotechnology, bacteria, attenuation, 030304 developmental biology, Pharmacology, 0303 health sciences, Antigen delivery, Immunotherapy, Active, antigen delivery, Immunotherapy, 030220 oncology & carcinogenesis, Commentary, immunotherapy
Abstract: International audience; Active immunotherapy based on live attenuated bacterial vectors has matured in terms of industrial development and develops through a combination of three phenomena. First, active immunotherapy that stimulates an antigen-specific cytotoxic T-cell immune response has become a reality after several years of work. Second, there is still a need to identify vectors that can deliver antigens to the cytosol of antigen-presenting cells in vivo. Third, the recent progress in the understanding of bacterial lifestyle and in developing genetic engineering tools has enabled the design of bioengineered bugs that are capable of delivering antigens. Here, we review the mechanisms by which clinical bacterial vectors deliver antigens into the cytosol of antigen-presenting cells and summarize the development strategy of the three identified firms in this field.
Published: 2012
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19. Antithrombin-Independent Effects of Heparins on Fibrin Clot Nanostructure

Author: Christelle Yeromonahos, Raphaël Marlu, Benoît Polack, François Caton, Laboratoire de rhéologie (LR), Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique de Grenoble (INPG)-Université Joseph Fourier - Grenoble 1 (UJF), TheREx, Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), inconnu, and Inconnu
Subjects: MESH: Heparin, Nanostructure, [PHYS.PHYS.PHYS-BIO-PH]Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], Nanotechnology, 030204 cardiovascular system & hematology, Fondaparinux, Fibrinogen, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, Fibrin, MESH: Thrombin, 03 medical and health sciences, 0302 clinical medicine, Thrombin, Fibrinolytic Agents, medicine, MESH: Fibrinolytic Agents, Humans, MESH: Thrombosis, MESH: Fibrin, Blood Coagulation, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, MESH: Humans, MESH: Spectrophotometry, biology, Heparin, Chemistry, Antithrombin, Thrombosis, [CHIM.MATE]Chemical Sciences/Material chemistry, [INFO.INFO-MO]Computer Science [cs]/Modeling and Simulation, 3. Good health, [CHIM.THEO]Chemical Sciences/Theoretical and/or physical chemistry, Spectrophotometry, MESH: Blood Coagulation, MESH: Fibrinogen, Biophysics, biology.protein, [PHYS.PHYS.PHYS-CHEM-PH]Physics [physics]/Physics [physics]/Chemical Physics [physics.chem-ph], Cardiology and Cardiovascular Medicine, Fibrinolytic agent, medicine.drug
Abstract: Objective— Because of the widespread clinical use of heparins, their effects on the enzymatic cascade are very well known. In contrast, little is known about the direct effect of heparins on the nanostructure of fibrin fibers, even though this nanostructure plays a major role in the mechanical strength and lysis of clots. This lack of reliable data can be correlated with the lack of a nonintrusive, quantitative method to determine this structure. We recently developed such a method that allows the simultaneous determination of the average fiber radius and the protein content using spectrometric data. In this study, we assessed the nanostructure of fibrin in a system composed of human thrombin and fibrinogen. Methods and Results— We provide quantitative evidence showing that both unfractionated heparin and low molecular weight heparin directly alter the nanostructure of fibrin fibers independent of their other actions on the coagulation cascade; as expected, the pentasaccharide fondaparinux has no effect. Conclusion— Our results show that in addition to the effect of heparin on the coagulation cascade, modifications of the fibrin nanostructure may also contribute to improved fibrinolysis.
Published: 2012
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20. Optimal epitope composition after antigen screening using a live bacterial delivery vector

Author: Jean-François Mayol, Yan Wang, Madiha Derouazi, Audrey Le Gouellec, Raphaël Marlu, Olivier Epaulard, Bertrand Toussaint, Benoît Polack, Nicolas Pasqual, Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Department of infectious diseases, CHU Grenoble, Departement des Effets Biologiques des Rayonnements, CRSSA, Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire d'Ingénierie des Macromolécules (LIM), Institut de biologie structurale (IBS - UMR 5075 ), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), TheREx, VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Grant from the French National Agency for Research. The first author was supported by the Swiss National Science Foundation., Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), and Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)
Subjects: MESH: Immunotherapy, medicine.medical_treatment, MESH: Amino Acid Sequence, MESH: Base Sequence, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, Applied Microbiology and Biotechnology, MESH: Cancer Vaccines, Epitope, MESH: Glioma, MESH: Recombinant Proteins, Epitopes, Mice, 0302 clinical medicine, Cancer immunotherapy, MESH: Genetic Vectors, MESH: Animals, bacteria, epitope, 0303 health sciences, MESH: Dendritic Cells, Glioma, Recombinant Proteins, 3. Good health, Intramolecular Oxidoreductases, Vaccination, antigen delivery system, 030220 oncology & carcinogenesis, Pseudomonas aeruginosa, MESH: Pseudomonas aeruginosa, MESH: Genetic Engineering, Female, Immunotherapy, Genetic Engineering, Biotechnology, MESH: DNA Primers, MESH: Cell Line, Tumor, MESH: Epitopes, Genetic Vectors, chemical and pharmacologic phenomena, Bioengineering, Biology, Vaccines, Attenuated, Cancer Vaccines, TTSS, 03 medical and health sciences, Antigen, MESH: Mice, Inbred C57BL, Immunity, Cell Line, Tumor, Report, MESH: Vaccines, Attenuated, medicine, Animals, Amino Acid Sequence, MESH: Mice, DNA Primers, 030304 developmental biology, cancer immunotherapy, Innate immune system, Base Sequence, T-cell receptor, Dendritic Cells, MESH: Bioengineering, MESH: Intramolecular Oxidoreductases, Virology, Mice, Inbred C57BL, CTL, TRP-2, MESH: T-Lymphocytes, Cytotoxic, MESH: Female, T-Lymphocytes, Cytotoxic
Abstract: International audience; Immunotherapeutic approaches, based on the generation of tumor-specific cytotoxic T-lymphocytes (CTL), are currently emerging as promising strategies of anti-tumor therapy. The potential use of attenuated bacteria as engineered vectors for vaccine development offers several advantages, including the stimulation of innate immunity. We developed an attenuated live bacterial vector using the type III secretion system (TTSS) of Pseudomonas aeruginosa to deliver in vivo tumor antigens. Using an inducible and rapid expression plasmid, vaccination with several antigens of different length and epitope composition, including TRp-2, gp100 and MUC18, was evaluated against glioma tumor cells. We observed similar CTL immunity and T-cell receptor (TCR) repertoire diversity with the vaccines, TRP2(125-243), TRP2L(125-376) and TRP2S(291-376). However, only immunization with TRP2L(125-376) induced significant anti-tumor immunity. Taken together, our data indicate the importance of the epitopes composition and/or peptide length of these peptides for inducing cytotoxic T-lymphocyte (CTL) mediated immunity. Characteristics that consistently improved anti-tumor immunity include: long peptides with immunodominant and cryptic CD8(+) epitopes, and strong CD4(+) Th epitopes. Our bacterial vector is versatile, easy-to-use and quick to produce. This vector is suitable for rapid screening and evaluation of antigens of varying length and epitope composition.
Published: 2010
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21. Optimization of a Type III Secretion System-Based Pseudomonas aeruginosa Live Vector for Antigen Delivery

Author: Carole Margerit, Madiha Derouazi, Olivier Epaulard, Didier Filopon, Raphaël Marlu, Benoît Polack, Bertrand Toussaint, Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), TheREx, VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), and Grants from the Association pour la Recherche contre le Cancer and from the Agence Nationale de la Recherche ('Emergence et Maturation de Projets de biotechnologie' BacVac 2007)
Subjects: [SDV.BIO]Life Sciences [q-bio]/Biotechnology, medicine.medical_treatment, Clinical Biochemistry, Gene mutation, MESH: Quorum Sensing, Median lethal dose, Mice, Immunology and Allergy, MESH: Animals, [INFO.INFO-BT]Computer Science [cs]/Biotechnology, 0303 health sciences, biology, Aroa, Quorum Sensing, Vaccine Research, 3. Good health, Bacterial vaccine, Protein Transport, Bacterial Vaccines, Pseudomonas aeruginosa, MESH: Pseudomonas aeruginosa, Toxicity, Female, MESH: Genes, Bacterial, MESH: Dose-Response Relationship, Immunologic, Microbiology (medical), MESH: Protein Transport, MESH: Cell Line, Tumor, Ovalbumin, Virulence Factors, Immunology, Dose-Response Relationship, Immunologic, Vaccines, Attenuated, Microbiology, Lethal Dose 50, 03 medical and health sciences, Antigen, MESH: Mice, Inbred C57BL, Cell Line, Tumor, MESH: Vaccines, Attenuated, medicine, Animals, MESH: Mice, MESH: Virulence Factors, 030304 developmental biology, MESH: Ovalbumin, 030306 microbiology, Lethal dose, Immunotherapy, biology.organism_classification, Mice, Inbred C57BL, MESH: Lethal Dose 50, Genes, Bacterial, MESH: Gene Deletion, MESH: Female, Gene Deletion, MESH: Bacterial Vaccines
Abstract: During the last few years, the use of type III secretion system-based bacterial vectors for immunotherapy purposes has been assessed in various applications. We showed that a type III secretion-based Pseudomonas aeruginosa vector delivering the ovalbumin (OVA) antigen induced an efficient specific CD8 + T-lymphocyte immune response against OVA-expressing cells. Because of the intrinsic toxicity of the vector, further virulence attenuation was needed. Therefore, we explored the effects of the deletion of quorum-sensing genes and the aroA gene toward toxicity and efficiency of the vector strain. The aroA mutation in our strain (making the strain auxotrophic for aromatic amino acids) conferred a strikingly reduced toxicity, with the bacterial lethal dose being more than 100 times higher than that of the parental strain. The quorum-sensing gene mutation alone was associated with a slightly reduced toxicity. In a prophylactic OVA-expressing melanoma mouse model, an OVA-delivering aroA -deficient mutant was the most efficient at a low dose (10 5 ), but dose enhancement was not associated with a greater immune response. The quorum-sensing-deficient strain was the most efficient at a mild dose (10 6 ), but this dose was close to the toxic dose. Combination of both mutations conferred the highest efficiency at an elevated dose (10 7 ), in agreement with the known negative effects of quorum-sensing molecules upon T-cell activation. In conclusion, we have obtained a promising immunotherapy vector regarding toxicity and efficiency for further developments in both antitumor and anti-infectious strategies.
Published: 2008
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22. EQOFIX: a combined economic and quality-of-life study of hemophilia B treatments in France

Author: Benoît, Polack, Thierry, Calvez, Hervé, Chambost, Chantal, Rothschild, Jenny, Goudemand, Ségolène, Claeyssens, Annie, Borel-Derlon, Isabelle, Bardoulat, Frédérique, Maurel, Marie-Christine, Woronoff-Lemsi, Nadine, Bouvet, Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Laboratoire d'hématologie, Université Pierre et Marie Curie - Paris 6 - UFR de Médecine Pierre et Marie Curie (UPMC), Université Pierre et Marie Curie - Paris 6 (UPMC), Institut Pierre Louis d'Epidémiologie et de Santé Publique (iPLESP), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service d'Hématologie pédiatrique, Hôpital de la Timone, Marseille, Assistance Publique - Hôpitaux de Marseille (APHM)- Hôpital de la Timone [CHU - APHM] (TIMONE), Service d'immuno-hématologie pédiatrique [CHU Necker], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Hôpital cardiologique, Université de Lille, Droit et Santé-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), Nutrition, inflammation et dysfonctionnement de l'axe intestin-cerveau (ADEN), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Sérine protéases et physiopathologie de l'unité neurovasculaire, Université de Caen Normandie (UNICAEN), Department of Health Economics and Outcome Research, IMS Health, La Défense, France, Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC (UMR INSERM 1098) (RIGHT), Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS BFC)-Université de Franche-Comté (UFC)
Subjects: Adult, Male, Pediatrics, medicine.medical_specialty, Adolescent, Immunology, Population, Hemorrhage, Disease, Hemophilia B, Cohort Studies, Factor IX, Quality of life, medicine, Humans, Immunology and Allergy, [INFO.INFO-BT]Computer Science [cs]/Biotechnology, Child, education, education.field_of_study, business.industry, Hematology, Middle Aged, 3. Good health, Clinical trial, Cohort, Costs and Cost Analysis, Quality of Life, Female, Observational study, France, business, medicine.drug, Cohort study
Abstract: BACKGROUND EQOFIX is a medicoeconomic study that analyzed the health-related quality of life (HRQoL) and costs of care of the moderate and severe forms of hemophilia B, treated on demand or by prophylaxis with either plasma-derived Factor IX (pdFIX) or recombinant FIX (rFIX). STUDY DESIGN AND METHODS The primary objectives were evaluations of the impact of hemophilia B on HRQoL and of the costs associated with its management. The secondary objectives were evaluations of the clinical efficacy and costs of care of pdFIX and rFIX. In this observational study we included and followed for 1 year severe and moderate hemophilia B patients without inhibitor. HRQoL was evaluated through generic and disease-specific questionnaires. Information on the health resources consumed was collected every 3 months. RESULTS The EQOFIX cohort was composed of 155 patients, including 51 children and 104 adults, with 114 having severe disease and 41 having moderate disease. The regimens were prophylactic for 61 and on demand for 94. Altogether, 78 were treated with rFIX and 77 with pdFIX. There was no difference in the QoL between the pdFIX and rFIX treatments. The extra cost of prophylaxis was €22,605 per bleeding event prevented. The consumption of FIX was 1.4-fold higher for the patients treated with rFIX than for the patients treated with pdFIX. CONCLUSION Our findings in a cohort composed of 25% of the French population of moderate and severe hemophilia B patients show, with similar clinical and HRQoL results, that treatment with rFIX is more expensive than treatment with pdFIX.
Published: 2015
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23. High level production of secreted proteins: Example of the human tissue inhibitor of metalloproteinases 1

Author: Bertrand Toussaint, Benoît Polack, Nicolas Mouz, L. Crombez, J.L. Lenormand, B. Marques, Candice Trocme, Groupe de Recherche et d'Etude du Processus Inflammatoire (GREPI), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), European Laboratory HumProTher, CHU Grenoble, Protein'eXpert SA, and Laboratoire d'Hématologie
Subjects: Biophysics, Biology, Matrix metalloproteinase, Kidney, Protein Engineering, Transfection, Biochemistry, Cell Line, law.invention, MESH: Recombinant Proteins, 03 medical and health sciences, 0302 clinical medicine, In vivo, law, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], Humans, Molecular Biology, 030304 developmental biology, 0303 health sciences, Tissue Inhibitor of Metalloproteinase-1, MESH: Humans, MESH: Transfection, MESH: Kidney, Cell Biology, Protein engineering, Chromatography, Ion Exchange, MESH: Chromatography, Ion Exchange, Recombinant Proteins, In vitro, MESH: Cell Line, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], 3. Good health, MESH: Protein Engineering, Genetic Enhancement, Secretory protein, MESH: Tissue Inhibitor of Metalloproteinase-1, Cell culture, 030220 oncology & carcinogenesis, Recombinant DNA, MESH: Genetic Enhancement
Abstract: International audience; The major difficulty for high-throughput screening of therapeutic protein candidates in experimental animal models of pathologies or for structural studies is their fast and efficient production. The tissue inhibitors of metalloproteinases (TIMPs) considered to play a role in many physiological and pathological processes, such as arthritis or cancer, by inhibiting matrix metalloproteinases or acting as signalling molecules, have always been produced with huge difficulties. We hereby propose a new method to overproduce human recombinant TIMP-1 by transient expression in HEK293E cells, followed by a one-step chromatography purification, yielding in only 2 weeks, dozens of milligrams of pure, stable, glycosylated and active protein for in vitro and in vivo studies. This easy to set up, rapid, and efficient method could be applied for any naturally secreted mammalian protein.
Published: 2005
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24. Combined sacB-based negative selection and cre-lox antibiotic marker recycling for efficient gene deletion in Pseudomonas aeruginosa

Author: Danièle Lamotte, Benoît Polack, Lauriane E. Quenee, Groupe de Recherche et d'Etude du Processus Inflammatoire (GREPI), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Laboratoire d'Hématologie, and CHU Grenoble
Subjects: Genetic Markers, MESH: Integrases, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, MESH: Selection (Genetics), Tetracycline, Genetic Vectors, Molecular Sequence Data, Cre recombinase, MESH: Gene Transfer Techniques, MESH: Base Sequence, MESH: Gene Targeting, MESH: Genetic Markers, Biology, medicine.disease_cause, Genome, General Biochemistry, Genetics and Molecular Biology, Microbiology, Viral Proteins, 03 medical and health sciences, Negative selection, Antibiotic resistance, Plasmid, MESH: Genetic Vectors, medicine, Selection, Genetic, [INFO.INFO-BT]Computer Science [cs]/Biotechnology, Gene, 030304 developmental biology, Genetics, 0303 health sciences, MESH: Molecular Sequence Data, Base Sequence, Integrases, 030306 microbiology, Pseudomonas aeruginosa, Gene Transfer Techniques, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, MESH: Viral Proteins, MESH: Gene Deletion, Gene Targeting, MESH: Pseudomonas aeruginosa, MESH: Genetic Engineering, Genetic Engineering, Gene Deletion, Biotechnology, medicine.drug
Abstract: International audience; The complete genome of the bacterial pathogen Pseudomonas aeruginosa has now been sequenced, allowing gene deletion, one of the most frequently used methods in gene function study, to be fully exploited. In this study, we combine the sacB-based negative selection system with a cre-lox antibiotic marker recycling method. This methodology allows allelic exchange between a target gene and a gentamicin cassette flanked by the two lox sequences. A tetracycline plasmid expressing the cre recombinase is then introduced in the mutant strain to catalyze the excision of the lox-flanked resistance marker. We demonstrate here the efficiency of the combination of these two methods in P. aeruginosa by successively deleting ExoS and ExoT, which are two genetically independent toxins of the type-three secretion system (TTSS). This functional cre-lox recycling antibiotic marker system can create P. aeruginosa strains with multiple mutations without modifying the antibiotic resistance profile when compared to the parental strain.
Published: 2005
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25. Epigenesis and Dynamic Similarity in Two Regulatory Networks in Pseudomonas Aeruginosa

Author: Janine Guespin-Michel, Benoît Polack, Adrien Richard, Jean-Paul Comet, Gilles Bernot, Christian Hulen, and Annabelle Merieau
Subjects: Transcription, Genetic, Pseudomonas aeruginosa, Applied Mathematics, General Medicine, Computational biology, Models, Theoretical, Biology, Bioinformatics, medicine.disease_cause, Formal methods, General Biochemistry, Genetics and Molecular Biology, Epigenesis, Genetic, Philosophy, Philosophy of biology, Identity (object-oriented programming), medicine, Dynamic similarity, Experimental methods, General Agricultural and Biological Sciences, General Environmental Science, Epigenesis
Abstract: Mucoidy and cytotoxicity arise from two independent modifications of the phenotype of the bacterium Pseudomonas aeruginosa that contribute to the mortality and morbidity of cystic fibrosis. We show that, even though the transcriptional regulatory networks controlling both processes are quite different from a molecular or mechanistic point of view, they may be identical from a dynamic point of view: epigenesis may in both cases be the cause of the acquisition of these new phenotypes. This was highlighted by the identity of formal graphs modelling these networks. A mathematical framework based on formal methods from computer science was defined and implemented with a software environment. It allows an easy and rigorous validation and certification of these models and of the experimental methods that can be proposed to falsify or validate the underlying hypothesis.
Published: 2004
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26. Surgery in severe factor XIII deficiency: report of a case of epilepsy neurosurgery and review

Author: A.L. Benabid, Minotti L, S. Garrel, A. Joannard, Benoît Polack, C. Barro, I. Wrobleski, P. Kahn, A. Koudsie, F. Blanc-Jouvan, G. Pernod, and B. Arnutti
Subjects: medicine.medical_specialty, medicine.diagnostic_test, business.industry, Magnetic resonance imaging, Hematology, General Medicine, Factor XIII, medicine.disease, Surgery, Epilepsy, Hemostasis, medicine, Factor XIII deficiency, Epilepsy surgery, Neurosurgery, business, Genetics (clinical), medicine.drug, Congenital disorder
Abstract: Summary. Factor XIII (FXIII) deficiency is a rare autosomal recessive congenital disorder of haemostasis, associated with a high risk of intracranial haemorrhage. Intracranial haemorrhage can result in neurological sequelae including seizure disorders. In some cases, medically intractable epilepsy led to epilepsy surgery. Little has been reported on the management of FXIII deficiency during surgery, and there is only a few data on the management, safety and efficacy of epilepsy surgery in the patients with haemostatic disorder. We report here an epilepsy neurosurgery in a case of severe FXIII deficiency.
Published: 2003
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27. Activation of the Pseudomonas aeruginosa Type III Secretion System Requires an Intact Pyruvate Dehydrogenase aceAB Operon

Author: Benoît Polack, Bertrand Toussaint, Rozen Leberre, Olivier Epaulard, Benoit Guery, Arjan de Groot, Ina Attree, and Denis Dacheux
Subjects: Pore Forming Cytotoxic Proteins, Cystic Fibrosis, Histidine Kinase, Neutrophils, Operon, Immunology, Mutant, Pyruvate Dehydrogenase Complex, Dihydrolipoyllysine-Residue Acetyltransferase, medicine.disease_cause, Microbiology, Type three secretion system, Rats, Sprague-Dawley, Bacterial Proteins, Phosphoprotein Phosphatases, Pneumonia, Bacterial, medicine, Animals, Humans, Pseudomonas Infections, Secretion, Cellular Microbiology: Pathogen-Host Cell Molecular Interactions, L-Lactate Dehydrogenase, biology, Pseudomonas aeruginosa, biochemical phenomena, metabolism, and nutrition, biology.organism_classification, Pyruvate dehydrogenase complex, Molecular biology, Rats, DNA-Binding Proteins, Disease Models, Animal, Mutagenesis, Insertional, Infectious Diseases, Pseudomonadales, Trans-Activators, Parasitology, Protein Kinases, Pseudomonadaceae
Abstract: Pseudomonas aeruginosa clinical cystic fibrosis isolate CHA was mutagenized with Tn 5 Tc to identify new genes involved in type III secretion system (TTSS)-dependent cytotoxicity toward human polymorphonuclear neutrophils. Among 25 mutants affected in TTSS function, 14 contained the insertion at different positions in the aceAB operon encoding the PDH-E1 and -E2 subunits of pyruvate dehydrogenase. In PDH mutants, no transcriptional activation of TTSS genes in response to calcium depletion occurred. Expression in trans of ExsA restored TTSS function and cytotoxicity.
Published: 2002
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28. Mechanisms of Improvement of Erythropoiesis with Low Dose of Deferasirox in Low Risk Myelodysplastic Syndromes

Author: P. Faure, C. Lefebvre, S. Ancelet, Sophie Park, C. Garrel, Jean-Yves Cahn, Benoît Polack, J. Arnaud, J.M. Moulis, Y. Wang, and M. Meunier
Subjects: Oncology, Cancer Research, medicine.medical_specialty, business.industry, Myelodysplastic syndromes, Low dose, Deferasirox, Hematology, medicine.disease, 03 medical and health sciences, 0302 clinical medicine, 030220 oncology & carcinogenesis, Internal medicine, Medicine, Erythropoiesis, business, 030215 immunology, medicine.drug
Published: 2017
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29. Preanalytical Recommendations of the ‘Groupe d’Etude sur l’Hémostase et la Thrombose’ (GEHT) for Venous Blood Testing in Hemostasis Laboratories

Author: Benoît Polack, Bernard Boneu, and Jean-François Schved
Subjects: Blood Specimen Collection, medicine.medical_specialty, Hematologic Tests, Third party, Clinical Laboratory Techniques, business.industry, Hematology, Guideline, Venous blood, Blood collection, Patient care, Specimen Handling, Surgery, Physiology (medical), Hemostasis, medicine, Humans, Blood Coagulation Tests, Intensive care medicine, business, Blood sampling
Abstract: Clinical laboratory investigation of hemostasis has considerably progressed during the last decade with the use of high-quality instruments and reagents. In order to transfer these improvements to the quality of patient care and to increase interand intra-laboratory reproducibility, more stringent requirements in the management of samples from blood collection to the validation of results are needed. The outcome of even the best clinical test in hemostasis is critically dependent upon the quality of the sample used. Therefore, the quality of blood sampling, the transport to the laboratory, sample preparation and storage are factors determining the test results. To minimize this significant source of variation, the implementation of good biological practice rules is compulsory. Both the National Committee for Clinical Laboratory Standards (NCCLS) and the European Concerted Action on Thrombosis have established and/ or reinforced such guidelines [1–5]. In addition, at least in France, the clinical pathologist is responsible for the whole preanalytical phase, including blood collection, even if it is carried out by a third party [6]. In this study, the criteria approved by the ‘Groupe d’Etude sur l’Hemostase et la Thrombose’ (GEHT) are summarized and updated [7].
Published: 2001
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30. Protein Delivery by Pseudomonas Type III Secretion System: Ex Vivo Complementation of p67phox-Deficient Chronic Granulomatous Disease

Author: Françoise Morel, Bertrand Toussaint, Marie Hélène Paclet, Sabrina Vergnaud, Danièle Lamotte, and Benoît Polack
Subjects: Herpesvirus 4, Human, Histidine Kinase, Recombinant Fusion Proteins, Biophysics, Virulence, Granulomatous Disease, Chronic, medicine.disease_cause, Biochemistry, Microbiology, Type three secretion system, Cytosol, Chronic granulomatous disease, Bacterial Proteins, NADPH oxidase complex, medicine, Humans, Molecular Biology, Cell Line, Transformed, B-Lymphocytes, NADPH oxidase, biology, Pseudomonas aeruginosa, Genetic Complementation Test, NADPH Oxidases, Biological Transport, Cell Biology, Phosphoproteins, medicine.disease, Fusion protein, biology.protein, Protein Kinases
Abstract: Bacterial type III secretion system drives the translocation of virulence factors into the cystosol of host target cells. In phagocytes and in Epstein-Barr virus immortalized B lymphocytes, NADPH oxidase generates O(-2) through an electron transfer chain the activity of which depends on the assembly of three, p67(phox), p47(phox) and p40(phox) cytosolic activating factors with Rac 1/2 and a membrane redox component, cytochrome b(558). In p67(phox) deficient chronic granulomatous disease (CGD) patients, p67-phox is missing and NADPH oxidase activity is abolished. ExoS is a virulence factor of Pseudomonas aeruginosa which is secreted via the type III secretion system: it was fused with p67(phox). Pseudomonas aeruginosa synthesized and translocated the hybrid ExoS-p67(phox) fusion protein into the cytosol of B lymphocytes via the type III secretion system. Purified ExoS-p67(phox) hybrid protein was as efficient as normal recombinant p67(phox) in cell-free reconstitution of NADPH oxidase activity. Therefore, ExoS-p67(phox) was transferred via the type III secretion system of Pseudomonas aeruginosa into the cytosol of B lymphocytes from a p67(phox)-deficient CGD patient and functionally reconstituted NADPH oxidase activity. In the complementation process, ExoS acted as a molecular courier for protein delivery: the reconstitution of an active NADPH oxidase complex suggests type III secretion system to be a new approach for cellular therapy.
Published: 2000
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31. A quels patients s'adresse le dosage des D-dimères ?

Author: Claire Barro, Michelle Fontaine, Benoît Polack, Jean-Luc Bosson, Patrick H. Carpentier, and Gilles Pernod
Subjects: Analytical Chemistry
Abstract: Resume Les D-dimeres plasmatiques, produits de degradation de la fibrine stabilisee, representent une aide diagnostique pour l'exclusion de l'embolie pulmonaire (EP) et de la thrombose veineuse profonde (TVP), evaluee chez les patients ambulatoires. Nous avons voulu determiner quelle etait la place du dosage des D-dimeres en milieu hospitalier pour l'ensemble des patients cliniquement suspects de thrombose veineuse profonde et/ou d'embolie pulmonaire. Nous rapportons les resultats d'une etude retrospective cas-temoins concernant 300 patients hospitalises, non selectionnes avant leur inclusion, ou le dosage des D-dimeres a ete effectue par Elisa. Les vrais negatifs s'elevent a 45 % chez les patients des urgences, ce qui confirme les resultats publies pour les patients ambulatoires, pour lesquels l'utilite des D-dimeres a ete demontree. Chez les malades hospitalises, nous demontrons l'utilite des D-dimeres pour les patients qui repondent aux trois criteres suivants: âge
Published: 1999
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32. Immunothérapie par vecteur bactérien vivant : utilisation du système de sécrétion de type III de Pseudomonas aeruginosa

Author: Madiha Derouazi, Bertrand Toussaint, L. Pelletier, Olivier Epaulard, and Benoît Polack
Subjects: medicine.medical_specialty, Pseudomonas aeruginosa, medicine.medical_treatment, Cancer, Immunotherapy, Biology, medicine.disease_cause, medicine.disease, biology.organism_classification, Microbiology, Infectious Diseases, Tropical medicine, Pseudomonadales, medicine, Secretion, Bacteria, Pseudomonadaceae
Published: 2008
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33. Effects of two sex steroids (17β estradiol and testosterone) on proliferation and clonal growth of the human monoblastic leukemia cell line, U937

Author: Jean Jacques Sotto, Marie-France Sotto, L. Kolodie, Anne Castinel, Benoît Polack, Francois Cousin, Martine Chauvet, and Pascal Mossuz
Subjects: Cancer Research, medicine.medical_specialty, Time Factors, medicine.drug_class, Cellular differentiation, Apoptosis, Biology, Internal medicine, medicine, Humans, Hydroxyurea, Cytotoxic T cell, Testosterone, Estrogen binding, Dose-Response Relationship, Drug, Estradiol, Cell growth, Cell Cycle, Cell Differentiation, U937 Cells, Hematology, Cell cycle, Androgen, Endocrinology, Oncology, Estrogen, Leukemia, Monocytic, Acute, Neoplastic Stem Cells, Cell Division
Abstract: We investigated the effects of two sex steroids (17 β estradiol and testosterone) on five human leukemia cell lines. We observed a statistically significant inhibition of proliferation, dose and time dependent, of the human monoblastic leukemia cell line U937. This inhibition was associated with a dose dependent decrease in the number of CFU-blasts in clonogenic cultures. Cytostatic effect was obtained with doses of 5 μM for estrogen and 10 μM for androgen and was not due to a non-specific cytotoxic effect, some cell viability remained high (>90%) even after 6 days of incubation. More accurately, we demonstrated that growth inhibition was associated with a cell cycle arrest, U937 cells accumulating in G2/M phase. This blockade was dose related with a maximum number of cells accumulating at day 4. Sensitivity of these cells to an S-phase specific agent (hydroxyurea) was not increased, suggesting that these cells were blocked in G2/M and did not undergo mitosis. Expression in U937 cells of high affinity nuclear receptors for estrogen and androgen was negative which was in favour of a type II estrogen binding site, mediated mechanism. Moreover, a small fraction of these cells underwent apoptosis or differentiation with about 12% apoptotic cells and a significant increase (more than 30%) of two myelomonocytic markers (CD13 and CD64). These results demonstrate that the proliferation of some leukemic cells may be inhibited by micromolar concentrations of sex steroids, independently of nuclear receptor expression. The main mechanism seems to be a block in cell cycle associated with modulation of apoptosis and differentiation. It provided additional evidence for the potential value of sex steroids and their analogues in the treatment of leukemias.
Published: 1998
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34. Optimized Factor V Gene Mutation Detection Using Buffy-Coat Direct PCR

Author: Benoît Polack, Pascal Mossuz, and Gilles Pernod
Subjects: biology, DNA Mutational Analysis, Drug Resistance, Factor V, DNA, Buffy coat, Gene mutation, Polymerase Chain Reaction, Molecular biology, General Biochemistry, Genetics and Molecular Biology, law.invention, Enzyme Activation, law, Mutation (genetic algorithm), medicine, biology.protein, Humans, Deoxyribonucleases, Type II Site-Specific, Gene, Protein C, Polymerase chain reaction, Biotechnology, medicine.drug
Published: 1997
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35. Role of Oxygen Radicals in Tissue Factor Induction by Endotoxin in Blood Monocytes

Author: Benoît Polack, Jacques Doussière, Claire Barro, and Gilles Pernod
Subjects: Lipopolysaccharide, Nitric Oxide, medicine.disease_cause, Monocytes, Thromboplastin, Nitric oxide, chemistry.chemical_compound, Tissue factor, Pyrrolidine dithiocarbamate, Superoxides, Physiology (medical), medicine, Humans, Enzyme Inhibitors, NADPH oxidase, biology, Superoxide, Monocyte, NADPH Oxidases, Drug Synergism, Free Radical Scavengers, Hematology, Endotoxins, medicine.anatomical_structure, Biochemistry, chemistry, biology.protein, Nitric Oxide Synthase, Reactive Oxygen Species, Oxidative stress
Abstract: In response to bacterial endotoxin (lipopolysaccharide, LPS) monocytes synthesize and express on their surface tissue factor (TF) which triggers the blood coagulation cascade. Since LPS stimulates active oxygen species production by these cells, we investigated the roles of superoxide anion and nitric oxide in the induction of TF in human blood monocytes. Scavengers of reactive oxygen intermediates such as N-acetyl cys-teine or pyrrolidine dithiocarbamate were able to block TF induction. In addition, inhibition of NADPH oxidase and/or NO synthase which are major sources of active oxygen species in phagocytes also blocked TF induction. The restoration of TF expression, in monocytes treated with inhibitors of reactive oxygen production, by NN’-dimethyl-γγ‘-dipyridylium dichloride and/or sodium nitrosylpentacyanoferrate (III), which generate respectively O-2 and NO, suggests that these two radicals participate in the induction of TF at the surface of blood monocytes stimulated by LPS.
Published: 1997
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36. Contents, Vol. 27, 1997

Author: D.-H. Ku, Shirou Fukuhara, Y. Katoh-Oishi, M. Kamiyama, B. Sampson, T. Yoshimi, Gui Lan Xie, J. Woody, H. Alder, Akikazu Takada, F. Peyron, S. J. H. Van Deventer, G. N. J. Tytgat, M.M. Levi, Tomohiko Taminato, N. Nagai, M. Dugal, V.W. Ing, A. Marchand, A. van der Ende, A. Rydzewski, D.R. Anderson, Yumiko Takada, Tetsumei Urano, F. Delolme, E. Zayed, R. Pajaro, Benoît Polack, Y.S. Arkel, H. M. Van Dullemen, Daan W. Hommes, and Shosaku Nomura
Subjects: Physiology (medical), Hematology
Published: 1997
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37. Protective Role of Platelets in Chronic (Balb/C) and Acute (CBA/J) Plasmodium berghei Murine Malaria

Author: F. Peyron, F. Delolme, and Benoît Polack
Subjects: biology, Hematology, biology.organism_classification, medicine.disease, Proinflammatory cytokine, BALB/c, Pathogenesis, Cerebral Malaria, Physiology (medical), parasitic diseases, Immunology, medicine, Plasmodium berghei, Tumor necrosis factor alpha, Platelet, Malaria
Abstract: The role of blood platelets in the pathogenesis of malaria remains unclear. In this study we investigated the role of experimentally caused thrombocytopaenia in chronic (Balb/C) and acute (CBA/J) Plαsmodium-berghei-induced murine malaria. A group of 30 Balb/C mice were rendered thrombocyto-paenic by injection of anti-mouse platelet serum given every 2 days from day 2 to day 8 after malaria infection. Compared with the control group, thrombocytopaenic mice showed a greater mortality. The Kaplan-Meier estimate of the risk of death was 4.37-fold higher in the thrombocytopaenic group. Parasitaemia and weight loss was in agreement with the protective effect of platelets. In the acute malaria model, the survival rate was less significant but the estimated risk of death was 1.7-fold higher in the treated group. These results suggesting a protective role of platelets in murine malaria are not in line with previous reports of the literature. Taken together our data suggest, however, that platelets, similarly to some inflammatory cytokines like TNF, play a dual role in the pathogenesis of malaria. Depending on experimental conditions, e.g. the time of onset of thrombocytopaenia, the beneficial role of platelets may outweigh the deleterious one.
Published: 1997
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38. Live-attenuated bacteria as a cancer vaccine vector

Author: Xavier Chauchet, Bertrand Toussaint, Audrey Le Gouellec, Yan Wang, Benoît Polack, TheREx, Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), and VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)
Subjects: Salmonella, Listeria, medicine.medical_treatment, Immunology, Biology, medicine.disease_cause, Cancer Vaccines, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, 03 medical and health sciences, 0302 clinical medicine, Listeria monocytogenes, Cancer immunotherapy, Antigen, Antigens, Neoplasm, Neoplasms, Drug Discovery, medicine, Humans, Vector (molecular biology), 030304 developmental biology, Pharmacology, 0303 health sciences, Clinical Trials as Topic, Drug Carriers, Vaccines, Synthetic, Vaccination, biology.organism_classification, Virology, 3. Good health, 030220 oncology & carcinogenesis, Molecular Medicine, Cancer vaccine, Adjuvant
Abstract: International audience; In the emerging field of active and specific cancer immunotherapy, strategies using live-attenuated bacterial vectors have matured in terms of academic and industrial development. Different bacterial species can be genetically engineered to deliver antigen to APCs with strong adjuvant effects due to their microbial origin. Proteic or DNA-encoding antigen delivery routes and natural bacterial tropisms might differ among species, permitting different applications. After many academic efforts to resolve safety and efficacy issues, some firms have recently engaged clinical trials with live Listeria or Salmonella spp. We describe here the main technological advances that allowed bacteria to become one of the most promising vectors in cancer immunotherapy.
Published: 2013
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39. A safe bacterial microsyringe for in vivo antigen delivery and immunotherapy

Author: Bertrand Toussaint, Mitra Ahmadi, Xavier Chauchet, Sandrine Martin, David Laurin, François Cretin, Alexis Broisat, Charlotte Genestet, Julien Verove, Candice Trocme, Joel Plumas, Catherine Ghezzi, Caroline Aspord, Yan Wang, Audrey Le Gouellec, Benoît Polack, Université Grenoble Alpes, Inserm, CEA, BIG-Biologie du Cancer et de l’Infection, Grenoble, France, Laboratoire d'Ingénierie des Macromolécules (LIM), Institut de biologie structurale (IBS - UMR 5075 ), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), TheREx, Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS), Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Biochimie et biophysique des systèmes intégrés (BBSI), Université Joseph Fourier - Grenoble 1 (UJF)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS), Radiopharmaceutiques biocliniques, Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM), Association nationale de prévention en alcoologie et addictologie, ANPAA30, Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Joseph Fourier - Grenoble 1 (UJF), and VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)
Subjects: [SDV.BIO]Life Sciences [q-bio]/Biotechnology, MESH: Immunotherapy, medicine.medical_treatment, MESH: Photosensitizing Agents, Epitopes, T-Lymphocyte, Active immunotherapy, CD8-Positive T-Lymphocytes, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, Epitope, MESH: Cancer Vaccines, Mice, Furocoumarins, Neoplasms, Drug Discovery, MESH: Animals, MESH: Neoplasms, [INFO.INFO-BT]Computer Science [cs]/Biotechnology, Cytotoxicity, MESH: Bacterial Secretion Systems, Bacterial Secretion Systems, MESH: Furocoumarins, 0303 health sciences, Antigen Presentation, Immunity, Cellular, Photosensitizing Agents, MESH: Dendritic Cells, MESH: CD8-Positive T-Lymphocytes, 3. Good health, Pseudomonas aeruginosa, MESH: Pseudomonas aeruginosa, Molecular Medicine, Original Article, Female, MESH: Antigens, Immunotherapy, MESH: Xenograft Model Antitumor Assays, MESH: Mutation, MESH: Cell Line, Tumor, Lymphoid Tissue, Heterologous, Biology, Vaccines, Attenuated, Cancer Vaccines, Microbiology, MESH: Immunity, Cellular, MESH: Epitopes, T-Lymphocyte, 03 medical and health sciences, Immune system, Antigen, In vivo, Cell Line, Tumor, MESH: Vaccines, Attenuated, medicine, Genetics, Animals, Humans, Antigens, Molecular Biology, MESH: Mice, 030304 developmental biology, Pharmacology, MESH: Humans, 030306 microbiology, Dendritic Cells, Xenograft Model Antitumor Assays, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Disease Models, Animal, Mutation, MESH: Antigen Presentation, MESH: Lymphoid Tissue, Cancer research, MESH: Disease Models, Animal, MESH: Female
Abstract: International audience; The industrial development of active immunotherapy based on live-attenuated bacterial vectors has matured. We developed a microsyringe for antigen delivery based on the type III secretion system (T3SS) of P. aeruginosa. We applied the "killed but metabolically active" (KBMA) attenuation strategy to make this bacterial vector suitable for human use. We demonstrate that attenuated P. aeruginosa has the potential to deliver antigens to human antigen-presenting cells in vitro via T3SS with considerable attenuated cytotoxicity as compared with the wild-type vector. In a mouse model of cancer, we demonstrate that this KBMA strain, which cannot replicate in its host, efficiently disseminates into lymphoid organs and delivers its heterologous antigen. The attenuated strain effectively induces a cellular immune response to the cancerous cells while lowering the systemic inflammatory response. Hence, a KBMA P. aeruginosa microsyringe is an efficient and safe tool for in vivo antigen delivery.
Published: 2013
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40. Successful use of recombinant factor VIIa for severe surgical liver bleeding in a 5 month-old baby

Author: M Cartal, C. Piolat, C. Barro, I. Wrobleski, J.F. Dyon, P. Andrini, Benoît Polack, G. Pernod, and C. Jacquier
Subjects: medicine.medical_specialty, Hepatoblastoma, biology, business.industry, medicine.medical_treatment, Hematology, General Medicine, medicine.disease, Surgery, law.invention, Recombinant factor VIIa, law, Surgical biopsy, Activated factor VII, biology.protein, Recombinant DNA, Medicine, Hepatectomy, business, Genetics (clinical)
Abstract: Summary. A 5 month-old baby developed non-ceasing intra-peritoneal bleeding after extensive surgical biopsy for an hepatoblastoma. A single recombinant activated factor VII injection following enlarged hepatectomy helped to resolve quickly this life-threatening haemorrhagic syndrome.
Published: 2004
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41. HDAC6 controls the kinetics of platelet activation

Author: Jin Wang, Anne Laure Vitte, Xiaodong Xi, Thérèse Rossini, Boubou Diagouraga, Karin Sadoul, Thierry Buchou, Benoît Polack, Saadi Khochbin, Patrick Matthias, Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Shanghai Institute of Hematology, Shanghai Jiao Tong University School of Medicine-Shanghai Rui Jin Hospital, Transduction du signal : signalisation calcium, phosphorylation et inflammation, Université Joseph Fourier - Grenoble 1 (UJF)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM), TheREx, Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Friedrich Miescher Institute for Biomedical Research (FMI), Novartis Research Foundation, and Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)
Subjects: MESH: Hydroxamic Acids, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, MESH: Amino Acid Sequence, MESH: Tubulin, 030204 cardiovascular system & hematology, Histone Deacetylase 6, Hydroxamic Acids, Microtubules, Biochemistry, MESH: Mice, Knockout, Mice, 0302 clinical medicine, Tubulin, Platelet, MESH: Animals, MESH: Histone Deacetylase Inhibitors, [INFO.INFO-BT]Computer Science [cs]/Biotechnology, Cells, Cultured, MESH: Blood Platelets, MESH: Cell Size, Mice, Knockout, 0303 health sciences, MESH: Microtubules, Acetylation, Hematology, 3. Good health, Cell biology, MESH: Acetylation, MESH: Cells, Cultured, Blood Platelets, Molecular Sequence Data, Immunology, Biology, Histone Deacetylases, 03 medical and health sciences, Microtubule, Animals, Humans, MESH: Platelet Activation, MESH: Cell Shape, Amino Acid Sequence, Platelet activation, Cell Shape, MESH: Mice, Cell Size, 030304 developmental biology, Tubulin deacetylation, MESH: Molecular Sequence Data, MESH: Humans, Cell Biology, HDAC6, Platelet Activation, Histone Deacetylase Inhibitors, MESH: Histone Deacetylases, Hemostasis, MESH: Protein Processing, Post-Translational, biology.protein, Protein Processing, Post-Translational
Abstract: HDAC6, a major cytoplasmic deacetylase, is shown here to fine-tune the kinetics of platelet activation, a process that must be precisely regulated to ensure hemostasis after blood vessel injury while preventing pathologic thrombus formation. The discoid shape of resting platelets in the circulation is maintained by several highly acetylated microtubules organized in a marginal band. During platelet activation, microtubules undergo major reorganizations, which contribute to the shape change of activating platelets. We show that, during these activation-induced shape changes, a dramatic HDAC6-mediated tubulin deacetylation takes place, followed by microtubule reacetylation in spread platelets. In addition, although HDAC6-controlled tubulin deacetylation is not required for platelet activation, the capacity of HDAC6 to prevent tubulin hyperacetylation influences the speed of platelet spreading. These results are particularly important in view of HDAC6 inhibitors being currently used in clinical trials and represent the first example of cell signaling by lysine acetylation in platelet biology.
Published: 2012
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42. Gla-domainless factor Xa: molecular bait to bypass a blocked tenase complex

Author: Benoît Polack, Raphaël Marlu, Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Thérapeutique Recombinante Expérimentale (TIMC-IMAG-TheREx), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), and TheREx
Subjects: MESH: Signal Transduction, MESH: Neoplasm Proteins, MESH: Factor Xa, [SDV]Life Sciences [q-bio], 030204 cardiovascular system & hematology, Pharmacology, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, MESH: Thrombin, 0302 clinical medicine, hemic and lymphatic diseases, ComputingMilieux_MISCELLANEOUS, 0303 health sciences, Chemistry, Antithrombin, Thrombin, [SDV.MHEP.HEM]Life Sciences [q-bio]/Human health and pathology/Hematology, Hematology, MESH: Lipoproteins, 3. Good health, Neoplasm Proteins, Cysteine Endopeptidases, Coagulation, Biochemistry, Factor Xa, medicine.drug, MESH: Half-Life, Half-Life, Protein Binding, Signal Transduction, Lipoproteins, Hemophilia A, Antibodies, Antithrombins, 03 medical and health sciences, Tissue factor, Tissue factor pathway inhibitor, Prothrombinase, medicine, MESH: Protein Binding, Humans, 030304 developmental biology, MESH: Humans, MESH: Antithrombins, MESH: Antibodies, MESH: Multiprotein Complexes, MESH: Hemophilia A, Multiprotein Complexes, Original Articles and Brief Reports, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology, Tenase, Discovery and development of direct thrombin inhibitors, MESH: Cysteine Endopeptidases
Abstract: International audience; BACKGROUND: Hemophilia is caused by deficiencies in coagulation factor VIII or IX, resulting in direct blockade of the intrinsic tenase complex and indirect blockade of the extrinsic tenase complex which is rapidly inhibited upon binding of factor Xa to tissue factor pathway inhibitor. We evaluated the ability of Gla-domainless factor Xa, a truncated form of factor Xa devoid of procoagulant properties, to bind to tissue factor pathway inhibitor and to alleviate the physiological inhibition of the extrinsic tenase. DESIGN AND METHODS: Using a thrombin generation assay triggered by a low concentration of tissue factor, we evaluated the ability of Gla-domainless factor Xa to restore blood coagulation in plasma from hemophilia A and B patients without and with inhibitors. We then compared its efficacy to generate thrombin to depletion of antithrombin or tissue factor pathway inhibitor by specific antibodies. Finally, we compared the kinetics of neutralization of factor Xa and Gla-domainless factor Xa by antithrombin and tissue factor pathway inhibitor. RESULTS: Gla-domainless factor Xa was able to restore thrombin generation in plasma samples from hemophiliacs. This effect was observed for plasma from hemophilia A patients without or with inhibitors and for plasma from hemophilia B patients. Gla-domainless factor Xa had a lower affinity than factor Xa for tissue factor pathway inhibitor whereas the affinities of both proteins for antithrombin were similar. Finally, despite a short half-life in plasma, the effect of Gla-domainless factor Xa on thrombin generation was sustained for at least 1 hour. CONCLUSIONS: As Gla-domainless factor Xa was able to restore thrombin generation in plasma from hemophilia patients, our results suggest that it may be an effective alternative to current treatments for hemophilia with or without an inhibitor.
Published: 2012
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43. Optimization of antitumor immunotherapy mediated by type III secretion system-based live attenuated bacterial vectors

Author: Audrey Le Gouellec, Benoît Polack, Hichem Chaker, Bertrand Toussaint, Yan Wang, Hoda Asrih, Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), TheREx, and VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)
Subjects: Cancer Research, MESH: Immunotherapy, MESH: Antigens, Neoplasm, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, Autoantigens, MESH: Cancer Vaccines, MESH: Gene Order, Type three secretion system, Mice, 0302 clinical medicine, MESH: Genetic Vectors, Neoplasms, Gene Order, Immunology and Allergy, MESH: Animals, MESH: Neoplasms, Vector (molecular biology), MESH: Treatment Outcome, 0303 health sciences, 3. Good health, Bacterial vaccine, Vaccination, Treatment Outcome, MESH: Autoantigens, 030220 oncology & carcinogenesis, MESH: Pseudomonas aeruginosa, Pseudomonas aeruginosa, Female, Immunotherapy, Immunology, Genetic Vectors, Biology, Vaccines, Attenuated, Cancer Vaccines, Cell Line, 03 medical and health sciences, Immune system, Antigen, MESH: Mice, Inbred C57BL, In vivo, Antigens, Neoplasm, MESH: Vaccines, Attenuated, Animals, Humans, MESH: Mice, 030304 developmental biology, Pharmacology, MESH: Humans, MESH: Cell Line, Mice, Inbred C57BL, Cancer vaccine, MESH: Female
Abstract: International audience; Recently, due to their effective ability to deliver antigen to antigen-presenting cells in vivo, type III secretion system-based attenuated bacterial vectors have increasingly attracted attention for their potential interest in cancer vaccine development. We have previously developed live attenuated Pseudomonas aeruginosa type III secretion system-based vectors to deliver in vivo tumor antigens. In this work, we improved the performance of these bacterial vectors through several approaches in different murine cancer models involving non-self-antigens or self-antigens. First, by modulating injection frequency and interval, bacterial vaccination-activated immune response could be enhanced and the in vivo therapeutic efficacy of bacterial vaccines could be improved. The optimized vaccination scheme induced long-lasting CD8+ T cells' response. Second, a dual antigen delivery pattern was successfully applied in our bacterial vectors. Compared with a single antigen delivery vector, biantigen delivery vectors demonstrated several advantages including better tumor rejection efficiency, simplicity of use, and safety. Third, 1 more attenuated mutant-CHA-OAL strain that is totally avirulent in mice was further adapted to grow in a chemically defined medium to comply with current good manufacturing processes. The poor infectivity of this new strain could be overcome by vaccinations at multiple loci, yielding an efficiently improved vaccination performance. Taken together, our results highlight the potential of our live attenuated P. aeruginosa vectors for applications in relevant clinical trials.
Published: 2012
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44. Molecular interface of S100A8 with cytochrome b558 and NADPH oxidase activation

Author: Minh Vu Chuong Nguyen, Marc-André Hograindleur, Françoise Morel, Athan Baillet, Marie-Hélène Paclet, Benoît Polack, Sylvie Berthier, Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Thérapeutique Recombinante Expérimentale (TIMC-IMAG-TheREx), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), 'Ministère de l'Enseignement Supérieur de la Recherche et Technologie, Paris', 'UFR de Médecine, Université Joseph Fourier, Grenoble', 'Région Rhône-Alpes, programme Emergence 2003 and ARCUS (Actions en Régions de Coopération Universitaire et Scientifique) France/Chine 2007-2008', CGD (Chronic Granulomatous Disease) research Trust 2006-2007, UK, 'Groupement des Entreprises Françaises dans la lutte contre le Cancer, délégation de Grenoble', 'Société Française de Rhumatologie', and 'Délégation à la Recherche Clinique et à l'Innovation, Centre Hospitalier Universitaire, Grenoble'
Subjects: Herpesvirus 4, Human, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Cytochrome, Neutrophils, lcsh:Medicine, NADPH Oxidase, MESH: Amino Acid Sequence, Plasma protein binding, Biochemistry, biophysics & molecular biology [F05] [Life sciences], MESH: Neutrophils, Biochemistry, MESH: Antibodies, Monoclonal, MESH: Recombinant Proteins, MESH: Protein Structure, Tertiary, Cytosol, MESH: Structure-Activity Relationship, 0302 clinical medicine, Protein structure, MESH: Cytosol, ExoS toxin, MESH: Cell-Free System, MESH: Calgranulin B, MESH: Calgranulin A, Lymphocytes, [INFO.INFO-BT]Computer Science [cs]/Biotechnology, Biochimie, biophysique & biologie moléculaire [F05] [Sciences du vivant], MESH: NADPH Oxidase, MESH: Bacterial Secretion Systems, lcsh:Science, Bacterial Secretion Systems, Peptide sequence, 0303 health sciences, Oxidase test, Multidisciplinary, NADPH oxidase, biology, Enzyme Classes, Antibodies, Monoclonal, Recombinant Proteins, Oxygen Metabolism, Enzymes, Protein Transport, Cross-Linking Reagents, 030220 oncology & carcinogenesis, Pseudomonas aeruginosa, MESH: Pseudomonas aeruginosa, Oxidoreductases, Protein Binding, Research Article, MESH: Enzyme Activation, MESH: Protein Transport, Hemeprotein, MESH: Cross-Linking Reagents, Molecular Sequence Data, Enzyme Regulation, Structure-Activity Relationship, 03 medical and health sciences, Escherichia coli, Calgranulin B, Humans, MESH: Cytochrome b Group, MESH: Protein Binding, Calgranulin A, Amino Acid Sequence, Protein Interactions, Biology, 030304 developmental biology, MESH: Humans, MESH: Molecular Sequence Data, Cell-Free System, lcsh:R, NADPH Oxidases, Proteins, MESH: Herpesvirus 4, Human, Cytochrome b Group, Fusion protein, Protein Structure, Tertiary, Regulatory Proteins, Enzyme Activation, Transmembrane Proteins, Metabolism, Enzyme Structure, biology.protein, MESH: Lymphocytes, lcsh:Q
Abstract: International audience; S100A8 and S100A9 are two calcium binding Myeloid Related Proteins, and important mediators of inflammatory diseases. They were recently introduced as partners for phagocyte NADPH oxidase regulation. However, the precise mechanism of their interaction remains elusive. We had for aim (i) to evaluate the impact of S100 proteins on NADPH oxidase activity; (ii) to characterize molecular interaction of either S100A8, S100A9, or S100A8/S100A9 heterocomplex with cytochrome b(558); and (iii) to determine the S100A8 consensus site involved in cytochrome b(558)/S100 interface. Recombinant full length or S100A9-A8 truncated chimera proteins and ExoS-S100 fusion proteins were expressed in E. coli and in P. aeruginosa respectively. Our results showed that S100A8 is the functional partner for NADPH oxidase activation contrary to S100A9, however, the loading with calcium and a combination with phosphorylated S100A9 are essential in vivo. Endogenous S100A9 and S100A8 colocalize in differentiated and PMA stimulated PLB985 cells, with Nox2/gp91(phox) and p22(phox). Recombinant S100A8, loaded with calcium and fused with the first 129 or 54 N-terminal amino acid residues of the P. aeruginosa ExoS toxin, induced a similar oxidase activation in vitro, to the one observed with S100A8 in the presence of S100A9 in vivo. This suggests that S100A8 is the essential component of the S100A9/S100A8 heterocomplex for oxidase activation. In this context, recombinant full-length rS100A9-A8 and rS100A9-A8 truncated 90 chimera proteins as opposed to rS100A9-A8 truncated 86 and rS100A9-A8 truncated 57 chimeras, activate the NADPH oxidase function of purified cytochrome b(558) suggesting that the C-terminal region of S100A8 is directly involved in the molecular interface with the hemoprotein. The data point to four strategic (87)HEES(90) amino acid residues of the S100A8 C-terminal sequence that are involved directly in the molecular interaction with cytochrome b(558) and then in the phagocyte NADPH oxidase activation.
Published: 2012
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45. Monocyte Tissue Factor Expression Induced by Plasmodium falciparum-Infected Erythrocytes

Author: Pierre Ambroise-Thomas, Benoît Polack, F. Santoro, L. Kolodie, François Peyron, Annick Luisy, and Gilles Pernod
Subjects: Erythrocytes, Plasmodium falciparum, Parasitemia, In Vitro Techniques, Biology, Fibrinogen, Monocytes, Thromboplastin, Tissue factor, Thrombin, parasitic diseases, medicine, Animals, Humans, Platelet, Malaria, Falciparum, Blood Coagulation, Monocyte, Hematology, medicine.disease, biology.organism_classification, Molecular biology, Blood Coagulation Factors, medicine.anatomical_structure, Coagulation, Immunology, medicine.drug
Abstract: SummaryMonocytes are active elements of the host response against Plasmodium falciparum. They are able to express tissue factor and trigger the extrinsic pathway of blood coagulation the activation of which remained unclear in malaria. Our aim was to assess the tissue factor expression of purified blood monocytes stimulated by cultured Plasmodium falciparum-infected erythrocytes. Malaria parasite induced an early generation of tissue factor with a peak between 8 and 12 h of stimulation. Maximum expression was observed for parasitemia ranging from 1 to 2%. Plasmodium falciparum culture supernatants had the same effect showing the existence of a soluble factor able to induce the tissue factor expression. These data, demonstrating an activation of the tissue factor pathway by the malaria parasite, emphasize thrombin generation. Therefore, thrombin could participate in malaria pathology either in the microcirculatory blockade via platelet and fibrinogen activation or as a mitotic.
Published: 1992
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46. Management of haemophilia A-inhibitor patients: clinical and regulatory perspectives

Author: Zera Tellier, Benoît Polack, Marie-Hélène André, LFB Biomédicaments, TheREx, Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), and VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)
Subjects: [SDV.BIO]Life Sciences [q-bio]/Biotechnology, MESH: Blood Coagulation Factor Inhibitors, 030204 cardiovascular system & hematology, Epitope, Immune tolerance, MESH: Antibodies, Neutralizing, 0302 clinical medicine, Isoantibodies, hemic and lymphatic diseases, MESH: Child, Haemophilia A, Immunology and Allergy, Medicine, [INFO.INFO-BT]Computer Science [cs]/Biotechnology, Child, Severe complication, MESH: Treatment Outcome, Clinical Trials as Topic, biology, Blood Coagulation Factor Inhibitors, Immunogenicity, General Medicine, MESH: Factor VIII, 3. Good health, Treatment Outcome, Child, Preschool, Antibody, congenital, hereditary, and neonatal diseases and abnormalities, MESH: Immune Tolerance, MESH: Clinical Trials as Topic, Regulatory issues, Haemophilia, Hemophilia A, 03 medical and health sciences, Immune Tolerance, Humans, Factor VIII, MESH: Humans, business.industry, Clinical management, MESH: Child, Preschool, FVIII inhibitors, medicine.disease, MESH: Hemophilia A, Antibodies, Neutralizing, MESH: Isoantibodies, Recombinant factor VIIa, Immunology, biology.protein, business, 030215 immunology
Abstract: International audience; Inhibitors to factor VIII (FVIII) are alloantibodies directed against epitopes able to neutralise FVIII procoagulant activity. They may render FVIII replacement therapy ineffective. They represent the most severe complication of haemophilia A. At least three mechanisms of FVIII neutralisation activity by anti-FVIII antibodies have been described: (1) steric hindrance; (2) recognition of neo-epitopes and (3) catalytic activity. The Nijmegen modification of the Bethesda is the recommended method for inhibitor surveillance. The occurrence of inhibitors is a relatively frequent and early event in previously untreated patients. Conversely, it is rare in previously treated patients. Therapeutic strategies for managing inhibitors include: inhibitor eradication, haemostatic management of bleeding episodes and/or surgery and supportive care. For high responding inhibitors, immune tolerance induction (ITI) is the strategy for achieving antigen-specific tolerance to FVIII. ITI success rate ranges commonly between 60% and 80%. For treatment of patients with high-titre, high-responding inhibitors, 'by-pass' therapy is generally recommended. Activated prothrombin complex concentrates represent the historically primary 'by-pass' treatment. Recombinant factor VIIa has also been widely used as a by-passing agent. Considering the small patient population, it has to be considered that full immunogenicity data cannot be collected premarketing authorisation. Thus, stringent follow-up of patients in the post-authorisation phase is required.
Published: 2009
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47. Should plasma homocysteine be used as a biomarker of venous thromboembolism? A case-control study

Author: Claire Barro, Véronique Ducros, Jean-Luc Bosson, Gilles Pernod, Jacqueline Yver, Marie-Dominique Desruet, Patrick H. Carpentier, Benoît Polack, Département de biologie intégrée, CHU Grenoble-Hôpital Michallon, Institut de biologie et chimie des protéines [Lyon] (IBCP), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Laboratoire Nutrition, Vieillissement et Maladies Cardiovasculaires, Université Joseph Fourier - Grenoble 1 (UJF), Laboratoire d'Hématologie, CHU Grenoble, Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), TheREx, and VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)
Subjects: Male, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Homocysteine, 030204 cardiovascular system & hematology, Gastroenterology, MESH: Venous Thromboembolism, chemistry.chemical_compound, 0302 clinical medicine, Risk Factors, Interquartile range, MESH: Risk Factors, Prevalence, Prospective Studies, 030212 general & internal medicine, [INFO.INFO-BT]Computer Science [cs]/Biotechnology, Ultrasonography, MESH: Aged, MESH: Middle Aged, biology, MESH: Methylenetetrahydrofolate Reductase (NADPH2), Venous Thromboembolism, Hematology, General Medicine, Middle Aged, MESH: Case-Control Studies, 3. Good health, Venous thrombosis, MESH: Hyperhomocysteinemia, MESH: Young Adult, Anesthesia, Female, Adult, Hyperhomocysteinemia, medicine.medical_specialty, Cobalamin, Young Adult, 03 medical and health sciences, Internal medicine, MESH: Polymorphism, Genetic, medicine, Humans, MESH: Homocysteine, Methylenetetrahydrofolate Reductase (NADPH2), MESH: Prevalence, Aged, Polymorphism, Genetic, MESH: Humans, business.industry, Case-control study, MESH: Biological Markers, MESH: Adult, Odds ratio, medicine.disease, MESH: Prospective Studies, MESH: Male, chemistry, Case-Control Studies, Methylenetetrahydrofolate reductase, biology.protein, business, MESH: Female, Biomarkers
Abstract: Mild or moderate hyperhomocysteinemia as a risk factor for venous thrombosis is still a matter of debate. The strength of this study is to bring a body of elements to evaluate whether hyperhomocysteinemia should be used as a biomarker for venous thromboembolism (VTE). These elements consist of a biological evaluation of several hematological risk factors, and an original control group made of patients with a negative Doppler ultrasonography. A total of 151 cases and 155 controls were included. Total plasma homocysteine level, MTHFR C677T polymorphism, inherited abnormalities of the natural anticoagulant system as well as plasma folate and cobalamin levels were determined. A total of 41 (27.2 %) of cases and only 9 (5.8%) of controls had at least one of the coagulation defects studied. No significant difference was observed for total homocysteine levels between the 2 groups: median (interquartile range) = 8.3 (7.2-10.8) μmol/L for cases and 8.4 (7-10.9) μmol/L for controls. We found significantly more plasma folates and/or cobalamin deficiencies in controls (18.3%) than in cases (8.6%). After adjustment for several variables significantly related to risk factors of VTE, hyperhomocysteinemia (>" xbd="2200" xhg="2176" ybd="1302" yhg="1257"/>13.2 μmol/L) was not found statistically associated with VTE: odds ratio 1.36 (95% confidence interval, 0.52-3.54). The prevalence of the homozygous 677TT polymorphism in the MTHFR gene was not increased in cases compared with controls. Mild or moderate hyperhomocysteinemia does not seem to be a strong determinant in VTE not only when the control group does not exclusively include healthy persons but also in investigated disease-free (thromboembolic disease) controls.
Published: 2009
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48. High-cell-density regulation of the Pseudomonas aeruginosa type III secretion system: implications for tryptophan catabolites

Author: Madiha Derouazi, Hichem Chaker, Didier Filopon, Stephanie Boullanger, Benoît Polack, Da-Kang Shen, Bertrand Toussaint, TheREx, Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Department of Microbiology and Parasitology, Shanghai Jiao Tong University School of Medicine-Shanghai Jiao Tong University School of Medicine, VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Centre de Recherches sur les Macromolécules Végétales (CERMAV), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), S. D. K. was supported by a PhD scholarship from the Agence Universitaire de la Francophonie, Universidad Michoacana de San Nicolas de Hidalgo, Michoacan, Mexico, University of Nottingham, UK, inconnu, and Inconnu
Subjects: MESH: Indoleacetic Acids, MESH: Protein Transport, MESH: Mutation, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Virulence Factors, MESH: Kynurenine, Tryptophan synthase, Biology, medicine.disease_cause, Microbiology, Virulence factor, Type three secretion system, 03 medical and health sciences, chemistry.chemical_compound, Bacterial Proteins, medicine, Transcriptional regulation, [INFO.INFO-BT]Computer Science [cs]/Biotechnology, MESH: Bacterial Proteins, Kynurenine, ComputingMilieux_MISCELLANEOUS, MESH: Tryptophan, 030304 developmental biology, MESH: Virulence Factors, Regulation of gene expression, 0303 health sciences, MESH: Gene Expression Regulation, Bacterial, Indoleacetic Acids, 030306 microbiology, Pseudomonas aeruginosa, Tryptophan, Gene Expression Regulation, Bacterial, biochemical phenomena, metabolism, and nutrition, bacterial infections and mycoses, 3. Good health, Protein Transport, Biochemistry, chemistry, Mutation, MESH: Pseudomonas aeruginosa, biology.protein, bacteria
Abstract: International audience; The Pseudomonas aeruginosa type III secretion system (T3SS) is known to be a very important virulence factor in acute human infections, but it is less important in maintaining chronic infections in which T3SS genes are downregulated. In vitro, the activation of T3SS expression involves a positive activating loop that acts on the transcriptional regulator ExsA. We have observed that in vivo T3SS expression is cell density-dependent in a manner that does not need known quorum-sensing (QS) signals. In addition, stationary-phase culture supernatants added to exponential-phase growing strains can inhibit T3SS expression. The analysis of transposon insertion mutants showed that the production of such T3SS-inhibiting signals might depend on tryptophan synthase and hence tryptophan, which is the precursor of signalling molecules such as indole-3-acetic acid (IAA), kynurenine and Pseudomonas quinolone signal (PQS). Commercially available tryptophan-derived molecules were tested for their role in the regulation of T3SS expression. At millimolar concentrations, IAA, 1-naphthalacetic acid (NAA) and 3-hydroxykynurenine inhibited T3SS expression. Inactivation of the tryptophan dioxygenase-encoding kynA gene resulted in a decrease in the T3SS-inhibiting activity of supernatants. These observations suggest that tryptophan catabolites are involved in the downregulation of T3SS expression in the transition from a low- to a high-cell-density state.
Published: 2008
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49. Tissue-type plasminogen activator has antiangiogenic properties without effect on tumor growth in a rat C6 glioma model

Author: Gilles Pernod, Benoît Polack, Richard J. Fish, N. Bolle, Egbert K. O. Kruithof, Bryan Simard, F. Solly, Groupe de Recherche et d’Étude du Processus Inflammatoire (TIMC-IMAG-GREPI), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Laboratory of Vascular Biology, Geneva University Hospital (HUG), Neurosciences précliniques, Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM), Thérapeutique Recombinante Expérimentale (TIMC-IMAG-TheREx), Techniques pour l'Evaluation et la Modélisation des Actions de la Santé (TIMC-IMAG-ThEMAS), Grant from the Groupement des Entreprises Franc¸aises pour la lutte contre le cancer (GEFLUC), Inserm U318, and anatomy-pathology laboratory ofGrenoble University Hospital
Subjects: Male, Cancer Research, MESH: Combined Modality Therapy, MESH: Tumor Burden, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Captopril, Angiogenesis, MESH: Angiostatins, Tissue Plasminogen Activator/genetics/metabolism/physiology, Angiotensin-Converting Enzyme Inhibitors, Tissue plasminogen activator, MESH: Glioma, angiogenesis, Mice, 0302 clinical medicine, Differentiation therapy, MESH: Genetic Vectors, MESH: Tissue Plasminogen Activator, ddc:576.5, MESH: Animals, [INFO.INFO-BT]Computer Science [cs]/Biotechnology, MESH: Lentivirus, ddc:616, Angiostatins/metabolism, 0303 health sciences, Angiostatin, integumentary system, Neovascularization, Pathologic, MESH: Angiotensin-Converting Enzyme Inhibitors, Glioma, Combined Modality Therapy, Immunohistochemistry, Angiotensin-Converting Enzyme Inhibitors/pharmacology/therapeutic use, Tumor Burden, Platelet Endothelial Cell Adhesion Molecule-1, Genetic Vectors/genetics, Gene Therapy/methods, Captopril/pharmacology/therapeutic use, 030220 oncology & carcinogenesis, Tissue Plasminogen Activator, Molecular Medicine, Tumor Burden/drug effects, medicine.drug, MESH: Xenograft Model Antitumor Assays, MESH: Cell Line, Tumor, MESH: Rats, Antigens, CD31/analysis, Blotting, Western, Genetic Vectors, Mice, Nude, 03 medical and health sciences, In vivo, Cell Line, Tumor, medicine, MESH: Mice, Nude, MESH: Blotting, Western, Animals, Molecular Biology, MESH: Mice, Angiostatins, 030304 developmental biology, business.industry, Glioma/drug therapy/pathology/therapy, Lentivirus/genetics, Lentivirus, MESH: Captopril, MESH: Immunohistochemistry, MESH: Antigens, CD31, Genetic Therapy, tissue-type plasminogen activator (tPA), medicine.disease, Xenograft Model Antitumor Assays, MESH: Male, Neovascularization, Pathologic/drug therapy/metabolism/therapy, Rats, Immunology, Cancer research, MESH: Gene Therapy, business, MESH: Neovascularization, Pathologic, Plasminogen activator, Ex vivo
Abstract: International audience; Tissue-type plasminogen activator (tPA) plays a major role in the fibrinolytic system. According to several reports, tPA may also have antiangiogenic properties, especially in combination with a free sulfhydryl donor (FSD). In the rat C6 glioma model, in vitro and in vivo tPA synthesis by glioma cells is enhanced by differentiation therapy. To address the antiangiogenic potential of tPA in this model, tPA was overexpressed in glioma tumors by ex vivo transduction of C6 cells with a lentiviral vector encoding tPA. The transduced cells were subcutaneously implanted into nude mice. Gene transfer allowed for efficient synthesis of tPA by the C6 tumors. Although the treatment of tPA+ tumor-bearing animals with the FSD captopril generated angiostatin in situ and reduced endothelial vascularization of the tumors, it had no effect on tumor growth. Alternative mechanisms could account for this lack of effect and consequently have important implications for vascular the treatment of glioblastoma.
Published: 2008
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50. High-yield production of secreted active proteins by the Pseudomonas aeruginosa type III secretion system

Author: Olivier Epaulard, Madiha Derouazi, Lauriane Quenee, Raphaël Marlu, Benoît Polack, Mathilde Guillaume, Bertrand Toussaint, Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), TheREx, VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), and Grants from the Agence Nationale de la Recherche and Vaincre la Mucoviscidose
Subjects: MESH: Protein Transport, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, medicine.disease_cause, Applied Microbiology and Biotechnology, Inclusion bodies, Microbiology, Type three secretion system, MESH: Membrane Transport Proteins, 03 medical and health sciences, Bacterial Proteins, medicine, [INFO.INFO-BT]Computer Science [cs]/Biotechnology, Escherichia coli, MESH: Bacterial Proteins, 030304 developmental biology, 0303 health sciences, Ecology, biology, 030306 microbiology, Membrane transport protein, Pseudomonas aeruginosa, Membrane Transport Proteins, biology.organism_classification, Physiology and Biotechnology, Transport protein, Protein Transport, Secretory protein, Biochemistry, Pseudomonadales, MESH: Pseudomonas aeruginosa, biology.protein, Food Science, Biotechnology
Abstract: The Escherichia coli system is the system of choice for recombinant protein production because it is possible to obtain a high protein yield in inexpensive media. The accumulation of protein in an insoluble form in inclusion bodies remains a major disadvantage. Use of the Pseudomonas aeruginosa type III secretion system can avoid this problem, allowing the production of soluble secreted proteins.
Published: 2008
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